Response to the IUPACIUPAP Joint Working Party Second Report 'On the Discovery of Elements
Responsetoclarificationletter2响应于澄清函2
Single Technology Appraisal – Bevacizumab in combination with oxaliplatin and either 5FU or capecitabine for the treatment of metastaticcolorectal cancerPART 2 – RESPONSE TO NON-PRIORITY ERG CLARIFICATION QUESTIONS• B15 Could you please clarify the following points regarding the economic analysisSection 7.2.9. Please provide 95% confidence intervals for the mean dose values in Table 29 p133 and mean number of cycles per month observed in Table 33 p137. In addition, please provide for each of the six treatment groups separately.Tabulated below are the mean dose and cycle durations with confidence intervals as requested.The cycle durations for the bevacizumab arms are based on the mean bevacizumab cycle durations and the cycle duration for all the other arms are based on theoxaliplatin cycle duration. Cycle duration was similar when using oxaliplatin doses as when using bevacizumab for the bevacizumab containing arms. (see footnote to table 2 below).The confidence intervals for the cycle durations were calculated based on a sample size (n) of the total number of cycles. This assumes independence between cycles, which may under estimate confidence intervals.In the economic model the mean per cycle dose was calculated by dividing the mean dose by the mean number of cycles. The updated table below shows the mean dose per each cycle calculated directly from the patient level data, which was calculated as part of the exercise for calculating the confidence intervals for the dose. Hence there is a slight variance between the mean values presented here and the mean values used in the economic model. The only variances however greater than 1% between the figures used in the model and those presented in the table below are for capecitabine and oxaliplatin. We can confirm that updating the model with the mean values presented below does not materially affect the results of the economic analysis; the ICER for B-XELOX vs XELOX+-P increases by only £24 and B-FOLFOX-6 vs FOLFOX-6+-P increases by only £46.2Twhen based on oxaliplatin doses• B16 Could you please clarify the following information in the appendices• Appendix E1. As the model submitted by the manufacturer is a cohort model the mean costs of treatment are appropriate. Please clarify whether the costs have been sampled using the quartiles described in table 51 on p182 and in table 52 rather than the standard error of the mean, which would be incorrect. • Appendix E3. The manufacturer’s submission states that a Beta Pert distribution was used to estimate uncertainty in adverse event costs. It is unclear whether the quartiles listed in Table 51 or the 50% and 150% of the mean were used as the low and high estimates. Please describe how the parameters for the beta pert distributions were calculated. Please also describe any assumptions made, including how the mode was estimated.Appendix E3. For the PSA a Beta (utility*1000, (1-utility)*1000) distribution was used to model the uncertainty in the utility values. Please use a Beta distribution that fits to the confidence intervals of the utility data.The model assumes a minimum and maximum value of 50% and 150% of the mean respectively for all adverse event costs and monthly progression free and does not use the inter-quartile range.The model has been amended with a beta distribution that fits the confidenceintervals of the utility data as requested. For the PFS T health state the s.e. was 0.02 it was assumed that this was the same standard error for the PFS PT health state. The standard error of the mean utility for the progressive disease health state has not been reported in the literature however the standard errors at each individual time point that the utility was measured in the source trial is available. The standard error of the estimated average utility at each time point is in the range of 0.02 – 0.03 or less. Given that there seems to be a small variability in average utility at each time pointone can assume that the standard error of the overall mean utility would be less than the standard error of the individual means at each time point. A standard error or 0.03 has been assumed in the PSA.Below are the results of the PSA based on the pooled analysis using all 6 arms of the NO16966 as per scenario 1 presented in appendix A of part 1 of our response to the clarification questions and utilizing the standard error to fit the distribution around the utility values as per the above request.。
重症肌无力
重症肌无力 Lambert-Eaton 综合征
Dep. of Neurology The 2nd Hospital Harbin Medical University
Neuromuscular Disorders
Myasthenia gravis ( MG)
Epidemiology
It is the prototypic neuromuscular disorders with an incidence of 80-200 per million and prevalence about 500 per million. In China, it is estimated that 0.6 million people were diagnosed as MG and most of them lives in the South of China. It had been a life-threatening disease before 1970’s, though nowadays the incidence of death has been greatly reduced to about 0.2%.
Definition
The diseases of neuromuscular junction (NMJ) describes a sets of disease caused by circulating factors such as neurotoxins or autoantibodies which bind with high affinity to specific proteins at the NMJ and disturb the neuromuscular transmission.
NCRP_151
Also true for laminated barriers:
Where for LOW ENERGY:
H Tot = H tr = W UT B1BmB2
d2
For HIGH ENERGY:and源自H cg = H n = 0
H Tot + H cg = 2.7
W UT B1BmB2 d2
3.3 Time Averaged Dose-Equivalent Rates When designing radiation shielding barriers it is usual to assume that the workload will be evenly distributed throughout the year. Therefore, it is reasonable to design a barrier to meet a weekly value equal to one-fiftieth of the annual shielding design goal (NCRP, 2004). However, further scaling the shielding design goal to shorter intervals is not appropriate and may be incompatible with the ALARA principle. Specifically, the use of a measured instantaneous dose-equivalent rate (IDR), with the accelerator operating at maximum output, does not properly represent the true operating conditions and radiation environment of the facility. It is more useful if the workload and use factor are considered together with the IDR when evaluating the adequacy of a barrier. For this purpose, the concept of time averaged dose equivalent rate (TADR) is used in this Report along with the measured or calculated IDR. The TADR is the barrier attenuated dose-equivalent rate averaged over a specified time or period of operation. TADR is proportional to IDR, and depends on values of W and U. There are two periods of operation of particular interest to radiation protection, the week and the hour.
PNAS-2017-Replyt...
LETTERREPLY TO SILBURT ET AL.:Concerning sterile inflammation following focused ultrasound and microbubbles in the brainZsofia I.Kovacs a,1,Scott R.Burks a,and Joseph A.Frank a,b,1We thank Silburt et al.(1)for their comments on ourarticle(2).The authors provide us with an opportunity toexpand on sterile inflammation in the brain induced bypulsed focused ultrasound(pFUS)(3).pFUS with micro-bubbles(MB)resulting in blood–brain barrier disruption(BBBD)is accompanied by plasma protein extravasa-tion into the extracellular space.Fundamentally,anyBBBD is a nonhomeostatic condition and leakage ofalbumin activates microglia and astrocytes(4).To appropriately contextualize our biological data(2),we must make a few technical notes.Silburt et al.(1)accurately point out that many techniques in theFUS field are plagued by lack of rigorous optimiza-tion.Accordingly,our study sought to use a relativelybenign but clinically relevant treatment strategy.TheFUS peak negative pressure was0.3MPa(in water),be-low limits for inertial cavitation and microhemorrhages(5,6).Silburt et al.(1)incorrectly claim we infused5-to10-fold more MB than McMahon et al.(7),who used2.4×108MB/kg of Definity,compared withthe2.0×108–3.2×108MB/kg of Optison used in our study(2).Furthermore,Silburt et al.(1)ignored sub-tleties relating to MB type,animal oxygenation status,and number of treatment targets.Definity’s disper-sity is more uniform than Optison’s,translating to agreater fraction of Definity’s population cavitatingunder similar sonication parameters.McDannold et al.(8)reported that BBBD is further hampered byinspiring100%O2,like our study,resulting from de-creased MB lifetimes in circulation(9).Our experi-mental parameters essentially reduced acousticcavitation effects relative to several other studies inthe literature,but resulted in BBBD accompanied bysterile inflammation nonetheless.Finally,we soni-cated nine regions in the frontal cortex,compared with four treatment regions in McMahon et al.(7). Many studies only sonicate a single target point, which limits detection of parenchymal abnormalities. The number of sonication points is an uninvestigated area of paramount importance if pFUS is to be used in pathologies like Alzheimer’s disease,which would re-quire sonicating large brain volumes.The molecular changes we(2)report and the tran-scriptomics reported in McMahon et al.(7)do not substantially overlap.Transcriptomic analyses fo-cused on the endothelium without considering shockwave effects on cells comprising the neuro-vascular unit and subsequent molecular changes. Proinflammatory interleukins and IFN-γwere not in-vestigated in McMahon et al.(7).The increased in-tercellular adhesion molecule-1(ICAM1)expression reported by us could be explained by additional target foci and the associated rapid cascade of cytokines(2). Vascular endothelial growth factor and erythopoietin are associated with increased ICAM(10)and BBBD, and are not strictly dependent on hypoxia,although previous studies demonstrate vasospasm during BBBD by pFUS(11).Our molecular profiling was not encyclo-pedic and unmeasured molecular factors could in-crease erythopoietin following FUS.Our study(2)did not contain long-term follow-up,so we cannot com-ment on Silburt et al.’s(1)claim of a lack of damage or regeneration in response to the BBBD and induced sterile inflammation.Our study(2)raises important complicating issues regarding FUS bioeffects.Substantial and rigorous evaluation of molecular and inflammatory changes from multiple sonications is required in other models (3)before the technique can be confidently moved into clinical trials.1Silburt J,Lipsman N,Aubert I(2017)Disrupting the blood–brain barrier with focused ultrasound:Perspectives on inflammation and regeneration.Proc Natl Acad Sci USA114:E6735–E6736.2Kovacs ZI,et al.(2017)Disrupting the blood-brain barrier by focused ultrasound induces sterile inflammation.Proc Natl Acad Sci USA 114:E75–E84.3Fung LK(2017)A sterile animal model for neuroinflammation?Sci Transl Med9:eaal4994.a Frank Laboratory,Radiology and Imaging Sciences,Clinical Center,National Institutes of Health,Bethesda,MD20892;andb National Institute ofBiomedical Imaging and Bioengineering,National Institutes of Health,Bethesda,MD20892Author contributions:Z.I.K.,S.R.B.,and J.A.F.wrote the paper.The authors declare no conflict of interest.1To whom correspondence may be addressed.Email:*********************or****************.gov./cgi/doi/10.1073/pnas.1711544114PNAS Early Edition|1of2L E T T E R4Shlosberg D,Benifla M,Kaufer D,Friedman A(2010)Blood-brain barrier breakdown as a therapeutic target in traumatic brain injury.Nat Rev Neurol6:393–403. 5McDannold N,Vykhodtseva N,Hynynen K(2008)Effects of acoustic parameters and ultrasound contrast agent dose on focused-ultrasound induced blood-brain barrier disruption.Ultrasound Med Biol34:930–937.6Treat LH,et al.(2007)Targeted delivery of doxorubicin to the rat brain at therapeutic levels using MRI-guided focused ultrasound.Int J Cancer121:901–907. 7McMahon D,Bendayan R,Hynynen K(2017)Acute effects of focused ultrasound-induced increases in blood-brain barrier permeability on rat microvascular transcriptome.Sci Rep7:45657.8McDannold N,Zhang Y,Vykhodtseva N(2017)The effects of oxygen on ultrasound-induced b-brain barrier disruption in mice.Ultrasound Med Biol43:469–475. 9Itani M,Mattrey RF(2012)The effect of inhaled gases on ultrasound contrast agent longevity in vivo.Mol Imaging Biol14:40–46.10Kim I,et al.(2001)Vascular endothelial growth factor expression of intercellular adhesion molecule1(ICAM-1),vascular cell adhesion molecule1(VCAM-1),and E-selectin through nuclear factor-kappa B activation in endothelial cells.J Biol Chem276:7614–7620.11Raymond SB,Skoch J,Hynynen K,Bacskai BJ(2007)Multiphoton imaging of ultrasound/Optison mediated cerebrovascular effects in vivo.J Cereb Blood Flow Metab27:393–403.2of2|/cgi/doi/10.1073/pnas.1711544114Kovacs et al.。
New cytochrome P450 1B1, 1C1, 2Aa, 2Y3, and 2K genes from Chinese
New cytochrome P4501B1,1C1,2Aa,2Y3,and 2K genes from Chinese rare minnow (Gobiocypris rarus ):Molecular characterization,basal expression and response of rare minnow CYP1s and CYP2s mRNA exposed to the AHR agonist benzo[a]pyrene qLilai Yuan a ,b ,Biping Lv a ,b ,Jinmiao Zha b ,⇑,Zijian Wang b ,Weimin Wang a ,Wei Li b ,Lifei Zhu ba College of Fisheries,Huazhong Agricultural University,Wuhan,ChinabState Key Laboratory of Environmental Aquatic Chemistry,Research Center for Eco-Environmental Sciences,Chinese Academy of Sciences,P.O.Box 2871,Beijing 100085,Chinah i g h l i g h t sNew CYP1B1,CYP1C1,CYP2K,CYP2Aa,and CYP2Y3gene from rare minnow were cloned. Basal expression patterns of the CYPs varied in these tissues of rare minnow. High induction of CYP1transcripts by benzo[a]pyrene was observed in rare minnow. The fish CYP2Y3mRNA can be induced by the AHR agonist benzo[a]pyrene.a r t i c l e i n f o Article history:Received 30January 2013Received in revised form 16April 2013Accepted 20April 2013Available online xxxx Keywords:Rare minnow CloneCytochrome P450Benzo[a]pyrene mRNA expressiona b s t r a c tCytochrome P450(CYP450)genes play an important role in catalyzing oxidative metabolism of toxicants.Recently,CYP1subfamily were discovered and reported in fish,however,little is known regarding the CYP2isoforms in fish.In the present study,the cDNA fragments of CYP 1B1and 1C1and CYP2Aa,2Y3,and 2K of rare minnow were cloned and exhibited a high amino acid sequence identity compared with their zebrafish orthologs.Basal expression showed CYP1C1and CYP 2Aa expression were observed in all eight tissues ana-lyzed (liver,gill,intestine,kidney,spleen,brain,skin,and muscle).CYP 1A,and 1B1expression was found in all tissues except for muscle and skin.However,CYP 2Y3was expressed in liver,spleen,intestine and mus-cle whereas CYP 2K in liver,kidney and intestine.4and 100l g L À1Benzo[a]pyrene (BaP)induced patterns showed that CYP 1A,1B1and 1C1expression in liver,gill,and intestine was strongly up-regulated (p <0.05).Furthermore,CYP 2Y3was strongly induced in liver from BaP treatments (p <0.05).The high induction on mRNA level of CYP1s and CYP 2Y3by BaP could be associated with catalyzing detoxification and indicated that CYP2s may also be potential biomarker to screen AHR agonist.The high responsiveness of CYP1and 2genes suggested Chinese rare minnow is feasible to screen and assess pollution with AHR agonist.Ó2013The Authors.Published by Elsevier Ltd.All rights reserved.1.IntroductionCytochrome P450enzymes are a large superfamily of heme-pro-teins,which are involved in the phase I biotransformation of manyendogenous and exogenous compounds.About 12,456CYP450s have been identified and found in animals,plants,fungi bacteria,and viruses (Nelson,2011).In mammals,the CYP1-4isoforms are mainly involved in the metabolism of xenobiotics such as drugs,chemical carcinogens (Goldstone et al.,2010).CYP genes are regu-lated by the constitutive androstane receptor (CAR),preg-nane Âreceptor (PXR),hepatocyte nuclear factor 4(HNF-4),and aryl hydrocarbon receptor (AHR)(Xu et al.,2005;Monostory and Pascussi,2008).In the previous studies,CYP1A,1B1,1C1,1C2,and 1D1genes in fish were discovered and identified (Zanette et al.,2009).Several studies have showed the CYP1genes are regu-lated by AHR in mammals as well as in fish (except fish CYP1D1)(Shimada et al.,2002;Lin et al.,2003;Gao et al.,2011).A number of isoforms of AHR including AHR1a,AHR1b,and AHR2in fish were reported,whereas AHR2has been proved to regulate the expres-0045-6535/$-see front matter Ó2013The Authors.Published by Elsevier Ltd.All rights reserved./10.1016/j.chemosphere.2013.04.064Abbreviations:CYP450,cytochrome P450;AHR,aryl hydrocarbon receptor;BaP,benzo[a]pyrene;PAHs,polycyclic aromatic hydrocarbons.q This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License,which per-mits non-commercial use,distribution,and reproduction in any medium,provided the original author and source are credited.⇑Corresponding author.Address:State Key Laboratory of Environmental Aquatic Chemistry,Research Center for Eco-Environmental Sciences,Shuangqing Rd.18,Haidian District,Beijing 100085,China.Tel.:+861062849197;fax:+861062923543.E-mail address:jmzha@ (J.Zha).sion of CYP1A in fish (Prasch et al.,2003;Timme-Laragy et al.,2007).The CYP2family is the largest and most diverse of the verte-brate CYPs,and plays an important role in mammalian drug metabolism (Goldstone et al.,2010;Kirischian et al.,2011).16,17,27,and 47CYP2isforms were reported in human,rabbit,opos-sum,and zebrafish,respectively (Kirischian et al.,2011).In mam-mals,CYP2S1is induced by dioxin and the induction is mediated by the AHR.Full complement of CYP genes were identified only in zebrafish (Goldstone et al.,2010).The CYP 2X1in channel catfish (Mosadeghi et al.,2007);CYP 2M1and CYP 2K1in rainbow trout (Yang et al.,1998);CYP 2Ps and CYP 2N in killifish (Yang et al.,1998);and CYP 2Y3,CYP 2C33-like and CYP 2P1-like in Atlantic cod (Olsvik et al.,2009)were identified.Phylogenetic tree analysis showed that CYP 2Y3and CYP 2Y4of zebrafish shared synteny with a cluster of CYP2genes including CYP2S1of human,whereas function of CYP2Y3and CYP2Y4in fish is unknown (Goldstone et al.,2010).Though some CYP2s were discovered and identified in fish,their function,tissue distribution,and induced patterns were not well-characterized.Polycyclic aromatic hydrocarbons (PAHs)are ubiquitous organic contaminants arising from incomplete combustion or pyrolysis of organic material.Exposure to PAHs has been associated with a vari-ety of toxicity,including developmental,immunological disorders,mutagenesis and carcinogenesis (Nicol et al.,1995;De Jong et al.,1999;Timme-Laragy et al.,2007).It is widely held that the under-standing of the toxicity caused by PAHs requires the study of AHR and its downstream genes including the CYP450enzymes (Mat-sumoto et al.,2007).In addition,a number of studies have linked CYP1A induction with PAH metabolites,DNA adducts,and tumor formation in fish (Wang et al.,2010),and the CYP1genes are in-volved in carcinogen bioactivation of PAHs including benzo[a]pyr-ene (BaP)(Ghanayem et al.,2000).BaP is one of the most studied environmentally relevant PAHs,and has been considering as an AHR agonist based on its ability to bind the receptor.BaP is most of-ten identified with cancer due to being a well-established chemical mutagen (Thompson et al.,2010).Based on this knowledge,BaP is suitable as a reference chemical to study responses of CYPs in fish.Most previous studies have reported that induced patterns of CYP1s in fish including zebrafish (Jönsson et al.,2007),killifish (Zanette et al.,2009),and three-spined stickle blacked (Gao et al.,2011)exposed to PCB126.Though CYP1A gene of rare minnow were identified in previous studies (Liu et al.,2008),the molecular characterization and basal expression and induced expression of other CYP1s,especial CYP2s,were not investigated.The aim of present study was to identify CYP 1B1,1C1and CYP2Aa,2Y3,2K genes in rare minnow and to characterize their basal expression pattern in eight tissues (liver,gill,intestine,kidney,spleen,brain,skin,and muscle)and,to investigate the BaP induced effects of the CYPs in liver,gill,and intestine of rare minnow,as these tissues (liver,gill,and intestine)are mainly involved with the uptake and metabolism of pollutants (Costa et al.,2012).The rare minnow is distributed mostly in the upstream region of the Yangtze River and in the Sichuan Province of China.It is consid-ered to be an appropriate species for the assessment of endocrine disrupting chemicals due to its small size,ease of culture,short life cycle and prolific egg production with high fertilization and hatch-ing rates (Zha et al.,2007;Li et al.,2009;Yang et al.,2010).2.Materials and methods 2.1.ChemicalsBaP (purity >98%)and acetone were purchased from Sigma (Chemical Co.,USA).Stock solution of BaP was prepared by dilution in acetone.2.2.Test fish and culture conditionsThe brood stock of rare minnow was raised in a flow through system with dechlorinated tap water (pH:7.2–7.6;hardness:44–61mg CaCO 3L À1;and temperature:25±1°C)with a photo period of 16:8h (light:dark)and has been used for testing chemicals in our laboratory for more than 10years (Zha et al.,2007;Li et al.,2009).Fish were fed a commercial food pellet (Trea,Germany)at a rate of 0.1%body weight per day and newly hatched brine shrimp (Artemia nauplii )two times daily.2.3.Cloning of cDNA fragments of CYP 1B1,1C1,2Aa,2Y3,and 2K by RT-PCRTotal RNA was isolated from the liver of rare minnow using SV Total RNA Isolation System following the manufacturer’s protocol (promega,USA).Then RNA samples were dissolved in ribonucle-ase-free water and stored at À80°C until the process of reverse-transcriptase polymerase chain reaction.The protocol of reverse-transcriptase polymerase chain reaction is according to previous report of our laboratory (Li et al.,2009).The reverse transcription reaction mixtures containing 10l L of to-tal RNA,2l L (0.05l g l L À1)of Oligo(dT)15,and diethylpyrocarbon-ate-treated water (a total volume of 12l L),were heated to 70°C for 5min and quickly chilled on ice.After cooling,50mM Tris–HCl buffer (pH 8.3),75mM KCl,10mM dithiothreitol,3mM MgCl 2,2mM deoxy-nucleotide triphosphate (10mM each),40units of RNAasin (RNAase inhibitor;Promega),and 200U of Moloney Mur-ine Leukemia Virus Reverse Transcriptase (Promega)were added to a total volume of 25l L and incubated for 1h at 37°C.Thereafter,the reaction mixture was heated to 70°C for 10min to inactivate the reverse transcription.To obtain cDNA fragments of the CYP genes (CYP1B1,1C1,and 2K),the primer pairs were designed based on highly conserved re-gions according to known sequences of other fish species available in GenBank;and the primer pairs of CYP 2Aa and 2Y3were de-signed based on zebrafish CYP 2Aa and 2Y3sequences,respectively (Table 1).The detailed information of the PCR experiment was as follows,each 50l L DNA amplification reaction contained polymer-ase chain reaction buffer (20mM Tris–HCl,pH 8.4,50mM KCl,1.5mM MgCl 2),200l mol dNTP,20pmol of each gene specific pri-mer,1l L of a 20-fold dilution of total cDNA,and 2.5U Taq DNA polymerase (Invitrogen).The thermocycle program included a denaturation step at 94°C (8min);35cycles of 94°C (30s),55°C (30s),and 72°C (30s);and a final elongation step at 72°C (10min).The amplification PCR products were analyzed by 1.5%agarose gel electrophoresis.The bands of expected size were ex-cised and purified using an Agarose Gel DNA Purification Kit Ver-sion 2.0(TaKaRa).The isolated fragments were inserted into pMD 19-T Vector (TaKaRa)and transformed into Escherichia coli (DH 5A ).The partial cDNAs were sequenced and compared by BLAST to the sequences available in GenBank.2.4.The experimental design for exposure and tissue distribution of the CYPsHealthy rare minnows (n =72),about 4month old and the off-spring from the same pair of brood stock,were used in this exper-iment.The body weights and lengths were 0.55±0.17g and 37.23±3.25mm,respectively.Reproductively immature fish were used to minimize the effects of gender and reproductive state dif-ferences on the expression of CYP450enzymes (Willett et al.,1997;Barber et al.,2007).The fish were exposed to various concentra-tions (4,100l g L À1)of BaP.A vehicle treatment containing acetone served as a control (Jönsson et al.,2007),the ratio of vehicle to water was 1:10,000(v/v).2L.Yuan et al./Chemosphere xxx (2013)xxx–xxxThe fish were acclimated to the exposure aquarium (3L)for 7days before initiate of the experiment.Then,the fish were ran-domly distributed into 3experimental groups,each group contained 3replicate aquariums and each replicate aquarium included 8fish.During the experiment,water temperature was maintained at 25±1°C and pH at 7.0±0.2.Fish were fed two times a day with newly hatched brine shrimp.After 12days exposure,the fish were sacrificed and the tissues (liver,gill and intestine)were excised and immediately frozen in liquid nitrogen and stored at À80°C.To determine tissue distribution of the CYP genes,another 15immature fish (4month old)were used,and the fish were randomly distributed into 3groups.Thereafter,the fish were sacrificed and the tissues (liver,gill,intestine,kidney,spleen,brain,skin,and mus-cle)were excised from each group (containing five replicates),and immediately frozen in liquid nitrogen and stored at À80°C.2.5.Determining the CYP genes expression by real-time PCRTo determine tissue distribution of the CYPs and assess their tran-scriptional effects following BaP exposure,real-time PCR was per-formed in a Mx3005P real-time quantitative polymerase chain reaction system (Stratagene,USA).The PCR reaction mixtures (25l L)consisted of Brilliant II SYBR Green QPCR master mix,300nM forward primer and 300nM reverse primer.The primer pairs used for real-time PCR were shown in Table 1.For each sample,gene expression was analyzed in triplicate with the following protocol:95°C for 8min and 40cycles of 30s at 95°C,30s at 57°C and 30s at 72°C.Melt curve analysis was performed on the PCR products at the end of each PCR run to ensure that a single product was amplified.The basal mRNA expression of the CYP genes was calculated by the 2ÀD Ct method (Schmittgen and Livak,2008)using b -actin as the reference gene.For fold-change in expression after exposure the 2ÀDD Ct method was used.The mean value of these triplicate mea-surements were used for calculations of mRNA expressions.2.6.Statistical analysisStatistical analyses were performed with the SPSS (version 13.0)and OriginPro (version 8.0).All quantitative data are expressed asthe mean ±S.E.of the mean (S.E.M.).The differences among tran-script levels of the CYPs in different tissues,and the means be-tween control and BaP exposure groups were evaluated using one-way ANOVA (p <0.05),followed by Dunnett’s test for multiple comparisons.3.Results3.1.Cloning and analysis of cDNA fragments of CYP 1B1,1C1,2Aa,2Y3,and 2KThe CYP 1B1,1C1,2Aa,2Y3,and 2K partial cDNAs were isolated and cloned from rare minnow.The partial fragments of these CYPs have been sequenced and submitted to GenBank.Accession num-bers are KC136245for CYP1B1,KC136246for CYP1C1,KC136248for CYP2Aa,KC165029for CYP2Y3,and KC136247for CYP2K.Sequence alignment and phylogenetic analysis revealed the pre-dicted amino acid sequences of the CYP1s in rare minnow have high similarities with those of other fish and mammalian species (Fig.1).For three CYP2s,the partial CYP2K amino acid sequences display high identity with other sequences of fish CYP2Ks (Fig.1).CYP2Y3and CYP2Aa share homology with mammalian CYP2Cs and CYP2Js,respectively (Fig.1).Moreover,the corresponding CYP2s in rare min-now and zebrafish display high degrees of amino acid sequence iden-tity.The predicted amino acid sequences of rare minnow share 86%,87%,,87%,75%,and 91%pair-wise identity with CYP1B1,CYP1C1,CYP2Aa2,CYP2Y3,and CYP2K19of zebrafish,respectively (Fig.1).3.2.Tissue distribution pattern of CYP1and CYP2isoformsThe tissue distribution patterns of CYP 1A,1B1,1C1,2Aa,2Y3,and 2K in eight different tissues (liver,gill,intestine,kidney,spleen,brain,skin,and muscle)in immature rare minnow were as-sessed by real-time RT-PCR.The results showed different tissue expression patterns for these CYPs (Fig.2).Expression of CYP1C1and 2Aa was observed in all eight organs,both CYP1A and CYP1B1transcripts were detected in all the tissues except for muscle and skin (Fig.2).However,the expression of CYP 2Y3and 2K was observed only in four organs (liver,muscle,Table 1Primer sequences used for cloning of CYP 1B1,1C1,2Y3,2K,and 2Aa,and quantification of CYP1and CYP2genes expression by real-time PCR in rare minnow.Gene Sequence (50?30)Product size (bp)Genbank accession no.Cloning CYP1B1F:CCACAGCCAACATCCAGA 239KC136245R:CATCCAGGGCATCACATC CYP1C1F:ATTGAGCACGGGAAAGAG 232KC136246R:GGACGAAACTGGTGAAGC CYP2K F:ACATTGTGCCCGTTGAGTC 266KC136247R:AGTTGTGAAAGCGGTAGCT CYP2Aa F:TCGGGTGTTAGAGGAAAGC 247KC136248R:CTCATTGGCGTTCTGTTTGA CYP2Y3F:TAGACATCACGCCATCTTAAG 367KC165029R:CGTCAGTGAAGGGAAGAGACT Real-time PCR b -actin F:CAGGGCGTGATGGTGGGGAT 226DQ539421R:GGTTGGCTTTGGGGTTGAG CYP1A F:ATCATCGGAAATGTGCTGG 170EU106660.1R:GAGAACTCCTCGCCCTGTT CYP1B1F:TTCTCCACAGCCAACATC 138R:CGACACCACTAAATACCG CYP1C1F:CGGGACAGGACACGATGT 157R:TCCAGGTATGCGAGGTTG CYP2Aa F:TGTTTGGAGATCGCTTCG 167R:ATTGGCGTTCTGTTTGAC CYP2K F:AATGGGAGAAACCAAACA 163R:TGAAGGAGGGAAGTAAAGA CYP2Y3F:CCAGAGACCACAGCACAACT 156R:GCGTCAGTGAAAGTAGGGACTL.Yuan et al./Chemosphere xxx (2013)xxx–xxx3Phylogenetic tree showing the relationships of rare minnow(A)CYP1B1,(B)CYP1C1,(C)CYP2Aa,(D)CYP2Y3,(E)CYP2K from other vertebrates.The phylogenetic was carried out by MEGA5on deduced amino acid sequences.Sequences were obtained from the NCBI GenBank database.Branch length is proportional to estimateddivergence across each branch.One thousand bootstrap replicates were performed and are expressed as percentages noted on each node.intestine,and spleen)and three organs(liver,kidney,and intestine), respectively(Fig.2).The expression of all six CYP genes(CYP1A,1B1, 1C1,2Aa,2Y3,and2K)was observed in liver and intestine,and all these genes transcription was detected in the kidney except the CYP2Y3.CYP1A shows the highest and lowest levels of mRNA expression in the liver and brain,respectively.However,CYP1B1 shows the highest level of mRNA expression in the brain,but the expression was low in liver.Expression level of CYP1C1was much higher in muscle and skin than the other organs.For the CYP2iso-forms,the highest levels of CYP2Y3,2K,and2Aa transcripts were ob-served in spleen,liver,and intestine,respectively(Fig.2).The basal levels of the six CYP1and CYP2genes were also com-pared within a tissue(liver,gill and intestine)and data are pre-sented as a percentage of the highest CYP transcript in each tissue(Fig.3).The liver showed a significantly higher expression of CYP2than of the CYP1genes(100%,24.07%,21.46%of CYP2Y3, 2Aa,and2K,respectively;12.46%,5.74%,1.08%of CYP1C1,1B1, and1A,respectively).In the liver,CYP2Y3was most highly ex-pressed and CYP1B1showed a weak expression.CYP1C1was induction pattern varied in these tissue.For CYP1A,the highest induction was observed in the liver and gill(33-and14-fold versus the control,Fig.4).CYP1B1showed highest induction in intestine(27-fold),while CYP1C1showed highest induction in gill(19-fold).For three CYP2genes,CYP2Y3only in liver was significantly in-duced from4and100l g LÀ1BaP treatments compared with thecontrols(p<0.05)(Fig.5).The level of CYP2Y3induction in liverby4and100l g LÀ1BaP were2.6-and6-fold over the control,respectively.However,no significantly induction of CYP2Aa,andCYP2K in all three tissues from BaP treatments was observed com-pared with the controls(Fig.5).4.Discussion4.1.Identification of new CYP1and CYP2genes in rare minnowIn our study,five previously unreported CYP genes in rare min-now were cloned and identified.The partial fragments of theseCYP1A,1B1,1C1,2Aa,2Y3,and2K in rare minnow shown as genes among tissues.The relative expressionvalue obtained by subtracting Ct of b-actin mRNA from Ct of the target mRNA).Data are presented astranscript.Data are shown as means±S.E.A statistical difference between groups at p<0.05(n=3,ANOVA)L.Yuan et al./Chemosphere xxx(2013)xxx–xxx5minnow and zebrafish displays high degree of amino acid sequence similarity.4.2.Basal expression of the CYP1and CYP2gene in adult rare minnowCYP genes play an important role to regulate physiological func-tions.In this study,different tissue expression profiles of these CYP1and2s have been observed in rare minnow.The transcripts of the CYP1s were detected almost in all the tissues examined, and the basal expression patterns varied in the tissues,indicating that the three CYP1s have tissue-specific regulation and possibly different functions,which was similar to those previous studies in otherfish(Jönsson et al.,2007;Zanette et al.,2009;Gao et al., 2011).Our results shows the highest and lowest levels of CYP1A mRNA expression were in the liver and brain,respectively,which largely agreed with previous studies with zebrafish(Jönsson et al.,2007),stickleback(Gao et al.,2011),and killifish(Zanette et al.,2009).However,in this study,the expression of CYP1B1in rare minnow was high in the brain and low in the liver.This may due to the different endogenous functions of CYP1A and CYP1B1. CYP1A has been proved to play a role in metabolizing endogenous AHR ligands,while CYP1B1can catalyze the formation of retinoic acid and may play a role in retinoic acid mediated patterning dur-ing embryogenesis(Gao et al.,2011).In previous studies,the tran-scripts of CYP1A and CYP1B1in mammals and otherfishes were detected in muscle or skin(Yengi et al.,2003;Lee et al.,2005;Er-dog˘an et al.,2011),whereas the transcripts of CYP1A and CYP1B1 in rare minnow were not detected(Fig.2).The species differences may account for this conflict results.The level of CYP1C1of rare minnow in muscle and skin were much higher than the otherof CYP1and CYP2transcript of rare minnow in a given tissue:liver,gill and intestine.Data are presented as a percentage of the highest are shown as means±S.E.A statistical difference between groups at p<0.05(n=3,ANOVA)is indicated by differences in the lettersinduction of CYP1A,1B1,and1C1in rare minnow liver,gill,and intestine following BaP(4,100l g LÀ1)exposure.Bars represent the relative versus control(acetone)fish.b-Actin was used for normalization.Values are presented as the mean±S.E.A statistical difference between indicated by differences in the letters above the bars.organs(Fig.2).The high mRNA level of CYP1C1in muscle and skin of rare minnow suggests that CYP1C1may play an important role in xenobiotic metabolism.For the CYP2s,the expression of CYP2K was detected only in three tissues(liver,kidney,and intestine),previous research has reported that the orthologous CYP2K6transcript was only detected in liver and ovary of zebrafish(Wang-Buhler et al.,2005),and the CYP2K1mRNA was mainly expressed in the liver and trunk kidney of rainbow trout(Buhler et al.,1994).Similar with CYP2K,the expression of CYP2Y3was observed only in liver,muscle,intestine, and spleen(Fig.2).Interestingly,the CYP2Aa was present in all the tissues analyzed similar with CYP1s.The expression of all six CYP genes was observed in liver,intes-tine and kidney,except no expression of CYP2Y3in kidney,these tissues are all located in the abdominal cavity of rare minnow.This could be linked with the role of these tissues in nutrient uptake and processing of body waste products, e.g.detoxification of endogenous metabolites and food derived pollutants(Jönsson et al.,2007;Zanette et al.,2009).4.3.Response of the CYP1s and CYP2s to BaPCYP1enzymes are prominent in metabolism of many toxicants including PAHs,and expression level is a major factor influencing the role of CYPs in substrate oxidation and effects in vivo(Jönsson et al.,2007).In the present study,all three CYP1genes transcrip-tion were significantly elevated in liver,gill and intestine of rare minnow exposed to BaP for12days(Fig.4).Previous studies have observed the elevated expression of the CYP1s(CYP1A,1B1and 1C1)by AHR agonist in various tissues offish(Jönsson et al., 2007,2010;Zanette et al.,2009;Gao et al.,2011;Dorrington et al.,2012).However,level of induction were significantly differ-ent among thosefish and rare minnow,e.g.about10-,200-,and 400-fold for CYP1A,1B1and1C1in killifish liver(Zanette et al., 2009).This difference might due to different species,chemicals, and exposure methods.Since the chemical was administered via intraperitoneal injection to killifish and via water to rare minnow, thus the difference in induction may due to different bioavailabil-ity as they followed different exposure methods(Jönsson et al., 2010).The BaP induction pattern of these CYP1s varied in the tissues of rare minnow,suggesting tissue-specific regulation and different function of these enzymes.Previous studies also showed different induction patterns of the CYP1s by AHR agonists(Zanette et al., 2009;Dorrington et al.,2012).It is widely held thatfish CYP1s are regulated by AHR,ligand-activated AHR dimerizes with the aryl hydrocarbon receptor nuclear translocator(ARNT)and binds to xenobiotic response elements(XREs),causing the induction of CYP1genes.The number of potential XREs upstream of the CYP1A genes is more than that in the CYP1B1and CYP1Cs in zebrafish,the number and position of XREs in the CYP1genes may influence the regulation of these genes(Zeruth and Pollenz,2005;Jönsson et al., 2007).In addition,in vitro study results in MCF-7cells indicated that the differential requirement for BRG-1(a co-activator protein) may be a consequence of different nucleosomal configurations over the CYP1A1and CYP1B1genes,furthermore,maybe related to their different degrees of induction by dioxin(Taylor et al., 2009).Therefore,the different induction of the CYP1s by BaP in rare minnow may due to the differential requirement of the co-activators.The CYP2family is the largest and most diverse of the verte-brate CYPs and plays an important role in mammalian drug metab-olism(Kirischian et al.,2011).In the present study,BaP significantly elevated the expression of CYP2Y3in rare minnow li-ver(Fig.5).Nonylphenol and bisphenol A exposure produced a sig-nificant reduction of CYP2Y3in Atlantic cod(Olsvik et al.,2009), however,the regulating mechanism is unclear.Previous study showed zebrafish CYP2Y3had a syntenic relationship to a cluster of CYP2genes including CYP2S1(Goldstone et al.,2010).Moreover, CYP2S1is induced by AHR agonist in mammals(Saarikoski et al., 2005),which was consistent with our result of CYP2Y3in rare min-now.Furthermore,to confirm the CYP2Y3is regulated by AHR in fish,further studies should be done to discover whether there are some XREs in this gene like the CYP1A gene.Despite this,the CYP2Y3gene may serve as a potential molecular biomarker of AHR agonist(e.g.BaP).CYP2Aa,2Y3,and2K in rare minnow liver,gill,and intestine following BaP(4,100l g LÀ1)exposure.Bars represent the (acetone)fish.b-Actin was used for normalization.Values are presented as the mean±S.E.A statistical difference by differences in the letters above the bars.5.ConclusionIn summary,we identified and cloned the partial cDNAs of pre-vious unreported CYP1B1,CYP1C1,CYP2Aa,CYP2Y3,and CYP2K in rare minnow.Basal expression of the CYP1s and the CYP2Aa were detected almost in all the tissues examined,while the expression of CYP2K and CYP2Y3were only detected in several tissues of rare minnow.BaP significantly induced the expression of the CYP1s in all the tissues examined,as well as the CYP2Y3in the liver.The ba-sal and the induced expression varied in these tissues,indicating tissue-specific regulation and different function of these enzymes in rare minnow.The strong induction of CYP2Y3mRNA level by BaP indicates that it may be a downstream gene of AHR and a po-tential molecular biomarker of AHR agonist.The responsiveness of these CYP genes in rare minnow means the fish is a suitable model to screen environmental AHR agonist (e.g.PAHs).AcknowledgementsThis work was supported by National Basic Research Program of China (2009CB421605);the National Natural Science Foundation of China (21007086);the National High-tech R&D Program (2012AA06A302).ReferencesBarber,D.S.,McNally,A.J.,Garcia-Reyero,N.,Denslow,N.D.,2007.Exposure to p,p 0-DDE or dieldrin during the reproductive season alters hepatic CYP expression in largemouth bass (Micropterus salmoides ).Aquat.Toxicol.81,27–35.Buhler,D.R.,Yang,Y.H.,Dreher,T.W.,Miranda,C.L.,Wang,J.L.,1994.Cloning andsequencing of the major rainbow trout constitutive cytochrome P450(CYP2K1):identification of a new cytochrome P450gene subfamily and its expression in mature rainbow trout liver and trunk kidney.Arch.Biochem.Biophys.312,45–51.Costa,J.,Reis-Henriques,M.A.,Castro,L.F.C.,Ferreira,M.,2012.Gene expressionanalysis of ABC efflux transporters,CYP1A and GST a in Nile tilapia after exposure to benzo(a)p.Biochem.Phys.C 155,469–482.De Jong,W.H.,Kroese, E.D.,Vos,J.G.,Van Loveren,H.,1999.Detection ofimmunotoxicity of benzo [a]pyrene in a subacute toxicity study after oral exposure in rats.Toxicol.Sci.50,214–220.Dorrington,T.,Zanette,J.,Zacchi,F.L.,Stegeman,J.J.,Bainy,A.C.D.,2012.Basal and 3-methylcholanthrene-induced expression of cytochrome P4501A,1B and 1C genes in the Brazilian guppy (Poecilia vivipara ).Aquat.Toxicol.125,106–113.Erdog˘an,O.,Ceyhun,S.B.,Ekinci, D.,Aksakal, E.,2011.Impact of deltamethrin exposure on mRNA expression levels of metallothionein A,B and cytochrome P4501A in rainbow trout muscles.Gene 484,13–17.Gao,K.,Brandt,I.,Goldstone,J.V.,Jönsson,M.E.,2011.Cytochrome P4501A,1B,and1C mRNA induction patterns in three-spined stickleback exposed to a transient and a persistent p.Biochem.Phys.C 154,42–55.Ghanayem,B.I.,Wang,H.,Sumner,S.,ing cytochrome P450gene knock-outmice to study chemical metabolism,toxicity,and carcinogenicity.Toxicol.Pathol.28,839–850.Goldstone,J.,McArthur,A.,Kubota,A.,Zanette,J.,Parente,T.,Jönsson,M.,Nelson,D.,Stegeman,J.,2010.Identification and developmental expression of the full complement of Cytochrome P450genes in Zebrafish.BMC Genom.11,643.Jönsson,M.E.,Orrego,R.,Woodin,B.R.,Goldstone,J.V.,Stegeman,J.J.,2007.Basaland 3,30,4,40,5-pentachlorobiphenyl-induced expression of cytochrome P4501A,1B and 1C genes in zebrafish.Toxicol.Appl.Pharm.221,29–41.Jönsson,M.E.,Gao,K.,Olsson,J.A.,Goldstone,J.V.,Brandt,I.,2010.Inductionpatterns of new CYP1genes in environmentally exposed rainbow trout.Aquat.Toxicol.98,311–321.Kirischian,N.,McArthur,A.G.,Jesuthasan,C.,Krattenmacher,B.,Wilson,J.Y.,2011.Phylogenetic and functional analysis of the vertebrate cytochrome p4502family.J.Mol.Evol.72,56–71.Lee,Y.M.,Williams,T.D.,Jung,S.O.,Lee,J.S.,2005.CDNA cloning and expression of acytochrome P4501A (CYP1A)gene from the hermaphroditic fish Rivulus marmoratus .Mar.Pollut.Bull.51,769–775.Li,W.,Zha,J.,Spear,P.A.,Li,Z.,Yang,L.,Wang,Z.,2009.Changes of thyroid hormonelevels and related gene expression in Chinese rare minnow (Gobiocypris rarus )during 3-amino-1,2,4-triazole exposure and recovery.Aquat.Toxicol.92,50–57.Lin,P.,Hu,S.W.,Chang,T.H.,2003.Correlation between gene expression of arylhydrocarbon receptor (AhR),hydrocarbon receptor nuclear translocator (Arnt),cytochromes P4501A1(CYP1A1)and 1B1(CYP1B1),and inducibility of CYP1A1and CYP1B1in human lymphocytes.Toxicol.Sci.71,20–26.Liu,Y.,Wang,J.,Wei,Y.,Zhang,H.,Dai,J.,2008.Molecular characterization ofcytochrome P4501A and 3A and the effects of perfluorooctanoic acid on their mRNA levels in rare minnow (Gobiocypris rarus )gills.Aquat.Toxicol.88,183–190.Matsumoto,Y.,Ide,F.,Kishi,R.,Akutagawa,T.,Sakai,S.,Nakamura,M.,Ishikawa,T.,Fujii-Kuriyama,Y.,Nakatsuru,Y.,2007.Aryl hydrocarbon receptor plays a significant role in mediating airborne particulate-induced carcinogenesis in mice.Environ.Sci.Technol.41,3775–3780.Monostory,K.,Pascussi,J.,2008.Regulation of drug-metabolizing humancytochrome P450s.Acta Chim.Slov.55,20.Mosadeghi,S.,Furnes, B.,Matsuo, A.Y.O.,Schlenk, D.,2007.Expression andcharacterization of cytochrome P4502X1in channel catfish (Ictalurus punctatus ).BBA Gen.Subjects 1770,1045–1052.Nelson,D.R.,2011.Progress in tracing the evolutionary paths of cytochrome P450.BBA Proteins Proteom.1814,14–18.Nicol, C.J.,Harrison,M.L.,Laposa,R.R.,Gimelshtein,I.L.,Wells,P.G.,1995.Ateratologic suppressor role for p53in benzo[a]pyrene–treated transgenic p53-deficient mice.Nat.Genet.10,181–187.Olsvik,P.A.,Lie,K.K.,Sturve,J.,Hasselberg,L.,Andersen,O.K.,2009.Transcriptionaleffects of nonylphenol,bisphenol A and PBDE-47in liver of juvenile Atlantic cod (Gadus morhua ).Chemosphere 75,360–367.Prasch,A.L.,Teraoka,H.,Carney,S.A.,Dong,W.,Hiraga,T.,Stegeman,J.J.,Heideman,W.,Peterson,R.E.,2003.Aryl hydrocarbon receptor 2mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish.Toxicol.Sci.76,138–150.Saarikoski,S.T.,Rivera,S.P.,Hankinson,O.,Husgafvel-Pursiainen,K.,2005.CYP2S1:a short review.Toxicol Appl.Pharm.207,62–69.Schmittgen,T.D.,Livak,K.J.,2008.Analyzing real-time PCR data by the comparativeCT method.Nat.Protoc.3,1101–1108.Shimada,T.,Inoue,K.,Suzuki,Y.,Kawai,T.,Azuma, E.,2002.Arylhydrocarbonreceptor-dependent induction of liver and lung cytochromes P4501A1,1A2,and 1B1by polycyclic aromatic hydrocarbons and polychlorinated biphenyls in genetically engineered C57BL/6J mice.Carcinogenesis 23,1199–1207.Smith,E.M.,2009.Cytochrome P450drug metabolism and protein induction andinhibition in fish liver microsomes.Open Access Dissertations and Thesis.Paper 4614.Taylor,R.T.,Wang,F.,Hsu,E.L.,Hankinson,O.,2009.Roles of coactivator proteins indioxin induction of CYP1A1and CYP1B1in human breast cancer cells.Toxicol.Sci.107,1–8.Thompson,E.D.,Burwinkel,K.E.,Chava,A.K.,Notch,E.G.,Mayer,G.D.,2010.Activityof Phase I and Phase II enzymes of the benzo [a]pyrene transformation pathway in zebrafish (Danio rerio )following waterborne exposure to p.Biochem.Phys.C 152,371–378.Timme-Laragy,A.R.,Cockman,C.J.,Matson,C.W.,Di Giulio,R.T.,2007.Synergisticinduction of AHR regulated genes in developmental toxicity from co-exposure to two model PAHs in zebrafish.Aquat.Toxicol.85,241–250.Wang,L.,Camus,A.C.,Dong,W.,Thornton,C.,Willett,K.L.,2010.Expression ofCYP1C1and CYP1A in Fundulus heteroclitus during PAH-induced carcinogenesis.Aquat.Toxicol.99,439–447.Wang-Buhler,J.,Lee,S.,Chung,W.,Stevens,J.,Tseng,H.,Hseu,T.,Hu,C.,Westerfield,M.,Yang,Y.,Miranda,C.,2005.CYP2K6from zebrafish (Danio rerio ):cloning,mapping,developmental/tissue expression,and aflatoxin B1activation by baculovirus expressed p.Biochem.Phys.C 140,207–219.Willett,K.L.,McDonald,S.J.,Steinberg,M.A.,Beatty,K.B.,Kennicutt,M.C.,Safe,S.H.,1997.Biomarker sensitivity for polynuclear aromatic hydrocarbon contamination in two marine fish species collected in Galveston Bay.Texas.Environ.Toxicol.Chem.16,1472–1479.Xu,C.,Li,C.Y.T.,Kong,A.N.T.,2005.Induction of phase I,II and III drug metabolism/transport by xenobiotics.Arch.Pharm.Res.28,249–268.Yang,Y.H.,Wang,J.L.,Miranda, C.L.,Buhler, D.R.,1998.CYP2M1:cloning,sequencing,and expression of a new cytochrome P450from rainbow trout liver with fatty acid (omega-6)-hydroxylation activity.Arch.Biochem.Biophys.352,271–280.Yang,L.,Zha,J.,Li,W.,Li,Z.,Wang,Z.,2010.Atrazine affects kidney and adrenalhormones (AHs)related genes expressions of rare minnow (Gobiocypris rarus ).Aquat.Toxicol.97,204–211.Yengi,L.G.,Xiang,Q.,Pan,J.,Scatina,J.A.,Kao,J.,Ball,S.E.,Fruncillo,R.,Ferron,G.,Roland Wolf,C.,2003.Quantitation of cytochrome P450mRNA levels in human skin.Anal.Biochem.316,103–110.Zanette,J.,Jenny,M.J.,Goldstone,J.V.,Woodin, B.R.,Watka,L.A.,Bainy, A.C.D.,Stegeman,J.J.,2009.New cytochrome P4501B1,1C2and 1D1genes in the killifish Fundulus heteroclitus:basal expression and response of five killifish CYP1s to the AHR agonist PCB126.Aquat.Toxicol.93,234–243.Zeruth,G.,Pollenz,R.S.,2005.Isolation and characterization of a dioxin-inducibleCYP1A1promoter/enhancer region from zebrafish (Danio rerio ).Zebrafish 2,197–210.Zha,J.,Wang,Z.,Wang,N.,Ingersoll, C.,2007.Histological alternation andvitellogenin induction in adult rare minnow (Gobiocypris rarus )after exposure to ethynylestradiol and nonylphenol.Chemosphere 66,488–495.8L.Yuan et al./Chemosphere xxx (2013)xxx–xxx。
responsetothetechnicalcheckresults
Dear Editor,Thank you for your useful comments and suggestions on the language and structure of our manuscript. We have modified the manuscript accordingly, and detailed corrections are listed below point by point:Language:1) In its current state the level of English throughout your manuscript does notmeet the journal's desired standard . Please check the manuscript and refine the language carefully.We have revised the whole manuscript carefully and tried to avoid any grammar or syntax error. We believe that the language is now acceptable for the review process.Technical:1)No abbreviations are allowed in the title.Abbreviation in the title has been replaced by the full name.2)No abbreviations are allowed in the keywords.Abbreviation in the keywords has been replaced by the full name.3)The manuscript source file has line numbers, but our system automatically addsline numbers to the PDF. Please save a version of the file locally that has noline numbers, and Edit submission to add this file and remove the incorrect one.The manuscript has been resubmitted to your journal. We look forward to your positive response.Line numbers in the manuscript source file has been deleted.Sincerely,Jiansheng Li。
Optik the Rights and Access information form
RIGHTS AND ACCESS Home | HelpFunding BodyOpen AccessLicensesRightsPublishing AgreementOpen Access FeePaymentOrder SummaryConfirmationArticle:Optical wave breaking and soliton trains generation due to cross-phase modulation in an optical fiber with quintic nonlinearityCorresponding author:Prof. Xianqiong ZhongE-mail address:xianqiongzhong@Journal:Optik - International Journal for Light and Electron OpticsOur reference:IJLEO56584PII:S0030-4026(15)01492-8DOI:10.1016/j.ijleo.2015.10.124SELECT YOUR FUNDING BODYElsevier has established agreements with the following funding bodies, which help authors comply with their funding body open access or public access policy.If the research reported was sponsored by multiple funding bodies in this list please contact your primary funding body to ensure the right funding body is selected.For more details see the Elsevier funding bodies pages.FUNDING BODY AGREEMENTSResearch reported in the article:received no funding or funding from source other than belowwas supported by Arthritis Research UKwas supported by Bloodwisewas funded by Breast Cancer Campaignwas supported by the BHF – British Heart Foundationwas funded by the CRUK – Cancer Research UKwas funded by the CSO – Chief Scientist Officewas supported by the UK DFID – Department for International Developmentwas funded by the DH – Department of Health UKwas funded by the Dunhill Medical Trustwas supported by European Molecular Biology Laboratorywas supported by European Research Councilwas supported by the MNDA – Motor Neurone Disease Associationwas funded by the US NIH – National Institutes of Healthwas performed by a US National Institutes of Health (NIH) employeewas supported by PUK - Parkinson's UKwas supported by a UK university with a prepayment agreement for AHRC-funded researchwas supported by a UK university with a prepayment agreement for BBSRC-funded researchwas supported by a UK university with a prepayment agreement for ESRC-funded researchwas supported by a UK university with a prepayment agreement for EPSRC-funded researchwas supported by a UK university with a prepayment agreement for MRC-funded researchwas supported by a UK university with a prepayment agreement for NERC-funded researchwas supported by a UK university with a prepayment agreement for STFC-funded researchsupported by a UK university with a prepayment agreement and not funded by an organisation listed herewas supported by the RCUK – AHRC – Arts & Humanities Research Councilwas supported by the RCUK – BBSRC – Biotechnology and Biological Sciences Research Councilwas supported by the RCUK – ESRC – Economic and Social Research Councilwas supported by the RCUK – EPSRC – Engineering and Physical Sciences Research Councilwas supported by the RCUK – MRC – Medical Research Council UKwas supported by the RCUK – NERC – Natural Environment Research Councilwas supported by the RCUK – STFC – Science and Technology Facilities Councilwas supported by Telethon (Italy)was supported by UNU-WIDERwas supported by Wellcome Trustwas supported by the World Bankwas supported by WHO - World Health Organizationwas conducted by an employee of WHO – World Health OrganisationPrivacy Policy Terms & Conditions HelpFunding BodyOpen AccessRightsPublishing AgreementOrder SummaryConfirmationArticle:Optical wave breaking and soliton trains generation due to cross-phase modulation in an optical fiber with quintic nonlinearityCorresponding author:Prof. Xianqiong ZhongE-mail address:xianqiongzhong@Journal:Optik - International Journal for Light and Electron OpticsOur reference:IJLEO56584PII:S0030-4026(15)01492-8DOI:10.1016/j.ijleo.2015.10.124FUNDING BODY OPEN ACCESS POLICYResearch reported in the article was not funded by any of the listed funding bodies.OPEN ACCESSThis is a hybrid journal which publishes both subscription and gold open access articles. Authors are free to choose below how they would like readers to access their research:No, I do not want to publish my article gold open access, and would like my final published article to be immediately available to all subscribers. If selected I will not have to pay a fee.Yes, I would like to publish my article gold open access, making my final published article freely available to everyone. If selected I will need to pay an open access fee.Privacy Policy Terms & Conditions HelpFunding BodyOpen AccessRightsPublishing AgreementOrder SummaryConfirmationElsevier GmbHArticle:Optical wave breaking and soliton trains generation due to cross-phase modulation in an optical fiber with quintic nonlinearityCorresponding author:Prof. Xianqiong ZhongE-mail address:xianqiongzhong@Journal:Optik - International Journal for Light and Electron OpticsOur reference:IJLEO56584PII:S0030-4026(15)01492-8DOI:10.1016/j.ijleo.2015.10.124SELECT YOUR STATUSI am the sole author of the manuscriptI am one author signing on behalf of all co-authors of the manuscriptPlease indicate which of the below applies to you and your co-authors:•We are all US Government employees and the Article is public domainand therefore the ‘Assignment of copyright’ clause does not apply•I am a US Government employee but some of my co-authors are not•I am not a US Government employee but some of my co-authors are•The work was performed by contractors of the US Government undercontract number:*•We are all UK Government employees electing to transfer copyright•We are all UK, Canadian or Australian Government employees andCrown Copyright is claimed•I am claiming Crown Copyright but some of my co-authors are notemployees of the UK, Canadian or Australian Government•I am not claiming Crown Copyright but some of my co-authors areemployees of the UK, Canadian or Australian Government•None of the aboveThe Article is a 'work made for hire' and I am signing as an authorizedrepresentative of my employerSIGNED ON BEHALF OF CORRESPONDING AUTHORPlease complete this section if you are not the corresponding author as listed above. A copy of the agreement will be sent to you and the corresponding author.DATA PROTECTION & PRIVACYI understand that staff of Elsevier GmbHthe and its affiliates worldwide will be contacting me concerning the publishing of the Article and occasionally for marketing purposes (unless, with respect to such marketing, I tick the box below).I do not wish to receive news, promotions and special offers about products and services of Elsevier GmbHthe and its affiliates worldwidePrivacy Policy Terms & Conditions HelpCopyright (c) 2015 Elsevier Ltd. All rights reserved.Cookies are set by this site. To decline them or learn more, visit our Cookies page.Funding BodyOpen AccessRightsPublishing AgreementOrder SummaryConfirmationElsevier GmbHArticle:Optical wave breaking and soliton trains generation due to cross-phase modulation in an optical fiber with quintic nonlinearityCorresponding author:Prof. Xianqiong ZhongE-mail address:xianqiongzhong@Journal:Optik - International Journal for Light and Electron OpticsOur reference:IJLEO56584PII:S0030-4026(15)01492-8DOI:10.1016/j.ijleo.2015.10.124YOUR STATUS•I am one author signing on behalf of all co-authors of the manuscript DATA PROTECTION & PRIVACY•I do wish to receive news, promotions and special offers about productsand services from Elsevier GmbH and its affiliated companies worldwide. ASSIGNMENT OF COPYRIGHTI hereby assign to Elsevier GmbH the copyright in the manuscript identified above (where Crown Copyright is claimed, authors agree to grant an exclusive publishing and distribution license) and any tables, illustrations or other material submitted for publication as part of the manuscript (the "Article") in all forms and media (whether now known or later developed), throughout the world, in all languages, for the full term of copyright, effective when the Article is accepted for publication. SUPPLEMENTAL MATERIALSWith respect to Supplemental Materials that I wish to make accessible either through a link in the Article or on a site or through a service of Elsevier GmbH, Elsevier GmbH shall be entitled to publish, post, reformat, index, archive, make available and link to such Supplemental Materials on anon-exclusive basis in all forms and media (whether now known or later developed) and to permit others to do so. "Supplemental Materials" shall mean additional materials that are not an intrinsic part of the Article, including but not limited to experimental data, e-components, encodings and software, and enhanced graphical, illustrative, video and audio material.REVERSION OF RIGHTSArticles may sometimes be accepted for publication but later rejected in the publication process, even in some cases after public posting in "Articles in Press" form, in which case all rights will revert to the author (see /locate/withdrawalpolicy).REVISIONS AND ADDENDAI understand that no revisions, additional terms or addenda to this Journal Publishing Agreement can be accepted without Elsevier GmbH's express written consent. I understand that this Journal Publishing Agreement supersedes any previous agreements I have entered into with Elsevier GmbH in relation to the Article from the date hereof.RETENTION OF RIGHTS FOR SCHOLARLY PURPOSES.I understand that I retain or am hereby granted (without the need to obtain further permission) the Retained Rights (see description below), and that no rights in patents, trademarks or other intellectual property rights are transferred to Elsevier GmbH.The Retained Rights include the right to use the Preprint , Accepted Manuscript and the Published Journal Article for Personal Use , Internal Institutional Use and for Scholarly Sharing .In the case of the Accepted Manuscript and the Published Journal Article the Retained Rights exclude Commercial Use (unless expressly agreed in writing by Elsevier GmbH), other than use by the author in a subsequent compilation of the author's works or to extend the Article to book length form or re-use by the author of portions or excerpts in other works (with full acknowledgment of the original publication of the Article).AUTHOR REPRESENTATIONS / ETHICS AND DISCLOSUREI affirm the Author Representations noted below, and confirm that I have reviewed and complied with the relevant Instructions to Authors, Ethics in Publishing policy, and Conflicts of Interest disclosure. Please note that some journals may require that all co-authors sign and submit Conflicts of Interest disclosure forms. I am also aware of the publisher's policies with respect to retractions and withdrawal (/locate/withdrawalpolicy).For further information see the publishing ethics pageat /publishingethics and the journal home page.Author representations•The Article I have submitted to the journal for review is original, has beenwritten by the stated authors and has not been published elsewhere.•The Article was not submitted for review to another journal while underreview by this journal and will not be submitted to any other journal.•The Article and the Supplemental Materials contain no libellous or otherunlawful statements and do not contain any materials that violate anypersonal or proprietary rights of any other person or entity.•I have obtained written permission from copyright owners for anyexcerpts from copyrighted works that are included and have creditedthe sources in the Article or the Supplemental Materials.•Except as expressly set out in this Journal Publishing Agreement, theArticle is not subject to any prior rights or licenses and, if my or any ofmy co-authors' institution has a policy that might restrict my ability togrant exclusive rights under this Journal Publishing Agreement, awritten waiver of that policy has been obtained.•If I am using any personal details or images of patients, research subjectsor other individuals, I have obtained all consents required by applicablelaw and complied with the publisher's policies relating to the use of suchimages or personal information.See /patientphotographs for furtherinformation.•Any software contained in the Supplemental Materials is free from viruses,contaminants or worms.•If the Article or any of the Supplemental Materials were prepared jointlywith other authors, I have informed the co-author(s) of the terms of thisJournal Publishing Agreement and that I am signing on their behalf astheir agent, and I am authorized to do so.For information on the publisher's copyright and access policies, pleasesee /copyright.For more information about the definitions relating to this agreement click here.I have read and agree to the terms of the Journal Publishing Agreement.1st November 2015T-copyright-v20/2015Funding BodyOpen AccessRightsPublishing AgreementOrder SummaryConfirmationArticle:Optical wave breaking and soliton trains generation due to cross-phase modulation in an optical fiber with quintic nonlinearityCorresponding author:Prof. Xianqiong ZhongE-mail address:xianqiongzhong@Journal:Optik - International Journal for Light and Electron OpticsOur reference:IJLEO56584PII:S0030-4026(15)01492-8DOI:10.1016/j.ijleo.2015.10.124ORDER SUMMARYFunding Body: Research reported in the article was not funded by any of the listed funding bodies.Open Access: No, I do not want to publish my article gold open access, and would like my final published article to be immediately available to all subscribers.。
responsetodr.marino
and every study needs to be critically analyzed before it is accepted,no matter what title it carries.Open discus-sion and criticism is not only good,it is necessary.I would hope that the editors will continue to address this problem in future issues.—J OSEPH T.M ARINO,MD6555Coyle AvenueCarmichael,California95608REFERENCES1.Smyth RL.Evidence-based pediatric pulmonary medicine:Howcan it help?Pediatr Pulmonol.1998;25:118–127.2.Bush A.Early treatment with dornase alfa in cystic fibrosis:Whatare the issues?Pediatr Pulmonol.1998;25:79–82.3.Konstan MW,Byard PJ,Hopper C,Davis PB.Effect of high-doseibuprofen in patients with cystic fibrosis.N Engl J Med.1995;332:848–854.4.Glantz SA.Primer of Biostatistics,4th Ed.New York:McGraw-Hill,1997:123.5.Garb JL.Understanding Medical Research:A Practitioner’sGuide.Boston:Little,Brown and Company,1996.6.Gehlbach SH.Interpreting The Medical Literature.3rd Ed.NewYork:McGraw-Hill,1993.7.Riegelman RV,Hirscu RP.Studying a Study and Testing a Test:How to Read the Health Science Literature.3rd Ed.Boston:Little, Brown and Company,1996.Response to Evidence-Based MedicineSir,Dr.Marino suggests that there is some conflict be-tween the views expressed by Bush in an editorial con-cerning dornase alfa in cystic fibrosis1and in my article on evidence-based medicine in the same issue of Pedi-atric Pulmonology.2The basis for this appears to be that Bush while stating that‘‘—dornase alfa has earned an established place in the treatment of CF on the basis of an impressive body of evidence’’also identified and dis-cussed the limitations of using FEV1as an outcome mea-sure in clinical trials in cystic fibrosis.I,on the other hand,highlighted the important findings of a randomized controlled trial of ibuprofen in cystic fibrosis,which showed that this therapy was associated with a slowerrate of decline in FEV1.3This was one of very manystudies from the whole spectrum of pediatric pulmonol-ogy,which I described in the article.I used the ibuprofen trial to illustrate the point that,although studies may be well designed and conducted,their findings are not al-ways implemented in clinical practice.My statements on this point were neutral and I did not,as Dr.Marino suggests,express‘‘disappointment’’at these findings. Bush’s editorial included a very useful discussion of outcome variables that may be used as an alternative toFEV1in clinical trials in cystic fibrosis patients.Thepoints he made are pertinent to all studies that evaluate interventions which may affect lung disease in cystic fibrosis.Clearly what matters to cystic fibrosis patients when considering such a therapy is the length of their life and the quality of that life,but,as Bush has pointed out, there are difficulties in measuring both these outcomes in cystic fibrosis patients.FEV1is,therefore,frequently used as a surrogate for mortality,but its limitations in that context are well recognized.4Dr.Marino’s argument is confused,because he seems to regard‘‘evidence-based medicine’’as a research study rather than as a process that contributes to clinical decision making.The third step in this process is to ‘‘evaluate(critically appraise)available evidence for its validity and usefulness.’’For randomized controlled tri-als such an appraisal includes assessing the strengths and weaknesses of study design and the applicability of their findings to the patient one is considering.In my article, I frequently made reference to the complexities of mak-ing decisions about individual patients,and Figure1il-lustrated some of the other important components in clinical decision making.Evidence-based medicine is not,as Dr.Marino sug-gests,a‘‘title’’that can be‘‘bestowed’’on some studies, but not others,but,rather,a process that aims to improve patient care by using a rigorous and systematic approach to identify,appraise,and,if appropriate,apply current best evidence to clinical decision making.—R OSALIND L.S MYTH,MA,MB,BS,MD,FRCPCHRespiratory UnitUniversity Department of Child HealthRoyal Liverpool Children’s HospitalLiverpool,United Kingdom REFERENCES1.Bush A.Early treatment with dornase alfa in cystic fibrosis:whatare the issues?Pediatr Pulmonol.1998;25:79–82.2.Smyth RL.Evidence-based pediatric pulmonary medicine:Howcan it help?Pediatr Pulmonol.1998;25:79–82.3.Konstan MW,Byard PJ,Hopper C,Davis PB.Effect of high-doseibuprofen in patients with cystic fibrosis.N Engl J Med.1995;332:848–854.4.Ramsey BW,Boat TF.Outcome measures for clinical trials incystic fibrosis:Summary of a Cystic Fibrosis Foundation Con-sensus Conference.J Pediatr.1994;124:177–192.Response to Dr.MarinoSir,there is no conflict between Dr.Smyth’s views on evidence-based medicine,1and specifically on the ibu-profen study,2and my own3on the use of rhDNase in mild CF lung disease.First,Dr.Marino is wrong in stat-ing that the same variable was measured in the two stud-ies:Konstan et al.2measured rate of change in lung func-tion over4years,and Geller et al.4measured changes in absolute lung function over14days.Second,evidence-based medicine involves critically weighing the quality of the evidence and the trade-off against possible risk, which was exactly the purpose of my annotation.Finally,232Letters to the Editorevidence-based medicine requires‘‘a bottom up ap-proach that integrates the best available evidence with individual clinical expertise and patient choice (x)ternal clinical evidence can inform,but can never re-place,individual clinical expertise.’’5It is certainly not news to me or to other protagonists of evidence-based medicine that‘‘each and every study needs to be ana-lyzed critically before it is accepted.’’—A NDREW B USH,MB,BS(HONS),MA,MD,FRCP,FRCPCH Department of Pediatric Respiratory MedicineRoyal Brompton HospitalSydney StreetLondon,SW36NP,United Kingdom REFERENCES1.Smyth RL.Evidence-based pediatric pulmonary medicine:Howcan it help?Pediatr Pulmonol.1998;25:118–127.2.Konstan MW,Byard PJ,Hopper C,David PB.Effect of high-doseibuprofen in patients with cystic fibrosis.N Engl J Med.1995;332:848–854.3.Bush A.Early treatment with dornase alfa in cystic fibrosis:Whatare the issues?Pediatr Pulmonol.1998;25:79–82.4.Geller DE,Eigen H,Fiel SB,Clark C,Lamarre AP,Johnson CA,Konstan MW,for the Dornase Alfa Nebulizer Group.Effect of smaller droplet size of dornase alfa on lung function in mild cystic fibrosis.Pediatr Pulmonol.1998;25:83–87.5.Sacket DL,Richardson WS,Rosenberg W,Haynes B.Evidence-based medicine.New York:Churchill Livingstone,1997.Letters to the Editor233。
successfulsecuki...
cceptedArticleThis article has been accepted for publication and undergone full peer review but has not been through thecopyediting, typesetting, pagination and proofreading process, which may lead to differences between thisversion and the Version of Record. Please cite this article as doi: 10.1111/jdv.14234Received Date: 11-Feb-2017Accepted Date: 21-Feb-2017Article Type: Letter to EditorSuccessful Secukinumab treatment of generalized pustular psoriasis anderythrodermic psoriasisDear Editor,Erythrodermic psoriasis (EP) and generalized pustular psoriasis (GPP) are the mostsevere forms of psoriasis, resulting in significant morbidity and even mortality 1.There are few evidence-based data on their treatment 2,3. Secukinumab, a fully humananti-IL-17A monoclonal antibody, demonstrated strong efficacy in treating moderateto severe plaque-type psoriasis 4,5.We evaluate Secukinumab in three patients with severe therapy-resistant psoriasis,one with GPP and two EP, with at least 2 regimen failures. After informed consent tothe off-label treatment, patients received the standard regimen: 300 mgsubcutaneously once weekly at weeks 0-4, followed by 300 mg every four weeks.Efficacy was evaluated as achievement of the psoriasis area and severity index (PASI)score 75% or 90% reduction from baseline.Case 1: a 66-year-old Caucasian male, with a 10-year history of stable plaque-typepsoriasis developed a generalized pustular eruption, affecting 95% of the bodysurface, associated with fever (38° C) and malaise. On admission, the patient had aneutrophilic leukocytosis, increased levels of liver enzymes and PCR. Histopathologyconfirmed a diagnosis of GPP. Initial treatment with cyclosporine (4mg/kg/day) wasAcceptedArticle stopped because of the occurrence of hypertension, and renal toxicity, while anti-TNF alpha treatment was given for 4 weeks with unsatisfactory results (Fig.1A). After the first week of Secukinumab treatment symptoms had rapidly improved, PASI-75 was achieved at week 4, PASI-90 at week 8 (Fig. 2A), and complete remission at week-12, further maintained at week 24.Case 2: a 28-year-old Caucasian male presenting with plaque-type psoriasis since the age of 19 years, underwent a sudden worsening after a flu-like episode, with development of extensive erythroderma (PASI 45). Treatment with cyclosporine 5 mg/kg/day for 1 month was ineffective, as well as anti-TNF alpha therapy performed for 4 weeks (Fig.1B). Standard dose secukinumab treatment achieved PASI-75 at week 4 and complete remission at week 8 (Fig. 2B), maintained until the last 24 week follow-up, without any side effect.Case 3: a 61-year-old Caucasian male suffering of heart disease and hypertension, with a 1-year history of palmoplantar psoriasis, suddenly developed a histologically confirmed erythrodermic psoriasis (PASI 41). Treatment with anti-TNF alpha was supplied for 4 weeks, without any improvement (Fig. 1C), while Secukinumab induced PASI-75 achievement at week 4 and complete clinical resolution at week 8 (Fig. 2C), maintained until the last follow-up at week 12, without any adverse events.There is no robust data on which to base treatment recommendations for EP and GPP 1-3. Among DMARDS, cyclosporine appears to be the first choice, as metrotrexate and acitretin usually work more slowly, while systemic steroids potentially trigger GPP, and expose to a severe rebound at tapering 1,2. Efficacy of TNF-alpha inhibitors in EP varies from 40% to 50%, with PASI75 improvement after 12-14 weeks of treatment 6.AcceptedArticle Successful control of GPP with adalimumab or etanercept was reported in small numbers of patients 2, while ustekinumab, an anti human IL-12/23 antibody and ixekinumab, an anti IL-17A antibody proved their efficacy both in EP and GPP 7,8. Finally, anakinra was also effective for GPP 2. However, given the lack of head-to-head trials, it is impossible to recommend one therapy over the other.To our knowledge, these are the first cases of EP remission after few Secukinumab administrations, with an excellent safety profile. As for GPP, there are only two reported cases of successful therapy with Secukinumab9,10. Our observations validly supports the rapid and safe clinical response to this drug. Further studies are warrant to confirm our data on secukinumab efficacy in EP and GPP.C. Mugheddu, L. Atzori, A. Lappi, M. Pau, S. Murgia, F. Rongioletti.Section of Dermatology, AOU of Cagliari - Department of Medical Science and Public Health; University of Cagliari, Italy.Corresponding Author:Dr.ssa Laura ATZORIClinica DermatologicaVia Ospedale 5409124 Cagliari, Italy.E-mail:****************Tel+390706092324Fax +390706092580AcceptedArticle Funding details: This work was supported by no grant or founds.Disclosure statementThe authors report no conflicts of interest.References1.Rosenbach M, Hsu S, Korman NJ, et al. National Psoriasis Foundation MedicalBoard. J Am Acad Dermatol 2010; 62:655-62.2.Robinson A, Van Voorhees As, Hsu S, et al. Treatment of pustular psoriasis:from the Medical Board of the National Psoriasis Foundation. J Am Acad Dermatol 2012; 67:279-88.3.Choon SE, Lai Nm, Mohammad NA,et al. Clinical profile, morbidity, andoutcome of adult-onset generalized pustular psoriasis: analysis of 102 cases seen in a tertiary hospital in Johor, Malaysia. Int J Dermatol 2014; 53:676-84.4.Lonnber AS, Zachariae C, Skov L. Targeting of interleukin-17 in the treatment ofpsoriasis. Clin Cosmet Investig Dermatol 2014; 7:251-9.ngley Rg, Elewski BE, Lebwohi M, et al. Secukinumab in plaque psoriasis-results of two phase 3 trials. N Engl J Med 2014; 371:326-38.6.Viguier M1, Pagès C, Aubin F, et al. Efficacy and safety of biologics inerythrodermic psoriasis: a multicentre, retrospective study.Br J Dermatol 2012;167:417-23.7.Santos-Juanes J, Coto-Segura P, Mas-Vidal A, Galache Osuna C. Ustekinumabinduces rapid clearing of erythrodermic psoriasis after failure of antitumornecrosis factor therapies. Br J Dermatol. 2010; 162:1144-6.AcceptedArticle8.Saeki H, Nakagawa H, Ishii T, et al. Efficacy and safety of open-labelixekizumab treatment in Japanese patients with moderate-to-severe plaquepsoriasis, erythrodermic psoriasis and generalized pustular psoriasis. J Eur AcadDermatol Venereol. 2015; 29:1148-55.9.Bohner A, Roenneberg S, Eyerich K, et al. Acute generalized pustular psoriasistreated with the IL-17° antibody secukinumab. JAMA Dermatol 2016; 152:482-84.10.Polesie S, Lidholm AG. Secukinumab in the treatment of generalized pustularpsoriasis: a case report. Acta Derm Venereol 2016; doi: 10.2340/00015555-2467.Legend to Figure:Fig.1: Case 1 of GPP (A); case 2 of EP (B), and case 3 of EP (C) before secukinumab treatment.Fig. 2: At week 8, PASI 90 achievement of Case 1 (A), complete remission of case 2 (B), and case 3 (C).AcceptedArticl eA。
Globally networked risks and how to respond
BOX 1
Risk, systemic risk and hyper-risk
According to the standard ISO 31000 (2009; /iso/ catalogue_detail?csnumber543170), risk is defined as ‘‘effect of uncertainty on objectives’’. It is often quantified as the probability of occurrence of an (adverse) event, times its (negative) impact (damage), but it should be kept in mind that risks might also create positive impacts, such as opportunities for some stakeholders.
Geogenic-PM10-exposure-exacerbates-responses-to-influenza-infection
Geogenic PM 10exposure exacerbates responses to in fluenza infectionHolly D.Clifford a ,b ,⁎,1,Kara L.Perks a ,1,Graeme R.Zosky a ,b ,ca Telethon Kids Institute,The University of Western Australia,Perth,Western Australia,Australiab Centre for Child Health Research,The University of Western Australia,Perth,Western Australia,Australia cSchool of Medicine,University of Tasmania,Hobart,Tasmania,AustraliaH I G H L I G H T S •Geogenic PM 10exposure exacerbates the response to a respiratory viral infection.•This exposure increases in flammation and viral load,and impairs lung function.•Iron content in the particles may be a driver of these responses.•This has important implications for lung health in communities in arid environments.a b s t r a c ta r t i c l e i n f o Article history:Received 28January 2015Received in revised form 4June 2015Accepted 1July 2015Available online 11July 2015Editor:D.Barcelo Keywords:Particle Geogenic In fluenza In flammation Lung function Viral titre IronParticulate matter (PM)exposure has been linked epidemiologically to exacerbations of lung disease,including respiratory infections.We investigated the effects of geogenic (earth-derived)PM 10(PM b 10μm diameter)on the response to a respiratory viral infection.Geogenic dust was sampled from four communities in arid environments in Western Australia.Adult female BALB/c mice were intranasally exposed to chronic doses of PM 10(10μg/day for 10days),and/or infected with in fluenza (A/Mem/1/71)virus.In flammation (cells,IL-6,IFN-γ)was measured in bronchoalveolar lavage.Lung mechanics were measured using the forced oscillation technique.Geogenic PM 10induced lung in flammation (neutrophils,macrophages)with additive effects in mice also infected with in fluenza.PM 10also modi fied the in fluenza-induced IL-6and IFN-γresponses.Geogenic PM 10increased airway resistance,and increased hysteresivity in those exposed to both insults.Viral titres were signi ficantly higher after PM 10exposure.Iron concentration was inversely associated with IFN-γand positively associated with viral titre and hysteresivity.Geogenic PM 10exposure increases in flammation,impairs lung function and increases viral load,exacerbating the response to respiratory viral infection.Iron in the particles may be a driver of these responses.This has important implications for respiratory health in communities exposed to high geogenic PM 10,such as those in arid environments.©2015Elsevier B.V.All rights reserved.1.BackgroundParticulate matter (PM)continues to be the aspect of air pollution that is most reliably associated with human disease;particularly PM with an aerodynamic diameter of b 10μm (PM 10)(Harrison and Yin,2000).Ambient PM 10concentrations have been associated epidemio-logically with increased hospitalisations for respiratory diseases,includ-ing lower respiratory tract infections (Lin et al.,2005;Xu et al.,2013).Most research has concentrated on the effects of urban PM which contains high levels of carbonaceous particles from exhaust emissions,and in fact,most national air quality standards for PM (Department of the Environment,2005;Environmental Protection Agency,2013;European Commission,2014)are based on urban data.Other studies have focused on occupational exposures (Hnizdo and Vallyathan,2003),however,there has been little research on PM 10from otherScience of the Total Environment 533(2015)275–282Abbreviations:PM 10,particulate matter with a diameter of b 10μm;MCh,methacholine;R aw ,airway resistance;G,tissue damping;H,tissue elastance;ICP-MS,inductively coupled plasma-mass spectrometry;ICP-OES,inductively coupled plasma-optical emission spectrometry;MMAD,mass median aerodynamic diameter;GSD,geo-metric standard deviation;VP-SFM,virus production serum-free medium;MDCK,Madin –Darby canine kidney cells;TGV,thoracic gas volume;FOT,forced oscillation tech-nique;I aw ,airway inertance;Zrs,respiratory system input impedance spectrum;BAL,bronchoalveolar lavage;PBS,phosphate buffered saline;ELISA,enzyme linked immuno-sorbent assay;MIP-2,macrophage in flammatory protein-2.⁎Corresponding author at:Telethon Kids Institute,100Roberts Road,Subiaco,Western Australia 6008,Australia.E-mail address:holly.clifford@.au (H.D.Clifford).1Joint firstauthors./10.1016/j.scitotenv.2015.07.0010048-9697/©2015Elsevier B.V.All rightsreserved.Contents lists available at ScienceDirectScience of the Total Environmentj o u r n a l h o me p a g e :ww w.e l s e v i e r.c o m /l o c a t e /s c i t o t e nvsources,such as particles from geogenic dusts.Many communities lo-cated in the arid regions of the world and regions exposed to prevailing winds that cross these regions(Esmaeil et al.,2014;Goudie,2014),are likely to be exposed to high geogenic PM10loads.Mineral dusts have been linked to many respiratory diseases from acute inflammatory reactions to chronic conditions that involve structural changes in the lung(Mossman and Churg,1998).Recently,dust storms have been associated with increased respiratory-related hospital admissions (Tam et al.,2012).However,information regarding the specific health effects of exposure to community-sampled geogenic dust is scarce.We have recently shown that acute exposure to geogenic PM10causes acute neutrophil-dominated inflammation in the lungs of mice(Zosky et al.,2014a)and that the magnitude of this acute inflammatory re-sponse and long-term deficits in lung function is associated with the concentration of iron(Fe)in the particles(Zosky et al.,2014b).However,in these studies we were focused on the impact of geogenic dust inhalation in an otherwise healthy lung.It has been shown in vitro that Fe may increase inflammatory mediators in the lung epithelium(Smith et al.,2000)and epidemiological data suggests that exposure to iron-laden geogenic dust can increase the risk of hospitalisation for respiratory infections(Mullan et al.,2006;South Australia Department of Health,2007).We aimed to determine wheth-er chronic,low-dose exposure to community-sampled geogenic PM10 exacerbates the response to a respiratory viral infection.2.Methods2.1.AnimalsEight week old female BALB/c mice(Animal Resource Centre, Murdoch,WA,Australia)were housed in a pathogen-free environment with a12h:12h light dark cycle,and provided with food and water ad libitum.All studies were approved by the Telethon Kids Institute Animal Ethics Committee and adhere to the guidelines of the National Health and Medical Research Council of Australia.2.2.Geogenic PM102.2.1.Sample collectionThe top2cm of a1m2area of surface soil was collected from four re-mote towns in arid environments across Western Australia.Newman (population~9,087)is close to several open cut iron ore extraction sites and Tom Price is primarily an iron ore mining town(population ~5460).Karratha(population~16,475)is a regional town that services local mining operations,while Kalgoorlie(population~30,841)is locat-ed adjacent to a large open cut gold mine.The PM10fraction was extracted as previously described(Ljung et al.,2011).Briefly,topsoil samples were collected and dry sieved to45μm.The45μm fraction was suspended in MilliQ water,vortexed and ultrasonically agitated to disperse the sample.Stoke's law was then used to separate the ~10μm fraction from the suspension.Aliquot were extracted from the suspension at the appropriate settling time and wetfiltered through a 10μm mesh.Filters were then dried overnight leaving the b10μm frac-tion for our in vivo experiments.We have previously shown that the physico-chemical attributes of this extracted PM10closely matches that of the airborne PM10from the same sites(Zosky et al.,2014a). 2.2.2.Physical and chemical characterisationThe chemical composition of the extracted PM10samples was deter-mined using inductively coupled plasma-mass spectrometry(ICP-MS; Chemistry Centre of W.A.,Australia)to obtain concentrations of Al,As, Cd,Co,Cr,Cu,Fe,Mn,Ni,Pb,U,and Zn,and inductively coupled plasma-optical emission spectrometry(ICP-OES;Perkin Elmer Optima 5300DV,Norwalk,CT)to measure Si.To obtain mass weighted esti-mates of the particle size distribution in the samples,2mL of the sample was aerosolised and drawn through an Andersen Cascade Impacter (Copley Scientific,Nottingham,UK)(Zosky et al.,2014a).The mass me-dian aerodynamic diameter(MMAD)and geometric standard deviation (GSD)of the particle sizes were calculated.Endotoxin levels were assessed using a limulus amebocyte lysate assay(GenScript,New Jersey, U.S.A.).2.2.3.In vivo exposure modelMice(n=10per group)were exposed intranasally,under light methoxyfluorane anaesthesia(Medical Development,Springfield,Vic, Australia),to10μg of geogenic particles from one of the four sites in 50μL of0.9%sodium chloride(Pfizer,Bentley,WA,Australia),daily for ten consecutive days.The100μg dose was chosen based on our previ-ous dose–response data(Zosky et al.,2014a)and prior work on diesel particles(Boylen et al.,2011;Larcombe et al.,2013)showing that parti-cle loads in macrophages are equivalent to those observed in human macrophages(Kulkarni et al.,2006).Control mice received50μL of saline alone.To prevent particle aggregation,all preparations were sonicated for30min.Six hours after the sixth exposure to geogenic particles,mice were exposed intranasally to104.5plaque forming units(pfu)of a recombi-nant mouse-adapted strain of Influenza A(Influenza A⁄Memphis⁄1⁄71(H3N1))in50μL of virus production serum-free medium(VP-SFM; Gibco,Mulgrave,Vic,Australia)under light methoxyfluorane anaesthe-sia.Control mice received the same volume of the supernatant of unin-fected Madin–Darby canine kidney(MDCK)cells diluted in VP-SFM.2.3.Lung physiology2.3.1.Animal preparationAt the peak of infection(4days post-infection),mice were anaesthetised by an intraperitoneal injection containing2mg/mL of xylazine and40mg/mL ketamine(Troy Laboratories,Glendenning, NSW,Australia),at a dose of0.01mL/g body weight.A tracheostomy was performed,a10mm length of tubing was inserted into the trachea, and the mouse was connected to a mechanical ventilator(HSE-Harvard MiniVent;Harvard Apparatus,Holliston,MA)and ventilated at400 breaths/min with a tidal volume of8mL/kg and2cm H2O positive end-expiratory pressure.2.3.2.Lung volumeThoracic Gas Volume(TGV)was measured as described previously (Jánosi et al.,2006;Zosky et al.,2008).Briefly,ventilation was stopped and the intercostal muscles were electrically stimulated at20V for1–2ms,to induce inspiratory efforts.Six measurements were taken over a6s period and,following correction for the impedance and thermal properties of the chamber,TGV was calculated using Boyle's Law (Jánosi et al.,2006).2.3.3.Lung mechanicsA modification of the forced oscillation technique(FOT)was used to measure lung mechanics(Hantos et al.,1992).An oscillatory signal con-taining nine frequencies(4–38Hz)was generated using a speaker and delivered to the tracheal cannula via a1m wavetube of known imped-ance.The respiratory system input impedance spectrum(Zrs)was then measured using a4-parameter model with constant-phase tissue im-pedance(Hantos et al.,1992),to obtain measures of airway resistance (R aw),airway inertance(I aw),tissue damping(G),tissue elastance (H)and hysteresivity(η).I aw is negligible after correcting for the trache-al cannula and is not reported.2.3.4.Methacholine challengeMethacholine responsiveness was measured as described previously (Zosky et al.,2004).Briefly,after the measurement of baseline Zrs,mice were challenged with a90s saline control aerosol delivered with an ultrasonic nebuliser(UltraNeb®,Devilbiss,Somerset,Pennsylvania). Five Zrs measurements were taken at1minute intervals before being276H.D.Clifford et al./Science of the Total Environment533(2015)275–282repeated with increasing concentrations of β-methacholine chloride (MCh;Sigma-Aldrich,MO,USA;0.001–30mg/mL).The peak response at each MCh dose was expressed as a percentage of the response to the saline aerosol.2.4.Lung in flammationUpon the completion of lung function measurements,mice were euthanased and their lungs were lavaged as described previously (Zosky et al.,2014a ).The bronchoalveolar lavage (BAL)fluid was used to measure total and differential cell counts via light microscopy and the levels of IL-6,IFN-γand MIP-2by ELISA (BD Biosciences,San Diego,CA,USA).2.5.Viral titreOn day 4post-infection,a separate subset of mice (n =9per group)were euthanased and lung tissue was collected under aseptic conditions.Lungs were homogenised in VP-SFM and lung viral titres (normalized to lung weight)were determined by plaque assay using con fluent MDCK cells,as previously described (Zosky et al.,2014b ).2.6.Data analysisStatistical analyses were performed using SigmaPlot (Version 12.0;SPSS Science,Chicago,IL,USA)and Stata (Version 10;StataCorp,Texas,USA).Outcomes were compared between groups by two-way ANOVA and Holm –Sidak post hoc tests.Data were transformed where necessary to satisfy the assumptions of homoscedasticity and normal distribution of the error terms.In order to explore the relationship be-tween the physico-chemical characteristics of the particles and the out-comes that were measured,we conducted multiple linear regression analyses for the outcomes in in fluenza/PM 10exposed mice where there was a primary effect of both particle exposure and in fluenza (additive effect:neutrophils,macrophages,baseline ƞ,R aw response to MCh)or where exposure to the particles modi fied the response to in fluenza (IL-6,IFN-γand viral titre)with the major physico-chemical characteristics of the particles as the independent variables ([Si],[Fe],[Al],MMAD and GSD).The responses in the in fluenza/PM 10exposed mice were expressed as a percentage of the responses in the in fluen-za/saline exposed mice.Data are presented as mean (SD)throughout.Table 1Physico-chemical characteristics of geogenic PM 10samples from Newman,Kalgoorlie,Karratha and Tom Price.NewmanTom Price Karratha Kalgoorlie MMAD (μm) 2.93 4.27 2.68 3.45GSD (μm)2.49 2.56 2.633.40Endotoxin (EU/50μL)93.203152.03614.06450.3Metal (mg·kg −1)Si 186,000188,000191,000170,000Al 84,00070,60070,80082,200Fe120,000100,00093,00060,000Al,aluminium;Fe,iron;GSD,geometric standard deviation;MMAD,mass median aerody-namic diameter;Si,silica.Fig.1.Cell counts of neutrophils (A)and macrophages (B)in bronchoalveolar lavage fluid in mice exposed to geogenic particles (grey bars)from Newman,Tom Price,Karratha and Kalgoorlie without in fluenza (media controls),or geogenic particles and in fluenza (black bars).*indicates difference p b 0.05compared with saline/media controls (white bars).#indicates p b 0.05compared with saline/in fluenza mice (striped bars).Data are mean(SD).Fig.2.Cytokine levels of IL-6(A)and IFN-γ(B)in bronchoalveolar lavage fluid in mice exposed to geogenic particles (grey bars)from Newman,Tom Price,Karratha and Kalgoorlie without in fluenza (media controls),or geogenic particles and in fluenza (black bars).*indicates difference p b 0.05compared with saline/media controls (white bars).&indicates p b 0.05compared with the particle/media exposed mice.Data are mean (SD).277H.D.Clifford et al./Science of the Total Environment 533(2015)275–282278H.D.Clifford et al./Science of the Total Environment533(2015)275–2823.Results3.1.Particle characterisationAll elements measured were detectable in all geogenic PM 10sam-ples.Three elements dominated (Si,Fe and Al),with the remaining ele-ments comprising b 0.1%of the metal content of the samples.Si was the most abundant element in all samples.Fe was the next abundant metal in Newman,Kalgoorlie and Tom Price,while Al was the second most abundant element in Karratha (Table 1).Endotoxin levels ranged from 93.2to 614.1EU/50μL (equivalent to 0.5–15.8ng/50μL).3.2.Body weightExposure to geogenic PM 10had no effect on body weight (p N 0.05for all pairwise comparisons).In fluenza infection caused a signi ficant decrease in body weight in control mice (saline exposed;p =0.01)and mice exposed to PM 10from Tom Price (p =0.01)and Karratha (p =0.04)but not mice exposed to PM 10from Newman (p =0.46)or Kalgoorlie (p =0.40).Mice in the control group (saline exposed)had the greatest weight loss (6.8%)in response to in fluenza infection suggestion that PM 10exposure did not exacerbate in fluenza induced weight loss (data not shown).3.3.In flammatory cellular in fluxExposure to PM 10from Newman (p =0.02),Kalgoorlie (p b 0.001)and Karratha (p b 0.001)resulted in a signi ficant in flux of neutrophils compared to the saline controls (Fig.1A).In fluenza infection also caused neutrophilia in all groups (p b 0.001).There was no signi ficant interac-tion (p =0.52)between PM 10and in fluenza exposure suggesting that there was an additive effect of the PM 10and in fluenza exposure on the neutrophil in flux.Particle exposure also signi ficantly increased the level of macrophages in mice exposed to PM 10from Newman and Kalgoorlie (p b 0.001)(Fig.1B),as did in fluenza in all groups (p b 0.001).Again,there was no signi ficant interaction between particle ex-posure and in fluenza infection (p =0.66),suggesting that these effects were additive.3.4.In flammatory cytokinesChronic exposure to particles alone did not affect the levels of IL-6or IFN-γ.However,these cytokines were signi ficantly increased in re-sponse to in fluenza infection when compared with the saline control (p b 0.001)(Fig.2).Exposure to particles modi fied the response to both IL-6(p =0.04)and IFN-γ(p =0.01)such that mice exposed to particles from Tom Price had a signi ficantly reduced in fluenza-induced IL-6response (p =0.68)while particles from Tom Price (p =0.53)and Kalgoorlie (p =0.08)had a signi ficantly reduced in fluenza-induced IFN-γresponse (Fig.2).MIP-2levels were not signi ficantly altered after exposure to particles or in fluenza (data not shown).3.5.Thoracic gas volumeTGV was signi ficantly lower in mice exposed to PM 10from Kalgoorlie (p b 0.001),Karratha (p =0.002)and Tom Price (p =0.017)compared with the saline control (Fig.3A).Conversely,mice infected with in fluenza infection tended to have a higher TGV than control mice (p =0.045)(Fig.3A).3.6.Baseline lung functionMice exposed to particles from Karratha (p b 0.001)and Tom Price (p b 0.001)exhibited an increase in R aw (Fig.3B).G and H were not af-fected by PM 10exposure (data not shown).In fluenza infection did not in fluence R aw (Fig.3B),however in fluenza was associated with in-creases in both G (p b 0.001)and H (p =0.010)(data not shown).Particle exposure had no signi ficant effect on hysteresivity (ƞ=G/H)nor did in fluenza by itself.However,in the presence of both in fluenza and PM 10there was a signi ficant increase in ƞ(p b 0.003).3.7.Responsiveness to methacholineResponses to methacholine (MCh)at the maximal dose of 100mg/mL were altered by exposure to geogenic particles.In mice ex-posed to PM 10from Kalgoorlie and Tom Price,the magnitudes of the change in R aw was increased relative to the saline control mice (p =0.01and p =0.03,respectively)(Fig.4A).Particle exposure did not af-fect G at the maximal MCh concentration (Fig.4B),however it did affect H but only in mice exposed to particles from Kalgoorlie (p =0.03)(Fig.4C).In fluenza was shown to have an effect on all lung function pa-rameters at the maximal dose of MCh (p =0.003,p b 0.001and p b 0.001for R aw ,G and H,respectively)(Fig.4).There was no signi fi-cant interaction between particles and in fluenza on responsiveness to MCh,suggesting that all responses were additive.3.8.Viral titreIn mice that were infected with in fluenza,those that were also exposed to geogenic particles from Newman or Tom Price had signi ficantly higher lung viral titres than those of control mice (p b 0.001and p =0.004,respectively;Fig.5).No virus was detected in the non-in fluenza infected (media control)mice.3.9.Regression analysesThere was no association between any of the outcomes selected for the regression analyses and MMAD,GSD or endotoxin content (data not shown).There was also no association between the concentrations of Si,Al and Fe and the in flux of neutrophils,macrophages,production of IL-6and response in R aw to MCh (Table 2).However,the concentra-tions of Si,Fe and Al were strongly associated with IFN-γ,viral titre and baseline hysteresivity (Tables 2,3).Speci fically,the concentration of Fe in the particles was positively associated with the baseline increase in hysteresivity (p =0.04)and viral titre (p =0.005)and inversely as-sociated with the production of IFN-γ(p =0.01)(Tables 2,3).In con-trast Al and Si were positively associated with IFN-γproduction and inversely associated with viral titre and hysteresivity (Tables 2,3).4.DiscussionExposure to geogenic PM has been linked to exacerbations of respi-ratory infections (Lin et al.,2005),including in fluenza (Xu et al.,2013).These studies have,however,been mostly restricted to cross-sectional epidemiological studies or anecdotal observations,and the speci fic ef-fects of this prevalent environmental exposure have not yet been fully elucidated.The present study suggests for the first time that repeated exposure to geogenic PM,depending on the composition of the parti-cles,can exacerbate respiratory viral infections such as in fluenza,with an increased in flammatory response,a de ficit in lung mechanics and in-creased viral load.Furthermore,the magnitude and nature of theFig.3.Thoracic gas volume (TGV)(A),baseline airway resistance (R aw )(B)and baseline hysteresivity (ƞ)(C)measured in mice exposed to geogenic particles (grey bars)from Newman,Tom Price,Karratha and Kalgoorlie without in fluenza (media controls)or geogenic particles and in fluenza (black bars).*indicates p b 0.05compared with saline/media controls (white bars),#indicates p b 0.05compared with saline/in fluenza mice (striped bars).Data are mean (SD).279H.D.Clifford et al./Science of the Total Environment 533(2015)275–282response varied depending on the origin of the PM 10,implicating Fe in contributing to these impaired responses.We have previously shown that a dose-dependent in flammatory re-sponse in the lung is induced in response to acute geogenic PM 10expo-sure (Zosky et al.,2014a );in this study we also demonstrated that chronic,low level exposure to geogenic PM 10induced a potent lung in-flux of neutrophils,macrophages and IL-6.Macrophages are clearly im-portant in the phagocytic response to inhaled particles (Warheit and Hartsky,1993)and neutrophils can cause lung damage through the pro-duction of reactive oxygen species in response to PM (Tao et al.,2003;van Berlo et al.,2010).IL-6has also been implicated in causing acute lung injury in the response to environmental pollutants (Yu et al.,2002).This in flammation may induce tissue damage and repair leading to remodelling of the lung and loss of function (Holgate et al.,2000).In-effective clearance of this PM from the airways could cause particle re-tention in lung tissues,resulting in a chronic,low-grade in flammatory response that may be pathogenically important in both the exacerba-tion as well as the progression of lung diseases (Ling and van Eeden,2009;Svartengren et al.,2004).This is consistent with the de ficits in lung function we observed in mice exposed to geogenic PM 10,suggest-ing that repeated low level exposure to these particles has detrimental impacts on lung health.In fluenza in mice caused a signi ficant increase in in flammation and de ficits in tissue mechanics,consistent with our previous studies (Larcombe et al.,2013;Ramsey et al.,2013).The large airways were affected by geogenic particle exposure,with an increase in airway resis-tance both at baseline and in response to a bronchoconstrictor.Previous studies have suggested that lifetime exposure to PM 10is associated with reduced FEV 1following bronchodilation,de ficits in lung function and reduced lung growth (Downs et al.,2007;Sunyer,2009).We also showed increased hysteresivity after PM 10exposure,suggesting that geogenic exposure with a secondary insult of viral infection results in inhomogeneities and structural changes in the lung (Lutchen and Gillis,1997).A signi ficant interaction was also identi fied with particle exposure and in fluenza infection in regards to hysteresivity,indicating that particles may amplify the increase in lung heterogeneity induced by in fluenza infection.Viral load in the lung was higher in those mice exposed to geogenic PM 10.These data are consistent with a study that showed an increase in viral load after acute exposure to diesel particulates during an existing in fluenza infection (Larcombe et al.,2013),however this is the first time this has been demonstrated with geogenic PM 10.These higher viral titres may represent alterations in viral replication and/or viral clearance from the lung and suggest that geogenic PM 10exposure may exacerbate the response to viral pathogens.The PM from the towns that we sampled had differing physico-chemical characteristics and the effects on in flammation and lung func-tion varied greatly from site to site,suggesting that particle characteris-tics such as metal content may be one of the driving factors of these responses.Particle characteristics have been shown to vary geographi-cally in our previous studies and in others (Harrison and Yin,2000;Mossman and Churg,1998;Zosky et al.,2014a ).Previous studies have linked speci fic trace metals to respiratory disease,and while many of these studies attribute the role of silicates on lung disease,numerous other inhaled metals can cause respiratory tract injury and sensitivity (Cook et al.,2005).Our data,in particular support a potential role for Fe in driving the adverse impacts of geogenic PM 10on lunghealth.Fig.4.Lung function parameters airway resistance (R aw )(A),tissue damping (G)(B)and tissue elastance (H)(C)in response to methacholine (MCh)at the maximal dose of 100mg/mL,as a %of saline,measured in mice exposed to geogenic particles (grey bars)from Newman,Tom Price,Karratha and Kalgoorlie without in fluenza (media controls),or geogenic particles and in fluenza (black bars).*indicates p b 0.05compared with sa-line/media controls (white bars).#indicates p b 0.05compared with saline/in fluenza mice (striped bars).Data are mean(SD).Fig.5.Lung viral titres (pfu/g lung)in mice infected with in fluenza and geogenic particles from Newman,Tom Price,Karratha and Kalgoorlie (black bars).#indicates p b 0.05com-pared with saline/in fluenza mice (striped bar).Data are mean (SD).280H.D.Clifford et al./Science of the Total Environment 533(2015)275–282Studies of iron ore miners have indicated that iron exposure in humans may lead to increased airway in flammation,respiratory symp-toms and obstructive lung diseases (Hedlund et al.,2004).Exposure to Fe in a community setting showed an association between high Fe expo-sure and hospitalisations for respiratory infections in children (South Australia Department of Health,2007).We have recently shown that the de ficit in lung mechanics induced by acute geogenic PM 10exposure is positively correlated with the concentration of Fe in the particles (Zosky et al.,2014b ).The present study results are consistent with this and underline the importance of determining the impact of metals in PM 10,particularly Fe.We showed that the higher the Fe concentra-tion in the particles,the lower the IFN-γresponse and subsequently,the higher the viral load in the lung and a greater de ficit in lung function.Iron has been shown to modulate IFN-γpreviously,with a dose-dependent inhibitory effect of Fe on the IFN-γsignal (Weiss et al.,1992).IFN-γis well known to directly inhibit viral replication and be critical in the antiviral immune response (Karupiah et al.,1993),includ-ing against in fluenza (Weiss et al.,2010).This higher viral load,and consequent prolonged in flammatory response in the lung,may lead to the greater impairment in lung function that we observed.The mecha-nisms underlying the impact of Fe (for example,oxidative stress path-ways)require further investigation.Some limitations should be noted.The particles were dominated by three metals.Nonetheless we were able to ascertain that responses were signi ficantly associated with certain particle characteristics.Future studies will include iron controls to further explore the contribution of iron.Furthermore,although we chose to measure some key players in PM toxicity,other elements that we did not measure may be contribut-ing to the responses that Fe content alone cannot explain;for example the surface structure of the particles.It should also be noted that there was limited evidence of a synergistic interaction between the exposures since most of the effects were additive.This may have been due to the magnitude of the effects of the individual exposures (particles or in flu-enza),which were relatively large in most cases,such that there was limited capacity for the system to increase the magnitude of the response.5.ConclusionsThis study demonstrates long-standing implications for communi-ties located in arid regions who are exposed chronically to geogenic PM 10.These results build on our previous work showing the effects of acute exposure,and re flect more real-world chronic exposures.Our data suggests that geogenic PM 10is likely to exacerbate the response to respiratory viral infection and it therefore should be given more con-sideration as an environmental factor with signi ficant impacts on com-munity respiratory health.Con flict of interestThe authors declare that they have no con flicts of interest.ReferencesBoylen,C.E.,Sly,P.D.,Zosky,G.R.,Larcombe,A.N.,2011.Physiological and in flammatoryresponses in an anthropomorphically relevant model of acute diesel exhaust particle exposure are sex and dose dependent.Inhal.Toxicol.23,906–917.Cook,A.G.,Weinstein,P.,Centeno,J.A.,2005.Health effects of natural dust:role of traceelements and compounds.Biol.Trace Elem.Res.103(1),1–15.Department of the Environment,2005.National standard for criteria air pollutants inAustralia Available:.au/atmosphere/airquality/publications/standards.html#fn .Downs,S.H.,Schindler,C.,Liu,L.J.,Keidel,D.,Bayer-Oglesby,L.,Brutsche,M.H.,et al.,2007.Reduced exposure to PM 10and attenuated age-related decline in lung function.N.Engl.J.Med.357(23),2338–2347.Environmental Protection Agency (EPA),2013.National ambient air quality standards forparticulate matter.Fed.Regist.78(10),3086–3287.Esmaeil,N.,Gharagozloo,M.,Rezaei,A.,Grunig,G.,2014.Dust events,pulmonary diseasesand immune system.Am.J.Clin.Exp.Immunol.3(1),20–29.European Commission,2014.Air quality standards Available:http://ec.europa.eu/environment/air/quality/standards.htm .Goudie,A.S.,2014.Desert dust and human health disorders.Environ.Int.63,101–113.Hantos,Z.,Daróczy,B.,Suki,B.,Nagy,S.,Fredberg,J.J.,1992.Input impedance and periph-eral inhomogeneity of dog lungs.J.Appl.Physiol.(1985)72(1),168–178.Harrison,R.M.,Yin,J.,2000.Particulate matter in the atmosphere:which particle proper-ties are important for its effects on health?Sci.Total Environ.249(1–3),85–101.Hedlund,U.,Järvholm,B.,Lundbäck,B.,2004.Respiratory symptoms and obstructive lungdiseases in iron ore miners:report from the obstructive lung disease in northern Sweden studies.Eur.J.Epidemiol.19(10),953–958.Hnizdo,E.,Vallyathan,V.,2003.Chronic obstructive pulmonary disease due to occupa-tional exposure to silica dust:a review of epidemiological and pathological evidence.Occup.Environ.Med.60,237–243.Holgate,S.T.,Lackie,P.,Wilson,S.,Roche,W.,Davies,D.,2000.Bronchial epithelium as akey regulator of airway allergen sensitization and remodeling in asthma.Am.J.Respir.Crit.Care Med.162(3Pt 2),S113–S117.Jánosi,T.Z.,Adamicza,A.,Zosky,G.R.,Asztalos,T.,Sly,P.D.,Hantos,Z.,2006.Plethysmo-graphic estimation of thoracic gas volume in apneicmice.J.Appl.Physiol.(1985)101(2),454–459.Karupiah,G.,Xie,Q.,Buller,R.M.L.,Nathan,C.,Duarte,C.,MacMicking,J.D.,1993.Inhibi-tion of viral replication by interferon-γinduced nitric oxide synthase.Science 261,1445–1448.Kulkarni,N.,Pierse,N.,Rushton,L.,Grigg,J.,2006.Carbon in airway macrophages andlung function in children.N.Engl.J.Med.355,21–30.Larcombe,A.N.,Foong,R.E.,Boylen,C.E.,Zosky,G.R.,2013.Acute diesel exhaust particleexposure increases viral titre and in flammation associated with existing in fluenza infection,but does not exacerbate de ficits in lung function.In fluenza Other Respir.Viruses 7(5),701–709.Lin,M.,Stieb,D.M.,Chen,Y.,2005.Coarse particulate matter and hospitalization forrespiratory infections in children younger than 15years in Toronto:a case-crossover analysis.Pediatrics 116(2),e235–e240.Table 2Regression analyses demonstrating the relationship between physico-chemical characteristics of geogenic PM 10and in flammatory and viral titre outcomes.NeutrophilsMacrophages IL-6IFN-γViral titre Metal βp βp βp βp βp[95%CI][95%CI][95%CI][95%CI][95%CI]Si 9.70.12 6.10.440.030.060.040.01−104.90.02[−2.7,22.1][−10.0,22.3][0.00,0.07][0.01,0.07][−188.0,−21.9]Al 8.80.0912.30.070.020.120.030.03−63.30.07[−1.6,19.1][−1.2,25.7][0.01,0.05][0.00,0.05][−132.0,5.4]Fe−2.70.14−0.10.97−0.010.05−0.010.0136.10.005[−6.3,0.9][−4.8,4.6][−0.02,0.00][−0.02,−0.01][11.8,60.4]Al,aluminium;Fe,iron;Si,silica.Table 3Regression analyses demonstrating the relationship between physico-chemical character-istics of geogenic PM 10and lung function outcomes.Baseline ƞ%R aw at 100mg·mL −1MCh Metal β(×10−5)p βp [95%CI](×10−5)[95%CI]Si −2.00.02−0.020.29[−3.6,−0.3][−0.07,0.02]Al −1.60.03−0.020.21[−3.0,−0.2][−0.06,0.02]Fe0.50.040.56[0.02,0.1][−0.01,0.02]ƞ,hysteresivity;CI,con fidence interval;Al,aluminium;Fe,iron;MCh,methacholine;Si,silica.281H.D.Clifford et al./Science of the Total Environment 533(2015)275–282。
微通道经皮肾镜取石术治疗输尿管结石的效果及其安全性评价
症肺炎的疗效及其对血清炎症因子水平及神经功能改善的影响[J].抗感染药学,2018,15(12):2186-2189.[15]刘金连.亚胺培南联合氨溴索治疗重症肺炎的疗效及对炎症因子的影响研究[J].中国现代药物应用,2022,16(8):139-141.[16]刘敬.低剂量甲强龙联合多巴胺在老年重症肺炎中的应用[J].智慧健康,2020,6(31):100-101.[17] CARR A C,SPENCER E,DIXON L,et al.Patients withcommunity acquired pneumonia exhibit depleted vitamin C status and elevated oxidative stress[J].Nutrients,2020,12(5):1318.[18] LIU W,MU X,WANG X,et al.Effects of comprehensivepulmonary rehabilitation therapy on pulmonary functions and blood gas indexes of patients with severe pneumonia[J].Experimental and Therapeutic Medicine,2018,16(3):1953-1957.[19] XUE M,DA C.The protective effect of Qingfei Huatan decoctionon pulmonary function and its influence on the immunological function in children with severe pneumonia[J].American Journal of Translational Research,2021,13(8):9404-9412.[20]葛燕萍,王绍谦,孔静.清肺化痰汤对重症肺炎(痰热壅肺证)患者中医证候积分、免疫功能及炎性因子的影响[J].中国中医急症,2022,31(8):1213-1216.(收稿日期:2022-12-27) (本文编辑:陈韵)①湖北省麻城市人民医院 湖北 麻城 438300通信作者:曾令浩微通道经皮肾镜取石术治疗输尿管结石的效果及其安全性评价张义木① 曾令浩①【摘要】 目的:探讨微通道经皮肾镜取石术(MPCNL)治疗输尿管结石的效果及其安全性评价。
人工肝治疗的临床实践与研究进展
人工肝治疗的临床实践与研究进展韩涛,张倩南开大学人民医院,天津市人民医院消化(肝病)科,天津 300121通信作者:韩涛,****************(ORCID: 0000-0003-4216-6968)摘要:人工肝支持系统是治疗肝衰竭的重要方法之一,近年来非生物型人工肝在肝衰竭救治中的作用越来越受到认可,在非肝衰竭疾病中的应用也日益广泛。
临床上需要综合多种因素,合理选择非生物型人工肝治疗的时机及模式,规范化、个体化、精准化治疗及不同模式的优化组合是人工肝临床应用的趋势。
生物型人工肝有关种子细胞来源、生物反应器等关键技术不断完善,且部分已经进入临床试验阶段。
尽管人工肝治疗的临床实践与研究已取得很大进展,但仍面临不少挑战。
如何通过技术创新与优化组合进一步提高其疗效与安全性,如何通过高质量的临床试验获得更高级别循证医学证据,仍是目前亟需解决的难题。
关键词:肝,人工;肝功能衰竭;治疗学Clinical practice and research advances in artificial liver support therapyHAN Tao,ZHANG Qian.(Department of Gastroenterology and Hepatology,Tianjin People’s Hospital,Tianjin Union Medical Center Affiliated to Nankai University, Tianjin 300121, China)Corresponding author: HAN Tao,****************(ORCID: 0000-0003-4216-6968)Abstract:Artificial liver support system is one of the important therapies for liver failure, and in recent years, the role of non-bioartificial liver support system in the treatment of liver failure has been gradually recognized, with wide application in non-liver failure diseases. In clinical practice,various factors should be considered to reasonably select the timing and mode of non-bioartificial liver support therapy,and standardized,individualized,and precise treatment and optimal combination of different modes are the trend of the clinical application of artificial liver support therapy. There have been constant improvements in the key techniques of bioartificial liver support system such as seed cell source and bioreactor, and some of them have entered the stage of clinical trial. Although remarkable progress has been made in the clinical practice and research of artificial liver support therapy,there are still many challenges,and it is urgently needed to solve the problems of how to further improve its efficacy and safety through technological innovation and combination optimization and how to obtain higher-level evidence-based medical evidence through high-quality clinical trials.Key words:Liver, Artificial; Liver Failure; Therapeutics肝衰竭是临床常见的因肝功能严重损伤引起的危重症,尤其是急性(亚急性)肝衰竭和慢加急性肝衰竭,病情进展快,病死率高。
Dose-Response
Classic Dose-Response Curve on Log – Log Coordinates
Non-Normal Distributions
Types of Exposure to Chemicals
Exposure may be classified as
Acute Exposure – This usually refers to a single exposure to a chemical. If repeated exposures are given they are given within a 24-hr period
Lethal dose (death) Toxic dose (ex. Liver injury) Effective dose (ex. Relief from itching)
Subchronic Testing
90 days is the most common test duration but 30 days to 90 days can be used Usually oral administration of the chemical via food; also implant Used to further characterize the specific organs affected by test compound after repeated administration of the chemical
Molecular Targets of Chemical Compounds
Receptors
TheQuickResponse...
Boise State UniversityScholarWorksLibrary Faculty Publications and Presentations The Albertsons Library4-1-2011The Quick Response (QR) Code: Graphic Potential for LibrariesMemo CordovaBoise State UniversityThis document was originally published by the Idaho Library Association in The Idaho Librarian. This work is provided under a Creative Commons Attribution License. Details regarding the use of this work can be found at:/licenses/by/3.0/./Font Size:Figure 1. 2D Progression from the stacked bar code method to the more information rich matrix method, aka QR Code.The QR code was developed by the Japanese company Denso Wave in the early 1990s, and was primarily used in the manufacturing sector to scan items in production lines. Soon after QR codes started showing up in Japanese retail and entertainment venues, and were adopted by a tech-savvy Japanese society.QR codes did not make a big impact in the United States until recently, thanks in part to the development of smartphones and other Web-ready devices and their accompanying corpus of applications. The private and commercial sectors have started using QR codes to promote, enhance, and engage this growing mobile demographic. Companies like Best Buy, Target, Nike and other retail businesses put QR codes in their advertisements to entice consumers to visit mobile-friendly product pages. For these types of businesses, QR codes are a low-cost, high-yield tool that can enhance a marketing campaign, and target tech-savvy consumer to visit mobile-friendly pages to showcase products or services. Because a QR code is a simple black and white image, one can print, paste, or embed a QR code just about anywhere. It’s a simple way to bridge the transmission of data from one point of access directly to the user’s mobile device via a scanner app and a camera.Figure 2. Best Buy makes weekly circular more interactive with mobile marketing.Create Your Ow n QR CodeThe process for making QR codes is fairly straightforward, and a good list of generators can be found at this web site: /qr-code-generators/. So what makes QR codes generators different? It depends what you want out of the code and what you want the end-user to experience. Some generators have basic differences: some are paid and some are free; some generators do not provide analytics (which measure visits to your website) or provide limited tracking data; some generators redirect scanned codes to a landing page not of your design (which is bad form from an end-user experience); some generators are the fly-by-night operation type, or their reliability is suspect; and some offer limited encoding abilities, meaning that only some types of codes can be generated.For QR codes to be an effective tool to optimize the online experience of your mobile user, here are some tips to take into consideration:1. QR codes are read by camera-ready devices in conjunction with a scanner or reader app. Provide a linksomewhere near the code where users can download a QR code reader/scanner.2. QR codes should link to specific websites, contact information, or a section of a larger body of information. Itshould be a direct static-to-mobile transaction, taking the user directly to the link encoded in the QR code without re-directs.3. Codes should be accompanied by contextual information—let the person scanning the code know where thecode will take them: a website, SMS, contact info, URL, YouTube link, etc.4. The code must be easy to scan: the bigger the code, the easier it is for the scanner to decipher the code. Thesmallest code should measure at least 1”x1”, but 2” x 2” is better.5. Leave at least a 1/8” of white space around the code. This makes it easier for the scanner application to visuallycue in to the code.6. For encoding links, a website like Bit.ly or TinyURL should be used to shorten the URL. The shorter the link, theless crowded the code will look.7. QR codes can be colorful and have embedded images, but simple black and white codes are easier for devicesto scan and are less error-prone.8. QR codes enable content to be accessed via a mobile device, so make the end destination a mobile-friendlypage.To learn more, visit my library guide about QR codes, where I suggest scanner applications for varying operating systems (OS) and best practices: “What is a QR Code?” /qrcodes.QR Code Use at Albertsons LibraryAt Albertsons Library I use a couple of QR code generators, depending on what I want to encode, but my favorites so far are Bit.ly and a QR code generator website developed by an IT professional named Kerem Erkam, located at/qr-code-and-2d-code-generator/.Bit.lyKnown for their simple, full-featured link shortener, Bit.ly offers a robust package that can track links you shorten using their service, and it can also generate a QR code based on shortened links. Bit.ly is my preferred QR code creator for several reasons. First, it’s free. Second, it provides a fairly robust dashboard for tracking links or finding out the number of times a QR code has been scanned. Third, I can customize shortened links to have names or titles instead of a random number-letter combination.If you give Bit.ly a try, I recommend creating an account, especially if you are going to create more than one QR code. Once you're logged in, simply shorten a link and Bit.ly will automatically generate a QR code for you. You can then save the QR code image that Bit.ly generates and embed, save, or print the QR code and post it as needed. Bit.ly will keep track of the number of times a link has been clicked and a QR code has been scanned.This screenshot below shows how Bit.ly’s dashboard details tracking data and provides a QR code specifically for a link I created for the library’s mobile website.Figure 3. Bit.ly Dashboard.Kerem Erkam’s WebsiteI use Bit.ly mainly to link to our library’s mobile site or other mobile-friendly links, so my use is focused on web-based, outbound URLs. However, when I need to create QR codes to encode other types of information, I use Mr. Erkam’s website. His “QR Code and 2D Code Generator” site provides an extensive array of codes (including Data Matrix, Aztec, and Micro codes) and encoding options, like maps, web links, calendar events, free text, SMS, and more. You can also change the color scheme of the code and shorten a link using some of the more popular link shorteners. Once you finish your selection and coding options a sizeable QR code is generated for you; from there you can right-click on the code and save it to your computer.Figure 4. Kerem Erkan’s QR Code and 2D Code Generator.QR Code Use in LibrariesA QR code is an image file that has information contained within. Because of its simple design and size, it can be printed on paper or displayed on a website. As long as the lines and squares of the code are sharp and well contrasted, they can be read by a scanner and its information disseminated effectively. QR codes can be placed pretty much anywhere and on anything, so deploying QR codes in your library is a question of preference. Libraries may wish to paste or display QR codes in any of these ways within their buildings:1. At the entrance of a library, linked to the library’s mobile website or a specific page on the mobile site (Hours,Circulation, reference phone number, etc.).2. Near the stairs or elevator, linked to a map or specific section of the library (floor plan, etc.).3. Near a new books display, linked to a collection of new books or takes the user to an RSS feed of newlypurchased library materials.4. Near the reference desk, linked to an email, text, or chat reference service.5. On a librarian’s door or window, linked to personal profile or subject guide.6. At the Circulation desk, linked to checkout procedures, fine information, etc.7. Near specific collection areas (children's, teens, career, reference, monthly book display, genealogy collection,etc.), linked to more information about the collection.8. Outside study or meeting rooms, linked to room policies.9. On library handouts, bookmarks, new database flyers, and promotional brochures and posters, linked to thelibrary main mobile site.Code Placement in Online Environments1. On the main library website and subpages, linked to the library’s mobile website or to a library’s social mediapresence (Twitter, Facebook, Flickr, etc.).2. Projected on digital frames and TV monitors in the library, linked to the library mobile site or relevant page.3. In library subject guides, librarian profiles, or staff web pages, linked to contact information for the individual.4. On subject-specific database pages, linked to librarian profiles or subject guides.One of the best lists I found on the use of QR codes to market your print and online presence is from the That’s Great blog, titled "101 Uses For Quick Response (QR) Codes: Creating Audience Engagement With The Next Killer US App."One can easily transfer the same conceptual promotional ideas to enhance the user experience with the manyresources available at your library. Another is Robin Ashford’s article in College & Research Libraries News titled, “QR codes and academic libraries.”QR codes are a simple tool to disseminate content to the hands of a growing mobile user demographic. A simple QR code strategy for your library is a low-cost and innovative way to interact with your users and provide them with easyaccess to library resources on their mobile device of choice.Figure 5. QR code linking to my profile on various social media sites.Works Cited"About 2D Code." (2010). Retrieved from /qrcode/aboutqr-e.htmlAshford, R. (2010). QR codes and academic libraries.College & Research Libraries News, 71 (10), 526-30. Retrieved from /content/71/10/526.fullBonsor, K. (2011). How augmented reality works. In HowStuffWorks. Retrieved from/augmented-reality1.htmCordova, M. (2011). What is a QR Code? In Albertsons Library Research Guides. Retrieved from/qrcodesErkan, K. (2011). QR Code and 2D Code Generator. Retrieved from /qr-code-and-2d-code-generator/IDC. (2011). Android Rises, Symbian^3 and Windows Phone 7 Launch as Worldwide Smartphone Shipments Increase The Idaho Librarian (ISSN: 2151-7738) is a publication of the Idaho Library Association.87.2% Year Over Year, According to IDC. Retrieved from /about/viewpressrelease.jsp?。
CHEMLOK TS701-43产品安全数据表说明书
USA SAFETY DATA SHEET3000000026721. CHEMICAL PRODUCT AND COMPANY IDENTIFICATIONProduct name: CHEMLOK TS701-43 Product Use/Class:AdhesiveLORD Corporation 111 LORD DriveCary, NC 27511-7923 USATelephone: 814 868-3180Non-Transportation Emergency: 814 763-2345 Chemtrec 24 Hr Transportation Emergency No.800 424-9300 (Outside Continental U.S. 703 527-3887)EFFECTIVE DATE: 05/16/20172. HAZARDS IDENTIFICATIONGHS CLASSIFICATION:Flammable liquids Category 2Acute toxicity Inhalation - Vapours Category 4Acute toxicity Inhalation - Dust and Mist Category 4 Skin corrosion/irritation Category 2Serious eye damage/eye irritation Category 2A Skin sensitization Category 1 Carcinogenicity Category 2Reproductive toxicity Category 2Specific target organ systemic toxicity (single exposure) Category 3Specific target organ systemic toxicity (single exposure) Category 1 Central nervous system, Kidney, Liver, Respiratory systemSpecific target organ systemic toxicity (repeated exposure) Category 2 Ears, Liver, KidneySpecific target organ systemic toxicity (repeated exposure) Category 1 Nervous system, Respiratory system, Central nervous system, Peripheral nervous systemHazardous to the aquatic environment - acute hazard Category 2Hazardous to the aquatic environment - chronic hazard Category 2GHS LABEL ELEMENTS: Symbol(s)Signal WordD ANGERHazard StatementsHighly flammable liquid and vapor. Harmful if inhaled. Causes skin irritation.Causes serious eye irritation.May cause an allergic skin reaction. Suspected of causing cancer.Suspected of damaging fertility or the unborn child. May cause harm to breast-fed children. May cause drowsiness or dizziness. May cause respiratory irritation.Causes damage to organs.(Central nervous system, Kidney, Liver, Respiratory system)May cause damage to organs through prolonged or repeated exposure.(Ears, Liver, Kidney)Causes damage to organs through prolonged or repeated exposure.(Nervous system, Respiratory system, Centralnervous system, Peripheral nervous system)Toxic to aquatic life.Toxic to aquatic life with long lasting effects.Precautionary StatementsPreventionKeep away from heat/sparks/open flames/hot surfaces. - No smoking.Ground/Bond container and receiving equipment.Use explosion-proof electrical/ventilating/lighting equipment.Use only non-sparking tools.Take precautionary measures against static discharge.Obtain special instructions before use.Do not handle until all safety precautions have been read and understood.Wear protective gloves/protective clothing/eye protection/face protection.Use personal protective equipment as required.Do not breathe dust/fume/gas/mist/vapors/spray.Wash thoroughly after handling.Do not eat, drink or smoke when using this product.Use only outdoors or in a well-ventilated area.Contaminated work clothing should not be allowed out of the workplace.Avoid release to the environment.ResponseIn case of fire: refer to section 5 of SDS for extinguishing media.Call a POISON CENTER or doctor/physician if you feel unwell.IF exposed: Call a POISON CENTER or doctor/physician.Specific treatment (see supplemental first aid instructions on this label).IF INHALED: Remove to fresh air and keep at rest in a position comfortable for breathing.IF ON SKIN (or hair): Remove/Take off immediately all contaminated clothing. Rinse skin with water/shower.If skin irritation or rash occurs: Get medical advice/attention.IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do.Continue rinsing.Collect spillage.StorageStore in a well-ventilated place. Keep cool.Store in a well-ventilated place. Keep container tightly closed.Store locked up.Disposal:Dispose of contents/container in accordance with waste/disposal laws and regulations of your country or particular locality.Other Hazards:This product contains component(s) which have the following warnings; however based on the GHS classification criteria of your country or locale, the product mixture may be outside the respective category(s).Acute: Vapor harmful; may affect the brain or nervous system causing dizziness, headache or nausea. Possibleirritation of the respiratory system can occur causing a variety of symptoms such as dryness of the throat, tightness of the chest, and shortness of breath. May cause central nervous system depression characterized by the followingprogressive steps: headache, dizziness, staggering gait, confusion, unconsciousness or coma. Phenolic novolac resins have been found to have weak skin sensitizing potential; they rarely cause allergic skin response. This productcontains a residual amount of a chemical substance that may cause an allergic skin and/or respiratory reaction.May be harmful if swallowed. Ingestion is not an expected route of entry in industrial or commercial uses.Chronic: May affect the gastrointestinal system. May affect the blood and blood-forming organs. Ethylbenzene has been classified by IARC as a possible human carcinogen (Group 2B) and reported by NTP to show clear evidence for carcinogenicity in animals.3. COMPOSITION/INFORMATION ON INGREDIENTSChemical Name CAS Number RangeMethyl ethyl ketone78-93-350 - 55%Xylene1330-20-720 - 25%Ethyl benzene100-41-4 5 - 10%Epoxy novolac resin PROPRIETARY 1 - 5%Any "PROPRIETARY" component(s) in the above table is considered trade secret, thus the specific chemical and its exact concentration is being withheld.4. FIRST AID MEASURESFIRST AID - EYE CONTACT: Flush eyes immediately with large amount of water for at least 15 minutes holding eyelids open while flushing. Get prompt medical attention.FIRST AID - SKIN CONTACT: Flush contaminated skin with large amounts of water while removing contaminated clothing. Wash affected skin areas with soap and water. Get medical attention if symptoms occur.FIRST AID - INHALATION: Move person to fresh air. Restore and support continued breathing. If breathing is difficult, give oxygen. Get immediate medical attention.FIRST AID - INGESTION: If swallowed, do not induce vomiting. Call a physician or poison control center immediately for further instructions. Never give anything by mouth if victim is rapidly losing consciousness, unconscious or convulsing.5. FIRE-FIGHTING MEASURESSUITABLE EXTINGUISHING MEDIA: Carbon Dioxide, Dry Chemical, Foam, Water FogUNSUITABLE EXTINGUISHING MEDIA: Not determined for this product.SPECIFIC HAZARDS POSSIBLY ARISING FROM THE CHEMICAL: Flammable liquid and vapor. Keep containers tightly closed. Isolate from heat, electrical equipment, sparks, open flame, and other sources of ignition. Closed containers may rupture when exposed to extreme heat. Use water spray to keep fire exposed containers cool. During a fire, irritating and/or toxic gases and particulate may be generated by thermal decomposition or combustion.SPECIAL PROTECTIVE EQUIPMENT AND PRECAUTIONS FOR FIRE-FIGHTERS: Wear full firefighting protective clothing, including self-contained breathing apparatus (SCBA). Water spray may be ineffective. If water is used, fog nozzles are preferable.6. ACCIDENTAL RELEASE MEASURESPERSONAL PRECAUTIONS, PROTECTIVE EQUIPMENT, AND EMERGENCY PROCEDURES: Remove all sources of ignition (flame, hot surfaces, and electrical, static or frictional sparks). Avoid contact. Avoid breathing vapors. Use self-contained breathing equipment.ENVIRONMENTAL PRECAUTIONS: Do not contaminate bodies of water, waterways, or ditches, with chemical or used container.METHODS AND MATERIALS FOR CONTAINMENT AND CLEANUP: Keep non-essential personnel a safe distance away from the spill area. Notify appropriate authorities if necessary. Avoid contact. Before attempting cleanup, refer to hazard caution information in other sections of the SDS form. Contain and remove with inert absorbent material and non-sparking tools.7. HANDLING AND STORAGEHANDLING: Keep closure tight and container upright to prevent leakage. Ground and bond containers when transferring material. Avoid skin and eye contact. Wash thoroughly after handling. Avoid breathing of vapor or spray mists. Do not handle until all safety precautions have been read and understood. Empty containers should not be re-used. Use with adequate ventilation. Because empty containers may retain product residue and flammable vapors, keep away from heat, sparks and flame; do not cut, puncture or weld on or near the empty container. Do not smoke where this product is used or stored.STORAGE: Do not store or use near heat, sparks, or open flame. Store only in well-ventilated areas. Do not puncture, drag, or slide container. Keep container closed when not in use. Refer to OSHA 29CFR Part 1910.106 "Flammable and Combustible Liquids" for specific storage requirements.INCOMPATIBILITY: Strong oxidizers, acids, bases, water.8. EXPOSURE CONTROLS/PERSONAL PROTECTIONCOMPONENT EXPOSURE LIMITChemical Name ACGIH TLV-TWA ACGIH TLV-STELOSHA PEL-TWAOSHA PEL-CEILINGSkinMethyl ethyl ketone200 ppm300 ppm590 mg/m3200 ppmN.E.N.A.Xylene100 ppm150 ppm435 mg/m3100 ppmN.E.N.A.Ethyl benzene20 ppm N.E.435 mg/m3100 ppmN.E.N.A.Epoxy novolac resin N.E.N.E.N.E. N.E.N.A.N.A. - Not Applicable, N.E. - Not Established, S - Skin DesignationEngineering controls: Sufficient ventilation in pattern and volume should be provided in order to maintain air contaminant levels below recommended exposure limits. Caution: Solvent vapors are heavier than air and collect in lower levels of the work area. Sufficient ventilation (using explosion-proof equipment) should be provided to prevent flammable vapor/air mixtures from accumulating.PERSONAL PROTECTION MEASURES/EQUIPMENT:RESPIRATORY PROTECTION: Use a NIOSH approved chemical/mechanical filter respirator designed toremove a combination of particulates and organic vapor if occupational limits are exceeded. For emergencysituations, confined space use, or other conditions where exposure limits may be greatly exceeded, use an approved air-supplied respirator. For respirator use observe OSHA regulations (29CFR 1910.134) or use in accordance with applicable laws and regulations of your country or particular locality.SKIN PROTECTION: Use neoprene, nitrile, or rubber gloves to prevent skin contact.EYE PROTECTION: Use safety eyewear including safety glasses with side shields and chemical goggles where splashing may occur.OTHER PROTECTIVE EQUIPMENT: Use disposable or impervious clothing if work clothing contamination is likely. Remove and wash contaminated clothing before reuse.HYGIENIC PRACTICES: Wash hands before eating, smoking, or using toilet facility. Do not smoke in anychemical handling or storage area. Food or beverages should not be consumed anywhere this product is handled or stored. Wash thoroughly after handling.9. PHYSICAL AND CHEMICAL PROPERTIESTypical values, not to be used for specification purposes.ODOR: Solvent VAPOR PRESSURE: N.D.APPEARANCE: Clear VAPOR DENSITY: Heavier than Air PHYSICAL STATE: Liquid LOWER EXPLOSIVE LIMIT: 1 %(V)FLASH POINT: 28 °F, -2 °C SetaflashClosed CupUPPER EXPLOSIVE LIMIT: 11.4 %(V)BOILING RANGE: 80 - 141 °C EVAPORATION RATE: Not Applicable AUTOIGNITION TEMPERATURE:N.D.DENSITY: 0.9 g/cm3 - 7.49 lb/gal DECOMPOSITION TEMPERATURE:N.D. VISCOSITY, DYNAMIC: ≥60 mPa.s @ 25 °C ODOR THRESHOLD: N.D.VISCOSITY, KINEMATIC: ≥67 mm2/s @ 25 °CSOLUBILITY IN H2O: Insoluble VOLATILE BY WEIGHT: 80.55 %pH: N.A.VOLATILE BY VOLUME: 88.54 %FREEZE POINT: N.D. VOC CALCULATED: 6.03 lb/gal, 729 g/l COEFFICIENT OF WATER/OIL N.D.DISTRIBUTION:LEGEND: N.A. - Not Applicable, N.E. - Not Established, N.D. - Not Determined10. STABILITY AND REACTIVITYHAZARDOUS POLYMERIZATION: Hazardous polymerization will not occur under normal conditions. STABILITY: Product is stable under normal storage conditions.CONDITIONS TO AVOID: High temperatures. Sources of ignition.INCOMPATIBILITY: Strong oxidizers, acids, bases, water.HAZARDOUS DECOMPOSITION PRODUCTS: Carbon dioxide, carbon monoxide, chlorine, hydrogen chloride, Phosgene11. TOXICOLOGICAL INFORMATIONEXPOSURE PATH: Refer to section 2 of this SDS.SYMPTOMS:Refer to section 2 of this SDS.TOXICITY MEASURES:Chemical Name LD50/LC50Methyl ethyl ketone Oral LD50: Rat2,483 mg/kgDermal LD50: Rabbit5,000 mg/kgInhalation LC50: Rat11700 ppm/4 hXylene Oral LD50: Rat3,500 mg/kgDermal LD50: Rabbit> 4,350 mg/kgInhalation LC50: Rat29.08 mg/l/4 hEthyl benzene Oral LD50: Rat3,500 mg/kgDermal LD50: Rabbit15,400 mg/kgInhalation LC50: Rat17.4 mg/l/4 hEpoxy novolac resin N.D.Germ cell mutagenicity: No classification proposedCarcinogenicity: Category 2 - Suspected of causing cancer.Components contributing to classification: Ethyl benzene.Reproductive toxicity: Category 2 - Suspected of damaging fertility or the unborn child. May cause harm to breast-fed children.Components contributing to classification: Xylene. Ethyl benzene. Toluene. Acetone.12. ECOLOGICAL INFORMATIONECOTOXICITY:Chemical Name EcotoxicityMethyl ethyl ketone Fish: Pimephales promelas3,130 - 3,320 mg/l96 h flow-throughInvertebrates: Daphnia magna> 520 mg/l48 hDaphnia magna5,091 mg/l48 hDaphnia magna4,025 - 6,440 mg/l48 h StaticXylene Fish: Pimephales promelas13.4 mg/l96 h flow-throughOncorhynchus mykiss2.661 - 4.093 mg/l96 h StaticOncorhynchus mykiss13.5 - 17.3 mg/l96 hLepomis macrochirus13.1 - 16.5 mg/l96 h flow-throughLepomis macrochirus19 mg/l96 hLepomis macrochirus7.711 - 9.591 mg/l96 h StaticPimephales promelas23.53 - 29.97 mg/l96 h StaticCyprinus carpio780 mg/l96 h semi-staticCyprinus carpio> 780 mg/l96 hPoecilia reticulata30.26 - 40.75 mg/l96 h StaticInvertebrates: water flea3.82 mg/l48 hGammarus lacustris0.6 mg/l48 hEthyl benzene Fish: Oncorhynchus mykiss11.0 - 18.0 mg/l96 h StaticOncorhynchus mykiss4.2 mg/l96 h semi-staticPimephales promelas7.55 - 11 mg/l96 h flow-throughLepomis macrochirus32 mg/l96 h StaticPimephales promelas9.1 - 15.6 mg/l96 h StaticPoecilia reticulata9.6 mg/l96 h StaticInvertebrates: Daphnia magna1.8 - 2.4 mg/l48 hPlants: Pseudokirchneriella subcapitata4.6 mg/l72 hPseudokirchneriella subcapitata> 438 mg/l96 hPseudokirchneriella subcapitata2.6 - 11.3 mg/l72 h StaticPseudokirchneriella subcapitata1.7 - 7.6 mg/l96 h StaticEpoxy novolac resin N.D.PERSISTENCE AND DEGRADABILITY:Not determined for this product.BIOACCUMULATIVE: Not determined for this product.MOBILITY IN SOIL: Not determined for this product.OTHER ADVERSE EFFECTS: Not determined for this product.13. DISPOSAL CONSIDERATIONSDISPOSAL METHOD: Disposal should be done in accordance with Federal (40CFR Part 261), state and local environmental control regulations. If waste is determined to be hazardous, use licensed hazardous waste transporter and disposal facility.14. TRANSPORT INFORMATIONUS DOT RoadDOT Proper Shipping Name: AdhesivesDOT Hazard Class: 3SECONDARY HAZARD: NoneDOT UN/NA Number: 1133Packing Group: IIEmergency Response Guide Number: 128IATA CargoPROPER SHIPPING NAME: AdhesivesDOT Hazard Class: 3HAZARD CLASS: NoneUN-NUMBER: 1133PACKING GROUP: IIEMS: 3LIMDGPROPER SHIPPING NAME: AdhesivesDOT Hazard Class: 3HAZARD CLASS: NoneUN-NUMBER: 1133PACKING GROUP: IIEMS: F-EThe listed transportation classification applies to non-bulk shipments. It does not address regulatory variations due to changes in package size, mode of shipment or other regulatory descriptors. For the most accurate shipping information, refer to your transportation/compliance department.15. REGULATORY INFORMATIONU.S. FEDERAL REGULATIONS: AS FOLLOWS:SARA SECTION 313This product contains the following substances subject to the reporting requirements of Section 313 of Title III of the Superfund Amendment and Reauthorization Act of 1986 and 40 CFR part 372.:Chemical Name CAS Number Weight % Less ThanXylene1330-20-725.0%Ethyl benzene100-41-410.0%TOXIC SUBSTANCES CONTROL ACT:INVENTORY STATUSThe chemical substances in this product are on the TSCA Section 8 Inventory.EXPORT NOTIFICATIONThis product contains the following chemical substances subject to the reporting requirements of TSCA 12(B) if exported from the United States:None16. OTHER INFORMATIONUnder HazCom 2012 it is optional to continue using the HMIS rating system. It is important to ensure employees have been trained to recognize the different numeric ratings associated with the HazCom 2012 and HMIS schemes.HMIS RATINGS - HEALTH: 2* FLAMMABILITY: 3 PHYSICAL HAZARD: 0* - Indicates a chronic hazard; see Section 2Revision: Section 2, Section 11Effective Date: 05/16/2017DISCLAIMERThe information contained herein is, to the best of our knowledge and belief, accurate. However, since the conditions of handling and use are beyond our control, we make no guarantee of results, and assume no liability for damages incurred by use of this material. It is the responsibility of the user to comply with all applicable federal, state and local laws and regulations.。
大修后文章直接录用
5. Response to comment: Point is entitled "stability and repeatability of ******s". Somequestions: Here, it is shown the dependence of SERS relative intensities of SC on time. Is it the same sample (******-adsorbate) that is stored and then SERS spectra are recorded at different times or instead, the same ****** is stored and then the adsorbate is added at different times and thereafter the SERS spectra are recorded Authors should clarify the procedure. Are different results expected with these two procedures Have been these two methods checked It is possible that only ******s stored without adsorbate are active for a longer timeExperimentally it is often observed that roughed surface in SERS produces molecular degradation and a comparation between Raman and SERS spectra is necessary to identify the molecular fundamental modes. Why the authors did not record Raman spectra of SC Raman spectra should be added to Fig. 5. The point should be because a characteristic of surface is explained and must follow point . The old becomes . Moreover, old epigrap****** 3.2.1. and should be removed and a unique point . should be presented instead and entitled " Biocompatibility of ******s. SERS spectra of ****** and ******". Also, epigraph must be removed and the text must follow to the new section .Response: We are very sorry for our unclear report in the section of .For the first question proposed by the Reviewer, our answer is as following:Yes, in the section of , the main intention is to display the stability and reproducibility of Agnanofilms. Fig. 6 (in the original manuscript, Fig. 5 in the revised manuscript) shows the dependence of SERS relative intensities of SC on time. In this section, the ******s are stored and then the adsorbate is added at different times and thereafter the SERS spectra are recorded. For the second question proposed by Reviewer, our answer is as following:As the Reviewer's good instruction, we have clarified the procedure in the revised manuscript.However, in this study, we have not studied the first procedure of the same sample (******-adsorbate) that is stored and then SERS spectra are recorded at different times. Thus, these two methods have not been checked. Take the time limit of the submission of the revised manuscript into account; it is difficult to carry out the first procedure in the revised manuscript. However, we appreciate for the Reviewers’ good advice earnestly. We will check these two procedures in future studies. For the third question proposed by Reviewer, our answer is as following: According to the Reviewer's good instruction, we have recorded Raman spectra of SC in solid state, as shown in Fig. 1(a) (Fig. 4(a) in revised manuscript). Meanwhile, the points and have also been re-written according to the instructions proposed by the Reviewer. (See the section of and 3.3 in the revised manuscript)图略spectrum of (a) solid SC. SERS spectra of ×10mol/L SC aqueous solutionadsorbed on (b) glass slide, (c) ******-Ag CNPs, and (d) ******, respectively.6. Response to comment: Sentence "the number of spectra for every condition is five" (page 9) or the number of spectra is 5 (Table 1 and 2), what does it mean Perhaps something like this: EachSERS spectrum is recorded 5 times in different points of the ****** surface?Response: As the Reviewer's good question, the sentence of “the number of spectra for everycondition is five" or “the number of spectra is 5 (Table 1 and 2)” in the original manuscr ipt is hard to be understood.This sentence means that each SERS spectrum is recorded 5 times in differpoints of the ****** surface. we have revised this sentence in the revised manuscript according to the Reviewer's advice.7. Response to comment: Epigrap****** of Table 1 and 2 should be revised. Corrections are indicatedin the manuscript.Response: According to the Reviewer's good instruction, we have revised the epigrap****** of Table1 and 2. The epigraph of Table 1 “Table 1 Preliminary assignations of the Raman bands (Mean ± ., n=5) for the SERS spectra of ******. The number of spectra is 5” is revised as“Assignments for the SERS bands (Mean ± ., n=5) of ****** (based on Refs. [25-30]).”The epigraph of Table 2 “Table 2 Observed wavenumbers (Mean ± ., n=5), assignments,and local coordinates of ******, excited at 785 nm. The number of spectra is 5. [35-37]” is revised as “Table 2Assignments for the SERS bands (Mean ± ., n=5) of ****** (based on Refs. [32-34]).”8. Response to comment: Figure 4 shows SERS spectra of ****** and ****** recorded on different Ag nanofilms. What do authors want to say This experiment is to check the reproducibility of the method Thus, it is better to use the word "reproducibility" and not "repeatability". This must be clarified in the text.Response: As the Reviewer's good advice, we should use the word “reproducibility” and not"repeatability". Figure 4 (in the original manuscript) shows SERS spectra of ****** and ****** recorded on the different ******s prepared under the same conditions. The authors want to display the reproducibility of the ******s prepared by this simple method. Thus, we have replaced theword "repeatability" by "reproducibility" in the revised manuscript.9. Response to comment: Uv-vis absorption spectrum of不******-protected Ag nanoparticules shows a maximum at 418 nm and at 785 nm the absorbance is zero (Figure 1b). Given that SERS spectra are recorded at 785 nm, I think that this excitation line is very far from the maximum to obtain a good signal. In fact, the Raman signal is very weak (Figure 3). Is it possible to obtain a better signal employing another excitation laser, for example, 514nm I mean, probably the 785nm line is better for ****** than for ******-protected Ag nanoparticules, but under other different excitation laser the ******-protected Ag nanoparticules could be a good substrate for ****** or ******. Have been checked different excitation laser linesResponse: We are very sorry for our negligence of the explanation for why SERS spectra of ******and ****** are recorded at 785 nm. In the studying of the SERS effect of ****** and ******, we think that the excitation with the 785 nm wavelength has a number of advantageous features compared to [a] other wavelengt******. A previous study has reported that a laser wavelength shorter than nm is known to enhance photodissociation and cause protein degradation even at a low power. However, the sample damage can be avoided using laser light of a longer wavelength. No paling effects were observed using laser light with wavelength ≥660 nm. In their study, degradation of the biological objects was observed when using nm excitation lasers. Meanwhile, it is known that when using 660 nm irradiation, for a laser power of 10mW and a diameter of 10 μm, 2the light intensity is up to 127 MW/m . In our system, the laser power is set at 65mW and the 2 3 2 diameter is 90μm. So the light intensity was ca. 10 MW/m(10 W/cm ), which is much smaller than that of 660 nm irradiation. Thus, in our studies, we employed a 65-mW, 785-nm diode laser to record the Raman and SERS spectra of ****** and ****** in order to avoid the photodissociation and degradation of the proteins.[a] G. J. Puppels, J. H. F. Olminkhof, G. M. J. Segers-Nolten, C. Otto, F. F. M. Demul, J. Greve. Laser Irradiation and Raman Spectroscopy of Single Living Cells and Chromosomes: Sample Degradation Occurs with nm but not with 660 nm Laser Light . Exp. Cell Res, 195 (1991) 361.Special thanks to you for your good comments.Reviewer 4#1. Response to comment: I'm not familiar with blood sample preparation but I wasn't able to extract the final concentration of ****** and ****** added to the ******-NPs and ******. This value has to be clearly reported in the text. Response: As the Reviewer suggested that we have calculated the final concentration of ****** and ****** added to the ******-protected ****** (******-Ag CNPs) and ****** in the revised manuscript. According our calculation, the final concentration of ****** and ****** added to ****** is ca.4.8 and %, respectively. However, take the dilution of the Ag colloid into account, the final concentration of ****** and ****** added to the ******-Ag CNPs is and %, respectively. All these concentrations have been added in the revised the final concentrations of ****** added to the ******-NPs and ****** are different, the quantity of ****** lighted by the laser spot based on ****** g) equals to (in order of magnitude) that based on ******-Ag CNP samples (ca. ×10g). Similarly, the quantity of ****** lighted by the laser spot based on ****** g) equals to (in order of magnitude) that based on ******-Ag CNPs (ca. ×10g). Take the SERS detection of ****** for example: For the SERS detection of ****** based on ******-Ag CNPs, the probe volume is considered a focal [b]“tube”with a waist diameter of 90 m and a depth of ca. 1cm. By using the concentration of ****** %) one can determine the quantity of ****** molecules contributing to the Raman intensity ( (1000+60) ×10×% × (45 ×10 )×1×10 / [(1000+60)×10 ] ~ ×10g). For the SERS detection of ****** based on ******, supposing that all the ****** molecules adsorbed on the surface of ****** evenly, one can determine the quantity of ****** molecules contributing to the Raman intensity (~ (50) ×10×% × (45 ×10 )/ (5 ×10 )~ ×10g).[b] L. Baia, K. Gigant, U. Posset, et al. Appl. Spectrosc, 56 (2002) 536.(2) Response to comment: In the experimental results section does not appear how the SERSmeasurements on ******-NPs were performed.Response: We are very sorry for our negligence of introduce how the SERS measurements on ******-Ag CNPs were performed in the experimental results section. As Reviewer suggested that we have made complementarities of this experimental process in the section of of the revised manuscript. (the section of , 16-20 lines)(3) Response to comment: A significant comparison between the SERS activities of ******-NPsand ****** substrates is not possible based on what reported in the paper. For instance, is thenumber of NPs lighted by the laser spot equals for ******-NP and ****** samples The authors have to make clear to the reader what they are comparing. Moreover, the normal Raman of ****** and ****** in solution and/or in solid state in fig. 3 would be helpful to understand the effect of the metal-analyte interaction.Response: We are very sorry for our unclear description of what the SERS activities of ******-NPsand ****** substrates are comparing. In Fig 3 of the original manuscript (Fig 8 in revisedmanuscript ), we mainly want to tell the reader the ******-Ag CNPs are unfit for the SERS detectionof ****** and ******, compared to the ******s prepared by electrostatic self-assembly with the******-Ag CNPs. That is, the SERS effect of ******-Ag CNPs for ****** and ****** is much weaker than that of ******. According to the Reviewer good question (“is the number of NPs lighted by the laser spot equals for ******-NP and ****** samples”), we have calculated the number of NPs lighted by the laser spot for ******-Ag CNPs and ******s. The calculation results show that the number of NPs lighted by the laser spot is not equal for ******-Ag CNPs and ******s. One can calculate the concentration of the ******-Ag CNPs in Ag colloid is ca. ×10 /m according to the electrochemical reaction in our work. For the SERS detection based on ******-Ag CNPs, the probe volume was considered a focal “tube” with a waist diameter of 90 m and a depth of ca. 1cm. By using the concentration of ******-Ag CNPs in Ag colloid ×10 /m ) one can determine the number of NPs lighted by the laser spot ×10 × (45×10 )×1×10 ~ ×10 ). For the SERS detection based on ******, by supposing that thedistribution of the NPs on the surface of ****** is uniform, one can determine the number of NPs lighted by the laser spot ( (45 ×10 )/ (200 ×10 /2)~×10). It can be seen that, the number of NPs lighted by the laser spot for ******-Ag CNPs is more than that for ******s. However, the experimental results show that the SERS effect of ****** and ****** on ******-Ag CNPs is much weaker than that on ******s, indicating that the SERS activity of ******s is better than that of ******-Ag CNPs. As the Reviewer suggested that, we have recorded the normal Raman scattering of ****** and ****** in solid state, as shown in Fig 2A(c) and Fig 2B(c), respectively. Very weak Raman signals are observed from solid ****** and ******. We know that Raman spectroscopy is a powerful tool that gives precise information on the vibration energies of molecules, and can provide the fingerprint for unique chemical identification. However, the Raman scattering cross section of most biologicalmacromolecules is extremely small, which causes the Raman signals of biological[c]macromolecules are very difficult to obtain . Thus, in this paper, we attempt to obtain new SERS substrates to solve this question. Such as the Ag nanofilms prepared by using e lectrostatic self-assembly with the ******-Ag CNPs in this work.图略Fig. 2. (a) Raman spectrum of (A) solid ****** and (B) solid ******. SERS spectra of (A) ****** and (B)****** solution adorbed on (b) ******-Ag CNPs and (c) ******, respectively.[c] ,, Appl. Phys. Lett. 91 (2005) 223105.(4) Response to comment: It is quite expected that larger EM enhancement occurson aggregated NPs adsorbed on the glass slide with respect to the un-aggregated ******-NPs. As far as I understood, the key point of the paper is the fabrication of ****** with interparticle regions of ca. 300 nm where the ****** and ****** molecules can be located, leading then to intense SERS signals. Therefore, this new substrate should be compared to other silver nanofilms which do not present these cavities, in order, also, to give some evidences to the previous hypothesis, which is a mere speculation based on the results reported in the article.Response: As the Reviewer suggested that we have checked the new substrate with another silvernanofilm which dose not present these cavities. Fig.3a (following) shows the SEM image of acommon ****** prepared using coupling agent of cysteamine and common Ag NPs on thesurface of the glass slide. It shows that the average size of the particles on the surface of this common ****** is ca.75 ± 5 nm, which is much smaller than that (200 ± 50 nm) of theparticles on the surface of the new ****** prepared using electrostatic self-assembly with******-Ag CNPs . Meanwhile, single layer of Ag nanoparticles is observed and no lots ofnano-scale regions are formed on the surface of the common ******. (following) shows the SERS spectra of ****** solution %, 50 L) on this common Ag nanofilm (Fig.4 a) and the new ****** with a diameter of 1 cm, respectively. It shows that the EM enhancement occurs on the common ****** is weaker than that on the new ******. Meanwhile, striking spectral differences are seen in SERS spectra at 812, and 1022 cm , indicating that the orientation of the adsorbed ****** molecules is different on these two ******s. We think that it is mainly related to the surface characteristics of this new Ag nanofilm. On the surface of the new ******, the average size of the aggregated particles is up to ca. 200 ± 50 nm, which is much larger than that of the common ******. It improves the adsorption ability of the ****** molecules onto the surface of the new ****** effectively. Meanwhile, lots of nano-scale regions with the size of ca. 300 ± 50 nm are formed between the adjacent nanoparticles on the surface of the new ******, which makes ****** molecules can be embedded in effectively. It implies that the presence of nano-scale regions on the surface of the new ****** is an important factor for SERS effect of biologicalmacromolecules.图略. SEM image of the ****** prepared by using coupling agent of cysteamine (a) and byusing electrostatic self-assembly with ******-Ag CNPs (b) on glass slide.图略Fig. 4. SERS spectra of ****** solution %) on (a) the common ****** prepared by usingcoupling agent of cysteamine and common Ag nanoparticles and (b) on the ****** prepared byusing electrostatic self-assembly with ******-Ag CNPs, respectively.(5) Response to comment: The SERS spectra showed in fig. 5c is doubtfully assigned to ******. I suggest to the authors to carry out control experiments so that to rule out the presence of impurities on their silver film. In the following paper is reported the SERS spectrum of Citrate anion on silver NPs: Title: Anomalous Raman bands appearing in surface-enhanced Raman spectra Author(s): Sanchez-Cortes, S; Garcia-Ramos, JV Source: JOURNAL OF RAMAN SPECTROSCOPY Volume: 29 Issue: 5 Pages: 365-371 Published: MAY 1998Response: We are very grateful for your providing of the reference above. We have read this reference carefully. As Sanchez-Cortes et al. reported that the ions and new molecular speciesresulting from the reduction of the metal will remain in the colloid system (especially in the citrate colloids), which has an obvious impact on the SERS of the analyte. Thus, in our present work, we employed the method of electrolysis to obtain ******-Ag CNPs, and employed the method of electrostatic self-assembly to obtain ******s. In our experiment of preparation of ******-Ag CNPs, ****** was not employed act as reducing agent in order to avoid the effect of citrate anion on the SERS of the analyte recorded on the surface of the new ****** the Reviewer’s good instruction, we have carried out a control experiment. In this control experiment, we have checked the Raman scattering on the surfaceof the new ********s prepared under the same conditions, as shown in following Fig .5. The experimental results show that the effect of the impurities on the surfaces of these ******s on the SERS of the analyte is negligible. Meanwhile, as reported by Sanchez-Cortes et al., the Raman spectrum of solid citrate is shown [d]in Fig. 6 a. We can see that the main SERS bands of solid citrate recorded by us (Fig. 7) are close to the Raman bands of solid citrate reported by Sanchez-Cortes et al.图略. Raman spectra of the surfaces of the different ******s (a, b, c, d) prepared usingelectrostatic self-assembly with ******-Ag CNPs.图略Fig. 6. Effect of excitation wavelength on the SERS background of an aggregated citrate colloid:(A) FT-Raman spectrum of solid citrate (λex=l 064 nm); (B) FT-Raman spectrum of aqueous citrate (2 M, λex=l 064 nm); (C) FT-SER spectrum of the aggregated colloid after addition of NO3(0.05 M) (λex=1064nm); (D) SERS spectrum of the same sam ple with excitation at nm.图略Fig. 7. SERS spectra of ****** based on ****** prepared by using electrostaticself-assembly with ******-Ag CNPs.[d] S. S. Corte and J. V. G. Ramos. Anomalous Raman Bands Appearing in Surface-EnhancedRaman Spectra. JOURNAL OF RAMAN SPECTROSCOPY. 29 (1998) 365.Special thanks to you for your good comments and suggestions!Dear Editors and Reviewers.We have tried our best to revise and improve the manuscript and made great changes in themanuscript according to the Reviwers′good comments. And here we did not list the changes butmarked in red in revised paper.We appreciate for Editors/Reviewers’ warm work earnestly, and hope that the corrections willmeet with approval.Once again, thank you very much for your comments and suggestions.We look forward to your information about my revised papers and thank you for your goodcomments.Yours sincerely,. Liu。
Response_to_Reviewer-第二次
………。
,Ph。
D。
ProfessorLaboratory of Plant Nutrition andEcological Environment Research,Huazhong Agricultural University,Wuhan, 430070, P。
R.ChinaE-mail:liugl0924@Aug。
1st, 2009RE:HAZMAT-D-09-00655R1Dear Editor,We thank the reviewers and editor for the time in closely viewing our manuscript。
We would like to thank the reviewers for giving us constructive suggestions which would help us both in English and in depth to improve the quality of the paper. Here we submit a new version, which has been modified according to the reviewers' suggestions. Efforts were also made to correct the mistakes and improve the English of the manuscript。
We mark all the changes in red in the revised manuscript。
Sincerely yours,………………,Ph.D。
Professor—-——--—-—-——----——————-—---—--——-—-----——----————-———————-—--——---—--—-——---—---—-—-—-—----—-—-—-—---—-The following is a point—to—point response to the two reviewers’ comments.Reviewer #1:General comments:Reviewer #1:Page 3, lines 49—50:Is it true that "…the thickness of application agriculture films in some regions in China is less than 0。
社区获得型肺炎英国指南
347Downloaded from at Library of Medical Center of Fudan University on April 21, 2014/348Bartlett et al.CID2000;31(August)is not available initially but is subsequently reported,changing to the antimicrobial agent that is most cost-effective,least toxic, and most narrow in spectrum is encouraged.Recommendations for treating patients who require empirical antibiotic selection are based on severity of illness,pathogen probabilities,resis-tance patterns of S.pneumoniae(the most commonly implicated etiologic agent),and comorbid conditions.The recommendation for outpatients is administration of a macrolide,doxycycline,orfluoroquinolone with enhanced ac-tivity against S.pneumoniae.For patients who are hospitalized, the recommendation is administration of afluoroquinolone alone or an extended-spectrum cephalosporin(cefotaxime or ceftriaxone)plus a macrolide.Patients hospitalized in the in-tensive care unit(ICU)should receive ceftriaxone,cefotaxime, ampicillin-sulbactam,or piperacillin-tazobactam in combina-tion with afluoroquinolone or macrolide.b-lactams,other than those noted,are not recommended.Intravenous antibiotics may be switched to oral agents when the patient is improving clin-ically,is hemodynamically stable,and is able to ingest drugs. Most patients show a clinical response within3–5days. Changes evident on chest radiographs usually lag behind the clinical response,and repeated chest radiography is generally not indicated for patients who respond.The failure to respond usually indicates an incorrect diagnosis;host failure;inappro-priate antibiotic;inappropriate dose or route of administration; unusual or unanticipated pathogen;adverse drug reaction;or complication,such as pulmonary superinfection or empyema. Prognosis.The most frequent causes of lethal community-acquired pneumonia are S.pneumoniae and Legionella.The most frequent reason for failure to respond is progression of pathophysiological changes,despite appropriate antibiotic treatment.Pneumococcal pneumonia.S.pneumoniae,the most com-mon identifiable etiologic agent of pneumonia in virtually all studies,accounts for about two-thirds of bacteremic pneumonia cases,and pneumococci are the most frequent cause of lethal community-acquired pneumonia.Management has been com-plicated in recent years by the evolution of multidrug resistance. b-lactams(amoxicillin,cefotaxime,and ceftriaxone)are gen-erally regarded as the drugs of choice,although pneumonia caused by resistant strains(MIC,у2m g/mL)may not respond as readily as pneumonia caused by more susceptible strains. The activity of macrolides and doxycycline or other b-lactams, including cefuroxime,is good against penicillin-susceptible strains but less predictable with strains that show reduced pen-icillin-susceptibility.Vancomycin,linezolid,and quinupristin/ dalfopristin are the only drugs with predictable in vitro activity. Fluoroquinolones are generally active against strains that are susceptible or resistant to penicillin,but recent reports indicate increasing resistance in selective locations that correlate with excessivefluoroquinolone use.Prevention.The major preventive measures are use of in-fluenza vaccine and use of pneumococcal vaccine,according to guidelines of the Advisory Council on Immunization Practicesof the Centers for Disease Control and Prevention(CDC). Performance indicators.Recommendations for perform-ance indicators include the collection of blood culture speci-mens before antibiotic treatment and the institution of anti-biotic treatment within8h of hospitalization,since both aresupported on the basis of evidence-based trials.Additional per-formance indicators recommended are laboratory tests for Le-gionella in patients hospitalized in the ICU,demonstration ofan infiltrate on chest radiographs of patients with an ICD-9 (International Classification of Diseases,9th edition)code for pneumonia,and measurement of blood gases or pulse oximetrywithin24h of admission.IntroductionLower respiratory tract infections are the major cause ofdeath in the world and the major cause of death due to infec-tious diseases in the United States.Recent advances in thefieldinclude the identification of new pathogens(Chlamydia pneu-moniae and hantavirus),new methods of microbial detection (PCR),and new antimicrobial agents(macrolides,b-lactamagents,fluoroquinolones,oxazolidinones,and streptogramins).Despite extensive studies,there are few conditions in medicinethat are so controversial in terms of management.Guidelinesfor management were published in1993by the American Tho-racic Society[1],the British Thoracic Society[2],and the Ca-nadian Infectious Disease Society[3],as well as the InfectiousDiseases Society of America(IDSA)in1998[4].The presentguidelines represent revised recommendations of the IDSA. Compared with previous guidelines,these guidelines are in-tended to reflect updated information,provide more extensive recommendations in selected areas,and indicate an evolutionof opinion.These therapeutic guidelines are restricted to com-munity-acquired pneumonia(CAP)in immunocompetentadults.Recommendations are given alphabetical ranking to reflecttheir strength and a Roman numeral ranking to reflect thequality of supporting evidence(table1).This is customary forquality standards from the IDSA[5].It should be acknowledgedthat no set of standards can be constructed to deal with themultitude of variables that influence decisions regarding site ofcare,diagnostic evaluation,and selection of antibiotics.Thus,these standards should not supplant good clinical judgement.EpidemiologyMagnitudeCAP is commonly defined as an acute infection of the pul-monary parenchyma that is associated with at least some symp-toms of acute infection,accompanied by the presence of anacute infiltrate on a chest radiograph or auscultatoryfindingsconsistent with pneumonia(such as altered breath sounds and/at Library of Medical Center of Fudan University on April 21, 2014/Downloaded fromCID2000;31(August)IDSA Guidelines for CAP in Adults349Table1.Categories for ranking recommendations in the therapeutic guidelines.Category DescriptionStrength of recommendationA Good evidence to support a recommendation for useB Moderate evidence to support a recommendation for useC Poor evidence to support a recommendationD Moderate evidence to support a recommendation against useE Good evidence to support a recommendation against useQuality of evidenceI Evidence from at least1randomized,controlled trialII Evidence from at least1well-designed clinical trial without randomizationIII Evidence from opinions of respected authorities,based on clinical experi-ence,descriptive studies,or reports of expert committeesor localized rales),in a patient not hospitalized or residing in a long-term-care facility forу14days before onset of symp-toms.Symptoms of acute lower respiratory infection may in-clude several(in most studies,at least2)of the following:fever or hypothermia,rigors,sweats,new cough with or without sputum production or change in color of respiratory secretions in a patient with chronic cough,chest discomfort,or the onset of dyspnea.Most patients also have nonspecific symptoms, such as fatigue,myalgias,abdominal pain,anorexia,and headache.Pneumonia is the sixth most common cause of death in the United States.From1979through1994,the overall rates of death due to pneumonia and influenza increased by59%(on the basis of ICD-9codes on death certificates)in the United States[6].Much of this increase is due to a greater proportion of persons agedу65years;however,age-adjusted rates also increased by22%,which suggests that other factors may have contributed to a changing epidemiology of pneumonia,includ-ing a greater proportion of the population with underlying med-ical conditions at increased risk of respiratory infection. Annually,2–3million cases of CAP result in∼10million physician visits,500,000hospitalizations,and45,000deaths in the United States[7,8].The incidence of CAP that requires hospitalization is estimated to be258persons per100,000pop-ulation and962per100,000persons agedу65years[8].Al-though mortality has ranged from2%to30%among hospi-talized patients in a variety of studies,the average is∼14%[9]. Mortality is estimated to be!1%for patients not hospitalized [9,10].The incidence of CAP is heavily weighted toward the winter months.Prognosis,Risk Stratification,and the Initial Site-of-Treatment DecisionKnowledge about the prognosis of a disease allows physi-cians to inform their patients about the expected natural history of an illness,the likelihood of potential complications,and the probability of successful treatment.Understanding the prog-nosis of CAP is of particular clinical relevance,since it ranges from rapid recovery from symptoms without functional im-pairment to serious morbid complications and death.The abil-ity to accurately predict medical outcomes in cases of CAP hasa major impact on management.The decision to hospitalize apatient or to treat him or her as an outpatient(figure1)isperhaps the single most important clinical decision made by physicians during the entire course of illness,which has directbearing on the location and intensity of laboratory evaluation,antibiotic therapy,and costs.The estimated total treatment costfor an episode of CAP managed in the hospital is$7500(USdollars)[11],120-fold higher than the cost of outpatient treatment.Numerous studies have identified risk factors for death incases of CAP[9,10,12].These factors were well-defined in thepre–penicillin era;studies of adults showed an increased riskwith alcohol consumption,increasing age,the presence of leu-kopenia,the presence of bacteremia,and radiographic changes[12].More recent studies have confirmed thesefindings[2,13–18].Independent associations with increased mortality havealso been demonstrated for a variety of comorbid illnesses,suchas active malignancies[10,16,19],immunosuppression[20,21], neurological disease[19,22,23],congestive heart failure[10,17,19],coronary artery disease[19],and diabetes mellitus[10,19,24].Signs and symptoms independently associated with in-creased mortality consist of dyspnea[10],chills[25],alteredmental status[10,19,23,26],hypothermia or hyperthermia[10,16,17,20],tachypnea[10,19,23,27],and hypotension(diastolic and systolic)[10,19,26–28].Laboratory and radiographicfindings independently asso-ciated with increased mortality are hyponatremia[10,19],hy-perglycemia[10,19],azotemia[10,19,27,28],hypoalbumi-nemia[16,19,22,25],hypoxemia[10,19],liver function test abnormalities[19],and pleural effusion[29].Infections due togram-negative bacilli or S.aureus,postobstructive pneumonia,and aspiration pneumonia are also independently associatedwith higher mortality[30].Despite our knowledge regarding the associations of clinical, laboratory,and radiographic factors and patient mortality,there is wide geographic variation in hospital admission ratesfor CAP[31,32].This variation suggests that physicians donot use a uniform strategy to relate the decision to hospitalizeto the prognosis.In fact,physicians often overestimate the riskof death for patients with CAP,and the degree of overesti-at Library of Medical Center of Fudan University on April 21, 2014/Downloaded from350Bartlett et al.CID2000;31(August)Figure1.Evaluation for diagnosis and management of community-acquired pneumonia,including site,duration,and type of treatment. b-Lactam:cefotaxime,ceftriaxone,or a b-lactam/b-lactamase inhibitor.Fluoroquinolone:levofloxacin,moxifloxacin,or gatifloxacin or another fluoroquinolone with enhanced antipneumococcal activity.Macrolide:erythromycin,clarithromycin,or azithromycin.CBC,complete blood cell count;ICU,intensive care unit.*Other tests for selected patients:see text,Diagnostic Evaluation:Etiology.**See table15for special considerations.mation is independently associated with the decision to hos-pitalize[30].Over the past10years,at least13studies have used multi-variate analysis to identify predictors of prognosis for patients with CAP[10,16–20,25–27,33–35].The Pneumonia PORT developed a methodologically sound clinical prediction rule that quantifies short-term mortality for patients with this illness [10].Used as a guideline,this rule may help physicians make decisions about the initial location and intensity of treatment for patients with this illness(table2).The Pneumonia PORT prediction rule was derived with 14,199inpatients with CAP;it was independently validated with 38,039inpatients with CAP and2287inpatients and outpatients prospectively enrolled in the Pneumonia PORT cohort study. With this rule,patients are stratified into5severity classes by means of a2-step process.In step1,patients are classified as risk class I(the lowest severity level)if they are agedр50years,have none of5important comorbid conditions(neoplastic dis-ease,liver disease,congestive heart failure,cerebrovascular dis-ease,or renal disease),and have normal or only mildly derangedvital signs and normal mental status.In step2,all patients whoare not assigned to risk class I on the basis of the initial historyand physical examinationfindings alone are stratified into clas-ses II–V,on the basis of points assigned for3demographicvariables(age,sex,and nursing home residence),5comorbidconditions(listed above),5physical examinationfindings(al-tered mental status,tachypnea,tachycardia,systolic hypoten-sion,hypothermia,or hyperthermia),and7laboratory or ra-diographicfindings(acidemia,elevated blood urea nitrogen, hyponatremia,hyperglycemia,anemia,hypoxemia,or pleuraleffusion;table3).Point assignments correspond with the fol-lowing classes:р70,class II;71–90,class III;91–130,class IV;and1130,class V.In the derivation and validation of this rule,mortality wasat Library of Medical Center of Fudan University on April 21, 2014/Downloaded fromCID2000;31(August)IDSA Guidelines for CAP in Adults351 parison of risk class–specific mortality rates in the derivation and validation cohorts.Risk class a (total points)MedisGroups MedisGroupsPneumonia PORT validation cohortderivation cohort validation cohort Inpatients Outpatients All patientsn Mortality,%n Mortality,%n Mortality,%n Mortality,%n Mortality,%I13720.430340.11850.55870.07720.1II(р70)24120.757780.62330.92440.44770.6III(71–90)2632 2.86790 2.8254 1.2720.03260.9IV(91–130)46978.513,1048.24469.04012.54869.3V(1130)308631.1933329.222527.110.022627.0 Total14,19910.238,03910.613438.09440.62287 5.2 NOTE.No statistically significant differences in overall mortality or mortality within risk class existed among patients in the MedisGroups derivation,MedisGroups validation,and overall Pneumonia Patient Outcome Research Team(PORT)validation cohorts(n denotes the no.of patients within each risk class in the derivation and validation cohorts).P values for the comparisons of mortality across risk classes are as follows:class I,;class II,;class III,;class IV,;and class V,.P p.22P p.67P p.12P p.69P p.09a Risk class I was determined by the absence of all predictors identified in step1of the prediction rule.Risk classes II–V were determined by a patient’s total risk score,which is computed by use of the point scoring system shown in table3.low for risk classes I–III(0.1%–2.8%),intermediate for class IV(8.2%–9.3%),and high for class V(27.0%–31.1%).Increases in risk class were also associated with subsequent hospitaliza-tion and delayed return to usual activities for outpatients and with rates of admission to the ICU and length of stay for inpatients in the Pneumonia PORT validation cohort.On the basis of these observations,Pneumonia PORT investigators suggest that patients in risk classes I or II generally are can-didates for outpatient treatment,risk class III patients are po-tential candidates for outpatient treatment or brief inpatient observation,and patients in classes IV and V should be hos-pitalized(table4).Estimates from the Pneumonia PORT cohort study suggest that these recommendations would reduce the proportion of patients receiving traditional inpatient care by 31%and that there would be a brief observational inpatient stay for an additional19%.The effectiveness and safety of applying the Pneumonia PORT prediction rule to the initial site of care for an indepen-dent population of patients with CAP have been examined with use of a modified version of the Pneumonia PORT prediction rule[36].Emergency room physicians were educated about the rule and were encouraged to treat those in risk classes I–III as outpatients,with close,structured follow-up and provision of oral clarithromycin at no cost to the patient,if desired.The outcomes for those treated at home during this intervention phase were compared with the outcomes for historical control subjects from the time period immediately preceding the intervention.During the intervention period,there were166eligible pa-tients classified as“low risk”for short-term mortality(risk classes I–III)for comparison with147control subjects.The percentage treated initially as outpatients was higher during the intervention period than during the control period(57%vs. 42%;relative increase of36%;).When initial plus sub-P p.01sequent hospitalization was used as the outcome measure,there was a trend toward more outpatient care during the interven-tion period,but the difference was no longer statistically sig-nificant(52%vs.42%;).None of those initially treatedP p.07in the outpatient setting during the intervention period diedwithin4weeks of presentation.A second multicenter controlled trial subsequently assessedthe effectiveness and safety of using the Pneumonia PORT pre-diction rule for the initial site-of-treatment decision[37].In thistrial,19emergency departments were randomly assigned eitherto continue conventional management of CAP or to implementa critical pathway that included the Pneumonia PORT predic-tion rule to guide the admission decision.Emergency room physicians were educated about the rule and were encouragedto treat those in risk classes I–III as outpatients with oral levo-floxacin.Overall,1743patients with CAP were enrolled in this6-month e of the prediction rule resulted in an18%reduction in the admission of low-risk patients(31%vs.49%;).Use of the rule did not result in an increase in mor-P p.013tality or morbidity and did not compromise patients’30-dayfunctional status.These studies support use of the PneumoniaPORT prediction rule to help physicians identify low-risk pa-tients who can be safely treated in the outpatient setting.The IDSA panel endorses thefindings of the PneumoniaPORT prediction rule,which identifies valid predictors for mor-tality and provides a rational foundation for the decision re-garding hospitalization.However,it should be emphasized thatthe PORT prediction rule is validated as a mortality predictionmodel and not as a method to triage patients with CAP.Newstudies are required to test the basic premise underlying the useof this rule in the initial site-of-treatment decision,so that pa-tients classified as“low risk”and treated in the outpatient set-ting will have outcomes equivalent to or better than those ofsimilar“low-risk”patients who are hospitalized.It is important to note that prediction rules are meant tocontribute to rather than to supersede physicians’judgment.Another limitation is that factors other than severity of illnessmust also be considered in determining whether an individualpatient is a candidate for outpatient care.Patients designatedas“low risk”may have important medical and psychosocial contraindications to outpatient care,including expected com-pliance problems with medical treatment or poor social supportat Library of Medical Center of Fudan University on April 21, 2014/Downloaded from352Bartlett et al.CID 2000;31(August)Table 3.Scoring system for step 2of the prediction rule:assignment to risk classes II–V .Patient characteristicPoints assignedaDemographic factor Age Male No.of years of age FemaleNo.of years of age Ϫ10Nursing home resident ϩ10Comorbid illnessesNeoplastic diseasebϩ30Liver diseasecϩ20Congestive heart failuredϩ10Cerebrovascular diseaseeϩ10Renal diseasefϩ10Physical examination findingAltered mental statusgϩ20Respiratory rate 130breaths/min ϩ20Systolic blood pressure !90mm Hg ϩ20Temperature !35ЊC or 140ЊC ϩ15Pulse 1125beats/minϩ10Laboratory or radiographic finding Arterial pH !7.35ϩ30Blood urea nitrogen 130mg/dL ϩ20Sodium !130mEq/L ϩ20Glucose 1250mg/dL ϩ10Hematocrit !30%ϩ10Arterial partial pressure of oxygen !60mm Hg hϩ10Pleural effusionϩ10aA total point score for a given patient is obtained by adding the patient’s age in years (age Ϫ10,for females)and the points for each applicable patient char-acteristic.Points assigned to each predictor variable were based on coefficients obtained from the logistic regression model used in step 2of the prediction rule.bAny cancer except basal or squamous cell cancer of the skin that was active at the time of presentation or diagnosed within 1year of presentation.cA clinical or histologic diagnosis of cirrhosis or other form of chronic liver disease such as chronic active hepatitis.dSystolic or diastolic ventricular dysfunction documented by history and physical examination,as well as chest radiography,echocardiography,Muga scanning,or left ventriculography.eA clinical diagnosis of stroke,transient ischemic attack,or stroke docu-mented by MRI or computed axial tomography.fA history of chronic renal disease or abnormal blood urea nitrogen and creatinine values documented in the medical record.gDisorientation (to person,place,or time,not known to be chronic),stupor,or coma.hIn the Pneumonia Patient Outcome Research Team cohort study,an oxygen saturation value !90%on pulse oximetry or intubation before admission was also considered abnormal.Table 4.Risk-class mortality rates.Risk class No.of points Validation cohortRecommended site of care No.of patientsMortality,%I —a30340.1Outpatient II р7057780.6Outpatient III 71–906790 2.8Outpatient or brief inpatient IV 91–13013,1048.2Inpatient V1130933329.2InpatientNOTE.Table is adapted from [10].aAbsence of predictors.at home.Ability to maintain oral intake,history of substance abuse,cognitive impairment,and ability to perform activities of daily living must be considered.In addition,patients may have rare conditions,such as severe neuromuscular disease or immunosuppression,which are not included as predictors in these prediction rules but increase the likelihood of a poor prognosis.Prediction rules may also oversimplify the way physicians interpret important predictor variables.For example,extreme alterations in any one variable have the same effect on risk stratification as lesser changes,despite obvious differences in clinical import (e.g.,a systolic blood pressure of 40mm Hg vs.one of 88mm Hg).Furthermore,such rules discount the cu-mulative importance of multiple simultaneous physiological de-rangements,especially if each derangement alone does not reach the threshold that defines an abnormal value (e.g.,systolicblood pressure of 90/40mm Hg,respiratory rate of 28breaths/min,and pulse of 120beats/min).Finally,prediction rules often neglect the importance of patients’preferences in clinical de-cision-making.This point is highlighted by the observation that the vast majority of low-risk patients with CAP do not have their preferences for site of care solicited,despite strong pref-erences for outpatient care [38].Role of Specific Pathogens in CAPProspective studies evaluating the causes of CAP in adults have failed to identify the cause of 40%–60%of cases of CAP and have detected у2etiologies in 2%–5%[2,7,26,39,40].The most common etiologic agent identified in virtually all studies of CAP is S.pneumoniae,which accounts for about two-thirds of all cases of bacteremic pneumonia cases [9].Other pathogens implicated less frequently include H.influenzae (most strains of which are nontypeable),Mycoplasma pneumoniae,C.pneumoniae,S.aureus,Streptococcus pyogenes,N.meningitidis,Moraxella catarrhalis,Klebsiella pneumoniae and other gram-negative rods,Legionella species,influenza virus (depending on the season),respiratory syncytial virus,adenovirus,parainflu-enza virus,and other microbes.The frequency of other etiol-ogies is dependent on specific epidemiological factors,as with Chlamydia psittaci (psittacosis),Coxiella burnetii (Q fever),Francisella tularensis (tularemia),and endemic fungi (histo-plasmosis,blastomycosis,and coccidioidomycosis).Comparisons of relative frequency of each of the etiologies of pneumonia are hampered by the varying levels of sensitivity and specificity of the tests used for each of the pathogens that they detect;for example,in some studies,tests used for legi-onella infections provide a much higher degree of sensitivity and possibly specificity than do tests used for pneumococcal infections.Thus,the relative contribution of many causes to the incidence of CAP is undoubtedly either exaggerated or un-derestimated,depending on the sensitivity and specificity of tests used in each of the studies.Etiology-Specific Diagnoses and the Clinical SettingNo convincing association has been demonstrated between individual symptoms,physical findings,or laboratory test re-sults and specific etiology [39].Even time-honored beliefs,suchat Library of Medical Center of Fudan University on April 21, 2014/Downloaded fromCID2000;31(August)IDSA Guidelines for CAP in Adults353Table5.Diagnostic studies for evaluation of community-acquired pneumonia.Baseline assessmentChest radiography to substantiate diagnosis of pneumonia,to detect associated lung diseases,to gain insightinto causative agent(in some cases),to assess severity,and as baseline to assess responseOutpatientsSputum Gram stain and culture for conventional bacteria are optionalInpatientsDetermination of complete blood cell and differential countsSerum creatinine,urea nitrogen,glucose,electrolyte,bilirubin,and liver enzyme valuesHIV serological status for persons aged15–54yearsO2saturation arterial blood gas values for selected patientsBlood cultures(ϫ2;before treatment)Gram stain and culture of sputum aTest for Mycobacterium tuberculosis,with acid-fast bacilli staining and culture for selected patients,especiallythose with cough for11mo,other common symptoms,or suggestive radiographic changesTest for Legionella in selected patients,including all seriously ill patients without an alternative diagnosis,es-pecially if aged140years,immunocompromised,or nonresponsive to b-lactam antibiotics,if clinicalfeatures are suggestive of this diagnosis,or in outbreak settingsThoracentesis with stain,culture,and determination of pH and leukocyte count differential(pleuralfluid)Alternative specimens to expectorated sputumAspirates of intubated patients,tracheostomy aspirates,and nasotracheal aspirates:manage as with expec-torated sputumInduced sputum:recommended for detection of M.tuberculosis or Pneumocystis cariniiBronchoscopy(see text under Special Considerations:Pnemococcal Pneumonia)Transtracheal aspiration:recommended only in cases of enigmatic pneumonia,to be done by personsskilled in the technique,preferably before antibiotic treatmentOptionalAdditional cytological or microbiological tests,as listed in table8,depending on clinical features,availableresources,underlying conditions,and/or epidemiological associations of the patientSerum:to be frozen and saved for serological analysis,if needed ba Should be deep-cough specimen obtained before antibiotic therapy.Gram stain should be interpreted by trainedpersonnel and culture done only if specimen is adequate by cytological criteria,except for Legionella and myco-bacteria.Consider diagnostic studies for endemic fungi and mycobacteria when clinical features suggest infectionwith these.For hospitalized patients with severe pneumonia or clinical features that suggest legionnaires’disease,perform culture and urinary antigen testing for Legionella.Inability to obtain specimens for diagnostic studiesshould not delay antibiotic treatment of acutely ill patients.b Serological tests would include those for Mycoplasma pneumoniae,Legionella pneumophila,Chlamydia pneu-moniae,or others(i.e.,viruses,Chlamydia psittaci,or Coxiella burnetii),depending on the circumstances.as the absence of productive cough or inflammatory sputum in pneumonia due to Mycoplasma,Legionella,or Chlamydia species,have not withstood close inspection.On the other hand, most comparisons have involved relatively small numbers of patients and have not evaluated the potential for separating causes by use of constellations of symptoms and physical findings.In one study,as yet unconfirmed,that compared patients identified in a prospective standardized fashion,a scoring sys-tem using5symptoms and laboratory abnormalities was able to differentiate most patients with legionnaires’disease from the other patients[41].A similar type of system has been devised for identifying patients with hantavirus pulmonary syndrome (HPS)[42].If validated,such scoring systems may be useful for identifying patients who should undergo specific diagnostic tests(which are too expensive to use routinely for all patients with CAP)and be empirically treated with specific antimicrobial drugs while test results are pending.Certain pathogens cause pneumonia more commonly among persons with specific risk factors.For instance,pneumococcal pneumonia is especially likely to occur in the elderly and in patients with a variety of medical conditions,including alco-holism,chronic cardiovascular disease,chronic obstructed air-way disease,immunoglobulin deficiency,hematologic malig-nancy,and HIV infection.However,outbreaks occur amongyoung adults under conditions of crowding,such as in armycamps or prisons.S.pneumoniae is second only to Pneumocystiscarinii as the most common identifiable cause of acute pneu-monia in patients with AIDS[43–45].Legionella is an oppor-tunistic pathogen;legionella pneumonia is rarely recognized inhealthy young children and young adults.It is an importantcause of pneumonia in organ transplant recipients and in pa-tients with renal failure and occurs with increased frequency inpatients with chronic lung disease,smokers,and possibly thosewith AIDS[46].Although M.pneumoniae historically has beenthought primarily to involve children and young adults,someevidence suggests that it causes pneumonia in healthy adultsof any age[8].There are seasonal differences in incidence of many of thecauses of CAP.Pneumonia due to S.pneumoniae,H.influenzae,and influenza occurs predominantly in winter months,whereasC.pneumoniae appears to cause pneumonia year-round.Al-though there is a summer prevalence of outbreaks of legion-naires’disease,sporadic cases occur with similar frequency dur-ing all seasons[8,46].Some studies suggest that there is noseasonal variation in mycoplasma infection;however,otherdata suggest that its incidence is greatest during the fall andwinter months[47].at Library of Medical Center of Fudan University on April 21, 2014/Downloaded from。
- 1、下载文档前请自行甄别文档内容的完整性,平台不提供额外的编辑、内容补充、找答案等附加服务。
- 2、"仅部分预览"的文档,不可在线预览部分如存在完整性等问题,可反馈申请退款(可完整预览的文档不适用该条件!)。
- 3、如文档侵犯您的权益,请联系客服反馈,我们会尽快为您处理(人工客服工作时间:9:00-18:30)。
Response to the IUPAC/IUPAP Joint Working Party Second Report‘On the Discovery of Elements 110-118’A. Marinov1, S. Gelberg1, D. Kolb2 and G. W. A. Newton31 Racah Institute of Physics, The Hebrew University of Jerusalem, Jerusalem 91904, Israel2 Department of Physics, University GH Kassel, 34109 Kassel, Germany3 Heron’s Reach, 382 Mossy Lea Road, Wrightington, Lancashire, WN6 9RZ, UKAbstract: Back in 1971 fission fragments were observed by us in Hg sourcesseparated from two CERN W targets that were irradiated with 24 GeVprotons. The masses of the fissioning species were measured and heavymasses like 272, 308 and 317-318 were found and interpreted as due to thesuperheavy element 112 with 160-161 neutrons and various molecules of it.Based on the measured mass of the produced superheavy nucleus cold fusionreactions like 88Sr + 184W →272112 and 86Sr + 186W →272112 were deduced.The ordinary heavy ion reaction 88Sr + 184W has been studied andcharacteristic X-rays of element 112 and a very high-energy αparticle incoincidence with a fission fragment have been observed. The data have beenstudied by the TWG and they were concerned about the question of thereaction mechanism since very large fusion cross sections, in the region of afew mb, have been deduced in the secondary reaction experiments. Thisquestion can now be answered in view of our recent discovery of long-livedsuper- and hyperdeformed isomeric states. The JWP did not accept ourarguments and this response answers their queries.The second report of the JWP regarding our Z = 112 work is based on their first report. For the sake of clarity we consider both reports in our response.Our claim for discovering element 112 has already been considered by the TWG [1]. It was based, in one set of experiments, on the observation of fission activities with measured appropriate masses1 in species which followed the chemistry of Hg, and like the latter electroplated without applying voltage on Cu and evaporated at about 300° C2. Furthermore, from the measured masses the cold fusion reaction 88Sr + 184W was deduced, and in another experiment this reaction has been studied and evidence for characteristic X-rays of element 1123 and for a very high energy (12.16 MeV) α-particle4in coincidence with a fission fragment were found. (According to the kinematics only the isotopes 271112 and 272112 could have been produced in this cold fusion reaction). Altogether we have found about 100 of these heavy Eka-Hg atoms, and the usual1 Recently, in the 3rd Int. Conf. on Exotic Nuclei and Atomic Masses, the recent data of the GSI and Dubna groups were presented. In the summary talk [21] given by Dr. C. N. Davids of ANL he mentioned that what he thinks is missing in their data are mass measurements. Such essential measurements were performed by us about 30 years ago and consistently interpreted back in 1984 [4].2 This low temperature eliminated any element with 90 ≤ Z ≤ 111.3 Photons with respective energies that are within 330 eV and 160 eV equal to the predictions for Kand Lβ4α1X-rays of element 112 have been seen in coincidence with low-energy particles [5 – 8], while the corresponding differences for adjacent elements are 4 keV and 800 eV respectively. The particles were assumed to be protons. Long-lived proton radioactivity has been observed by us. (See down below and Ref. [13]).4 The highest known today.background in the measurements was zero. To the best of our knowledge such a quality of data has never been obtained in the discovery of other superheavy elements, where the identification was usually based on just a few atoms. The data were described in Refs. [2-8].As mentioned above these data have been considered by the TWG in 1992 [1]. Generally speaking their attitude toward our work was positive. The TWG “…judges the experiments so interesting that it will express hope that they will get a follow-up” (quote from the minutes of the TWG meeting in Mogilany-Krakow 1-5 July 1991), or: “In your beautiful work which has been carried on during so many years, you have found, I think, many unexpected results…” (quote from a letter of Professor M. Lefort, a member of the TWG, to A. Marinov of March 20, 1991). They did not question the validity of our experimental data, but they were concerned about the problem of the reaction mechanism. Even then they pointed out that in both experiments "…this possibility cannot be definitely dismissed", or "…cannot be said to be impossible. Further work is needed". At that time only a partial answer could have been given by us to this question. First, we have claimed that from the experimental point of view it was shown that actinides, and in particular 236Am and 236Bk (in isomeric states), have been produced by secondary reactions in the same W target [9]. If 3.1 x 105 atoms of 236Am and 4.4 x 104 atoms of 236Bk have been produced in the target then the production of about 500 atoms of ~272112 is possible. This is particularly so since hot fusion reactions with large competition with fission are responsible for the production of the actinides while very cold fusion with much less fission is responsible for the production of element 112. Secondly, it was shown [4] that in the secondary reactions the projectiles are fragments that were produced just within about 5 x 10-14s before interacting with another W nucleus in the target. During this short time they are still at high excitation energy and quite deformed. Deformations have a very strong effect on the fusion cross sections between heavy nuclei as demonstrated by the well-known sub-barrier fusion effect [10,11] and seen in Fig. 4 of Ref. [7]. In addition, it was pointed out to the TWG that, like in the actinides [9], perhaps a long-lived isomeric state, rather than the ground state, was produced in element 112. However, at that time we did not have any clue about the character of the isomeric state, and whether it might help to explain the large cross section.It should be mentioned that these arguments have been considered favorably by the TWG. In the above mentioned letter sent to A. Marinov by Professor M. Lefort on March 20, 1991, he said: “The possibility of producing long lived isomeric states in neutron deficient very heavy nuclei is indeed reasonable, as well as the hypothesis of production of highly excited deformed fragments as possible projectiles”.Continuing our research, at the time when the first JWP was established we were able to propose a more complete answer to the cross section problem. Based on our discovery [12,13] of the existence of long-lived high-spin superdeformed isomeric states,5it was suggested that similar isomeric states were formed in element 112 (as well as in the actinides). It is clear that much less penetration and dissipation is needed in order to produce the compound nucleus in the superdeformed shape rather than in its normal states. Hence much less or even no extra-push energy is needed and the fusion cross section is expected to be much larger.5 The evidence for the existence of these isomeric states is based on the observation of a relatively low energy and very enhanced alpha particle group where the enhancement is in accord with penetrability calculations fora superdeformed to superdeformed transition, and the alpha particles themselves are in coincidence with superdeformed band gamma ray transitions, and on the observation of long-lived proton radioactivities.In their first report [14] the JWP did not follow the TWG but rather questioned the validity of the data themselves. They say (under 112; 04 [14]): “The situation pertaining to these collaboration results has not changed substantially since the TWG judgment. If anything, it has become weaker because independent attempts to duplicate the process of fusion with secondary residues from high-energy proton irradiations of heavy targets have failed to find yields of elements more than a half dozen atomic numbers greater than that of the target (71Ka01, 73Ba01, 73Ge01) rather than the three dozen or more invoked by Marinov et al.”This statement is not correct: As mentioned above, in a study that was performed over about 15 years [9], 236Am and 236Bk (in long-lived isomeric states) were identified in the W target. These are 21 and 23 atomic numbers greater than that of the target. Besides, the TWG report was issued in 1992, and it is logically impossible to claim that since then the situation became weaker because of the above quoted papers that were published about twenty years earlier, and were known to the TWG. As a matter of fact, the quoted papers were already been dealt with by us back in 1984 [4] and it was shown that they could not prove the point made by the JWP.In the second assessment of the JWP first report (under 112; 08) [14] addressing our discovery of the long-lived superdeformed isomeric states which can explain how the production of element 112 by us could be possible, they say: “These two papers continue to press arguments for the existence of very long-lived isomeric states of actinides and transactinides and of very high fusion cross-sections for their formation, each several orders of magnitude beyond current understanding. These extraordinary phenomena are, in part, necessary for the acceptance of the collaborations’ interpretation. The JWP remained unmoved.”This harsh verdict goes against the judgment of the TWG as expressed in the last above quotation from Prof. Lefort letter to A. Marinov of March 20, 1991.It seems that there are clear inconsistencies between the assessments of the TWG and of the JWP. Since our first submission to the JWP of our claim for priority in discovering element 112, further work was carried out by us regarding the existence of the long-lived isomeric states. The results have been summarized in two comprehensive papers [15,16]6and have been submitted to the JWP for their second report [19]. In [16], strong evidence for the existence of a long-lived high-spin hyperdeformed isomeric state is given. It is based on the observation of a 13 orders of magnitude retarded (40 d ≤t1/2 ≤2.1 y) high-energy α-particle group of about 8.6 MeV in coincidence with superdeformed band γ-ray transitions, where the energy of the α-particles fits with theoretical predictions for a III min→II min transition. (In addition several more long-lived superdeformed isomeric states were observed. A summary of all the new transitions is given in Table 3 of Ref. [16]). In [15] a quantitative interpretation is given to both the low energies and the enhanced lifetimes of the unidentified α-particles seen in various actinide sources separated from the CERN W target in terms of II min → II min and III min → III min transitions.Thus, long-lived isomeric states exist not only in the second minimum of the potential but also in the third minimum, and the evidence for this is based on 16 α - γ coincidence events where the γ-rays fit beautifully as superdeformed band transitions (Fig. 11 in Ref. [16]), and on several hundreds of low energy α-particles (Figs. 4-5, in Ref. [15]) where the background in these measurements is about zero.6 Short summaries of these papers are given in Refs. [17,18].Like in the first report, the attitude of the JWP towards our work remains negative in its second report as well [19]. Their assessment is essentially the same as described above in their first report.As has already been mentioned above our claim for the existence of very long-lived isomeric states of actinides and transactinides, of very high fusion cross sections for their formation, is not “beyond traditional understanding”. Long-lived isomeric states have been produced by the 16O + 197Au [12,13] and 28Si + 181Ta [16] reactions, and also by secondary reactions, in 236Am and 236Bk[9] and in heavier actinide nuclei [15]. Superdeformed long-lived high spin isomeric states have also been predicted by Nilsson et al. back in 1969 [20], and similar effects of this type can exist also in the hyperdeformed region.7 There is no reason to assume that long-lived high spin states could not be produced in the superheavy element region as well. The long measured half-life of the observed fission activity of several weeks and the large observed fusion cross sections indicate that this is indeed the case. Regarding the argument of the JWP about the fusion cross sections, we have already mentioned that the combined effect of having deformed fragments (“projectiles”) and producing the compound nucleus in the super- or hyperdeformed isomeric state increases the fusion cross section by many orders of magnitude.The JWP further writes: “As indirect evidence, their discovery of long-lived 236Bk and 236Am more than a decade ago is frequently cited in their papers, yet the several existing compendia of isotopes do not acknowledge the existence of these species.”The evidence for the existence of the long-lived isomeric states in 236Bk and 236Am is based on measuring during about 15 years the decay curve and half-life of a 5.76 MeV α-particle group from radioactive sources which followed the specific chemistry of Bk and Am, respectively [9]. Identification of an isotope on the basis of its α-decay energy and half-life is a standard procedure. In the “Table of Isotopes” by R. B. Firestone et al. there is a reference to our work of Ref. [9]. For both 236Am and 236Bk our work “87Ma21” is quoted under “Populating Reactions and Decay Modes”. For 236Bk it is also mentioned that the type of the reaction is W(p,x). Our results regarding the isomeric states in 236Am and 236Bk and their formation via secondary reaction in a W target irradiated with 24-GeV protons are also quoted in the “Evaluated Nuclear Structure Data File” (ENSDF) by F. Orlando.The JWP further writes: “The collaboration results include mention of observing long-lived proton-decay, of deformed spallation products undergoing secondary fusion reactions, and of hyperdeformed shapes any of which significant topics by themselves should have attracted studies by other groups years ago. Yet this has not occurred.”All our measurements repeated themselves several times and their statistical significance has been checked. We are not responsible for other groups’ research programs.The JWP further writes: “The collaboration’s arguable use of forceful expressions such as “overwhelming evidence”, “clear and proven”, and “impossible to refute” is neither convincing nor swaying. Extraordinary intriguing phenomena, not much selective in their measured character, are, in part, necessary for the acceptance of the collaborations' interpretations of their7 As shown in [15], due to calculations of Howard and Möller [24], hyperdeformed states actually could be the true ground states in the heavy actinide nuclei and in the superheavy element region.data. The Joint Working Party needs much more to be able to relinquish its deeply felt unease that the tautological rationalization of the Marinov et al. measurements remains inadequate.”We do not think that we exaggerate when we describe our data with the above quoted expressions. Observation of fission fragments from sources that followed the chemistry of Hg [2,3], measuring the masses [4] of the fissioning nuclei and characteristic X rays of element 112 [5-8], are indeed, to our mind, very convincing. We also think the same about our discovery of the long-lived super- and hyperdeformed isomeric states [12,13,15,16] and that it is not justified to ignore them. In addition, as was mentioned above, a coincidence event between a 12.16 MeV α particle and a fission fragment has been seen in the study of the heavy ion 88Sr + 184W reaction [5 – 8] where, from the kinematics point of view, only 271112 and 272112 isotopes could have been produced in the reaction. This measurement is of the correlated type as the events studied by the GSI group, except that in our case the background was zero (Fig. 3 in [6] and [7] and Figs. 7 and 8 in [8]), and the coincidence time was 1 µs as compared to correlation times of milliseconds to tens of seconds. Why the GSI experiment is considered by the JWP as producing “high-quality data with plausible interpretation” and ours is completely ignored?Finally in the SUMMARY OF JWP01 CONCLUSIONS [19] it is written: “Also, despite efforts by the Marinov et al. collaboration using atypical studies in conjunction with speculative theory to re-enforce their claim to element 112, we maintain that the results of secondary interactions involving hyperdeformed long-lived products of long lifetime and high production probability remain unconvincing curiosities, all aspects of which warrant more selective investigation.” Nothing in our theoretical explanations is speculative: standard penetration calculations including deformation up to super- and hyperdeformation have been used by us.Already the TWG expressed that: “The possibility of producing long lived isomeric states in neutron deficient very heavy nuclei is indeed reasonable, as well as the hypothesis of production of highly excited deformed fragments as possible projectiles” (the above mentioned letter of Professor M. Lefort from 1991). The several weeks-long measured lifetime of the fission activity in the Hg sources shows that a long-lived isomeric state was produced in element 112. Long-lived super- and hyperdeformed isomeric states have been discovered by us using the 16O + 197Au [12,13] and 28Si + 181Ta [16] reactions, and it was shown (Fig. 8 in Ref. 15) that the production of a very heavy compound nucleus in a super- or hyperdeformed isomeric state is much more probable then its production in a normal state. Also the increased cross section as a result of deformations of the projectile and/or target is a well-known effect. The measured long lifetime of Z=112 and the large fusion cross section strongly indicate that super- or hyperdeformation is involved.It also seems to us that the JWP should have said what is more selective than fission fragments from separated Hg sources followed by mass measurements of the fissioning nuclei, characteristic X-rays of element 112 and a coincidence measurement between an αparticle and a fission fragment, determination of isotopes according to their chemical behavior and α-particle energies and lifetimes. Or, what is more selective than enhanced α-particles where the enhancement fits with penetrability calculations for superdeformed to superdeformed transition and the α-particles themselves are in coincidence with superdeformed band γ-ray transitions. Or, what is more selective than abnormally high energy and very retarded α-particles in coincidence with superdeformed γ rays, where the high energy is in accord with predictions for hyperdeformed tosuperdeformed transition, or, in addition, low energy and very enhanced α-particle groups, where both the energies and the enhancements fit with hyperdeformed to hyperdeformed transitions.Before concluding let us comment on a question that was raised recently, namely, is it justified to expect that element 112 will act like Hg, since some relativistic calculations indicate that it might show properties more like a noble gas [22].First let us mention that already in 1971 we took into account the possibility that element 112 may be more volatile than Hg. (Similar to the fact that Hg is more volatile than Cd.)8 Secondly, fact is that the fission fragments were seen preferentially in the Hg sources and not in the Au, Tl and Pb sources [23], and that they also were seen in the mass separator experiments, where a Hg source first was electroplated on Cu without applying any voltage and then was evaporated in the ion source at a low temperature which eliminated any element with 90 ≤ Z ≤ 111. It is clear that the measured fission activity basically followed the chemistry of Hg, otherwise one would have readily lost it in the complex chemical procedure. Only Z=112 including relativistic effects as has been calculated by Pershina et al. [22] is similar enough to do this.9 Therefore, our chemistry did isolate element 112 and no other. It is essential that in our case the chemical separation was done on Hg and element 112 at various oxidation states and not at an elemental state like in [25,26] where one is basically sensitive to volatility and adsorption properties of an element. Furthermore, as mentioned in footnote 9 and is important for our experimental procedure, according to the relativistic calculations [22] the binding energies of Z=112 and Hg on Cu are almost the same. In summary let us mention a point of principle. One has to distinguish between the experimental evidence for the existence of an element and the understanding of the way of its production. Already in ancient times people knew and admired when they held a piece of gold in their hand. They of course did not have any idea about supernova explosions and how the gold or the heavy elements in general were produced. In the modern history of physics one may recall that when the continuous spectrum of βrays was discovered, people were so confused that they were even willing to give up the conservation law of energy. Yet, no one had doubt about the very existence of the continuous β spectrum itself.In this respect our discovery of element 112 was done back in 1971 [2,3] by observing the fission fragments in Hg sources and by measuring the fissioning masses. The understanding of the masses of the fissioning nuclei in 1984 [4] strongly support the original results. The measured masses led to a reasonable hypothesis about the fusion reactions that took place in the W targets, and by a study of a similar but ordinary heavy ion reaction evidence for characteristic X-rays of element 8 For instance, in the measurement of the energies of the fission fragments [2,4] the source was cooled to liquid nitrogen temperature to avoid its evaporation.9 It is shown that the pand s1/2 level energy distance even increases from Hg to element 112 and thus make1/2partial p1/2 occupancy as improbable as in Hg. Occupation of the p1/2 shell by at least one electron (Z=113 and higher) or at least one electron less in the s-d shell (Z below 112) would drastically change the chemical and physical properties. E.g. from the experience with s-d atomic level structures in the periodic table we see thatthe evaporation temperatures stay high as long as the s-d shell is not closed, with a strong drop for the closedd10s2 configuration as in Hg and Z=112. On the other hand the ns1/2 and (n-1)d5/2 which were quite apart in Hg are now almost energetically degenerate, because the 7s1/2 orbital becomes more bound due to a directrelativistic effect and the 6d5/2 less bound due to indirect relativistic effects (stronger shielding of the nuclear charge by the relativistically enhanced deeper binding of the s1/2 and p1/2 orbitals). The less bound 6d5/.2 makesit more reactive and the more bound 7s1/2 makes it more noble than in Hg. These two effects may compensateand make chemical behaviors of Hg and element 112 similar. (E.g. according to [22] the binding energies (D c)of Hg and element 112 on Cu are almost the same). However the chemistry of Hg cannot be similar to that ofany other superheavy element in this Z region.112 and for a very high energy α particle in coincidence with a fission fragment was obtained [5-8]. Our discovery of the long-lived super- and hyperdeformed isomeric states [12,13,15,16] enables us also to understand, in a fully consistent manner, the production of element 112 in both the secondary and the ordinary heavy ion reactions. Based on all this evidence we believe that element 112 has been discovered by us back in 1971.[1] Discovery of the Transfermium Elements,R. C. Barber, N. N. Greenwood, A. Z. Hrynkiewics, Y. P. Jeannin, M. Lefort, M. Sakai, I. Uleuhla, A. H. Wapstra and D. H. Wilkinson, Prog. Part. Nucl. Phys., Vol. 29, pp. 453-530 (1992).[2] Evidence for the Possible Existence of a Superheavy Element with Atomic Number 112, A. Marinov, C. J. Batty, A. I. Kilvington, G. W. A. Newton, V. J. Robinson and J. H. Hemingway, Nature, Vol. 229, pp. 464-467 (1971).[3] Spontaneous Fission Previously Observed in a Mercury Source, A. Marinov, C. J. Batty, A. I. Kilvington, J. L. Weil, A. M. Friedman, G. W. A. Newton, V. J. Robinson, J. H. Hemingway and D. S. Mather, Nature, Vol. 234, pp. 212-215 (1971).[4] Consistent Interpretation of the Secondary-Reaction Experiments in W Targets and Prospects for Production of Superheavy Elements in Ordinary Heavy-Ion Reactions, A. Marinov, S. Eshhar, J. L. Weil and D. Kolb, Phys. Rev. Lett., Vol. 52, pp. 2209-2212 (1984); Vol. 53, p. 1120 (1984) (E).[5] Observation of Three Coincidence Events in a Superheavy Element Search via the 88Sr + 184W Reaction, A. Marinov, S. Gelberg, H. Folger, D. Kolb and W. Oelert, 8th Int’l. Conf. On Atomic Masses and Fundamental Constants, AMCO-8, Jerusalem, Israel (1991) L-46. (Unfortunately the AMCO-8 conference was cancelled due to the tension in the Persian Gulf).[6] Study of Long-Lived Superheavy Elements in W targets via Secondary and Direct Heavy-Ion Reactions, A. Marinov, Proc. Int’l Symp. Structure and Reactions of Unstable Nuclei, Eds. K. Ikeda and Y. Suzuki, Niigata, Japan (1991) pp.317-324.[7] The evidence for production of the superheavy element with Z=112 via secondary and direct heavy ion reactions A. Marinov, S. Gelberg and D. Kolb, Inst. Phys. Conf. Ser. No. 132, Sixth Int'l Conf. On Nuclei far from Stability & Ninth Int'l. Conf. On Atomic Masses and Fundamental Constants, Bernkastel-Kues, Germany, Eds. R. Neugart and A. Wöhr (1992) pp. 437-442.[8] The Evidence for Element Z=112 Produced via Secondary and Direct Heavy-Ion Reactions A. Marinov, S. Gelberg and D. Kolb, Int'l. School-Seminar on Heavy Ion Physics, Eds. Y. Oganessian, Y. Penionzhkevich and R. Kalpakchieva, Dubna, Russia (1993) Vol. 1, pp.162-171.[9] Evidence for Long-Lived Isomeric States in Neutron-Deficient 236Am and 236Bk Nuclei, A. Marinov, S. Eshhar and D. Kolb, Phys. Letters B, Vol. 191, pp. 36-40 (1987)[10] Fusion of 16O + 148,150,152,154Sm at Sub-Barrier Energies, R. G. Stokstad, Y. Eisen, S. Kaplanis, D. Pelte, U. Smilansky and I. Tserruya, Phys. Rev. C, Vol. 21, pp. 2427-2435 (1980).[11] Nuclear Deformation and Sub-Barrier Fusion Cross Sections, A. Iwamoto and P.Möller, Nucl. Phys. A, Vol. 605, pp. 334-358 (1996) and references therein.[12] Discovery of Strongly Enhanced Low Energy Alpha Decay of a Long-LivedIsomeric State Obtained in the 16O + 197Au Reaction at 80 MeV, probably toSuperdeformed Band, A. Marinov, S. Gelberg and D. Kolb, Mod. Phys. Lett. A, Vol. 11, pp.861-869 (1996).[13] Evidence for Long-Lived Proton Decay Not Far From the β-Stability ValleyProduced by the 16O + 197Au Reaction at 80 MeV, A. Marinov, S. Gelberg and D. Kolb,Mod. Phys. Lett. A, Vol. 11, pp. 949-956 (1996).[14] On the Discovery of the Elements 110-112 P. J. Karol, H. Nakahara, B. W. Petley andE. Vogt, Pure Appl. Chem., Vol. 73, pp. 959-967 (2001).[15] Strongly Enhanced Low Energy Alpha-Particle Decay in Heavy Actinide Nuclei andLong-Lived Superdeformed and Hyperdeformed Isomeric States A. Marinov, S. Gelberg, and D. Kolb, Int. J. Mod. Phys. E, Vol. 10, pp. 209-236 (2001).[16] Discovery of Long-Lived Shape Isomeric States which Decay by Strongly RetardedHigh-Energy Particle Radioactivity, A. Marinov, S. Gelberg and D. Kolb, Int. J. Mod. Phys.E, Vol.10, pp.185-208 (2001).[17] Super- and Hyper-Deformed Isomeric States and Long-Lived SuperheavyElements, A. Marinov, S. Gelberg, D. Kolb and J. L. Weil, Proc. 2d Int. Conf., Fission andProperties of Neutron-Rich Nuclei, St. Andrews, Scotland (1999)Eds. J. H. Hamilton, W. R. Phillips and H. K. Carter (World Scientific, Singapore, 2000) pp. 231-234.[18] Abnormal Radioactive Decays out of Long-Lived Super- and Hyper-DeformedIsomeric States, A. Marinov, S. Gelberg and D. Kolb, APH NS Heavy Ion Physics, Vol. 13, pp.133-137 (2001).[19] On the Claims for Discovery of Elements 110, 111, 112, 114, 116, and 118,P. J. Karol, H. Nakahara, B. W. Petley, and E. Vogt, Pure Appl. Chem., Vol. 75, pp. 1601-1611 (2 003).[20] On a New Type of Fission-Isomeric State, S.G. Nilsson, G. Ohlén, C. Gustafson andP. Möller, Phys. Lett. Vol. 30B, pp. 437-440 (1969).[21] Concluding Remarks, C. N. Davids, Proc. 3rd Int’l. Conf. on Exotic Nuclei and Atomic Masses, ENAM2001, Hämeenlinna, Finland, Eds. J. Äystö et al., pp. 499-502 (2001).[22]Intermetallic Compounds of the Heaviest Elements: the Electronic Structure andBonding of Dimers of Element 112 and its Homolog Hg, V. Pershina, B. Bastug, T. Jacob,B. Fricke and S. Varga, Chem. Phys. Lett. Vol. 365, pp. 176-183 (2002) and referencestherein.[23] Study of Au, Tl and Pb Sources Separated from Tungsten Targets that wereIrradiated with 24-GeV Protons. Indications for the Possible Production of Superheavy Elements, A. Marinov, S. Eshhar and B. Alspector, Proc. Int'l. Symp. On Superheavy Elements, Lubbock, Texas, Ed. M. A. K. Lodhi (1978) pp. 81-88.[24] Calculated Fission Barriers, Ground-State Masses, and Particle SeparationEnergies for Nuclei with 76 ≤ Z ≤ 100 and 140 ≤ N ≤ 184, W. M. Howard and P. Möller, Atom. Data Nucl. Data Tables Vol. 25, pp. 219-285 (1980).[25] Chemical Identification and Chemical Properties of Element 112, A. B.Yakushevet al., Radiochimica Acta Vol. 91, pp. 433-440 (2003).[26] Indication for a Gaseous Element 112, S. Sovena for a Univ. Bern –PSI –Univ. Mainz–TUM – LBNL – UCB – IMP – collaboration, Jahresbericht der Kernchemie, Paul Scherrer Institute, Switzerland 2004.。