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Supplementary Fig. S3 from Relevance of the Fanconi anemia pathway in the response of human cells to trabectedin

Trabectedin (Yondelis; ET-743) is a potent anticancer drug that binds to DNA by forming a covalent bond with a guanine in one strand and one or more hydrogen bonds with the opposite strand. Using a fluorescence-based melting assay, we show that one single trabectedin-DNA adduct increases the thermal stability of the double helix by >20°C. As deduced from the analysis of phosphorylated H2AX and Rad51 foci, we observed that clinically relevant doses of trabectedin induce the formation of DNA double-strand breaks in human cells and activate homologous recombination repair in a manner similar to that evoked by the DNA interstrand cross-linking agent mitomycin C (MMC). Because one important characteristic of this drug is its marked cytotoxicity on cells lacking a functional Fanconi anemia (FA) pathway, we compared the response of different subtypes of FA cells to MMC and trabectedin. Our data clearly show that human cells with mutations in FANCA, FANCC, FANCF, FANCG, or FANCD1 genes are highly sensitive to both MMC and trabectedin. However, in marked contrast to MMC, trabectedin does not induce any significant accumulation of FA cells in G2-M. The critical relevance of FA proteins in the response of human cells to trabectedin reported herein, together with observations showing the role of the FA pathway in cancer suppression, strongly suggest that screening for mutations in FA genes may facilitate the identification of tumors displaying enhanced sensitivity to this novel anticancer drug. [Mol Cancer Ther 2008;7(5):1309–18]

Trabectedin (Yondelis) is a potent antitumor drug that has the unique characteristic of killing cells by poisoning the DNA nucleotide excision repair (NER) machinery. The basis for the NER-dependent toxicity has not yet been elucidated but it has been proposed as the major determinant for the drug's cytotoxicity. To study the in vivo mode of action of trabectedin and to explore the role of NER in its cytotoxicity, we used the fission yeast Schizosaccharomyces pombe as a model system. Treatment of S. pombe wild-type cells with trabectedin led to cell cycle delay and activation of the DNA damage checkpoint, indicating that the drug causes DNA damage in vivo. DNA damage induced by the drug is mostly caused by the NER protein, Rad13 (the fission yeast orthologue to human XPG), and is mainly repaired by homologous recombination. By constructing different rad13 mutants, we show that the DNA damage induced by trabectedin depends on a 46–amino acid region of Rad13 that is homologous to a DNA-binding region of human nuclease FEN-1. More specifically, an arginine residue in Rad13 (Arg961), conserved in FEN1 (Arg314), was found to be crucial for the drug's cytotoxicity. These results lead us to propose a model for the action of trabectedin in eukaryotic cells in which the formation of a Rad13/DNA-trabectedin ternary complex, stabilized by Arg961, results in cell death. (Cancer Res 2006; 66(16): 8155-62)

Supplementary Table S1, Methods and Materials from Cross-Talk between Nucleotide Excision and Homologous Recombination DNA Repair Pathways in the Mechanism of Action of Antitumor Trabectedin

Supplementary Figure S2 from Cross-Talk between Nucleotide Excision and Homologous Recombination DNA Repair Pathways in the Mechanism of Action of Antitumor Trabectedin

BACKGROUND AND AIMS Frailty is associated with a higher rate of complications and mortality after kidney transplantation (KT). Physical frailty phenotype (PFP) is the most used frailty instrument among KT recipients, it comprises five criteria and classifies patients as frail if they have ≥3. We aimed to determine the impact of scoring 1 PFP criterion (mild frailty) on transplant outcomes. METHOD We have prospectively followed 449 KT candidates evaluated for frailty by PFP at the time of inclusion on the KT waiting list. Patients were classified as follows: 0, robust; 1, pre-frail-1; 2, pre-frail-2; and ≥3, frail. A prospective longitudinal study was performed in all patients in the cohort who underwent KT. Clinical outcomes and survival after transplantation according to frailty status at listing was assessed. RESULTS About 71.3% of total listed cohort resulted to be pre-frail (one criterion 41%, two criteria 19.8%) or frail (10.5%). Disparities were observed between sexes, with 5.2% of men and 21.9% of women being frail (P CONCLUSION Frailty is frequent in KT candidates, more frequent in women than men. Frail patients have less chances to receive a KT. KT candidates are frequently listed with one (out of five) frailty criterion, and this has an independent impact on patient survival after KT. A pre-transplant clinical framework for multimodal prehabilitation interventions to mitigate the effects of frailty and poor fitness after KT may be justified even in mildly pre-frail candidates.

A total of 1345 specimens belonging to 58 different species of wild fish and seafood from the western Mediterranean Sea were analyzed to assess total mercury levels and to estimate which species meet the EU recommendations for human consumption (0.5 μg g-1 ww) in all cases. All fish species were caught off the Mediterranean coasts and intended for human consumption. All specimens were collected from local markets located in Spain, Italy and France that sell fish caught by local fishermen (Eivissa, Menorca, Mallorca, Alacant, L'Ampolla, Ametlla de Mar, Marseille, Genoa, Civitavecchia, Alghero) at different time periods. Mercury concentrations were measured by thermal decomposition-gold amalgamator-atomic absorption spectrometry. Only thirteen species were found that did not exceed 0.5 μg g-1 ww in any specimen analyzed. These safe species were sardines (Sardina pilchardus), anchovies (Engraulis encrasicolus), blue whiting (Micromesistius poutassou), picarel (Spicara smaris), blackspot seabream (Pagellus bogaraveo), gilthead seabream (Sparus aurata), pearly razorfish (Xyrichtys novacula), surmullet (Mullus surmuletus), painted comber (Serranus scriba), brown meagre (Sciaena umbra), salema (Sarpa salpa), common dolphinfish (Coryphaena hippurus) and squid (Loligo vulgaris). These species occupy different trophic levels, have different lengths and average weights, but show a low mercury concentration than others living in the same environments. Potential human consumption of these species as sole source of fish would imply estimated weekly intakes representing between 49% and 70% of the recommended provisional tolerable weekly intake of methylmercury in the worst case. Health authorities should pay specific attention to species that do not meet EU thresholds and make appropriate precautionary health recommendations, especially for pregnant women and children., This research was supported by Neurosome, a H2020 Marie Skłodowska-Curie Actions project (grant agreement number 766251), and PARC (HLTH-2021-ENVHLTH-3:101057014) from the European Commission.

We performed a systematic sampling and analysis of airborne SARS-CoV-2 RNA in different hospital areas to assess viral spread. Systematic air filtration was performed in rooms with COVID-19 infected patients, in corridors adjacent to these rooms, to rooms of intensive care units, and to rooms with infected and uninfected patients, and in open spaces. RNA was extracted from the filters and real-time reverse transcription polymerase chain reaction was performed using the LightMix Modular SARS-CoV-2 E-gene. The highest occurrence of RNA was found in the rooms with COVID-19 patients (mean 2600 c/m3) and the adjacent corridor (mean 4000 c/m3) which was statistically significant more exposed (p < 0.01). This difference was related to the ventilation systems. As is commonly found in many hospitals, each of the rooms had an individual air inlet and outlet, while in the corridors these devices were located at the distance of every four rooms. There was a significant transfer of viruses from the COVID-19 patients’ rooms to the corridors. The airborne SARS-CoV-2 RNA in the corridors of ICUs with COVID-19 patients or care rooms of uninfected patients were ten times lower, averages 190 c/m3 and 180 c/m3, respectively, without presenting significant differences. In all COVID-19 ICU rooms, patients were intubated and connected to respirators that filtered all exhaled air and prevented virus release, resulting in significantly lower viral concentrations in adjacent corridors. The results show that the greatest risk of nosocomial infection may also occur in hospital areas not directly exposed to the exhaled breath of infected patients. Hospitals should evaluate the ventilation systems of all units to minimize possible contagion and, most importantly, direct monitoring of SARS-CoV-2 in the air should be carried out to prevent unexpected viral exposures., Financial support from the European Union Project EDCMET (H2020-HEALTH/0490–825762) is acknowledged.

The aim of this study was to know the prevalence and severity of COVID-19 in patients treated with long-term macrolides and to describe the factors associated with worse outcomes. A cross-sectional study was conducted in Primary Care setting. Patients with macrolides dispensed continuously from 1 October 2019 to 31 March 2020, were considered. Main outcome: diagnosis of coronavirus disease-19 (COVID-19). Secondary outcomes: symptoms, severity, characteristics of patients, comorbidities, concomitant treatments. A total of 3057 patients met the inclusion criteria. Median age: 73 (64–81) years, 55% were men, 62% smokers/ex-smokers, 56% obese/overweight. Overall, 95% of patients had chronic respiratory diseases and four comorbidities as a median. Prevalence of COVID-19: 4.8%. This was in accordance with official data during the first wave of the pandemic. The most common symptoms were respiratory: shortness of breath, cough, and pneumonia. Additionally, 53% percent of patients had mild/moderate symptoms, 28% required hospital admission, and 19% died with COVID-19. The percentage of patients hospitalized and deaths were 2.6 and 5.8 times higher, respectively, in the COVID-19 group (p &lt, 0.001). There was no evidence of a beneficial effect of long-term courses of macrolides in preventing SARS-CoV-2 infection or the progression to worse outcomes in old patients with underlying chronic respiratory diseases and a high burden of comorbidity.

Resumen Objetivo Analizar la incidencia de la enfermedad del coronavirus 19 (COVID-19) en pacientes consumidores cronicos de hidroxicloroquina. Pacientes y metodos Estudio multicentrico observacional retrospectivo transversal en Areas de Salud de Castilla La-Mancha y distritos sanitarios de Andalucia. De los 4.451 participantes incluidos en el primer reclutamiento se seleccionaron 3.817 sujetos con datos validos. La variable principal del estudio ha sido la presencia o ausencia de infeccion por la COVID-19 por diagnostico clinico, serologico o por reaccion en cadena de la polimerasa. Se registraron variables sociodemograficas, clinicas y tratamientos y comorbilidades concomitantes. Resultados Ciento sesenta y nueve (4,45%) pacientes presentaron infeccion por la COVID-19, de los cuales fallecieron 12 (7,1%) y 32 (18,9%) requirieron ingreso hospitalario. La enfermedad respiratoria previa se relaciono con la infeccion por la COVID-19 (p Conclusion No existe relacion entre consumo cronico de hidroxicloroquina e incidencia de la COVID-19.

To analyze the incidence of Covid-19 in patients who are chronic users of hydroxychloroquine.Cross-sectional retrospective observational multicenter study in health areas and districts from Castilla La-Mancha and Andalucia. Of the 4451 participants included in the first recruitment, 3817 with valid data were selected. The main variable of the study is the presence or absence of Covid-19 infection by clinical, serological or polymerase chain reaction diagnosis. Sociodemographic and clinical variables and treatment and concomitant comorbidities were recorded.169 (4,45%) patients had Covid-19 infection, of which 12 (7.1 %) died and 32 (18.9%) required hospital admission. Previous respiratory pathology was related to Covid-19 infection (There is no relationship between chronic use of hydroxychloroquine and the incidence of Covid-19.Analizar la incidencia de la enfermedad del coronavirus 19 (COVID-19) en pacientes consumidores crónicos de hidroxicloroquina.Estudio multicéntrico observacional retrospectivo transversal en Áreas de Salud de Castilla La-Mancha y distritos sanitarios de Andalucía. De los 4.451 participantes incluidos en el primer reclutamiento se seleccionaron 3.817 sujetos con datos válidos. La variable principal del estudio ha sido la presencia o ausencia de infección por la COVID-19 por diagnóstico clínico, serológico o por reacción en cadena de la polimerasa. Se registraron variables sociodemográficas, clínicas y tratamientos y comorbilidades concomitantes.Ciento sesenta y nueve (4,45%) pacientes presentaron infección por la COVID-19, de los cuales fallecieron 12 (7,1%) y 32 (18,9%) requirieron ingreso hospitalario. La enfermedad respiratoria previa se relacionó con la infección por la COVID-19 (p 0,05). La maculopatía aparece en un 5,3% de los pacientes y está relacionada significativamente con la dosis de hidroxicloroquina consumida (p 0,05).No existe relación entre consumo crónico de hidroxicloroquina e incidencia de la COVID-19.

The composition of polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs), hexachlorobenzene (HCB), pentachlorobenzene (PeCB), and organophosphate flame retardants (OPFRs) present in the gas-phase fraction of the atmosphere of Barcelona was analyzed during the SARS-CoV-2 coronavirus disease (COVID-19) lockdown and prior to this period. The changes in daily concentrations of CO, NO, NO2, O3 and particulate matter smaller than 10 μm (PM10) were considered for comparison. Bayesian analysis considering serial dependencies and seasonality showed statistically significant decreases of CO, NO, NO2, and PM10 (between −28 and −76%) and O3 increases (+45%) during lockdown. However, the lockdown concentration decreases of PeCB (−90.5%, from 8.5 to 0.8 pg m–3), HCB (−79%, 25.5–5.4 pg m–3) and some PAHs, such as benz[a]anthracene (−87%, 120–17 pg m–3) and pyrene (−81%, 3,500–680 pg m–3), were even stronger. The PAH depletion ranged between −68 and −87% that could be primarily associated with the strong reduction of traffic mobility during this period (−80%). Besides traffic reduction, the observed air quality improvements could be related to lower generation of solid urban residues (−25%) and the subsequent decrease of urban waste incineration (between −25 and −28%). Tributyl phosphate also showed a reduction in concentration during lockdown but the other OPFRs were seemingly not affected by this restriction, possibly as a result of the uniform release from the emission sources, e.g., construction material, industrial applications, and household products., This work was supported by the Spanish Ministry of Science and Innovation (Projects: CUANTOX CTM2015-71832-P and INTEMPOL PGC2018-10228-B-I00). RP also acknowledges financial support from the Spanish Ministry of Science and Innovation (BES-2016-076339).

Some patients in the community receive a high burden of antibiotics. We aimed at describing the characteristics of these patients, antibiotics used, and conditions for which they received antibiotics. We carried out a cross‐sectional study. Setting: Thirty Health Primary Care Areas from 12 regions in Spain, covering 5,960,191 inhabitants. Patients having at least 30 packages of antibacterials for systemic use dispensed in 2017 were considered. Main outcome measures: Prevalence of antibiotic use, conditions for which antibiotics were prescribed, clinical characteristics of patients, comorbidities, concomitant treatments, and microbiological isolates. Patient’s average age was 70 years; 52% were men; 60% smokers/ex‐smokers; 54% obese. Overall, 93% of patients had, at least, one chronic condition, and four comorbidities on average. Most common comorbidities were cardiovascular and/or hypertension (67%), respiratory diseases (62%), neurological/mental conditions (32%), diabetes (23%), and urological diseases (21%); 29% were immunosuppressed, 10% were dead at the time of data collection. Patients received three antibiotic treatments per year, mainly fluoroquinolones (28%), macrolides (21%), penicillins (19%), or cephalosporins (12%). Most frequently treated conditions were lower respiratory tract (infections or prophylaxis) (48%), urinary (27%), and skin/soft tissue infections (11%). Thirty‐five percent have been guided by a microbiological diagnosis, being Pseudomonas aeruginosa (30%) and Escherichia coli (16%) the most frequent isolates. In conclusion, high antibiotic consumers in the community were basically elder, with multimorbidity and polymedication. They frequently received broad‐spectrum antibiotics for long periods of time. The approach to infections in high consumers should be differentiated from healthy patients receiving antibiotics occasionally., Patients receiving a high burden of antibiotics in the community in Spain.

The Mediterranean Sea is a water body in which the concentration of mercury is much higher than in the other world seas and oceans. Most inputs of this metal originate from the general atmospheric fallout. However, in this semi-enclosed environment there are specific sources that should be identified to understand the causes of the high toxicity by this metal. A significant proportion of Mediterranean fish devoted to human consumption is above the mercury threshold set by the European Community as suitable for human consumption. The proportion is even much larger if the recommended World Health Organization threshold is considered. Oily fish is known for containing mercury concentrations above these thresholds. Lean fish has been investigated in much fewer cases. The present study is devoted to this second fish type that constitutes a substantial component of human diet. Thus, the study of mercury and methylmercury in fish from local fishermen marketed in diverse Mediterranean sites has provided information on the exposure of diverse populations to this metal and has afforded a description of the Mediterranean areas that have received highest mercury spills.1350 commercial seafood samples from the Western Mediterranean Sea were collected (Feb 2014-July 2019) in several sites such as Mallorca, Menorca, Eivissa, Alacant (Spain), Marseille (France), Genoa, Alguer, Civitavecchia (Italy). Samples from Egypt and the Atlantic Ocean (Senegal, Mauritania coasts) were also taken for comparison. Fish species were selected considering the most consumed by the population.Comparison of the mercury concentrations in the specimens of the same fish species collected at different sites revealed where are the hot spots of introduction of the excess of this metal in comparison to the atmospheric fallout and allowed the identification of the source processes.The fish species were grouped in three trophic levels, those feeding on plankton (first), on small fish and crustaceans (second) and on fish and cephalopods (third).A considerable number of the analyzed fish species exceeded the maximum levels proposed by the European legislation, such as dusky grouper (100% of the examined specimens), common dentex (65%), conger (45%), common sole (38%), hake (26%) and angler (15%), among others. Representation of the Hg concentrations vs. weight of each specimen from the third trophic level showed a significant positive correlation, r = 0.78 (p < 0.01).The average THg intake due to fish consumption, 0.61 µg/g ww, involved Hg estimated weekly intakes of 5.7 µg/kg bw for children aged 7-12 years and 4.4 µg/kg bw for adults. These values were higher than the provisional tolerable weekly intakes for total Hg intake recommended by FAO/WHO, 4 µg/kg bw, 140% and 110%, respectively.

Inhalable particulate matter (size p-Xylene) and n-Pentane (5–15 μg/m3). ƩBTEX concentrations were, This work is supported by the Spanish Ministry of Economy, Industry and Competitiveness with FEDER funds (BUSAIR CGL2016-79132-R) and by CSIC with the “Proyecto Intramural: Exposición a compuestos organicos volatiles en el interior de autobuses de transporte publico urbano” (201730I003). We are grateful to Transports Metropolitans de Barcelona (TMB) for their co-operation throughout the whole project. Funding from the Generalitat de Catalunya (AGAUR 2017 SGR41) is also acknowledged.

Vehicle interior air quality (VIAQ) was investigated inside 14 diesel/non-diesel taxi pairs operating simultaneously and under normal working conditions over six weekday hours (10.00–16.00) in the city of Barcelona, Spain. Parameters measured included PM 10 mass and inorganic chemistry, ultrafine particle number (N) and size, lung surface deposited area (LDSA), black carbon (BC), CO 2 , CO, and a range of volatile organic compounds (VOCs). Most taxi drivers elected to drive with windows open, thus keeping levels of CO 2 and internally-generated VOCs low but exposing them to high levels of traffic-related air pollutants entering from outside and confirming that air exchange rates are the dominant influence on VIAQ. Median values of N and LDSA (both sensitive markers of VIAQ fluctuations and likely health effects) were reduced to around 10 4 #/cm 3 and < 20 µm 2 /cm 3 respectively under closed conditions, but more than doubled with windows open and sometimes approached 10 5 #/cm 3 and 240 µm 2 /cm 3 . In exceptional traffic conditions, transient pollution peaks caused by outside infiltration exceeded N = 10 6 #/cm 3 and LDSA= 1000 µm 2 /cm 3 . Indications of self-pollution were implicated by higher BC and CO levels, and larger UFP sizes, measured inside diesel taxis as compared to their non-diesel pair, and the highest concentrations of CO (>2 ppm) were commonly associated with older, high-km diesel taxis. Median PM 10 concentrations (67 µg/m 3 ) were treble those of urban background, mainly due to increased levels of organic and elemental carbon, with source apportionment calculations identifying the main pollutants as vehicle exhaust and non-exhaust particles. Enhancements in PM 10 concentrations of Cr, Cu, Sn, Sb, and a “High Field Strength Element” zircon-related group characterised by Zr, Hf, Nb, Y and U, are attributed mainly to the presence of brake-derived PM. Volatile organic compounds display a mixture which reflects the complexity of traffic-related organic carbon emissions infiltrating the taxi interior, with 2-methylbutane and n-pentane being the most abundant VOCs, followed by toluene, m-xylene, o-xylene, 1,2,4-trimethylbenzene, ethylbenzene, p-xylene, benzene, and 1,3,5-trimethylbenzene. Internally sourced VOCs included high monoterpene concentrations from an air freshener, and interior off-gassing may explain why the youngest taxi registered the highest content of alkanes and aromatic compounds. Carbon dioxide concentrations quickly climbed to undesirable levels (>2500 ppm) under closed ventilation conditions and could stay high for much of the working day. Taxi drivers face daily occupational exposure to traffic-related air pollutants and would benefit from a greater awareness of VIAQ issues, notably the use of ventilation, to encourage them to minimise possible health effects caused by their working environment. © 2019 The Author(s), This work was supported by the ACS Foundation contributing to the dissemination of good environmental practices and environmental protection activities, the Spanish Ministry of Economy and Competitiveness and FEDER funds within the I+D Project CGL2016–79132 (BUSAIR), and the IMPROVE LIFE project ( LIFE13ENV/ES/000263 ). We thank the Barcelona Metropolitan Taxi Institute (IMET) for their enthusiastic help and collaboration in this project.

Background and aims: Lack of achievement of secondary prevention objectives in patients with ischaemic heart disease remains an unmet need in this patient population. We aimed at evaluating the six-month efficacy of an intensive lipid-lowering intervention, coordinated by nurses and implemented after hospital discharge, in patients hospitalized for an ischaemic heart disease event. Methods: Randomized controlled trial, in which a nurse-led intervention including periodic follow-up, serial lipid level controls, and subsequent optimization of lipid-lowering therapy, if appropriate, was compared with standard of care alone in terms of serum lipid-level control at six months after discharge. Results: The nurse-led intervention was associated with an improved management of low-density lipoprotein (LDL) cholesterol levels compared with standard of care alone: LDL cholesterol levels ⩽100 mg/dL were achieved in 97% participants in the intervention arm as compared with 67% in the usual care arm ( p value Conclusions: Our findings highlight the opportunity that nurse-led, intensive, post-discharge follow-up plans may represent for achieving LDL cholesterol guideline-recommended management objectives in patients with ischaemic heart disease. These findings should be replicated in larger cohorts.

Background: Swimming in pools is a healthy activity that entails exposure to disinfection by-products (DBPs), some of which are irritant and genotoxic. Objectives: We evaluated exposure to DBPs during swimming in a chlorinated pool and the association with short-term changes in genotoxicity and lung epithelium permeability biomarkers. Methods: Non-smoker adults (N = 116) swimming 40 min in an indoor pool were included. We measured a range of biomarkers before and at different times after swimming: trihalomethanes (THMs) in exhaled breath (5 min), trichloroacetic acid (TCAA) in urine (30 min), micronuclei in lymphocytes (1 h), serum club cell protein (CC16) (1 h), urine mutagenicity (2 h) and micronuclei in reticulocytes (4 days in a subset, N = 19). Several DBPs in water and trichloramine in air were measured, and physical activity was extensively assessed. We estimated interactions with polymorphisms in genes related to DBP metabolism. Results: Median level of chloroform, brominated and total THMs in water was 37.3, 9.5 and 48.5, μg/L, respectively, and trichloramine in air was 472.6 μg/m3. Median exhaled chloroform, brominated and total THMs increased after swimming by 10.9, 2.6 and 13.4, μg/m3, respectively. Creatinine-adjusted urinary TCAA increased by 3.1 μmol/mol. Micronuclei in lymphocytes and reticulocytes, urine mutagenicity and serum CC16 levels remained unchanged after swimming. Spearman correlation coefficients showed no association between DBP exposure and micronuclei in lymphocytes, urine mutagenicity and CC16. Moderate associations were observed for micronuclei in reticulocytes and DBP exposure. Conclusions: The unchanged levels of the short-term effect biomarkers after swimming and null associations with personal estimates of exposure to DBPs suggest no measurable effect on genotoxicity in lymphocytes, urine mutagenicity and lung epithelium permeability at the observed exposure levels. The moderate associations with micronuclei in reticulocytes require cautious interpretation given the reduced sample size. © 2019 The Authors, We are grateful to all volunteers and the swimming pool facility. This work was funded by the EU 7th Framework Programme EXPOSOMICS Project (grant agreement 308610 ). We thank the Parc de Salut Mar Biobank (MARBiobanc) for providing support to the project. MARBiobanc is supported by grants from Instituto de Salud Carlos III / FEDER ( RD09/0076/00036 and PT13/0010/0005 ) and the “Xarxa de Bancs de tumors” sponsored by Pla Director d'Oncologia de Catalunya (XBTC). ISGlobal is a member of the CERCA Programme, Generalitat de Catalunya. We appreciate the work by Kate Jones (Health & Safety Laboratory, UK) for urinary TCAA analysis.

The changes of particles and organic pollutants in indoor atmospheres as consequence of vaping with electronic cigarettes have been analyzed. Changes in the composition of volatile organic compounds (VOCs) in exhaled breath of non-smoking volunteers present in the vaping environments have also been studied. The exposure experiments involved non-vaping (n = 5) and vaping (n = 5) volunteers staying 12 h together in a room (54 m

The outdoor atmospheric distributions of chlorinated volatile organic compounds (VOCs) from locations receiving the emissions of a chlor-alkali plant have been studied. Trichloroethylene and tetrachloroethylene (medians 2.4 μg/m3 and 1.7 μg/m3, respectively) were the most abundant compounds, which was in accordance with the production processes from these installations. The concentrations of trichlorofluoromethane, median 1.6 μg/m3, are rather similar to the average levels described in general in the troposphere and cannot be attributed to this specific source. Several by-products involving dichloroacetylene, carbon tetrachloride, hexachloroethane, hexachlorobutadiene, trans-1H-pentachloro-1,3-butadiene, 2H-pentachloro-1,3-butadiene, cis-1H-pentachloro-1,3-butadiene, tetrachloro- and trichloro-butadienes and hexachlorobenzene were also identified. Some of these compounds, e.g. carbon tetrachloride, chloroform and tetrachloroethane, could also have been manufactured during some periods. The occurrence of these manufactured compounds and by-products in the atmosphere could also reflect, at least in part, volatilization during the extraction of previously discharged chlor-alkali residues developed within the environmental restoration program of the Flix water reservoir. In this respect, the tri-, tetra- and pentachloro-1,3-butadienes could also originate from microbial transformation in the solid deposits accumulated in the water reservoir which were volatilized after extraction. Among all identified VOCs, trichloroethylene showed the highest health risks considering the measured airborne concentrations and the WHO and USEPA recommendations.

aServicio de Neumologia, Hospital Universitari Vall d’Hebron, Barcelona, Spain bCIBER de Enfermedades Respiratorias (CIBERES), Spain cUnidad de Neumologia, Servicio de Medicina Interna, Hospital de Requena, Valencia, Spain dServicio de Neumologia, Hospital Clinico San Carlos, Madrid, Spain eCentro de Salud Francia, Direccion Asistencial Oeste, Madrid, Spain fServicio de Medicina Interna, Hospital Universitari Mutua de Terrassa, Terrasa, Barcelona, Spain gCentro de Salud Lucena I, Lucena, Cordoba, Spain hServicio de Neumologia, Hospital San Pedro de Alcantara, Caceres, Spain iCentro de Salud Menasalbas, Toledo, Spain jServicio de Urgencias, Hospital General Universitario Reina Sofia, Murcia, Spain kServicio de Urgencias, Hospital General Yague, Burgos, Spain lMedicina Fisica y Rehabilitacion, Parc de Salut Mar, Grupo de Investigacion en Rehabilitacion, Institut Hospital del Mar d’Investigacions Mediques, Universitat Autonoma de Barcelona, Universitat Internacional de Catalunya, Barcelona, Spain mUnidad de Fisioterapia Respiratoria, Hospital Universitario de Gran Canaria Dr. Negrin, Facultad de Ciencias de la Salud, Universidad de Las Palmas de Gran Canaria, Spain nServicio de Medicina Fisica y Rehabilitacion, Hospital Germans Trias i Pujol, Universitat Autonoma de Barcelona, Badalona, Barcelona, Spain nServicio de Medicina Fisica y Rehabilitacion, Hospital Parc Tauli, Universitat Autonoma de Barcelona, Sabadell, Barcelona, Spain oCentro de Salud Gandhi, Madrid, Spain pServicio de Medicina Interna, Hospital Royo Vilanova, Zaragoza, Spain qFundacion Caubet-Cimera FISIB Illes Balears, Bunyola, Baleares, Spain rServicio de Neumologia, Hospital Universitario de la Princesa, Instituto de Investigacion Sanitaria Princesa (IP), Madrid, Spain

Source contributions of organic aerosol (OA) are still not fully understood, especially in terms of quantitative distinction between secondary OA formed from anthropogenic precursorsvs.that formed from natural precursors. In order to investigate the OA origin, a field campaign was carried out in Barcelona in summer 2013, including two periods characterized by low and high traffic conditions. Volatile organic compound (VOC) concentrations were higher during the second period, especially aromatic hydrocarbons related to traffic emissions, which showed a marked daily cycle peaking during traffic rush hours, similarly to black carbon (BC) concentrations. Biogenic VOC (BVOC) concentrations showed only minor changes from the low to the high traffic period, and their intra-day variability was related to temperature and solar radiation cycles, although a decrease was observed for monoterpenes during the day. The organic carbon (OC) concentrations increased from the first to the second period, and the fraction of non-fossil OC as determined by14C analysis increased from 43% to 54% of the total OC. The combination of14C analysis and Aerosol Chemical Speciation Monitor (ACSM) OA source apportionment showed that the fossil OC was mainly secondary (>70%) except for the last sample, when the fossil secondary OC only represented 51% of the total fossil OC. The fraction of non-fossil secondary OC increased from 37% of total secondary OC for the first sample to 60% for the last sample. This enhanced formation of non-fossil secondary OA (SOA) could be attributed to the reaction of BVOC precursors with NOxemitted from road traffic (or from its nocturnal derivative nitrate that enhances night-time semi-volatile oxygenated OA (SV-OOA)), since NO2concentrations increased from 19 to 42 μg m−3from the first to the last sample.

A large collection of structural snapshots along a full catalytic cycle of Escherichia coli thioredoxin reductase (TrxR) has been generated and characterized using a combination of theoretical methods. Molecular models were built starting from the available X-ray crystallographic structures of dimeric wild-type TrxR in the flavin-oxidizing conformation and a C135S TrxR mutant enzyme in a flavin-reducing conformation “trapped” by a cross-link between Cys138 of TrxR and Cys32 of C35S mutant thioredoxin (Trx). The transition between these two extreme states, which is shown to be reproduced in a normal mode analysis, as well as natural cofactor binding and dissociation, were simulated for the wild-type species using unrestrained and targeted molecular dynamics following docking of oxidized Trx to reduced TrxR. The whole set of simulations provides a comprehensive structural framework for understanding the mechanism of disulfide reduction in atomic detail and identifying the most likely intermediates that facilitate entry of NADPH and exit of NADP+. The crucial role assigned to Arg73 and Lys36 of Trx in substrate binding and complex stabilization was ascertained when R73G, R73D, and K36A site-directed mutants of Trx were shown to be impaired to different extents in their ability to be reduced by TrxR. On the basis of previous findings and the results reported herein, E. coli TrxR appears as a beautifully engineered molecular machine that is capable of synchronizing cofactor capture and ejection with substrate binding and redox activity through an interdomain twisting motion. Proteins 2010. © 2009 Wiley-Liss, Inc.

The different steps of the dehalogenation reaction carried out by LinB on three different substrates have been characterized using a combination of quantum mechanical calculations and molecular dynamics simulations. This has allowed us to obtain information in atomic detail about each step of the reaction mechanism, that is, substrate entrance and achievement of the near-attack conformation, transition state stabilization within the active site, halide stabilization, water molecule activation and subsequent hydrolytic attack on the ester intermediate with formation of alcohol, and finally product release. Importantly, no bias or external forces were applied during the whole procedure so that both intermediates and products were completely free to sample configuration space in order to adapt to the plasticity of the active site and/or search for an exit. Differences in substrate reactivity were found to be correlated with the ease of adopting the near-attack conformation and two different exit pathways were found for product release that do not interfere with substrate entrance. Additional support for the different entry and exit pathways was independently obtained from an examination of the enzyme's normal modes.

9 pages, 2 figures, 5 tables.-- PMID: 17574674 [PubMed].-- Printed version published Oct 2007., We aimed to analyse the influence of cholesterol and triglycerides, and of tobacco, coffee and alcohol consumption upon correlations between serum concentrations of organochlorine compounds (OCs) in patients with exocrine pancreatic cancer (EPC). Incident cases of EPC diagnosed in eastern Spain were prospectively identified (N=144). OCs were analysed by high-resolution gas chromatography with electron-capture detection. A strong correlation was observed between hexachlorobenzene (HCB) and β-hexachlorocyclohexane (β-HCH) (Spearman's ρ=0.758). β-HCH showed ρ>0.4 with p, p′-DDT, p, p′-DDE, PCB138 and PCB153 (all p, Supported by research grants from ‘Red temática de investigación cooperativa de centros en Cáncer’ (C03/10), ‘Red temática de investigación cooperativa de centros en Epidemiología y salud pública’ (C03/09), and CIBER de Epidemiología, Instituto de Salud Carlos III, Ministry of Health. Madrid.

A search for suppressors of the carnitine/acylcarnitine translocase (CACT) deficiency in Aspergillus nidulans permitted the identification of the suaE7 mutation, mapping at a new translational suppressor (suaE) gene. The suaE gene is essential in A. nidulans and encodes the eukaryotic release factor 1 (eRF1). The suaE7 mutation suppresses two acuH alleles (acuH13 and acuH31), both carrying nonsense mutations in the CACT encoding gene that involve the replacement of a CAG (Gln) codon with a premature TAG stop codon. In contrast, the suaE7 gene does not suppress the acuH20 amber nonsense mutation involving a TGG-->TAG change. The phenotype associated to the suaE7 mutation strictly resembles that of mutants at the suaA and suaC genes, two translational suppressor genes previously identified, suggesting that their gene products might functionally interact in translation termination. Sequencing of the suaE7 gene allowed the identification of a mutation in the domain 2 of the omnipotent class-1 eukaryotic release factor involving the Gly265Ser substitution in the A. nidulans eRF1. This mutation creates a structural context unfavourable for normal eRF binding that allows the misreading of stop codons by natural suppressor tRNAs, such as the tRNAs(Gln). Structural analysis using molecular modelling of A. nidulans eRF1 domain 2 bearing the G265S substitution and computer simulation results suggest that this mutation might impair the necessary conformational changes in the eRF1 to optimally recognize the stop codon and simultaneously interact with the peptidyl transferase centre of the 60S ribosomal subunit.

A method for the rapid analysis of volatile organic compounds (VOCs) in smoke from tobacco and electronic cigarettes and in exhaled breath of users of these smoking systems has been developed. Both disposable and rechargeable e-cigarettes were considered. Smoke or breath were collected in Bio-VOCs. VOCs were then desorbed in Tenax cartridges which were subsequently analyzed by thermal desorption coupled to gas chromatography-mass spectrometry. The method provides consistent results when comparing the VOC compositions from cigarette smoke and the equivalent exhaled breath of the smokers. The differences in composition of these two sample types are useful to ascertain which compounds are retained in the respiratory system after tobacco cigarette or e-cigarette smoking.Strong differences were observed in the VOC composition of tobacco cigarette smoke and exhaled breath when comparing with those of e-cigarette smoking. The former involved transfers of a much larger burden of organic compounds into smokers, including benzene, toluene, naphthalene and other pollutants of general concern. e-Cigarettes led to strong absorptions of propylene glycol and glycerin in the users of these systems. Tobacco cigarettes were also those showing highest concentration differences between nicotine concentrations in smoke and exhaled breath. The results from disposable e-cigarettes were very similar to those from rechargeable e-cigarettes. © 2015 Elsevier B.V., We thank the Spanish Ministry of Economy and Competitiveness for the Expo-Cov (CTM2012-39298) fellowship. Financial support from the EU projects HEALS (FP7-ENV-2013- 603946) and CROME (LIFE12 ENV/GR/001040) is acknowledged.

Background: Swimming in pools entails a considerable exposure to carcinogenic and irritant disinfection by-products (DBPs). Biomarkers of lung epithelium permeability (serum CC16), genotoxicity (mi...

This study describes the relationship between physical activity and intake of trihalomethanes (THMs), namely chloroform (CHCl3), bromodichloromethane (CHCl2Br), dibromochloromethane (CHClBr2) and bromoform (CHBr3), in individuals exposed in two indoor swimming pools which used different disinfection agents, chlorine (Cl-SP) and bromine (Br-SP). CHCl3 and CHBr3 were the dominant compounds in air and water of the Cl-SP and Br-SP, respectively. Physical exercise was assessed from distance swum and energy expenditure. The changes in exhaled breath concentrations of these compounds were measured from the differences after and before physical activity.A clear dependence between distance swum or energy expenditure and exhaled breath THM concentrations was observed. The statistically significant relationships involved higher THM concentrations at higher distances swum. However, air concentration was the major factor determining the CHCl3 and CHCl2Br intake in swimmers whereas distance swum was the main factor for CHBr3 intake. These two causes of THM incorporation into swimmers concurrently intensify the concentrations of these compounds into exhaled breath and pointed to inhalation as primary mechanism for THM uptake. Furthermore, the rates of THM incorporation were proportionally higher as higher was the degree of bromination of the THM species. This trend suggested that air-water partition mechanisms in the pulmonary system determined higher retention of the THM compounds with lower Henry's Law volatility constants than those of higher constant values. Inhalation is therefore the primary mechanisms for THM exposure of swimmers in indoor buildings. © 2015 Elsevier Inc., Financial supportfromPISCINA(SAF2005-07643-CO3-02), Expo-Cov (CTM2012-39298)andConsolider-Ingenio(CSD2007-00067)ProjectsfromtheSpanishMinistriesofResearchandDe- velopmentandEconomyandCompetitivenessand(LIFE12ENV/GR/001040)andHEALS(FP7-ENV-2013-603946)fromtheEuropean Unionisacknowledged.WealsothanktheINMACooperativeResearchNetwork.CLthankstheCSIC-BancodeSantander agreementforaPhDfellowship.

Trabectedin and its N12-demethylated analogue ET729 bind covalently to the central guanine of selected DNA triplets. Although both drugs equally target several sites, including AGA, we show that covalent modification of CGA is only achieved by ET729. By means of molecular dynamics simulations of the precovalent complexes, we explain in atomic detail how such a simple structural modification brings about this notable change in the DNA-binding selectivity profiles of these two drugs.

The major structural determinant of the preference to bind to CpG binding sites on DNA exhibited by the natural quinoxaline bis-intercalators echinomycin and triostin A, or the quinoline echinomycin derivative, 2QN, is the 2-amino group of guanine (G). However, relocation of this group by means of introduction into the DNA molecule of the 2-aminoadenine (=2,6-diaminopurine, D) base in place of adenine (A) has been shown to lead to a drastic redistribution of binding sites, together with ultratight binding of 2QN to the sequence DTDT. Also, the demethylated triostin analogs, TANDEM and CysMeTANDEM, which bind with high affinity to TpA steps in natural DNA, bind much less tightly to CpI steps, despite the fact that both adenosine and the hypoxanthine-containing nucleoside, inosine (I), provide the same hydrogen bonding possibilities in the minor groove. To study both the increased binding affinity of 2QN for DTDT relative to GCGC sites and the remarkable loss of binding energy between CysMeTANDEM and ICIC compared with ATAT, a series of thermodynamic integration free energy simulations involving conversions between DNA base pairs have been performed. Our results demonstrate that the electrostatic component of the stacking interactions between the heteroaromatic rings of these compounds and the bases that make up the intercalation sites plays a very important role in the modulation of their binding affinities.

Yondelis (trabectedin) is an antitumor ecteinascidin that binds covalently to the 2-amino group of the central guanine in the minor groove of selected DNA pyrimidine-G-G and purine-G-C triplets. Chromomycin A3 is an aureolic acid derivative that binds noncovalently to the DNA minor groove in G/C-rich triplet sites as a metal-chelated dimer. Despite their different binding modes, the cytotoxicity profiles of these two drugs, as assessed in the COMPARE analysis carried out by the National Cancer Institute on data from 60 human tumor cell lines, are highly correlated (Pearson's correlation coefficient of 0.96). We now report that in an oligonucleotide containing the "natural bending element" TGGCCA, the structural distortions inflicted by the tail-to-tail bonding of two trabectedin molecules to adjacent target sites on opposing strands are strikingly similar to those observed in a crystal containing d(TTGGCCAA)2 and two bound chromomycin A3 molecules arranged in a head-to-tail orientation in the minor groove. In both complexes, the double helix is characterized by being considerably unwound and possessing a notably widened minor groove. Binding of the drugs to this sequence could be favored by the distinct bends at each of the TpG steps that are already present in the free oligonucleotide. Simultaneous drug binding to the two strands in the manner described here is proposed to stabilize the helical structure of duplex DNA to prevent or hamper strand separation and stall replication and transcription forks.

Didemnins and tamandarins are closely related marine natural products with potent inhibitory effects on protein synthesis and cell viability. On the basis of available biochemical and structural evidence and results from molecular dynamics simulations, a model is proposed that accounts for the strong and selective binding of these compounds to human elongation factor eEF1A in the presence of GTP. We suggest that the p-methoxyphenyl ring of these cyclic depsipeptides is inserted into the same pocket in eEF1A that normally lodges either the 3' terminal adenine of aminoacylated tRNA, as inferred from two prokaryotic EF-Tu.GTP.tRNA complexes, or the aromatic side chain of Phe/Tyr-163 from the nucleotide exchange factor eEF1Balpha, as observed in several X-ray crystal structures of a yeast eEF1A:eEF1Balpha complex. This pocket, which has a strong hydrophobic character, is formed by two protruding loops on the surface of eEF1A domain 2. Further stabilization of the bound depsipeptide is brought about by additional crucial interactions involving eEF1A domain 1 in such a way that the molecule fits snugly at the interface between these two domains. In the GDP-bound form of eEF1A, this binding site exists only as two separate halves, which accounts for the much greater affinity of didemnins for the GTP-bound form of this elongation factor. This binding mode is entirely different from those seen in the complexes of the homologous prokaryotic EF-Tu with kirromycin-type antibiotics or the cyclic thiazolyl peptide antibiotic GE2270A. Interestingly, the set of interactions used by didemnins to bind to eEF1A is also distinct from that used by eEF1Balpha or eEF1Bbeta, thus establishing a competition for binding to a common site that goes beyond simple molecular mimicry. The model presented here is consistent with both available biochemical evidence and known structure-activity relationships for these two classes of natural compounds and synthetic analogues and provides fertile ground for future research.

The antitumor ecteinascidin ET743 has been shown to inhibit the transcriptional activation of a number of genes at nanomolar concentrations. Cell sensitivity to subnanomolar concentrations of the drug has also been shown to specifically depend on the transcription-coupled nucleotide excision repair system. ET743 is known to bind covalently to the minor groove of a DNA double helix in regions comprising selected sets of three consecutive base pairs. Following alkylation of a central guanine, the minor groove is widened and the DNA is bent toward the major groove. We have previously shown that in the resulting adduct the DNA triplet containing the covalently modified guanine bears a strong resemblance to a DNA triplet recognized by a C(2)H(2) zinc finger. We now expand this earlier finding and use simulation methods to show that head-to-tail binding of three ET743 molecules to three adjacent optimal binding sites stabilizes a DNA structure whose conformation is intermediate between A- and B-form DNA. Furthermore, despite the increase in roll at the sites of covalent attachment, no net curvature is apparent in this complex due to cancellation of the localized bends over virtually one turn of the helix. Both observations are in good analogy to findings in zinc finger-DNA complexes. Triplets are virtually superimposable both directly and upon shifting the register one base pair. In this latter case, the central guanine in a triplet alkylated by ET743 corresponds to the third nucleic base in the triplet recognized by a zinc finger of transcription factors such as EGR1 or Sp-1. The DNA conformation found in the ET743-DNA complex is also strongly reminiscent of an RNA-DNA hybrid, as found in the RNA polymerase II elongation complex. The possible biological implications of these findings in relation to the antitumor action of ET743 are discussed.

Hexachlorobenzene (HCB) is an organochlorine compound widespread in the environment, highly lipophilic, that accumulates in biological systems. It has been suggested that it should be classified as a dioxin-like compound. Newborns are exposed to organochlorine compounds across the placenta and through breastfeeding. Although HCB is one of the most common organochlorine compounds, the transplacental transference of HCB from mother to fetus during pregnancy has been scarcely documented. This study reports the levels of HCB, dichlorodiphenyl trichloroethane (DDT) and its metabolite p,p'DDE, polychlorinated biphenyls (PCBs), and beta-hexachlorocyclohexane (beta-HCH) in 72 maternal blood samples at delivery and in 69 cord blood samples, from which 62 corresponded to mother infant pairs born between May 1997 and September 1999 in a rural area highly exposed to HCB. Results show that all newborns presented detectable levels of HCB, PCBs, and p,p'DDE, and, to a lesser extent, of beta-HCH, the HCB levels being the highest. The geometric mean of HCB was 1.1 ng/ml, ranging from 0.3 to 5.7 ng/ml. Concentrations of HCB levels in cord blood (log ng/ml) were positively associated with concentrations in maternal blood (log ng/ml) (coefficient = 0.45, P < 0.01). Gestational age was not associated with the transplacental transfer of HCB. Maternal p,p'DDE and beta-HCH levels were also associated with newborn levels, but levels of PCBs were not. We conclude that HCB, similar to other organochlorinated compounds, has a transplacental transfer.

The anthranilic acid diamides represent the most recent class of nonpeptide CCK(1) receptor (CCK(1)-R) antagonists. Herein we describe the second phase of the anthranilic acid C-terminal optimization using nonproteinogenic amino acids containing a phenyl ring in their side chain. The Homo-Phe derivative 2 (VL-0797) enhanced 12-fold the affinity for the rat CCK(1)-R affinity and 15-fold for the human CCK(1)-R relative to the reference compound 12 (VL-0395). The eutomer of 2 (6) exhibited a nanomolar range affinity toward the human CCK(1)-R and was at least 400-fold selective for the CCK(1)-R over the CCK(2)-R. Molecular docking in the modeled CCK(1)-R and its validation by site-directed mutagenesis experiments showed that the 6 binding site overlaps that occupied by the C-terminal bioactive region of the natural agonist CCK. Owing to their interesting properties, new compounds provided by this study represent a solid basis for further advances aimed at synthesis of clinically valuable CCK(1)-R antagonists.

This first study of trihalomethanes (THMs) in swimming pools using bromine agents for water disinfection under real conditions shows that the mixtures of these compounds are largely dominated by bromoform in a similar process as chloroform becomes the dominant THM in pools disinfected with chlorine agents. Bromoform largely predominates in air and water of the pool installations whose concentration changes are linearly correlated. However, the air concentrations of bromoform account for about 6–11% of the expected concentrations according to theoretical partitioning defined by the Henry law. Bromoform in exhaled air of swimmers is correlated with the air concentrations of this disinfectant by-product in the pool building. Comparison of the THM exhaled air concentrations between swimmers and volunteers bathing in the water without swimming or standing in the building outside the water suggest that physical activity enhance exposure to these disinfectant by-products. They also indicate that in swimming pools, besides inhalation, dermal absorption is a relevant route for the incorporation of THMs, particularly those with lower degree of bromination. Keywords: Water disinfection, In-door swimming pools, Trihalomethanes, Bromoform, Exhaled air pollutants, Volatile organic compounds

A method for the estimation of the human intake of trihalomethanes (THMs), namely chloroform, bromodichloromethane, dibromochloromethane and bromoform, during showering and bathing is reported. The method is based on the determination of these compounds in exhaled breath that is collected by solid adsorption on Tenax using a device specifically designed for this purpose. Instrumental measurements were performed by automatic thermal desorption coupled to gas chromatography with electron capture detection. THMs in exhaled breath samples were determined during showering and swimming pool attendance. The levels of these compounds in indoor air and water were also determined as reference for interpretation of the exhaled breath results. The THM concentrations in exhaled breath of the volunteers measured before the exposure experiments showed a close correspondence with the THMs levels in indoor air where the sampler was located. Limits of detection in exhaled breath were dependent on THM analytes and experimental sites. They ranged between 170 and 710 ng m−3 in the swimming pool studies and between 97 and 460 ng m−3 in the showering studies. Application of this method to THMs determination during showering and swimming pool activities revealed statistically significant increases in THMs concentrations when comparing exhaled breath before and after exposure., GRACCIE Consolider Ingenio Network (CSD2007-00067) and PISCINA Project (SAF2005-07643-CO3-02) from the Spanish Ministry of Science and Innovation is acknowledged.

8 pages, 4 pages.-- PMID: 19797353 [PubMed].-- Printed version published Nov 2009., While KRAS activation is a fundamental initiating event in the aetiopathogenesis of pancreatic ductal adenocarcinoma (PDA), environmental factors influencing the occurrence and persistence of KRAS mutations remain largely unknown. The objective was to test the hypothesis that in PDA there are aetiopathogenic relationships among concentrations of some organochlorine compounds (OCs) and the mutational status of the KRAS oncogene, as well as among the latter and coffee intake. Incident cases of PDA were interviewed and had blood drawn at hospital admission (N = 103). OCs were measured by high-resolution gas chromatography with electron capture detection. Cases whose tumours harboured a KRAS mutation had higher concentrations of p,p′-dichlorodiphenyltrichloroethane (DDT), p,p′-dichlorodiphenyldichloroethene (DDE) and polychlorinated biphenyls (PCBs) 138, 153 and 180 than cases with wild-type KRAS, but differences were statistically significant only for p,p′-DDT and PCBs 138 and 153. The association between coffee intake and KRAS mutations remained significant (P-trend, Government of Catalonia (2009 SGR 1350); ‘Red temática de investigación cooperativa de centros en Cáncer’ (C03/10); ‘Red temática de investigación cooperativa de centros en Epidemiología y salud pública’ (C03/09); CIBER de Epidemiología y Salud Pública, Instituto de Salud Carlos III, Madrid, Government of Spain.

Knowledge is scant on the relationships between pathophysiologic processes common during cancer progression and changes in blood concentrations of organochlorine compounds (OCs). To analyze the influence of tumor stage, cancer symptoms, and time of blood extraction on serum concentrations of OCs in exocrine pancreatic cancer (EPC). Subjects were 144 incident cases of EPC prospectively recruited in eastern Spain. Blood was drawn and face-to-face interviews with patients were conducted during hospital admission. Information on signs and symptoms was obtained from medical records and patient interviews. OCs were analyzed by high-resolution gas chromatography with electron-capture detection. General linear models were applied to analyze log-transformed OCs corrected for total lipids. Lower concentrations of six of the seven OCs analyzed (p,p′-DDE, three polychlorinated biphenyls, hexachlorobenzene, and β-hexachlorocyclohexane) were observed in patients with cholestatic syndrome (jaundice, hypocholia, and choluria). The constitutional syndrome increased only p,p′-DDT. The lowering effect of the cholestatic syndrome was stronger than the increasing effect of the constitutional syndrome (fatigue, anorexia, and weight loss), except for p,p′-DDT. When symptoms were considered, stage had only weakly inverse relationships with OC levels. The effects of symptoms on p,p′-DDE, p,p′-DDT, and the three PCBs remained significant after adjusting by the interval from blood extraction to first symptom of EPC, and even when further adjusting by stage. Restriction or adjustment by stage and timing of blood draw may be insufficient to prevent biases associated with cancer progression. Symptoms may enable investigators to assess disease-induced changes in lipophilic exposure biomarkers.

Introduction: When studying the effects of organochlorine compounds (OCs) on human health it is common to correct serum concentrations of OC by total lipids (TL). However, the relationship between serum OCs and serum TL is far from established in many diseases, including several cancers. Our aim was to analyze the relationship between serum OC and TL in patients with pancreatic ductal adenocarcinoma (PDA), and to explore several alternatives to perform the OC lipid correction. Methods: Incident cases of PDA were interviewed and had blood drawn soon around hospital admission (n=144). Serum concentrations of OCs were analysed by high-resolution gas chromatography with electron-capture detection. Results: Most patients with high TL had moderate or low concentrations of OCs. By contrast, the variability of OC values among patients with normal TL was large. Correlations were of a similar magnitude between OC and TL and between OC and total cholesterol; while these correlations were weak (all Spearman's ρ

International audience; Given the importance of G-protein-coupled receptors as pharmacological targets in medicine, efforts directed at the understanding the molecular mechanism by which pharmacological compounds regulate their activity is of paramount importance. Here, we investigated at an atomic level the mechanism of inverse agonism and partial agonism of two high affinity, high selectivity very similar non-peptide ligands of the cholecystokinin-2 receptor (CCK2R) which differ by the absence or presence of a methyl group on their indole moiety. Using in silico, site-directed mutagenesis and pharmacological experiments, we demonstrated that these functionally different activities are due to differing anchoring modes of the two compounds to a residue of helix II (Thr-2.61) in the inactive state of the CCK2R. The binding mode of the inverse agonist allows the ligand to interact through its phenyl moiety with a key amino acid for CCK2R activation (Trp-6.48), preventing rotation of helix VI and, thus, CCK2R activation, whereas the partial agonist binds deeper into the binding pocket and closer to helix V, so that CCK2R activation is favored. This study on the molecular mechanism of ligand action opens the possibility of target-based optimization of G protein-coupled receptor non-peptide ligands.