How clean is your ice machine? Revealing microbial amplification and presence of opportunistic pathogens in hospital ice-water machines.

BACKGROUND: Ice machines in healthcare facilities have been suspected and even linked to outbreaks and pseudo-outbreaks. Guidelines exist for maintenance of these devices but there is no clear independent infection control standard, and little is known about their microbial contamination. AIM: To evaluate the microbial contamination, amplification, and presence of opportunistic pathogens in ice-water machines in a healthcare facility. METHODS: Concentrations of general microbial indicators (heterotrophic plate counts (HPC), total and intact cells), faecal indicators (enterococci) and opportunistic pathogens (Pseudomonas aeruginosa, non-tuberculous mycobacteria (NTM), Candida spp.) were measured in 36 ice-water machines on patient wards of a 772-bed hospital. Profile sampling was performed on five ice-water machines and adjacent faucets to identify sites of microbial proliferation. FINDINGS: Candida spp. were found in half of ice-water samples while enterococci and P. aeruginosa were present in six and 11 drain inlets respectively. NTM were measured in all ice-water samples and 35 out of 36 biofilms. Pre-filters and ice machines are sites for additional amplification: NTM densities were on average 1.3 log10 higher in water of ice machine flushed 5 min compared to flushed adjacent tap water. CONCLUSION: Ice machine design needs to be adapted to reduce microbial proliferation. The absence of correlation between HPC densities (current microbial indicators) and NTM concentrations suggests a need for cleaning efficiency indicators better correlated with opportunistic pathogens. Cleaning and disinfection guidelines of ice machines in healthcare facilities need to be improved, especially when ice is given to the most vulnerable patients, and NTM may be an efficiency indicator.

Arterial Stiffness in a Cohort of Young People Living With Perinatal HIV and HIV Negative Young People in England.

Background: Antiretroviral therapy (ART) has increased life expectancy and consequently the risk of cardiovascular disease (CVD) in adults living with HIV. We investigated the levels and predictors of arterial stiffness in young people (YP) living with perinatal HIV (PHIV) and HIV negative YP in the Adolescents and Adults Living with Perinatal HIV (AALPHI) study. Methods: AALPHI was a prospective study evaluating the impact of HIV infection and exposure to ART on YP living with PHIV (aged 13-21 years) who had known their HIV status for at least 6 months, and HIV negative YP (aged 13-23 years) who either had a sibling, friend or parent living with HIV. Participants were enrolled from HIV clinics and community services in England. Two hundred and thirteen PHIV and 65 HIV negative YP (42% siblings of PHIV) had pulse wave velocity (PWV) measurements taken (Vicorder software) from the supra-sternal notch to the middle of the thigh cuff, at their second interview in the study between 2015 and 2017. Average PWV was calculated from the three closest readings (≥3 and ≤ 12 m/s) within 0.6 m/s of each other. Linear regression examined predictors of higher (worse) PWV, including age, sex, HIV status and height as a priori, ethnicity, born outside UK/Ireland, alcohol/nicotine/drug use, weight, waist-to-hip-ratio, mean arterial pressure (MAP), caffeine 2 h before PWV and nicotine on day of PWV. A separate PHIV model included CD4, viral load, years taking ART and ART regimen. Findings: One hundred and twenty eight (60%) PHIV and 45 (69%) HIV negative YP were female (p = 0.18), with median (IQR) age 18 (16, 20) and 18 (16, 21) years (p = 0.48) respectively. Most PHIV were taking a combination of three ART drugs from two classes. There was a trend toward higher (worse) mean PWV in the PHIV group than the HIV negative group [unvariable analysis 6.15 (SD 0.83) m/s vs. 5.93 (0.70) m/s, respectively, unadjusted p = 0.058], which was statistically significant in the multivariable analysis [adjusted p (ap) = 0.020]. In multivariable analysis being male (ap = 0.002), older age (ap < 0.001), higher MAP (ap < 0.001) and nicotine use on day of measurement (ap = 0.001) were also predictors of higher PWV. The predictors were the same in the PHIV model. Interpretation: By late adolescence PHIV had worse PWV in comparison to HIV negative peers, and traditional risk factors for CVD (higher arterial pressure, being male and older age) were associated with higher PWV values. Regular detailed monitoring of cardiovascular risk factors should become standard of care for every young person with PHIV worldwide.

Molecular mechanism of thiamine pyrophosphate import into mitochondria: a molecular simulation study.

The import of thiamine pyrophosphate (TPP) through both mitochondrial membranes was studied using a total of 3-µs molecular dynamics simulations. Regarding the translocation through the mitochondrial outer membrane, our simulations support the conjecture that TPP uses the voltage-dependent anion channel, the major pore of this membrane, for its passage to the intermembrane space, as its transport presents significant analogies with that used by other metabolites previously studied, in particular with ATP. As far as passing through the mitochondrial inner membrane is concerned, our simulations show that the specific carrier of TPP has a single binding site that becomes accessible, through an alternating access mechanism. The preference of this transporter for TPP can be rationalized mainly by three residues located in the binding site that differ from those identified in the ATP/ADP carrier, the most studied member of the mitochondrial carrier family. The simulated transport mechanism of TPP highlights the essential role, at the energetic level, of the contributions coming from the formation and breakage of two networks of salt bridges, one on the side of the matrix and the other on the side of the intermembrane space, as well as the interactions, mainly of an ionic nature, formed by TPP upon its binding. The energy contribution provided by the cytosolic network establishes a lower barrier than that of the matrix network, which can be explained by the lower interaction energy of TPP on the matrix side or possibly a uniport activity.

Lipid composition and salt concentration as regulatory factors of the anion selectivity of VDAC studied by coarse-grained molecular dynamics simulations.

The voltage-dependent anion channel (VDAC) is a mitochondrial outer membrane protein whose fundamental function is to facilitate and regulate the flow of metabolites between the cytosol and the mitochondrial intermembrane space. Using coarse-grained molecular dynamics simulations, we investigated the dependence of VDAC selectivity towards small inorganic anions on two factors: the ionic strength and the lipid composition. In agreement with experimental data we found that VDAC becomes less anion selective with increasing salt concentration due to the screening of a few basic residues that point into the pore lumen. The molecular dynamics simulations provide insight into the regulation mechanism of VDAC selectivity by the composition in the lipid membrane and suggest that the ion distribution is differently modulated by POPE compared to the POPC bilayer. This occurs through the more persistent interactions of acidic residues located at both rims of the ß-barrel with head groups of POPE which in turn impact the electrostatic potential and thereby the selectivity of the pore. This mechanism occurs not only in POPE single component membranes but also in a mixed POPE/POPC bilayer by an enrichment of POPE over POPC lipids on the surface of VDAC. Thus we show here that computationally-inexpensive coarse-grained simulations are able to capture, in a semi-quantitative way, essential features of VDAC anion selectivity and could pave the way toward a molecular level understanding of metabolite transport in natural membranes.

Neural correlates of the healthiness evaluation processes of food labels.

OBJECTIVES: This fMRI study evaluated the cognitive mechanisms and the cerebral substrates when evaluating the healthiness of food products from nutritional information displayed either with a traffic light (TL) system, a colored nutritional label, or with a guideline daily amount (GDA) system, a numeric label. We postulated that TL label would recruit emotional processes and activation of subjacent cerebral regions (e.g. insula and amygdala). On the contrary, the nutritional information presented in a GDA label, would recruit, due to its numeric format and higher complexity, supplementary cognitive processes and activation of related brain regions (e.g. middle and superior frontal as well as parietal cortices). METHODS: We examined 50 healthy participants during an evaluation task on the healthiness of real food products from nutritional information only. Per total, 60 food products nutritional labels have been presented, with either colored (TL) or numeric (GDA) nutritional information and three levels of complexity of nutritional information. RESULTS: In line with our predictions, evaluations based on GDA recruited prefrontal and parietal regions reported for analytic processes. Contrary to our predictions, the same network has been recruited when evaluations were based on TL. Finally, we found significant correlation between response time and the superior parietal lobule in the GDA condition. DISCUSSION: Our results suggested that TL did not have an effect on the used strategy compared to GDA, based on calculation and arithmetic processes. Correlations between response time and brain activations suggested a significant involvement of the arithmetic mechanisms in the evaluation of food healthiness.

The distribution dynamics and desorption behaviour of mobile pharmaceuticals and caffeine to combined sewer sediments.

Pharmaceuticals are discharged to the environment from wastewater resource recovery facilities, sewer overflows, and illicit sewer connections. To understand the fate of pharmaceuticals, there is a need to better understand their sorption dynamics to suspended sediments (SS) and settled sediments (StS) in sewer systems. In this study, such sorption dynamics to both SS and StS were assessed using a batch equilibrium method under both static and dynamic conditions. Experiments were performed with natively occurring and artificially modified concentrations of sewer pharmaceuticals (acetaminophen, theophylline, carbamazepine, and a metabolite of carbamazepine) and caffeine. Differences in apparent distribution coefficients, Kd,app, between SS and StS were related to differences in their organic carbon (OC) content, and the practice of artificially modifying the concentration. Kd,app values of modified contaminant concentrations and high OC sediments were substantially higher. Pseudo-second order desorption rates for these mobile compounds were also quantified. Successive flushing events to simulate the addition of stormwater to sewer networks revealed that aqueous concentrations would not necessarily decrease, because the added water will rapidly return to equilibrium concentrations with the sediments. Sorption and desorption kinetics must be considered in addition to dilution, to avoid underestimating the influence of dilution on concentrations of pharmaceuticals discharged to the environment.

Analysis of steroid hormones and their conjugated forms in water and urine by on-line solid-phase extraction coupled to liquid chromatography tandem mass spectrometry.

BACKGROUND: In recent years, endocrine disrupting compounds (EDCs) have been found in rivers that receive significant inputs of wastewater. Among EDCs, natural and synthetic steroid hormones are recognized for their potential to mimic or interfere with normal hormonal functions (development, growth and reproduction), even at ultratrace levels (ng L(-1)). Although conjugated hormones are less active than free hormones, they can be cleaved and release the unconjugated estrogens through microbial processes before or during the treatment of wastewater. Due to the need to identify and quantify these compounds, a new fully automated method was developed for the simultaneous determination of the two forms of several steroid hormones (free and conjugated) in different water matrixes and in urine. RESULTS: The method is based on online solid phase extraction coupled with liquid chromatography and tandem mass spectrometry (SPE-LC-MS/MS). Several parameters were assessed in order to optimize the efficiency of the method, such as the type and flow rate of the mobile phase, the various SPE columns, chromatography as well as different sources and ionization modes for MS. The method demonstrated good linearity (R(2) > 0.993) and precision with a coefficient of variance of less than 10 %. The quantification limits vary from a minimum of 3-15 ng L(-1) for an injection volume of 1 and 5 mL, respectively, with the recovery values of the compounds varying from 72 to 117 %. CONCLUSION: The suggested method has been validated and successfully applied for the simultaneous analysis of several steroid hormones in different water matrixes and in urine.

Wastewater micropollutants as tracers of sewage contamination: analysis of combined sewer overflow and stream sediments.

A sensitive method was developed to measure the sediment concentration of 10 wastewater micropollutants selected as potential sanitary tracers of sewage contamination and include: nonsteroidal anti-inflammatory drugs (acetaminophen - ACE and diclofenac - DIC), an anti-epileptic drug (carbamazepine - CBZ), a ß-blocker (atenolol - ATL), a stimulant (caffeine - CAF), a bronchodilator (theophylline - THEO), steroid hormones (progesterone - PRO and medroxyprogesterone - MedP), an artificial sweetener (aspartame - APM) and personal care products (N,N-diethyl-3-methylbenzamide - DEET). Natural sediments (combined sewer overflow and stream sediments) were extracted by ultrasonic-assisted extraction followed by solid-phase extraction. Analyses were performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) using atmospheric pressure chemical ionisation in positive mode (APCI+) with a total analysis time of 4.5 min. Method detection limits were in the range of 0.01 to 15 ng g(-1) dry weight (dw) for the compounds of interest, with recoveries ranging from 75% to 156%. Matrix effects were observed for some compounds, never exceeding |±18%|. All results displayed a good degree of reproducibility and repeatability, with relative standard deviations (RSD) of less than 23% for all compounds. The method was applied to an investigation of stream and combined sewer overflow sediment samples that differed in organic carbon contents and particle size distributions. Acetaminophen, caffeine and theophylline (as confounded with paraxanthine) were ubiquitously detected at 0.13-22 ng g(-1) dw in stream bed sediment samples and 98-427 ng g(-1) dw in combined sewer overflow sediment samples. Atenolol (80.5 ng g(-1) dw) and carbamazepine (54 ng g(-1) dw) were quantified only in combined sewer overflow sediment samples. The highest concentrations were recorded for DEET (14 ng g(-1) dw) and progesterone (11.5 ng g(-1) dw) in stream bed and combined sewer overflow sediment samples, respectively. The ratio of concentration to its limit of detection (C : LOD) in sediments for a subset of compounds were compared to their C : LOD in water. In waters with a large capacity for dilution relative to fecal sources, the C : LOD ranges in sediments were greater than in water. Thus monitoring programs for fecal source tracking using wastewater micropollutants should consider sediment sampling, particularly for waters with highly diluted sources of fecal contamination.

Exposure of young children to household water lead in the Montreal area (Canada): the potential influence of winter-to-summer changes in water lead levels on children's blood lead concentration.

Drinking water represents a potential source of lead exposure. The purpose of the present study was to estimate the magnitude of winter-to-summer changes in household water lead levels (WLLs), and to predict the impact of these variations on BLLs in young children. A study was conducted from September, 2009 to March, 2010 in 305 homes, with a follow-up survey carried out from June to September 2011 in a subsample of 100 homes randomly selected. The first 1-L sample was drawn after 5 min of flushing, followed by a further 4 consecutive 1-L samples after 30 min of stagnation. Non-linear regression and general linear mixed models were used for modelling seasonal effects on WLL. The batchrun mode of Integrated Exposure Uptake Biokinetic (IEUBK) model was used to predict the impact of changes in WLL on children's blood lead levels (BLLs). The magnitude of winter-to-summer changes in average concentrations of lead corresponded to 6.55 µg/L in homes served by lead service lines (LSL+ homes) and merely 0.30 µg/L in homes without lead service lines. For stagnant samples, the value reached 10.55 µg/L in 'LSL+ homes' and remained very low (0.36 µg/L) in 'LSL- homes'. The change in the probability of BLLs ≥5 µg/dL due to winter-to-summer changes in WLL was increased from <5% (in winter) to about 20% (in summer) in children aged 0.5-2 years. The likelihood of having BLLs ≥5 µg/dL in young children during warm months was reduced by at least 40% by flushing tap-water.

Repeat antenatal HIV testing in the third trimester: a study of feasibility and maternal uptake rates.

OBJECTIVES: The study aimed to assess the feasibility and acceptability of third-trimester antenatal HIV testing within our service after two cases of HIV seroconversion in pregnancy were noted in 2008. North American Guidelines recommend universal third-trimester HIV testing in areas with an HIV prevalence of more than 1 per 1000. The HIV prevalence rate in our area is 3.01 per 1000. METHODS: Pregnant women prior to 28 weeks of gestation were recruited at booking between 1 September 2008 and 31 August 2009 and offered an additional third-trimester HIV test. Consent was obtained and testing was performed by hospital and community midwives. Information was entered into a modified existing electronic maternity database. A qualitative e-mail survey of midwives investigated barriers to participation in the study. RESULTS: A total of 4134 women delivered; three (< 0.1%) declined first-trimester testing. Twenty-two women (0.5%) tested HIV positive, of whom six were newly diagnosed. Overall, 2934 of 4134 women (71%) were offered and accepted a third-trimester HIV test and had results available. Data were unavailable for 195 women (4.7%). A total of 663 of 4131 women (16%) were not offered a third-trimester test. Of 3273 women documented as having been offered a test, 3177 (97.1%) accepted. There were no positive third-trimester tests. Forty of 50 (80%) midwives surveyed responded with questionnaire feedback and cited lack of national policy and extra workload as barriers to performing third-trimester testing. CONCLUSIONS: Third-trimester testing was feasible and consent rates were high in those offered repeat testing. Third-trimester testing has the potential to prevent paediatric HIV infection and universal testing should be considered in high-prevalence areas.

Using a novel sol-gel stir bar sorptive extraction method for the analysis of steroid hormones in water by laser diode thermal desorption/atmospheric chemical ionization tandem mass spectrometry.

A new coating material was used for a stir bar sorptive extraction (SBSE) method coupled to a high throughput sample analysis technique. This allowed for a simple procedure for fast determinations of eight steroid hormones (estriol, estradiol, ethynylestradiol, estrone, progesterone, medroxyprogesterone, levonorgestrel, northindrone) in water. Sample pre-treatment was performed using an in-house SBSE method based on a polydimethylsiloxane/phenyltrimethylsiloxane/ß-cyclodextrin sol-gel material. The analytes were desorbed by liquid extraction prior to their analysis by laser diode thermal desorption/atmospheric pressure chemical ionization coupled to tandem mass spectrometry (LDTD-APCI-MS/MS). Several parameters, including ionic strength, volume and time of extraction as well as volume and time of desorption, were investigated to maximize extraction efficiency by SBSE in aqueous solutions. The in-house stir bar showed good reproducibility and could be used for at least 50 extractions without affecting analytical performance. The recoveries of the spiked steroid hormones ranged from 55% to 96% in all water matrices studied (HPLC grade water, tap water and raw wastewater). Only one compound showed poor recovery values (<2% for estriol) in all matrices. The method detection limits (MDLs) in real matrices were within the range of 0.1-0.3 µg L(-1) except for estriol at 48 µg L(-1). The extraction performance of the in-house SBSE for the eight selected hormones was also compared with that of a commercially-available stir bar coated with polydimethylsiloxane (PDMS). This novel stir bar coating could prove to be useful method for the detection and quantification of trace levels of steroid hormones.

Identification of key residues for the binding of glucagon to the N-terminal domain of its receptor: an alanine scan and modeling study.

Glucagon plays an essential role in the glycemia maintenance during fasting, but also aggravates hyperglycemia in diabetic patients. A series of analogues of glucagon were synthesized replacing each amino acid of the C-terminal region (residues 15-29) with alanine. The residues affecting the binding to the glucagon receptor are found to be located on one face of the glucagon helix. Several 3-dimensional models of the N-terminal domain of the glucagon receptor in complex with its ligand peptide were built and used to analyze the peptide-receptor interface in terms of the nature of the peptide residues and the interactions they form with the receptor. The models suggest that glucagon keeps its native helical structure upon binding, and that a large part of the interface formed with the receptor is hydrophobic. We find that in the C-terminal region, F22, V23, M27, and D15 are the most important residues for peptide binding. They bury a large portion of their solvent accessible surface area and make numerous interactions with the receptor mainly of the hydrophobic type.

Sensitivity analysis of some critical factors affecting simulated intrusion volumes during a low pressure transient event in a full-scale water distribution system.

Intrusion events caused by transient low pressures may result in the contamination of a water distribution system (DS). This work aims at estimating the range of potential intrusion volumes that could result from a real downsurge event caused by a momentary pump shutdown. A model calibrated with transient low pressure recordings was used to simulate total intrusion volumes through leakage orifices and submerged air vacuum valves (AVVs). Four critical factors influencing intrusion volumes were varied: the external head of (untreated) water on leakage orifices, the external head of (untreated) water on submerged air vacuum valves, the leakage rate, and the diameter of AVVs' outlet orifice (represented by a multiplicative factor). Leakage orifices' head and AVVs' orifice head levels were assessed through fieldwork. Two sets of runs were generated as part of two statistically designed experiments. A first set of 81 runs was based on a complete factorial design in which each factor was varied over 3 levels. A second set of 40 runs was based on a latin hypercube design, better suited for experimental runs on a computer model. The simulations were conducted using commercially available transient analysis software. Responses, measured by total intrusion volumes, ranged from 10 to 366 L. A second degree polynomial was used to analyze the total intrusion volumes. Sensitivity analyses of both designs revealed that the relationship between the total intrusion volume and the four contributing factors is not monotonic, with the AVVs' orifice head being the most influential factor. When intrusion through both pathways occurs concurrently, interactions between the intrusion flows through leakage orifices and submerged AVVs influence intrusion volumes. When only intrusion through leakage orifices is considered, the total intrusion volume is more largely influenced by the leakage rate than by the leakage orifices' head. The latter mainly impacts the extent of the area affected by intrusion.

Monitoring of potentially toxic cyanobacteria using an online multi-probe in drinking water sources.

Toxic cyanobacteria threaten the water quality of drinking water sources across the globe. Two such water bodies in Canada (a reservoir on the Yamaska River and a bay of Lake Champlain in Québec) were monitored using a YSI 6600 V2-4 (YSI, Yellow Springs, Ohio, USA) submersible multi-probe measuring in vivo phycocyanin (PC) and chlorophyll-a (Chl-a) fluorescence, pH, dissolved oxygen, conductivity, temperature, and turbidity in parallel. The linearity of the in vivo fluorescence PC and Chl-a probe measurements were validated in the laboratory with Microcystis aeruginosa (r(2) = 0.96 and r(2) = 0.82 respectively). Under environmental conditions, in vivo PC fluorescence was strongly correlated with extracted PC (r = 0.79) while in vivo Chl-a fluorescence had a weaker relationship with extracted Chl-a (r = 0.23). Multiple regression analysis revealed significant correlations between extracted Chl-a, extracted PC and cyanobacterial biovolume and in vivo fluorescence parameters measured by the sensors (i.e. turbidity and pH). This information will help water authorities select the in vivo parameters that are the most useful indicators for monitoring cyanobacteria. Despite highly toxic cyanobacterial bloom development 10 m from the drinking water treatment plant's (DWTP) intake on several sampling dates, low in vivo PC fluorescence, cyanobacterial biovolume, and microcystin concentrations were detected in the plant's untreated water. The reservoir's hydrodynamics appear to have prevented the transport of toxins and cells into the DWTP which would have deteriorated the water quality. The multi-probe readings and toxin analyses provided critical evidence that the DWTP's untreated water was unaffected by the toxic cyanobacterial blooms present in its source water.

Structure-activity relationship analyses of glycyrrhetinic acid derivatives as anticancer agents.

Cancer cell resistance to kinase inhibitors and targeted agents, acquisition of a multidrug-resistant (MDR) phenotype and/or intrinsic resistance to apoptosis prevent effective treatment in about 50% of solid cancers in adults, and the percentage is even higher in children. Glycyrrhetinic acid (GA) and some of its derivatives may offer hope in combating cancer types associated with poor prognoses. Some GA derivatives are indeed able to target both the proteasome and peroxisome proliferator-activated receptors (PPARs), two proteins that play major roles in cancer cell biology but are not related to MDR and/or apoptosis-related resistance phenotypes.

Use of in vivo phycocyanin fluorescence to monitor potential microcystin-producing cyanobacterial biovolume in a drinking water source.

The source water of a drinking water treatment plant prone to blooms, dominated by potential microcystin-producing cyanobacteria, was monitored for two seasons in 2007-2008. In the 2008 season, the median value for potential microcystin-producing cyanobacterial biovolume was 87% of the total phytoplankton biovolume in the untreated water of the plant. Depth profiles taken above the plant's intake identified three sampling days at high risk for the contamination of the plant's raw water with potentially toxic cyanobacteria. Chlorophyceae and Bacillariophyceae caused false positive values to be generated by the phycocyanin probe when cyanobacteria represented a small fraction of the total phytoplanktonic biovolume present. However, there was little interference with the phycocyanin probe readings by other algal species when potential microcystin-producing cyanobacteria dominated the phytoplankton of the plant's untreated water. A two-tiered method for source water monitoring, using in vivo phycocyanin fluorescence, is proposed based on (1) a significant relationship between in vivo phycocyanin fluorescence and cyanobacterial biovolume and (2) the calculated maximum potential microcystin concentration produced by dominant Microcystis sp. biovolume. This method monitors locally-generated threshold values for cyanobacterial biovolume and microcystin concentrations using in vivo phycocyanin fluorescence.

[Conceptualization of the preventive theories]. / Concepts théoriques en prévention: heurs et malheurs.

In front of the explosion of the costs of the curative care, the conceptualization of the preventive theories and methodologies, and the shown efficiency of the actions of prevention, this one acquires with evolving time a growing importance and a development desired with the eyes of the doctors, but also of the patients, policy decision makers and insurers. This article intends to approach and clarify the positive contributions but also the multiple limits of the preventive steps.

Ozone oxidation of pharmaceuticals, endocrine disruptors and pesticides during drinking water treatment.

This study investigates the oxidation of pharmaceuticals, endocrine disrupting compounds and pesticides during ozonation applied in drinking water treatment. In the first step, second-order rate constants for the reactions of selected compounds with molecular ozone (k(O3)) were determined in bench-scale experiments at pH 8.10: caffeine (650+/-22M(-1)s(-1)), progesterone (601+/-9M(-1)s(-1)), medroxyprogesterone (558+/-9M(-1)s(-1)), norethindrone (2215+/-76M(-1)s(-1)) and levonorgestrel (1427+/-62M(-1)s(-1)). Compared to phenolic estrogens (estrone, 17beta-estradiol, estriol and 17alpha-ethinylestradiol), the selected progestogen endocrine disruptors reacted far slower with ozone. In the second part of the study, bench-scale experiments were conducted with surface waters spiked with 16 target compounds to assess their oxidative removal using ozone and determine if bench-scale results would accurately predict full-scale removal data. Overall, the data provided evidence that ozone is effective for removing trace organic contaminants from water with ozone doses typically applied in drinking water treatment. Ozonation removed over 80% of caffeine, pharmaceuticals and endocrine disruptors within the CT value of about 2 mg min L(-1). As expected, pesticides were found to be the most recalcitrant compounds to oxidize. Caffeine can be used as an indicator compound to gauge the efficacy of ozone treatment.

DD-ligases as a potential target for antibiotics: past, present and future.

DD-ligases catalyze the synthesis of the D-Ala-D-Ala and D-Ala-D-Ser dipeptides or the D Ala-D-Lac depsipeptide in an early step of peptidoglycan synthesis. Their function is essential for bacterial growth and specific to bacteria, making them attractive targets for the development of novel antibiotics. This review examines the biochemical and structural features of these enzymes and presents the main families of inhibitors described so far. Over the last 20 years, 7 structures of DD-ligases have been solved by X-ray crystallography, giving a detailed view of the general topology of the active site and of the residues in the catalytic pocket that play a central role in substrate recognition. This has paved the way to the rational design of inhibitors, which can be classified as (i) analogues of substrates, (ii) analogues of the product of the reaction, (iii) analogues of the transition state, and (iv) original scaffolds discovered by screening or by rational computer-aided design. The three first strategies have led to molecules that are polar by nature and have therefore poor access to their cytosolic target. The fourth one is potentially most promising as it yields more diverse structures. The most active molecules show affinity constants in the microM range, but microbiological evaluation remains scarce (typical MIC 1-8 mg/L for the tested compounds). These data strongly suggest targeting DD-ligases is a promising approach for discovery of new antibiotics. Future research should, however, aim at finding more potent inhibitors endowed with the appropriate pharmacokinetic properties that ensure access to their intracellular target.

Analysis of natural and synthetic estrogenic endocrine disruptors in environmental waters using online preconcentration coupled with LC-APPI-MS/MS.

This paper describes a fully automated online method for solid-phase extraction coupled with liquid chromatography and tandem mass spectrometry using atmospheric pressure ionization (LC-LC-APPI-MS/MS) to simultaneously detect selected dissolved natural and synthetic hormones at concentrations as low as 5 ng/L from aqueous matrices. The method shows excellent performance for the direct analysis of water and wastewater with respect to calibration curve linearity, analytic specificity, sensitivity, and carryover, as well as overall method accuracy and precision. With a runtime of 15 min, a minimum of 200 samples were analyzed using a single online solid-phase extraction (SPE) column without noticeable differences in system performance. Because of the ruggedness and simplicity of this system, generic methods can be easily developed and applied for the high-throughput analysis of a wide variety of compounds without the need to resort to laborious offline SPE sample preparation.