Impact of saponification and silver-nitrate purification on lacustrine alkenone distributions and alkenone-based indices.

A growing interest in lacustrine alkenones as a proxy for continental paleotemperature reconstructions accompanied important methodological improvements over the past two decades. New gas chromatography (GC) columns were used for alkenone analysis, that drastically improved alkenone separation, especially for freshwater lakes. However, these recent advances are sometimes not sufficient in separating compounds that interfere with alkenones in the resulting chromatograms and concurrently, new chemical procedures were implemented to further clean up the samples. Here we investigate the impact of two clean-up procedures, saponification and silver-nitrate purification, on alkenone distribution, alkenone quantification, and C37 alkenone-based indices, including the U37K index. The silver-nitrate purification modified the C37 alkenone distribution and thus the C37 alkenone-based indices, especially the U37K index, in 6 out of the 9 studied samples by further retaining alkenones with more double bonds. These changes would result on an average error of 3 °C in reconstructed temperatures. Saponification also influenced the C37 alkenone distribution mainly by removing co-eluting compounds, thereby improving the quality of the results. Both saponification and purification resulted in the reduction of the C37 alkenone concentration by almost half. Clean-up steps should thus be used carefully, paying particular attention to any change in alkenone distribution and concentration. Limiting the use of additional clean-up steps reduces the risk of modifying the alkenone distribution.

North Atlantic cooling triggered a zonal mode over the Indian Ocean during Heinrich Stadial 1.

Abrupt changes in the Atlantic meridional overturning circulation (AMOC) are thought to affect tropical hydroclimate through adjustment of the latitudinal position of the intertropical convergence zone (ITCZ). Heinrich Stadial 1 (HS1) involves the largest AMOC reduction in recent geological time; however, over the tropical Indian Ocean (IO), proxy records suggest zonal anomalies featuring intense, widespread drought in tropical East Africa versus generally wet but heterogeneous conditions in the Maritime Continent. Here, we synthesize proxy data and an isotope-enabled transient deglacial simulation and show that the southward ITCZ shift over the eastern IO during HS1 strengthens IO Walker circulation, triggering an east-west precipitation dipole across the basin. This dipole reverses the zonal precipitation anomalies caused by the exposed Sunda and Sahul shelves due to glacial lower sea level. Our study illustrates how zonal modes of atmosphere-ocean circulation can amplify or reverse global climate anomalies, highlighting their importance for future climate change.

Discovery and Optimization of Potent and Orally Available CTP Synthetase Inhibitors for Use in Treatment of Diseases Driven by Aberrant Immune Cell Proliferation.

Herein, we report the discovery of a first-in-class chemotype 2-(alkylsulfonamido)thiazol-4-yl)acetamides that act as pan-selective inhibitors of cytidine 5'-triphosphate synthetase (CTPS1/2), critical enzymes in the de novo pyrimidine synthesis pathway. Weak inhibitors identified from a high-throughput screening of 240K compounds have been optimized to a potent, orally active agent, compound 27, which has shown significant pharmacological responses at 10 mg/kg dose BID in a well-established animal model of inflammation.

Tracking the legacy of early industrial activity in sediments of Lake Zurich, Switzerland: using a novel multi-proxy approach to find the source of extensive metal contamination.

Historical industrial activities at the Horn Richterwil, on the shore of Lake Zurich (Switzerland), caused widespread metal contamination on land and in the adjacent lake sediments. This study provides an estimation of the age and source of the contamination by using XRF core scanning, ICP-OES, and Hg-AFS for quantitative measurements of trace metals and MC-ICP-MS for the stable isotope analysis of mercury. Radiometric dating ([Formula: see text]Cs, [Formula: see text]Pb, and Pu dating) of two proximal cores and varve chronology in a distal core suggest two different contaminations, one stemming from around 1960 (Zn, Cd) and an earlier one from 1880 (Cr, Cu, Hg, Pb, Sn). The XRF data suggest two different contamination pathways: one by landfill of contaminated soil and another one by industrial wastewater effluents. Maximum concentrations found within all samples are in the range of per mil (dry weight) for Cr, Cu, Hg, Pb, Sn, and Zn and lie within the top 10 cm of the sediment cores. The analysis of the mercury isotopic composition ([Formula: see text]Hg and [Formula: see text]Hg) shows a significantly different signature for one of the cores, indicating a second mercury source. We could not identify the exact source or process leading to the isotopic fractionation of mercury, but the isotopic data confirm two different sources.

Potential 2-Aminoindane Fatality Invalidated by Careful Mass Spectrometric Analysis.

We describe herewith the case of a patient presenting to the emergency department for worsening ear-nose-throat symptoms. As chemsex was evocated by the family, patient's serum was submitted to a new psychoactive substances screening. After a simple liquid-liquid extraction, serum was injected on a high-resolution mass spectrometer using quite usual conditions (C18 column, gradient mode with acidic buffer, methanol and acetonitrile). An almost perfect match with 2-aminoindane (2-AI) was observed considering that the precursor ion was present in the sample but absent in the commercial library. Literature concerning 2-AI is sparse, and further investigations were undertaken. After injection of the reference standard, a small retention time shift has been observed (0.3 min) between the standard and the sample. The case was only closed while spiking the sample with the standard, giving rise to two distinct peaks. As a result, 2-AI was then considered as absent from the sample and death was attributed only to infection. Moreover, a rapid liquid chromatography-tandem mass spectrometry method dedicated to 2-AI was developed. It generated the same false-positive result highlighted by significant differences observed in ion ratios (2.37 for the sample versus 6.62 for the neat standard).

Deposit-feeding worms control subsurface ecosystem functioning in intertidal sediment with strong physical forcing.

Intertidal sands are global hotspots of terrestrial and marine carbon cycling with strong hydrodynamic forcing by waves and tides and high macrofaunal activity. Yet, the relative importance of hydrodynamics and macrofauna in controlling these ecosystems remains unclear. Here, we compare geochemical gradients and bacterial, archaeal, and eukaryotic gene sequences in intertidal sands dominated by subsurface deposit-feeding worms (Abarenicola pacifica) to adjacent worm-free areas. We show that hydrodynamic forcing controls organismal assemblages in surface sediments, while in deeper layers selective feeding by worms on fine, algae-rich particles strongly decreases the abundance and richness of all three domains. In these deeper layers, bacterial and eukaryotic network connectivity decreases, while percentages of clades involved in degradation of refractory organic matter, oxidative nitrogen, and sulfur cycling increase. Our findings reveal macrofaunal activity as the key driver of biological community structure and functioning, that in turn influence carbon cycling in intertidal sands below the mainly physically controlled surface layer.

Eutrophication as a driver of microbial community structure in lake sediments.

Lake sediments are globally important carbon sinks. Although the fate of organic carbon in lake sediments depends significantly on microorganisms, only few studies have investigated controls on lake sedimentary microbial communities. Here we investigate the impact of anthropogenic eutrophication, which affects redox chemistry and organic matter (OM) sources in sediments, on microbial communities across five lakes in central Switzerland. Lipid biomarkers and distributions of microbial respiration reactions indicate strong increases in aquatic OM contributions and microbial activity with increasing trophic state. Across all lakes, 16S rRNA genes analyses indicate similar depth-dependent zonations at the phylum- and class-level that follow vertical distributions of OM sources and respiration reactions. Yet, there are notable differences, such as higher abundances of nitrifying Bacteria and Archaea in an oligotrophic lake. Furthermore, analyses at the order-level and below suggest that changes in OM sources due to eutrophication cause permanent changes in bacterial community structure. By contrast, archaeal communities are differentiated according to trophic state in recently deposited layers, but converge in older sediments deposited under different trophic regimes. Our study indicates an important role for trophic state in driving lacustrine sediment microbial communities and reveals fundamental differences in the temporal responses of sediment Bacteria and Archaea to eutrophication.

Analytical validation of a quantitative method for therapeutic drug monitoring on the Alinity®c Abbott.

OBJECTIVE: The aim of this study was to evaluate the analytical performance of the Alinity®c Abbott compared to the Architect® immunoassay system for the determination of drugs having a narrow therapeutic index. METHODS: Valproic acid, amikacin, gentamicin, phenobarbital and vancomycin were analyzed using Particle-Enhanced Turbidimetric Inhibitor Immunoassay (Petinia), phenytoin and theophylline were analyzed using an immunoenzymatic method and a colorimetric method was performed to quantify lithium. The methods were validated according to the total error approach. Seven validation standards were analyzed in quintuplet during four days to establish the limits of the methods. Dilution integrity and interferences (hemolysis and high concentrations of bilirubin and lipids) were also tested. Depending on the analyte, the results obtained for twenty to forty patients on the Alinity® were compared to those obtained on the Architect®. RESULTS: The bias and the coefficients of variation for repeatability and for intermediate precision were lower than 15% for all drugs. Accuracy profiles were acceptable (acceptance limits fixed at 30%) in the validated ranges. The lower limits of quantification (LLOQ) were similar to those determined by Abbott except for gentamicin for which we determined a LLOQ at 1.22 mg/L while Abbott determined it at 0.5 mg/L. All assays diluted linear and analyte concentrations were not affected by interferences. Concentrations obtained for real samples on the Alinity®c are comparable to those obtained on the Architect®ci. CONCLUSIONS: The analytical validation of a method suitable for therapeutic drug monitoring of drugs on the Alinity®c meets the requirements of European Medicines Agency.

Validation of an UHPLC/DAD method for the determination of cannabinoids in seized materials: Analysis of 213 samples sold in Belgian CBD shops.

PURPOSE: Nowadays, (-)-cannabidiol (CBD) is gaining popularity for the treatment of various problems and can be found easily in many stores in Belgium. However, such product must comply with the law: if the total tetrahydrocannabinol (THC) content [(-)-Δ9-tetrahydrocannabinol + (-)-Δ9-tetrahydrocannabinolic acid A (THC-A)] is higher than 0.2%, it is considered as narcotic by the Belgian legislation. In this context, we have developed a method to quantify major cannabinoids (THC, THC-A, CBD, cannabidiolic acid, cannabigerolic acid, cannabigerol and cannabinol) in plant material. METHODS: After drying, a liquid-liquid extraction was performed on plant materials, followed by dilutions. Extracts were analyzed by ultra-high-performance liquid chromatography combined with a photodiode array detector. Mobile phases consisted of methanol and 0.1% formic acid in water applied in a 16-minute gradient mode. After validating the method, it was applied to 213 samples seized by the police in CBD shops. RESULTS: The method fulfilled the criteria in terms of specificity, calibration curve, precision, trueness and dosing range. Total THC content ranged from 0.14 to 1.17% (median 0.38%) with 110 samples exceeding the Belgian legal threshold of 0.2%. The amounts measured in the samples varied greatly, some were 6 times below the amount labelled on the packaging, others showed a concentration 4 times higher than stated on the package. Same strain also showed concentration differences from shop to shop. CONCLUSION: Our method was successfully validated and applied to samples seized in CBD shops. Half of the products exceeded the Belgian legal threshold. THC and CBD concentrations discrepancies showed that products sold in CBD shops are not pharmaceutical grade.

Earthquake Impact on Active Margins: Tracing Surficial Remobilization and Seismic Strengthening in a Slope Sedimentary Sequence.

Strong earthquakes at active ocean margins can remobilize vast amounts of surficial slope sediments and dynamically strengthen the margin sequences. Current process understanding is obtained from resulting event deposits and low-resolution shear strength data, respectively. Here we directly target a site offshore Japan where both processes are expected to initiate, that is, at the uppermost part (15 cm) of a sedimentary slope sequence. Based on a novel application of short-lived radionuclide data, we identified, dated, and quantified centimeter-scale gaps related to surficial remobilization. Temporal correlation to the three largest regional earthquakes attest triggering by strong earthquakes (M w >8). Also, extremely elevated shear strength values suggest a strong influence of seismic strengthening on shallow sediments. We show that despite enhanced slope stability by seismic strengthening, earthquake-induced sediment transport can occur through surficial remobilization, which has large implications for the assessment of turbidite paleoseismology and carbon cycling at active margins.

Death following consumption of MDAI and 5-EAPB.

5-(2-ethylaminopropyl)benzofuran (5-EAPB) and 5,6-methylenedioxy-2-aminoindane (MDAI) are two new psychoactive substances (NPS) exhibiting MDMA-like properties. In this paper, we report the case of a 28-years old man, known as drug addict, found dead at home, with two unidentified powders next to him. External examination by the forensic pathologist was unremarkable but no autopsy was performed. Powders, blood and urine (which were the only samples available) were submitted to general unknown screening by high pressure liquid chromatography with a diode array detector (HPLC-DAD) and ultra high pressure liquid chromatography with a time-of-flight detector (UPLC-TOF-MS), after liquid-liquid extraction for biological samples, or simple dilution for powders. Analysis revealed 68% of MDAI in one powder and 87% of 5-EAPB in the other one. Significant levels of the same substances were found in blood (MDAI: 2.09 mg/L and 5-EAPB: 6.45 mg/L). The cause of death was therefore attributed to the consumption of these NPS since screening for other drugs of abuse and for alcohol was negative (oxazepam was found in urine only). 5-methylaminopropylbenzofuran (5-MAPB) and 5-aminopropylbenzofuran (5-APB) were also found in blood (0.089 and 0.546 mg/L, respectively) and urine (1.00 and 4.88 mg/L, respectively). In addition to the inherent complexity of NPS identification by itself, another analytical difficulty in this case was the identification of the EAPB positional isomer. Our routine screening methods were not able to distinguish the positional isomer, but an additional classical gas chromatography technique was able to make the distinction. Anyway, in our case, this issue was simplified thanks to the availability of a relatively pure powder that was analyzed by nuclear magnetic resonance (NMR).

Improving the Accuracy of Flow Cytometric Quantification of Microbial Populations in Sediments: Importance of Cell Staining Procedures.

The accuracy of flow cytometric (FCM) quantifications of microbial populations in sediments varies with FCM settings, cell extraction and staining protocols, as well as sample types. In the present study, we improve the accuracy of FCM for enumerating microorganisms inhabiting diverse lake and marine sediment types based on extensive tests with FCM settings, extraction buffer chemical compositions, cell separation methods, and staining procedures. Tests on the FCM settings, (e.g., acquisition time, rates of events) and salinity of extraction solutions show minor impacts on FCM enumerations and yields of cell extraction, respectively. Existing methods involving hydrofluoric acid (HF) treatment to release sediment-attached cells into solution prove effective on both marine and freshwater samples. Yet, different staining techniques (direct staining of cell extracts, staining of membrane-filtered cell extracts) produce clear differences in cell number estimates. We demonstrate that, while labor-intensive membrane-staining generates high cell staining efficiency and accurate cell counts that are consistent across FCM and epifluorescence microscopy-based (EFM) quantification methods, accurate cell counts determined by more time- and labor-efficient direct staining require consideration of dye concentration, sample dilution, and lithology. Yet, good agreement between the two staining methods can be achieved through sample-specific adjustments of dye concentrations and sample dilutions during direct staining. We thus present a complete protocol for FCM-based cell quantification, that includes all steps from the initial sample fixation to the final enumeration, with recommendations for buffer compositions, direct and membrane-based staining procedures, and the final FCM assay. This protocol is versatile, accurate, and reliable, as is evident from good agreement with cell quantifications by EFM and quantitative polymerase chain reaction (qPCR) of 16S rRNA genes across a wide range of sedimentary sample types.

Arrival order and release from competition does not explain why haplochromine cichlids radiated in Lake Victoria.

The frequent occurrence of adaptive radiations on oceanic islands and in lakes is often attributed to ecological opportunity resulting from release from competition where arrival order among lineages predicts which lineage radiates. This priority effect occurs when the lineage that arrives first expands its niche breadth and diversifies into a set of ecological specialists with associated monopolization of the resources. Later-arriving species do not experience ecological opportunity and do not radiate. While theoretical support and evidence from microbial experiments for priority effects are strong, empirical evidence in nature is difficult to obtain. Lake Victoria (LV) is home to an exceptional adaptive radiation of haplochromine cichlid fishes, where 20 trophic guilds and several hundred species emerged in just 15 000 years, the age of the modern lake that was preceded by a complete desiccation lasting several thousand years. However, while about 50 other lineages of teleost fish also have established populations in the lake, none of them has produced more than two species and most of them did not speciate at all. Here, we test if the ancestors of the haplochromine radiation indeed arrived prior to the most competent potential competitors, 'tilapias' and cyprinids, both of which have made rapid radiations in other African lakes. We assess LV sediment core intervals from just before the desiccation and just after refilling for the presence of fossil fish teeth. We show that all three lineages were present when modern LV began to fill with water. We conclude that the haplochromines' extraordinary radiation unfolded in the presence of potentially competing lineages and cannot be attributed to a simple priority effect.

Homogenization of lake cyanobacterial communities over a century of climate change and eutrophication.

Human impacts on biodiversity are well recognized, but uncertainties remain regarding patterns of diversity change at different spatial and temporal scales. Changes in microbial assemblages are, in particular, not well understood, partly due to the lack of community composition data over relevant scales of space and time. Here, we investigate biodiversity patterns in cyanobacterial assemblages over one century of eutrophication and climate change by sequencing DNA preserved in the sediments of ten European peri-Alpine lakes. We found species losses and gains at the lake scale, while species richness increased at the regional scale over approximately the past 100 years. Our data show a clear signal for beta diversity loss, with the composition and phylogenetic structure of assemblages becoming more similar across sites in the most recent decades, as have the general environmental conditions in and around the lakes. We attribute patterns of change in community composition to raised temperatures affecting the strength of the thermal stratification and, as a consequence, nutrient fluctuations, which favoured cyanobacterial taxa able to regulate buoyancy. Our results reinforce previous reports of human-induced homogenization of natural communities and reveal how potentially toxic and bloom-forming cyanobacteria have widened their geographic distribution in the European temperate region.

Site-, Technique-, and Time-Related Aspects of the Postmortem Redistribution of Diazepam, Methadone, Morphine, and their Metabolites: Interest of Popliteal Vein Blood Sampling.

Sampling site, technique, and time influence postmortem drug concentrations. In 57 cases, we studied drug concentration differences as follows: subclavian vein-dissection/clamping versus blind stick, femoral vein-dissection/clamping versus blind stick, right cardiac chamber, and popliteal vein-dissection and clamping only. Cases were distributed in group #1 (all cases with both techniques), group #2 (dissection/clamping), and group #3 (blind stick). Sampled drugs were diazepam, methadone, morphine, and their metabolites. To assess PMR, mean concentrations and ratios were calculated for each group. Time-dependent variations of blood concentrations and ratios were also assessed. Results indicate that site, method, and time may influence postmortem distribution interpretation in different ways. Popliteal blood seems less subject to PMR. In conclusion, our study is the first to evaluate concurrently three main aspects of PMR and confirms that the popliteal vein may represent a site that is more resistant to the changes seen as a result of PMR.

Loss of treatment benefit when heroin-assisted treatment is stopped after 12 months.

PURPOSE: In 2013, during a recent heroin-assisted treatment trial, participants in heroin-assisted treatment (HAT) decreased significantly more their street heroin use than participants in oral methadone treatment. After the trial, HAT was discontinued. To examine whether the treatment benefits were sustained three months after the trial, the use of street heroin by the participants was analyzed in a follow-up study. RESULTS: At the follow-up assessment, street heroin use increased in the experimental group. The two groups no longer showed a significant difference (p=0.55) in the level of street heroin use. CONCLUSION: A predetermined and forced end of HAT was followed by a significant increase in the level of street level use.

Efficacy of Heroin-assisted Treatment In Belgium: A Randomised Controlled Trial.

BACKGROUND/AIMS: Heroin-assisted treatment (HAT) can improve the condition of heroin addicts still using street heroin after a methadone treatment. In Belgium, a new trial compared the efficacy of a HAT to existing methadone maintenance treatment. METHODS: In this randomised controlled trial, HAT was limited to 12 months. Participants were assessed every 3 months. They were responders if they showed improvement on the level of street heroin use, health or criminal involvement. RESULTS: 74 participants were randomised in the trial. The experimental group (n = 36) counted 30% of responders more than the control group (n = 38) at each assessment point (p < 0.05), except at 12 months where the difference (11%) was no longer significant (p = 0.35). Still, after 12 months, participants in the experimental group reported significantly greater improvements (p < 0.05) than the control group on the level of street heroin use and on the level of physical and mental health. Both groups reported significantly less criminal acts after 12 months (p < 0.001), but with no significant difference between the groups. CONCLUSIONS: This trial confirms the short-term efficacy of HAT for severe heroin addicts, who already failed methadone treatment.

Estimation of the Time Interval between the Administration of Heroin and the Sampling of Blood in Chronic Inhalers.

To develop a model for estimating the time delay between last heroin consumption and blood sampling in chronic drug users. Eleven patients, all heroin inhalers undergoing detoxification, were included in the study. Several plasma samples were collected during the detoxification procedure and analyzed for the heroin metabolites 6-acetylmorphine (6AM), morphine (MOR), morphine-6-glucuronide (M6G) and morphine-3-glucuronide (M3G), according to a UHPLC/MSMS method. The general linear mixed model was applied to time-related concentrations and a pragmatic four-step delay estimation approach was proposed based on the simultaneous presence of metabolites in plasma. Validation of the model was carried out using the jackknife technique on the 11 patients, and on a group of 7 test patients. Quadratic equations were derived for all metabolites except 6AM. The interval delay estimation was 2-4 days when only M3G present in plasma, 1-2 days when M6G and M3G were both present, 0-1 day when MOR, M6G and M3G were present and <2 h for all metabolites present. The 'jackknife' correlation between declared and actual estimated delays was 0.90. The overall precision of the delay estimates was 8-9 h. The delay between last heroin consumption and blood sampling in chronic drug users can be satisfactorily predicted from plasma heroin metabolites.

[Analytical validation of a chromatographic method dedicated to search and identify natural and semi-synthetic opiates]. / Validation analytique d'une méthode chromatographique destinée à rechercher et à identifier les opiacés naturels ou (semi) synthétiques.

The identification of a product absorbed by an opiate consumer is sometimes problematic since there is no specific biomarker for all molecules. We developed an ultra-high pressure liquid chromatography coupled to tandem mass spectrometry technique which allows the identification and the quantification of 25 opiates in plasma. The sample preparation consists in a solid-phase extraction on Oasis MCX cartridges (Waters). The method has been validated according to FDA criteria completely for 21 substances and with some reservations for the remaining 4 analytes. This method has been applied to 80 patients treated at the University Hospital of Liege for whom the screening of opiates was positive. The identification of the product consumed was effective in 86% of cases.

Simultaneous determination of some phthalate metabolites, parabens and benzophenone-3 in urine by ultra high pressure liquid chromatography tandem mass spectrometry.

Phthalates, parabens and 2-hydroxy-4-methoxybenzophenone or benzophone-3 are thought to act as endocrine disrupting chemicals, being able to disrupt the endocrine balance and therefore able to lead to some hormonal diseases. Numerous large-scale biomonitoring studies have detected the biomarkers of these compounds in more than 75% of the general population. To assess the exposure to these chemicals, we developed an analytical method based on a Solid Phase Extraction (SPE) prior to ultra high pressure liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for the simultaneous measurement of seven phthalate metabolites (monobenzyl phthalate, mono-n-butyl phthalate, mono-iso-butyl phthalate, mono-2-ethylhexyl phthalate, mono-2-ethyl-5-hydroxyhexyl phthalate, mono-2-ethyl-5-oxohexyl phthalate, monoethyl phthalate), four parabens (methyl paraben, ethyl paraben, n-propyl paraben, n-butyl parabens) and benzophenone-3 in human urine. The distinction between unconjugated, glucuro- and sulfoconjugated forms was achieved using different enzymatic hydrolyses. The whole procedure was validated according to the total error approach, and was demonstrated to be linear (regression coefficient ranging from 0.987 to 0.998) and accurate (inter and intra assay precision <17.71%, relative bias <5.87%) in the dosing range of concentrations. The limits of quantification (LOQs) obtained ranged between 0.30 and 1.23ng/ml depending on the analyte. The reliability of the method was proven in passing successfully the German External Quality Assessment Scheme (G-EQUAS). Moreover, the urine from 25 volunteers were analyzed for the determination of glucuro-, sulfo- and free species separately. Phthalate metabolites, parabens and benzophenone-3 were positively detected in almost all urine samples, with detection rates ranging from 40 to 100%. Levels measured ranged from