Search

Teratogen-induced congenital diaphragmatic hernia (CDH) rat models are commonly used to study the pathophysiology. We have created a new and reliable surgically induced diaphragmatic hernia (DH) model to obtain a purely mechanical DH rat model, and avoid the confounding teratogen-induced effects on the lung development. Fetal DH was surgically created on fetuses at E18.5 and harvested at E21.5 in rats. Four groups were evaluated (n = 16): control (CONT), control exposed to Nitrofen (CONT NIT), DH surgically created (DH SURG), and CDH Nitrofen (CDH NIT). Body weight, total lung weights, and their ratio (BW, TLW, and TLBR) were compared. Air space (AS), parenchyma (PA), total protein, and DNA contents were measured to verify lung hypoplasia. Medial wall thickness (MWT) of pulmonary arterioles was also analyzed. DH SURG showed significant hypoplasia (decreased in total protein and DNA) vs CONT (p

A distinction between the homogeneous and heterogeneous catalytic mechanisms of a myriad of Pd catalytic systems for the Suzuki-Miyaura reaction has been made based on soluble metal complexes or he...

The results of a kinetic study on the character (fast or slow) of a step of alkene activation in the catalytic cycle of the Mizoroki–Heck reaction with unreactive aryl chlorides in the presence of a so-called ligand-free catalytic system based on palladium are presented. An analysis of integral kinetic curves of the consumption of two competing alkenes in the reaction with aryl chloride showed that alkenes participate in a fast step of the catalytic cycle.

Neurodegenerative diseases are characterised by progressive damage to the nervous system including the selective loss of vulnerable populations of neurons leading to motor symptoms and cognitive decline. Despite millions of people being affected worldwide, there are still no drugs that block the neurodegenerative process to stop or slow disease progression. Neuronal death in these diseases is often linked to the misfolded proteins that aggregate within the brain (proteinopathies) as a result of disease-related gene mutations or abnormal protein homoeostasis. There are two major degradation pathways to rid a cell of unwanted or misfolded proteins to prevent their accumulation and to maintain the health of a cell: the ubiquitin–proteasome system and the autophagy–lysosomal pathway. Both of these degradative pathways depend on the modification of targets with ubiquitin. Aging is the primary risk factor of most neurodegenerative diseases including Alzheimer’s disease, Parkinson’s disease and amyotrophic lateral sclerosis. With aging there is a general reduction in proteasomal degradation and autophagy, and a consequent increase of potentially neurotoxic protein aggregates of β-amyloid, tau, α-synuclein, SOD1 and TDP-43. An often over-looked yet major component of these aggregates is ubiquitin, implicating these protein aggregates as either an adaptive response to toxic misfolded proteins or as evidence of dysregulated ubiquitin-mediated degradation driving toxic aggregation. In addition, non-degradative ubiquitin signalling is critical for homoeostatic mechanisms fundamental for neuronal function and survival, including mitochondrial homoeostasis, receptor trafficking and DNA damage responses, whilst also playing a role in inflammatory processes. This review will discuss the current understanding of the role of ubiquitin-dependent processes in the progressive loss of neurons and the emergence of ubiquitin signalling as a target for the development of much needed new drugs to treat neurodegenerative disease.

Three-component cross-coupling in the system aryl halide–arylacetylene–alkene in the presence of simplest ligand-free palladium catalysts gave products of both 1+1+1-coupling and cross-dimerization of arylacetylene with alkene. The possibility of controlling the reaction selectivity has been demonstrated.

Most drugs and xenobiotics are metabolized in the liver. Amongst others, different cytochrome P450 (CYP) enzymes catalyze the metabolic conversion of foreign compounds, and various transport proteins are engaged in the excretion of metabolites from the hepatocytes. Inter-species and inter-individual differences in the hepatic levels and activities of drug-metabolizing enzymes and transporters result from genetic as well as from environmental factors, and play a decisive role in determining the pharmacokinetic properties of a compound in a given test system. To allow for a meaningful comparison of results from metabolism studies, it is, therefore, of utmost importance to know about the specific metabolic properties of the test systems, especially about the levels of metabolic enzymes such as the CYPs. Using a targeted proteomics approach, we, therefore, compared the hepatic levels of important CYP enzymes and transporters in different experimental systems in vivo and in vitro, namely Wistar rats, C57/Bl6 mice, mice humanized for the two xeno-sensing receptors PXR (pregnane-X-receptor) and CAR (constitutive androstane receptor), mice with human hepatocyte-repopulated livers, human HepaRG hepatocarcinoma cells, primary human hepatocytes, and human liver biopsies. In addition, the effects of xenobiotic inducers of drug metabolism on CYP enzymes and transporters were analyzed in selected systems. This study for the first time presents a comprehensive overview of similarities and differences in important drug metabolism-related proteins among the different experimental models. Electronic supplementary material The online version of this article (10.1007/s00204-020-02939-4) contains supplementary material, which is available to authorized users.

Background Nuclear magnetic resonance (NMR) spectroscopy allows triglycerides to be subclassified into 14 different classes based on particle size and lipid content. We recently showed that these subfractions have differential associations with cardiovascular disease events. Here we report the distributions and define reference interval ranges for 14 triglyceride-containing lipoprotein subfraction metabolites. Methods Lipoprotein subfractions using the Nightingale NMR platform were measured in 9073 participants from four cohort studies contributing to the UCL-Edinburgh-Bristol consortium. The distribution of each metabolite was assessed, and reference interval ranges were calculated for a disease-free population, by sex and age group (65 years), and in a subgroup population of participants with cardiovascular disease or type 2 diabetes. We also determined the distribution across body mass index and smoking status. Results The largest reference interval range was observed in the medium very-low density lipoprotein subclass (2.5th 97.5th percentile; 0.08 to 0.68 mmol/L). The reference intervals were comparable among male and female participants, with the exception of triglyceride in high-density lipoprotein. Triglyceride subfraction concentrations in very-low density lipoprotein, intermediate-density lipoprotein, low-density lipoprotein and high-density lipoprotein subclasses increased with increasing age and increasing body mass index. Triglyceride subfraction concentrations were significantly higher in ever smokers compared to never smokers, among those with clinical chemistry measured total triglyceride greater than 1.7 mmol/L, and in those with cardiovascular disease, and type 2 diabetes as compared to disease-free subjects. Conclusion This is the first study to establish reference interval ranges for 14 triglyceride-containing lipoprotein subfractions in samples from the general population measured using the nuclear magnetic resonance platform. The utility of nuclear magnetic resonance lipid measures may lead to greater insights for the role of triglyceride in cardiovascular disease, emphasizing the importance of appropriate reference interval ranges for future clinical decision making.

Xenobiotica-metabolizing enzyme (XME) induction is a relevant biological/biochemical process vital to understanding the toxicological profile of xenobiotics. Early recognition of XME induction potential of compounds under development is therefore important, yet its determination by traditional XME activity measurements is time consuming and cost intensive. A proof-of-principle study was therefore designed due to the advent of faster and less cost-intensive methods for determination of enzyme protein and transcript levels to determine whether two such methods may substitute for traditional measurement of XME activity determinations. The results of the study show that determination of enzyme protein levels by peptide group-specific immunoaffinity enrichment/MS and/or determination of gene expression by NanoString nCounter may serve as substitutes for traditional evaluation methodology and/or as an early predictor of potential changes in liver enzymes. In this study, changes of XME activity by the known standard XME inducers phenobarbital, beta-naphthoflavone and Aroclor 1254 were demonstrated by these two methods. To investigate the applicability of these methods to demonstrate XME-inducing activity of an unknown, TS was also examined and found to be an XME inducer. More specifically, TS was found to be a phenobarbital-type inducer (likely mediated by CAR rather than PXR as nuclear receptor), but not due to Ah receptor-mediated or antioxidant response element-mediated beta-naphthoflavone-type induction. The results for TS were confirmed via enzymatic activity measurements. The results of the present study demonstrate the potential applicability of NanoString nCounter mRNA quantitation and peptide group-specific immunoaffinity enrichment/MS protein quantitation for predicting compounds under development to be inducers of liver XME activity.

Background Dysfunction of mitochondrial energy generation may contribute to neurodegeneration, leading to synaptic loss in Parkinson's disease (PD). The objective of this study was to find cross-sectional and longitudinal changes in PET markers of synaptic vesicle protein 2A, sigma 1 receptor, and mitochondrial complex 1 in drug-naive PD patients. Methods Twelve early drug-naive PD patients and 16 healthy controls underwent a 3-Tesla MRI and PET imaging to quantify volume of distribution of [11 C]UCB-J, [11 C]SA-4503, and [18 F]BCPP-EF for synaptic vesicle protein 2A, sigma 1 receptor, and mitochondrial complex 1, respectively. Nine PD patients completed approximately 1-year follow-up assessments. Results Reduced [11 C]UCB-J volume of distribution in the caudate, putamen, thalamus, brain stem, and dorsal raphe and across cortical regions was observed in drug-naive PD patients compared with healthy controls. [11 C]UCB-J volume of distribution was reduced in the locus coeruleus and substantia nigra but did not reach statistical significance. No significant differences were found in [11 C]SA-4503 and [18 F]BCPP-EF volume of distribution in PD compared with healthy controls. Lower brain stem [11 C]UCB-J volume of distribution correlated with Movement Disorder Society Unified Parkinson's Disease Rating Scale part III and total scores. No significant longitudinal changes were identified in PD patients at follow-up compared with baseline. Conclusions Our findings represent the first in vivo evidence of mitochondrial, endoplasmic reticulum, and synaptic dysfunction in drug-naive PD patients. Synaptic dysfunction likely occurs early in disease pathophysiology and has relevance to symptomatology. Mitochondrial complex 1 and sigma 1 receptor pathology warrants further investigations in PD. Studies in larger cohorts with longer follow-up will determine the validity of these PET markers to track disease progression. © 2020 The Authors. Movement Disorders published by Wiley Periodicals, Inc. on behalf of International Parkinson and Movement Disorder Society.

Perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) are man-made chemicals that are used for the fabrication of many products with water- and dirt-repellent properties. The toxicological potential of both substances is currently under debate. In a recent Scientific Opinion, the European Food Safety Authority (EFSA) has identified increased serum total cholesterol levels in humans as one major critical effect being associated with exposure to PFOA or PFOS. In animal studies, both substances induced a decrease of serum cholesterol levels, and the underlying molecular mechanism(s) for these opposed effects are unclear so far. In the present study, we examined the impact of PFOA and PFOS on cholesterol homoeostasis in the human HepaRG cell line as a model for human hepatocytes. Cholesterol levels in HepaRG cells were not affected by PFOA or PFOS, but both substances strongly decreased synthesis of a number of bile acids. The expression of numerous genes whose products are involved in synthesis, metabolism and transport of cholesterol and bile acids was strongly affected by PFOA and PFOS at concentrations above 10 µM. Notably, both substances led to a strong decrease of CYP7A1, the key enzyme catalyzing the rate-limiting step in the synthesis of bile acids from cholesterol, both at the protein level and at the level of gene expression. Moreover, both substances led to a dilatation of bile canaliculi that are formed by differentiated HepaRG cells in vitro. Similar morphological changes are known to be induced by cholestatic agents in vivo. Thus, the strong impact of PFOA and PFOS on bile acid synthesis and bile canalicular morphology in our in vitro experiments may allow the notion that both substances have a cholestatic potential that is connected to the observed increased serum cholesterol levels in humans in epidemiological studies. Electronic supplementary material The online version of this article (10.1007/s00204-020-02732-3) contains supplementary material, which is available to authorized users.

In this study, the performance of an In2O3/ZrO2 catalyst for hydrogenation of CO2 to methanol is reported in the presence of typical impurities of industrial CO2 feed gas streams. Experiments were performed with defined amounts of SO2, H2S, NO2, NH3 and hydrocarbons purposely added to the reaction mixture under industrially relevant conditions (225–275 °C, 50 bar total pressure, H2 : CO2 = 3 : 1). Comparative studies – with and without added impurities – further relate the observed effects to the behaviour of the commercial Cu/ZnO/Al2O3 methanol catalyst under identical treatments. Elemental analysis was used to determine the amount of sulphur, nitrogen and carbon residing on the catalyst after poisoning and after methanol synthesis. Moreover, temperature programmed desorption (TPD), temperature programmed reduction (TPR) and X-ray photoelectron spectroscopy (XPS) allowed identifying potential catalyst poisons formed on the catalyst surface. This work discloses that H2S and SO2 impurities led to drastic deactivation of the In2O3/ZrO2 catalyst (due to indium sulfide and -sulphate formation), while NH3, NO2 and hydrocarbon impurities have no major effect on the methanol production.

The aryl halide-arylacetylene-arylboronic acid three-component coupling in the presence of a Pd catalyst containing no organic ligands is described. The reaction routes that take place in the reaction system are shown to include the consecutive carbopalladation of two arylacetylene molecules, leading to unsaturated polyaromatic compounds. The possibility of controlling the alternative reaction routes is demonstrated.

This paper presents the results of a kinetic study on the step of activation of aryl iodides and aryl bromides under conditions of ligand-free catalysis in the direct arylation of indole at a C–H bond performed using an analysis of the integral kinetic curves of consumption of the competing aryl halides. In the course of an experiment under the conditions of competition between a pair of aryl halides having different reactivity, an acceleration in the consumption of a less active substrate was observed, which coincided in time with the almost complete consumption of a more active one. The experimental data unambiguously indicated a rapid character of the stage of oxidative addition with the participation of not only reactive aryl iodides but also nonactivated aryl bromides.

The kinetic approach for distinguishing between homogeneous and heterogeneous catalysis mechanisms by the analysis of differential selectivity was applied for direct C–H arylation of an indole with aryl halides under “ligandless” conditions. The differential selectivity of competing arylation of an indole with two aryl iodides or bromides and the differential regioselectivity of a parallel formation of C2- and C3-regioisomers of arylated indoles were studied by the simple method of phase trajectories needed for raw kinetic data only. The results obtained indicated that the selectivities strongly depended on the type (soluble or insoluble) and concentration of the Pd precursor. Considering the pathways of Pd transformations under the reaction conditions, this indicates a substantial contribution of catalysis occurring on the surface of metallic Pd (including nanoparticles).

The results of a comparative study of phosphine-containing and phosphine-free catalytic systems of the Heck reaction using aromatic carboxylic anhydrides as arylating agents are presented. It was demonstrated that the patterns of diff erential selectivity of the reaction under competition of two aromatic anhydrides or two alkenes are independent of the presence of a tertiary phosphine additive in the system. It was established that palladium complexes with no phosphine ligands in their coordination sphere are catalytically active at the step of activation of aromatic carboxylic anhydride and alkene. The patterns of diff erential selectivity for regioisomers of arylated products provide the evidence of the participation of phosphine-containing anionic palladium complexes in the regioselectivity-determining step of the catalytic cycle. The data obtained are in agreement with the phosphine involvement in the catalyst transformations proceeding outside the main catalytic cycle.

Background: Patients with congenital diaphragmatic hernia (CDH) have a short postnatal period of ventilatory stability called the honeymoon period, after which changes in pulmonary vascular reactivity result in pulmonary hypertension. However, the mechanisms involved are still unknown. The aim of this study was to evaluate mechanical ventilation's effect in the honeymoon period on VEGF, VEGFR-1/2 and eNOS expression on experimental CDH in rats.Materials and Methods: Neonates whose mothers were not exposed to nitrofen formed the control groups (C) and neonates with left-sided defects formed the CDH groups (CDH). Both were subdivided into non-ventilated and ventilated for 30, 60, and 90 min (n = 7 each). The left lungs (n = 4) were evaluated by immunohistochemistry of the pulmonary vasculature (media wall thickness), VEGF, VEGFR-1/2 and eNOS. Western blotting (n = 3) was performed to quantify the expression of VEGF, VEGFR-1/2 and eNOS.Results: CDH had lower biometric parameters than C. Regarding the pulmonary vasculature, C showed a reduction in media wall thickness with ventilation, while CDH presented reduction with 30 min and an increase with the progression of the ventilatory time (honeymoon period). CDH and C groups showed different patterns of VEGF, VEGFR-1/2 and eNOS expressions. The receptors and eNOS findings were significant by immunohistochemistry but not by western blotting, while VEGF was significant by western blotting but not by immunohistochemistry.Conclusion: VEGF, its receptors and eNOS were altered in CDH after mechanical ventilation. These results suggest that the VEGF-NO pathway plays an important role in the honeymoon period of experimental CDH.

Graphene-induced energy transfer (GIET) was recently introduced for the precise localization of fluorescent molecules along the optical axis of a microscope. GIET is based on near-field energy transfer from an optically excited fluorophore to a single sheet of graphene. As a proof-of-concept, we demonstrated its potential by determining the distance between the two leaflets of supported lipid bilayers (SLBs) with sub-nanometer accuracy. Here, we use GIET imaging for three-dimensional reconstruction of the mitochondrial membrane architecture. We map two quasi-stationary states of the inner and outer mitochondrial membranes before and during adenosine tri-phosphate (ATP) synthesis. We trigger the ATP synthesis state in vitro by activating mitochondria with precursor molecules. Our results demonstrate that the inner membrane (IM) approaches the outer membrane (OM) while the outer membrane (OM) does not show a measurable change in average axial position upon activation. As a result, the inter-membrane space (IM-OM distance) is reduced by ∼2 nm upon activation of the mitochondria. This direct experimental observation of the subtle dynamics of mitochondrial membranes and the change in inter-membrane distance induced by ATP synthesis is relevant for our understanding of the physical functioning of mitochondria.

Despite the recent development of vaccines and monoclonal antibodies preventing SARS-CoV-2 infection, treating critically ill COVID-19 patients still remains a top goal. In principle, drug repurposing – the use of an already existing drug for a new indication – could provide a shortcut to a treatment. However, drug repurposing is often very speculative due to lack of clinical evidence. We report here on a methodology to find and test drug target candidates for drug repurposing. Using UK Biobank data, we matched critically ill COVID-19 cases with healthy controls and screened for significant differences in 33 blood cell types, 30 blood biochemistries, and body mass index. Significant differences in traits that have been associated with critically ill COVID-19 status in prior literature, such as alanine aminotransferase, body mass index, C-reactive protein, and neutrophil cell count, were further investigated. In-depth statistical analysis of COVID-19 associated traits and their genetics using regression modeling and propensity score stratification identified cyclin-dependent kinase 6 (CDK6) as a more promising drug target for the selective treatment of critically ill COVID-19 patients than the previously reported interleukin 6. Four existing CDK6 inhibitors -- abemaciclib, ribociclib, trilaciclib, and palbociclib -- have been approved for the treatment of breast cancer. Clinical evidence for CDK6 inhibitors in treating critically ill COVID-19 patients has been reported. Further clinical investigations are ongoing.

In real life, organisms are exposed to complex mixtures of chemicals at low concentration levels, whereas research on toxicological effects is mostly focused on single compounds at comparably high doses. Mixture effects deviating from the assumption of additivity, especially synergistic effects, are of concern. In an adverse outcome pathway (AOP)-guided manner, we analyzed the accumulation of triglycerides in human HepaRG liver cells by a mixture of eight steatotic chemicals (amiodarone, benzoic acid, cyproconazole, flusilazole, imazalil, prochloraz, propiconazole and tebuconazole), each present below its individual effect concentration at 1-3 mu M. Pronounced and significantly enhanced triglyceride accumulation was observed with the mixture, and similar effects were seen at the level of pregnane-X-receptor activation, a molecular initiating event leading to hepatic steatosis. Transcript pattern analysis indicated subtle pro-steatotic changes at low compound concentrations, which did not exert measurable effects on cellular triglycerides. Mathematical modeling of mixture effects indicated potentially more than additive behavior using a model for compounds with similar modes of action. The present data underline the usefulness of AOP-guided in vitro testing for the identification of mixture effects and highlight the need for further research on chemical mixtures and harmonization of data interpretation of mixture effects.

Mixed chain phospholipids containing a saturated fatty acid at sn1 and a polyunsaturated fatty acid in sn2 are common in the specialized biological membranes prevalent in neural, retinal and organ tissues. Particularly important are mixed lipids containing palmitic or stearic acid and arachidonic or docosahexaenoic acid. Gradient temperature Raman spectroscopy (GTRS) applies the temperature gradients utilized in differential scanning calorimetry to Raman spectroscopy, providing a straightforward technique to identify molecular rearrangements and phase transitions. Herein we utilize GTRS for 1-18:0, 2-20:4n-6 PC; 1-18:0 2-22:6n-3 PC; and 1-18:0, 2-18:0 PC from −80 to 50 °C temperatures. 20 Mb three-dimensional data arrays with 0.2 °C increments and first/second derivatives allowed detailed vibrational mode assignment and analysis. Samples were analyzed neat and with molecular hydration. Previously reported phase transitions for hydrated 18:0−20:4PC and 18:0−22:6PC and numerous spectral differences resulting from hydration and the double bond structure were clearly observed. Molecular models showed that the addition of minimal water molecules results in significant structural differences compared to the neat molecules; 18:0−22:6PC is strikingly compact with water when viewed from the hydrophilic end. This precise Raman data cannot be observed in typically utilized fully hydrated vesicle samples, however the improved GTRS will allow for more precise analysis in fully hydrated vesicles because the underlying modes in the unavoidably broadened spectra can be identified.

Background:DNA methylation may be one of the mechanisms by which alcohol consumption is associated with the risk of disease. We conducted a large-scale, cross-sectional, genome-wide DNA methylation association study of alcohol consumption and a longitudinal analysis of repeated measurements taken several years apart.Methods:Using the Illumina Infinium HumanMethylation450 BeadChip, DNA methylation measures were determined using baseline peripheral blood samples from 5,606 adult Melbourne Collaborative Cohort Study (MCCS) participants. For a subset of 1,088 of them, these measures were repeated using blood samples collected at follow-up, a median of 11 years later. Associations between alcohol intake and blood DNA methylation were assessed using linear mixed-effects regression models adjusted for batch effects and potential confounders. Independent data from the LOLIPOP (N=4,042) and KORA (N=1,662) cohorts were used to replicate associations discovered in the MCCS.Results:Cross-sectional analyses identified 1,414 CpGs associated with alcohol intake at P-7, 1,243 of which had not been reported previously. Of these 1,243 novel associations, 1,078 were replicated (PConclusion:Our study indicates that, for middle-aged and older adults, alcohol intake is associated with widespread changes in DNA methylation across the genome. Longitudinal analyses showed that the methylation status of alcohol-associated CpGs may change with changes in alcohol consumption.

While real-life exposure occurs to complex chemical mixtures, toxicological risk assessment mostly focuses on individual compounds. There is an increasing demand for in vitro tools and strategies for mixture toxicity analysis. Based on a previously established set of hepatotoxicity marker genes, we analyzed mixture effects of non-cytotoxic concentrations of different pesticides in exposure-relevant binary mixtures in human HepaRG hepatocarcinoma cells using targeted transcriptomics. An approach for mixture analysis at the level of a complex endpoint such as a transcript pattern is presented, including mixture design based on relative transcriptomic potencies and similarities. From a mechanistic point of view, goal of the study was to evaluate combinations of chemicals with varying degrees of similarity in order to determine whether differences in mechanisms of action lead to different mixtures effects. Using a model deviation ratio-based approach for assessing mixture effects, it was revealed that most data points are consistent with the assumption of dose addition. A tendency for synergistic effects was only observed at high concentrations of some combinations of the test compounds azoxystrobin, cyproconazole, difenoconazole, propiconazole and thiacloprid, which may not be representative of human real-life exposure. In summary, the findings of our study suggest that, for the pesticide mixtures investigated, risk assessment based on the general assumption of dose addition can be considered sufficiently protective for consumers. The way of data analysis presented in this paper can pave the way for a more comprehensive use of multi-gene expression data in experimental studies related to mixture toxicity.

Drug development of cholesteryl ester transfer protein (CETP) inhibition to prevent coronary heart disease (CHD) has yet to deliver licensed medicines. To distinguish compound from drug target failure, we compared evidence from clinical trials and Mendelian randomization (MR) results. Findings from meta-analyses of CETP inhibitor trials (≥ 24 weeks follow-up) were used to judge between-compound heterogeneity in treatment effects. Genetic data were extracted on 190+ pharmacologically relevant outcomes; spanning 480,698-21,770 samples and 74,124-4,373 events. Drug target MR of protein concentration was used to determine the on-target effects of CETP inhibition and compared to that of PCSK9 modulation. Fifteen eligible CETP inhibitor trials of four compounds were identified, enrolling 79,961 participants. There was a high degree of heterogeneity in effects on lipids, lipoproteins, blood pressure, and clinical events. For example, dalcetrapib and evacetrapib showed a neutral effect, torcetrapib increased, and anacetrapib decreased cardiovascular disease (CVD); heterogeneity p-value < 0.001. In drug target MR analysis, lower CETP concentration (per μg/ml) was associated with CHD (odds ratio 0.95; 95%CI 0.91; 0.99), heart failure (0.95; 95%CI 0.92; 0.99), chronic kidney disease (0.94 95%CI 0.91; 0.98), and age-related macular degeneration (1.69; 95%CI 1.44; 1.99). Lower PCSK9 concentration was associated with a lower risk of CHD, heart failure, atrial fibrillation and stroke, and increased risk of Alzheimer’s disease and asthma. In conclusion, previous failures of CETP inhibitors are likely compound related. CETP inhibition is expected to reduce risk of CHD, heart failure, and kidney disease, but potentially increase risk of age-related macular disease.

[Image: see text] Proteinase-activated receptor 2 (PAR2) is a class A G protein-coupled receptor whose activation has been associated with inflammatory diseases and cancer, thus representing a valuable therapeutic target. Pathophysiological roles of PAR2 are often characterized using peptidic PAR2 agonists. Peptidic ligands are frequently unstable in vivo and show poor bioavailability, and only a few approaches toward drug-like nonpeptidic PAR2 ligands have been described. The herein-described ligand 5a (IK187) is a nonpeptidic PAR2 agonist with submicromolar potency in a functional assay reflecting G protein activation. The ligand also showed substantial β-arrestin recruitment. The development of the compound was guided by the crystal structure of PAR2, when the C-terminal end of peptidic agonists was replaced by a small molecule based on a disubstituted phenylene scaffold. IK187 shows preferable metabolic stability and may serve as a lead compound for the development of nonpeptidic drugs addressing PAR2.

Although oral drug delivery is preferred by patients, it is not possible for proteins because the gastrointestinal tract is not sufficiently permeable. To enable the non-toxic oral uptake of protein drugs, we investigated plant-based foods as intestinal permeation enhancers, hypothesizing that compounds found in food would be well-tolerated by the gastrointestinal tract. Following a screen of over 100 fruits, vegetables, herbs, and fungi, we identified strawberry as a potent enhancer of macromolecular permeability in vitro and in mice. Natural product chemistry techniques identified pelargonidin, an anthocyanidin, as the active compound. In mice, insulin was orally administered with pelargonidin to induce sustained pharmacodynamic effects with doses as low as 1 U/kg and bioactivity of over 100% relative to the current gold standard of subcutaneous injection. Pelargonidin-induced permeability was reversible within two hours of treatment, and one month of daily dosing did not adversely affect mice as determined by weight tracking, serum concentrations of inflammatory markers, and tight junction gene expression. Results underscore the utility of plant-based foods in biomedical applications and demonstrate pelargonidin as an especially potent enhancer for the oral delivery of biologics.

A kinetic study of reaction of alkene arylation by anhydrides of aromatic acids (modified Mizoroki–Heck reaction) was carried out using differential selectivity as the main measured parameter. The results obtained under conditions of competition between a pair of alkenes or a pair of aromatic anhydrides point to the occurrence of the reaction via a homogeneous mechanism of catalysis analogously to a conventional variant of reaction with aryl halides as arylating reagents. The hypothesis that the active catalyst is homogeneous agrees with the results of kinetic studies of processes of catalyst formation and deactivation.

Background: Folate and other one-carbon metabolism nutrients are essential to enable DNA methylation to occur, but the extent to which their dietary intake influences methylation in adulthood is unclear. Objective: We assessed associations between dietary intake of these nutrients and DNA methylation in peripheral blood, overall and at specific genomic locations. Design: We conducted a cross-sectional study using baseline data and samples from 5186 adult participants in the Melbourne Collaborative Cohort Study (MCCS). Nutrient intake was estimated from a food-frequency questionnaire. DNA methylation was measured by using the Illumina Infinium HumanMethylation450 BeadChip array (HM450K). We assessed associations of intakes of folate, riboflavin, vitamins B-6 and B-12, methionine, choline, and betaine with methylation at individual cytosine-guanine dinucleotides (CpGs), and with median (genome-wide) methylation across all CpGs, CpGs in gene bodies, and CpGs in gene promoters. We also assessed associations with methylation at long interspersed nuclear element 1 (LINE-1), satellite 2 (Sat2), and Arthrobacter luteus restriction endonuclease (Alu) repetitive elements for a subset of participants. We used linear mixed regression, adjusting for age, sex, country of birth, smoking, energy intake from food, alcohol intake, Mediterranean diet score, and batch effects to assess log-linear associations with dietary intake of each nutrient. In secondary analyses, we assessed associations with low or high intakes defined by extreme quintiles. Results: No evidence of log-linear association was observed at P

Differential selectivity of the direct arylation of indole with aryl halides under competing and noncompeting conditions with a varying set of reaction parameters was determined using phase trajectories. The results described herein allow for conclusions to be drawn regarding the character of active complexes (cationic, neutral, or anionic) as well as realization of the indole electrophilic substitution in the catalytic cycle using the ligand-free catalytic system.

Fluorescent protein based pH sensors are a useful tool for measuring protein trafficking through pH changes associated with endo-and exocytosis. However, commonly used pH sensing probes are ubiquitously expressed with their protein of interest throughout the cell, hindering the ability to focus on specific trafficking pools of proteins. We developed a family of excitation-ratiometric, activatable pH responsive tandem dyes, consisting of a pH sensitive Cy3 donor linked to a fluorogenic malachite green acceptor. These cell-excluded dyes are targeted and activated upon binding to a genetically expressed fluorogen activating protein, and are suitable for selective labeling of surface proteins for analysis of endocytosis and recycling in live cells using both confocal and superresolution microscopy. Quantitative profiling of the endocytosis and recycling of tagged β2-adrenergic receptor (B2AR) at a single vesicle level revealed differences among B2AR agonists, consistent with more detailed pharmacological profiling.

Specialized pro-resolving mediators provide promising targets for new drugs and natural products. Much work has been accomplished on the structure/ function of the lipoxygenase and cyclooxygenase enzymes but not on the substrates. A better visualization of three-dimensional lipid structures will allow increased refinement of the interactions that produce the pro-resolving mediators, and lead to improvements in synthetic pathways. We present systematic analysis of oleic (18:1n-9), linoleic (18:2n-6), alpha-linolenic (18:3n-3), arachidonic (20:4n-6), docosapentaenoic (22:5n-3), and docosahexaenoic (22:6n-3) acids. Continuous gradient temperature Raman spectroscopy (GTRS) applies the temperature gradients utilized in differential scanning calorimetry to Raman spectroscopy. GTRS can identify and differentiate specific carbon chain sites, finally allowing Raman analysis to explain why the long-chain polyunsaturated fatty acids (LC-PUFA) exhibit such extreme functional differences despite minimal changes in chemical structure. Detailed vibrational analysis of the important frequency ranges 1450 - 1200 cm-1 (includes CH2 bending and twisting) and 1750 - 1425 cm-1 (includes C=C stretching and C-C stretching plus H-C in-plane rocking) shows for the first time that each molecule has its own characteristic set of modes with only some redundancy/commonality. The number and frequency of modes correlates with three-dimensional molecular structure, not the degree of unsaturation. The high degree of specificity of lipoxygenase and cyclooxygenase enzymes should be reconsidered in light of the fact that individual sites on the polyunsaturated fatty acid chain are nonequivalent, and each LC-PUFA molecule has an individual, specific three dimensional structure incorporating torsion.

The role of ionic Pd species in the catalytic systems used for cross-coupling reactions has been revised in recent years, mainly utilizing in situ analytical techniques to detect catalyst species in the reaction mixture. The possible inclusion of phosphine molecules in the active Pd complexes is still debated. We have carried out a comparative study on the differential selectivity of the Mizoroki–Heck reaction with competition between two aryl halides or two alkenes to elucidate the active Pd species in different types of catalytic systems, including those favoring the formation of ionic complexes. The sensitivity of the reaction differential selectivity of two competing aryl halides to the additives of tertiary phosphine and halide salts points to the catalytic activity of anionic Pd(0) complexes containing at least one phosphine ligand when the phosphine-containing catalytic systems are used. The changes of the reaction differential selectivity of two competing alkenes to phosphine and halide salt allow to propose the entrance of both phosphine and halide ions into the coordination sphere of active Pd(II) complexes possessing anionic character. In addition, evidence for the effect of phosphine ligand and salt additives on the composition of the active Pd complexes that determine the regioselectivity of the reaction is presented. The data obtained indicate the decisive role of anionic Pd species in the Mizoroki–Heck reaction using both phosphine-free and phosphine-containing catalytic systems.

Many streets in Europe and the US are lined with horse chestnut trees. However, its fruit is inedible and poorly utilized. Therefore, its effective utilization has been a big issue. For recycling of horse chestnut seed, it was separated into “sarcocarp and endodermis” and “outer theca”; each part was finely ground for analysis of its functions. Ground horse chestnut seed showed excellent antibacterial activity toward Staphylococcus aureus, Escherichia coli, and Streptococcus mutans. Antibacterial activity was particularly strong against E. coli, which is a Gram-negative bacterium. The “outer theca” part showed a higher antibacterial activity than did the “sarcocarp and endodermis” part. It was found that the polyphenol content of the “outer theca” of the horse chestnut fruit was approximately three times that of the “sarcocarp and endodermis”; these polyphenols are believed to play a role in the seed’s antioxidation capacity. The ground outer theca of horse chestnut seed showed oxygen radical absorbance capacity (ORAC) of 531 mmol Trolox equivalent (TE)/g. Antioxidant activity was assumed to contribute to the antibacterial activity. In other words, antioxidant activity was assumed to damage the peptidoglycan layer of the bacterial cell wall. For possible recycling of horse chestnut seed, compounded paper containing the ground outer theca blended with pulp fibers was prepared. In this study, compounded paper containing ground “outer theca” was prepared using a traditional Washi paper-making method. The compounded paper was found to retain high deodorizing activity of the ground theca. Furthermore, although mechanical properties deteriorated as the proportion of the ground outer theca increased, these parameters remained adequate for practical applications. The compounded paper may be expected to have a wide range of applications as a highly functional paper with high gas permeability, deodorant activity, and antibacterial activity.

Since the 1950's, the broad-spectrum, organochlorine insecticide endosulfan (6,7,8, 9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3-benzodioxathiepine-3-oxide) has been used on numerous crops. Due to its persistence, bioaccumulation, long-range transport, and adverse effects to human health and ecosystems, it was officially identified as a persistent organic pollutant (POP) in 2011. The last uses in the United States were phased out in 2016. Endosulfan consists of two diastereomers, α-endosulfan and β-endosulfan, and while the α-isomer exists as two asymmetrical, twist-chair enantiomers which interchange, the β-isomer is a symmetrical-chair conformation. In addition, the β-isomer was found to isomerize to the α-isomer. Gradient Temperature Raman Spectroscopy (GTRS) enables identification the molecular sites at which bending vibrational modes becomes twisting or wagging vibrational modes. Previous studies using GTRS and chemical calculations afforded evidence for specific bond movements and the irreversibility of the isomerization mechanism. However, not all of the vibrational modes observed in the spectra could be explained. Thus, new analyses of the GTRS data were conducted to examine the effects of torsional forces on the bond movement, which allowed for the identification of all the peaks. These newly-identified torsional forces provide further confirmation of the isomerization mechanism and its irreversibility. Finally, this isomerization explains why β-endosulfan is rarely detected in the atmosphere.

Subsurface inspection of food and agricultural products is challenging for optical-based sensing techniques due to complex interactions between light and heterogeneous or layered samples. In this study, a method for subsurface food inspection was presented based on a newly developed line-scan spatially offset Raman spectroscopy (SORS) technique. A 785 nm point laser was used as a Raman excitation source. The line-shape SORS data from the sample was collected in a wavenumber range of 0–2815 cm −1 using a detection module consisting of an imaging spectrograph and a CCD camera. Two layered samples, one by placing a 1 mm thick plastic sheet cut from original container on top of cane sugar and the other by placing a 5 mm thick carrot slice on top of melamine powder, were created to test the subsurface food inspection method. For each sample, a whole set of SORS data was acquired using one CCD exposure in an offset range of 0–36 mm (two sides of the incident laser point) with a spatial interval of 0.07 mm. Raman spectra from the cane sugar under the plastic sheet and the melamine powder under the carrot slice were successfully resolved using self-modeling mixture analysis (SMA) algorithms, demonstrating the potential of the technique for authenticating foods and ingredients through packaging and evaluating internal food safety and quality attributes. The line-scan SORS measurement technique provides a rapid and nondestructive method for subsurface inspection of food safety and quality.

Both surface and subsurface food inspection is important since interesting safety and quality attributes can be at different sample locations. This paper presents a multipurpose line-scan Raman platform for food safety and quality research, which can be configured for Raman chemical imaging (RCI) mode for surface inspection and spatially offset Raman spectroscopy (SORS) mode for subsurface inspection. In the RCI mode, macro-scale imaging was achieved using a 785 nm line laser up to 24 cm long with a push-broom method. In the SORS mode, a 785 nm point laser was used and a complete set of SORS data was collected in an offset range of 0–36 mm with a spatial interval of 0.07 mm using one CCD exposure. The RCI and SORS modes share a common detection module including a dispersive imaging spectrograph and a CCD camera, covering a Raman shift range from −674 to 2865 cm−1. A pork shoulder and an orange carrot were used to test large-field-of-view (230 mm wide) and high-spatial-resolution (0.07 mm/pixel) settings of the RCI mode for food surface evaluation. Fluorescence-corrected images at selected Raman peak wavenumbers were used to view Raman-active analytes on the whole sample surfaces (e.g., fat on the pork shoulder and carotenoids over the carrot cross section). Also, three layered samples, which were created by placing carrot slices with thicknesses of 2, 5, and 8 mm on top of melamine powder, were used to test the SORS mode for subsurface food evaluation. Raman spectra from carrot and melamine were successfully resolved for all three layered samples using self-modeling mixture analysis. The line-scan Raman imaging and spectroscopy platform provides a new tool for surface and subsurface inspection for food safety and quality.

Docosahexaenoic acid (DHA, 22:6n-3) is exclusively utilized in fast signal processing tissues such as retinal, neural and cardiac. N-3 docosapentaenoic acid (n-3DPA, 22:5n-3), with just one less double bond, is also found in the marine food chain yet cannot substitute for DHA. Gradient temperature Raman spectroscopy (GTRS) applies the temperature gradients utilized in differential scanning calorimetry (DSC) to Raman spectroscopy, providing a straightforward technique to identify molecular rearrangements that occur near and at phase transitions. Herein we apply GTRS and both conventional and modulated DSC to n-3DPA and DHA from -100 to 20°C. Three-dimensional data arrays with 0.2°C increments and first derivatives allowed complete assignment of solid, liquid and transition state vibrational modes. Melting temperatures n-3DPA (-45°C) and DHA (-46°C) are similar and show evidence for solid-state phase transitions not seen in n-6DPA (-27°C melt). The C6H2 site is an elastic marker for temperature perturbation of all three lipids, each of which has a distinct three dimensional structure. N-3 DPA shows the spectroscopic signature of saturated fatty acids from C1 to C6. DHA does not have three aliphatic carbons in sequence; n-6DPA does but they occur at the methyl end, and do not yield the characteristic signal. DHA appears to have uniform twisting from C6H2 to C12H2 to C18H2 whereas n-6DPA bends from C12 to C18, centered at C15H2. For n-3DPA, twisting is centered at C6H2 adjacent to the C2-C3-C4-C5 aliphatic moiety. These molecular sites are the most elastic in the solid phase and during premelting.

We previously discovered a method to estimate the groundwater mean residence time using the changes in the enantiomeric ratio of metolachlor ethanesulfonic acid (MESA), (2-[(2-ethyl-6-methylphenyl)(2-methoxy-1-methylethyl)amino]-2-oxoethanesulfonic acid), a metabolite of the herbicide metolachlor. However, many grab samples would be needed for each watershed over an extended period, and this is not practical. Thus, we examined the use of a polar organic chemical integrative sampler (POCIS) deployed for 28 days combined with a modified liquid chromatography-mass spectrometry LC-MS/MS method to provide a time-weighted average of the MESA enantiomeric ratio. POCISs equipped with hydrophilic-lipophilic-balanced (HLB) discs were deployed at five sites across the United States where metolachlor was used before and after 1999 and compared the effectiveness of the POCIS to capture MESA versus grab samples. In addition, an in situ POCIS sampling rate (Rs) for MESA was calculated (0.15 L/day), the precision of MESA extraction from stored POCIS discs was determined, and the effectiveness of HLB to extract MESA was examined. Finally, using molecular modeling, the influence of the asymmetric carbon of metolachlor degradation on the MESA enantiomeric ratio was predicted to be negligible. Results of this work will be used in projects to discern the groundwater mean residence times, to evaluate the delivery of nitrate-N from groundwater to surface waters under various soil, agronomic, and land use conditions, and to examine the effectiveness of conservation practices.

We report the first experimental evidence of spontaneous electron emission from a homonuclear dimer anion through direct measurements of ${\text{Ag}}_{2}^{\ensuremath{-}}\ensuremath{\rightarrow}{\text{Ag}}_{2}+{e}^{\ensuremath{-}}$ decays on milliseconds and seconds timescales. This observation is very surprising as there is no avoided crossing between adiabatic energy curves to mediate such a process. The process is weak, yet dominates the decay signal after 100 ms when ensembles of internally hot ${\mathrm{Ag}}_{2}^{\ensuremath{-}}$ ions are stored in the cryogenic ion-beam storage ring, DESIREE, for 10 s. The electron emission process is associated with an instantaneous, very large reduction of the vibrational energy of the dimer system. This represents a dramatic deviation from a Born-Oppenheimer description of dimer dynamics.

Starting from the β-adrenoceptor agonist isoprenaline and beta-blocker carvedilol, we designed and synthesized three different chemotypes of agonist/antagonist hybrids. Investigations of ligand-mediated receptor activation using bioluminescence resonance energy transfer biosensors revealed a predominant effect of the aromatic head group on the intrinsic activity of our ligands, as ligands with a carvedilol head group were devoid of agonistic activity. Ligands composed of a catechol head group and an antagonist-like oxypropylene spacer possess significant intrinsic activity for the activation of Gαs, while they only show weak or even no β-arrestin-2 recruitment at both β1- and β2-AR. Molecular dynamics simulations suggest that the difference in G protein efficacy and β-arrestin recruitment of the hybrid ( S)-22, the full agonist epinephrine, and the β2-selective, G protein-biased partial agonist salmeterol depends on specific hydrogen bonding between Ser5.46 and Asn6.55, and the aromatic head group of the ligands.

Yellow turmeric (Curcuma longa) is widely used for culinary and medicinal purposes, and as a dietary supplement. Due to the commercial popularity of C. longa, economic adulteration and contamination with botanical additives and chemical substances has increased. This study used FT-IR spectroscopy for identifying and estimating white turmeric (Curcuma zedoaria), and Sudan Red G dye mixed with yellow turmeric powder. Fifty replicates of yellow turmeric&mdash, Sudan Red mixed samples (1%, 5%, 10%, 15%, 20%, 25% Sudan Red, w/w) and fifty replicates of yellow turmeric&mdash, white turmeric mixed samples (10%, 20%, 30%, 40%, 50% white turmeric, w/w) were prepared. The IR spectra of the pure compounds and mixtures were analyzed. The 748 cm&minus, 1 Sudan Red peak and the 1078 cm&minus, 1 white turmeric peak were used as spectral fingerprints. A partial least square regression (PLSR) model was developed for each mixture type to estimate adulteration concentrations. The coefficient of determination (R2v) for the Sudan Red mixture model was 0.97 with a root mean square error of prediction (RMSEP) equal to 1.3%. R2v and RMSEP for the white turmeric model were 0.95 and 3.0%, respectively. Our results indicate that the method developed in this study can be used to identify and quantify yellow turmeric powder adulteration.

We analyzed the unsaturated fatty acids oleic (OA, 18:1n-9) and linoleic (LA, 18:2n-3), and a 3:1 LA:OA mixture from −100 to 50 °C with continuous gradient temperature Raman spectroscopy (GTRS). The 20 Mb three-dimensional data arrays with 0.2 °C increments and first/second derivatives allowed rapid, complete assignment of solid, liquid, and transition state vibrational modes. For OA, large spectral and line width changes occurred in the solid state γ to α transition near −4 °C, and the melt (13 °C) over a range of only 1 °C. For LA, major intensity reductions from 200 to 1750 cm−1 and some peak shifts marked one solid state phase transition at −50 °C. A second solid state transition (−33 °C) had minor spectral changes. Large spectral and line width changes occurred at the melt transition (−7 °C) over a narrow temperature range. For both molecules, melting initiates at the diene structure, then progresses towards the ends. In the 3:1 LA:OA mixture, some less intense and lower frequencies present in the individual lipids are weaker or absent. For example, modes assignable to C8 rocking, C9H–C10H wagging, C10H–C11H wagging, and CH3 rocking are present in OA but absent in LA:OA. Our data quantify the concept of lipid premelting and identify the flexible structures within OA and LA, which have characteristic vibrational modes beginning at cryogenic temperatures.

We report an advanced analysis protocol that allows to quantitatively study the course of step-growth reactions by size exclusion chromatography on the example of the depolymerization of a Diels–Alder polymer based on a furane/maleimide couple at elevated temperatures. Frequently occurring issues of molar mass calibrations and overlap of monomer with solvent signals are addressed for determining reliable molar masses. Thereby, even kinetic parameters (e.g., rate coefficients) can be derived that otherwise would require performing additional spectroscopic experiments. Our results confirm first-order behavior of the rDA reaction with an activation energy of 33 kJ mol–1.

Raman spectroscopy has proven to be a reliable method for detection of chemical contaminants in food ingredients and products. To detect each contaminant particle in a food sample, it is important to determine the effective depth of penetration of the laser through the food sample and the corresponding laser intensity required for the penetration. It is also important to determine the effective spatial resolution needed to detect each contaminant particle in the mixture. This study examined the depth of penetration of a 785 nm laser through tapioca starch and wheat flour when using three different laser intensities. Melamine, known to exhibit identifiable Raman spectral peaks, was selected as a subsurface reference material for determining the depth of laser penetration through the food powder samples. The food powders were layered in five depths between 1 and 5 mm over a Petri dish packed with melamine. It was observed that the 785 nm laser could penetrate to 3 mm depth in starch; however, the penetration depth was limited to 2 mm in flour. These depths were achieved using laser intensities of 200 and 100 mW for starch and flour, respectively. The selected depth and laser intensity parameters were next used to examine the effective spatial resolution required for detection of maleic acid in starch and benzoyl peroxide in flour, which was selected to be 0.5 mm. Finally, an experiment was conducted to demonstrate the use of these parameters for quantitative Raman imaging-based detection of these contaminants prepared in mixtures at 0.1%, 0.3%, and 0.5% (w/w) concentrations.

Two 28-day feeding studies were performed in male rats to investigate combination effects of azole fungicides in a broad dose range. Following separate administration of cyproconazole, epoxiconazole, prochloraz, propiconazole, and tebuconazole at five dose levels, the first three compounds were selected to be administered in two different mixtures at three dose levels including very low doses. Here we present the data obtained by clinical observations, pathology, histopathology, clinical chemistry and haematology. The liver was the common main target organ of all compounds and their mixtures. In addition, epoxiconazole exhibited an effect on the adrenals. Furthermore, food consumption and efficiency and body weight (gain) were affected. Adverse effects of the combinations were observed at dose levels at which the individual substances caused similar effects. No evidence of adverse effects was found at dose levels below the previously established NOAELs. Our findings indicate that the concept of dose additivity appears sufficiently protective for risk assessment of the fungicides examined. Besides toxicological testing, tissue residues of the azole compounds in liver, testis and kidney were determined revealing remarkable differences following administration of the single substances and of the mixtures.

Tonantzitlolone A, a diterpene isolated from the Mexican plant Stillingia sanguinolenta, shows cytostatic activity. Both the natural product tonantzitlolone A and its synthetic enantiomer induce monoastral spindle formation in cell experiments which indicates inhibitory activity on kinesin-5 mitotic motor molecules. These inhibitory effects on kinesin-5 could be verified in in vitro single-molecule motility assays, where both tonantzitlolones interfered with kinesin-5 binding to its cellular interaction partner microtubules in a concentration-dependent manner, yet with a larger effect of the synthetic enantiomer. In contrast to kinesin-5 inhibition, both tonantzitlolone A enantiomers did not affect conventional kinesin-1 function; hence tonantzitlolones are not unspecific kinesin inhibitors. The observed stronger inhibitory effect of the synthetic enantiomer demonstrates the possibility to enhance the overall moderate anti-proliferative effect of the lead compound tonantzitlolon A by chemical modification.

To investigate the effect of combined prenatal treatment with retinoic acid (RA) and tracheal occlusion (TO) on the pulmonary vascular morphology and expression of vascular endothelial growth factors (VEGF) and its receptors in a rat model of congenital diaphragmatic hernia (CDH). Rats were given nitrofen at 9 days of gestation followed by no treatment (CDH), RA (CDH + RA), TO (CDH + TO), or both (CDH + RA + TO) (n = 16). We measured the median wall thickness of the pulmonary arterioles (MWT) and analyzed the expression of VEGF and its receptors (VEGFR1 and VEGFR2). Compared to control animals, CDH had increased MWT (44 ± 15 vs. 58 ± 7; p

Herein, the independent differential selectivity in the Suzuki-Miyaura reaction of two competing arylboronic acids with regard to the nature of the aryl halide (both halide and aryl moieties) was established. This pattern cannot be realized via the “textbook” mechanism of the reaction, which suggests the interaction of boron-containing species with an ArPdX intermediate (product of the aryl halide catalytic activation). The data obtained indicate the participation of two distinct palladium species in the aryl halide and arylboronic acid activations in the catalytic cycle of biaryl formation, which are in accordance with the so-called cooperative mechanism.