Acetylcholinesterase is involved in the termination of impulse transmission by rapid hydrolysis of the
neurotransmitter acetylcholine in numerous cholinergic pathways in the central and peripheral ...nervous systems. The
enzyme inactivation, induced by various inhibitors, leads to acetylcholine accumulation, hyperstimulation of nicotinic and
muscarinic receptors, and disrupted neurotransmission. Hence, acetylcholinesterase inhibitors, interacting with the
enzyme as their primary target, are applied as relevant drugs and toxins. This review presents an overview of toxicology
and pharmacology of reversible and irreversible acetylcholinesterase inactivating compounds. In the case of reversible
inhibitors being commonly applied in neurodegenerative disorders treatment, special attention is paid to currently
approved drugs (donepezil, rivastigmine and galantamine) in the pharmacotherapy of Alzheimer’s disease, and toxic
carbamates used as pesticides. Subsequently, mechanism of irreversible acetylcholinesterase inhibition induced by
organophosphorus compounds (insecticides and nerve agents), and their specific and nonspecific toxic effects are
described, as well as irreversible inhibitors having pharmacological implementation. In addition, the pharmacological
treatment of intoxication caused by organophosphates is presented, with emphasis on oxime reactivators of the inhibited
enzyme activity administering as causal drugs after the poisoning. Besides, organophosphorus and carbamate insecticides
can be detoxified in mammals through enzymatic hydrolysis before they reach targets in the nervous system.
Carboxylesterases most effectively decompose carbamates, whereas the most successful route of organophosphates
detoxification is their degradation by corresponding phosphotriesterases.
Are uterine fluid-derived extracellular vesicles (UF-EVs) a 'liquid biopsy' reservoir of biomarkers for real-time monitoring of endometrial status?
The transcriptomic cargo of UF-EVs reflects the RNA ...profile of the endometrial tissue as well as changes between the non-receptive and the receptive phase, possibly supporting its use for a novel endometrial receptivity test.
EVs have been previously isolated from uterine fluid, where they likely contribute to the embryo-endometrium crosstalk during implantation. Based on a meta-analysis of studies on endometrial tissue implantation-associated genes and the human exosomes database, 28 of the 57 transcripts considered as receptivity markers refer to proteins present in human exosomes. However, the specific transcriptomic content of receptive phase UF-EVs has yet to be defined.
Two experimental series were set up. First, we simultaneously sequenced RNA species derived from paired UF-EVs and endometrial tissue samples collected from physiologically cycling women. Second, we analyzed RNA species of UF-EVs collected during the non-receptive (LH + 2) and receptive (LH + 7) phase of proven fertile women and from the receptive (LH + 7) phase of a population of women undergoing ART and transfer of euploid blastocysts.
For paired UF-endometrial tissue sampling, endometrial tissue biopsies were obtained with the use of a Pipelle immediately after UF collection performed by lavage of the endometrial cavity. Overall, n = 87 UF samples were collected and fresh-processed for EV isolation and total RNA extraction, while western blotting was used to confirm the expression of EV protein markers of the isolated vesicles. Physical characterization of UF-EVs was performed by Nanoparticle Tracking Analysis. To define the transcriptomic cargo of UF-EV samples, RNA-seq libraries were successfully prepared from n = 83 UF-EVs samples and analyzed by RNA-seq analysis. Differential gene expression (DGE) analysis was used to compare RNA-seq results between different groups of samples. Functional enrichment analysis was performed by gene set enrichment analysis with g:Profiler. Pre-ranked gene set enrichment analysis (GSEA) with WebGestalt was used to compare RNA-seq results with the gene-set evaluated in a commercially available endometrial receptivity array.
A highly significant correlation was found between transcriptional profiles of endometrial biopsies and pairwise UF-EV samples (Pearson's r = 0.70 P < 0.0001; Spearman's ρ = 0.65 P < 0.0001). In UF-EVs from fertile controls, 942 gene transcripts were more abundant and 1305 transcripts less abundant in the LH + 7 receptive versus the LH + 2 non-receptive phase. GSEA performed to evaluate concordance in transcriptional profile between the n = 238 genes included in the commercially available endometrial receptivity array and the LH + 7 versus LH + 2 UF-EV comparison demonstrated an extremely significant and consistent enrichment, with a normalized enrichment score (NES)=9.38 (P < 0.001) for transcripts up-regulated in LH + 7 in the commercial array and enriched in LH + 7 UF-EVs, and a NES = -5.40 (P < 0.001) for transcripts down-regulated in LH + 7 in the commercial array and depleted in LH + 7 UF-EVs. When analyzing LH + 7 UF-EVs of patients with successful versus failed implantation after transfer of one euploid blastocyst in the following cycle, we found 97 genes whose transcript levels were increased and 64 genes whose transcript levels were decreased in the group of women who achieved a pregnancy. GSEA performed to evaluate concordance in transcriptional profile between the commercially available endometrial receptivity array genes and the comparison of LH + 7 UF-EVs of women with successful versus failed implantation, demonstrated a significant enrichment with a NES = 2.14 (P = 0.001) for transcripts up-regulated in the commercial array in the receptive phase and enriched in UF-EVs of women who conceived, and a not significant NES = -1.18 (P = 0.3) for transcripts down-regulated in the commercial array and depleted in UF-EVs. In terms of physical features, UF-EVs showed a homogeneity among the different groups analyzed except for a slight but significant difference in EV size, being smaller in women with a successful implantation compared to patients who failed to conceive after euploid blastocyst transfer (mean diameter ± SD 205.5± 22.97 nm vs 221.5 ± 20.57 nm, respectively, P = 0.014).
Transcriptomic data were deposited in NCBI Gene Expression Omnibus (GEO) and can be retrieved using GEO series accession number: GSE158958.
Separation of RNA species associated with EV membranes might have been incomplete, and membrane-bound RNA species-rather than the internal RNA content of EVs-might have contributed to our RNA-seq results. Also, we cannot definitely distinguish the relative contribution of exosomes, microvesicles and apoptotic bodies to our findings. When considering patients undergoing ART, we did not collect UFs in the same cycle of the euploid embryo transfer but in the one immediately preceding. We considered this approach as the most appropriate in relation to the novel, explorative nature of our study. Based on our results, a validation of UF-EV RNA-seq analyses in the same cycle in which embryo transfer is performed could be hypothesized.
On the largest sample size of human EVs ever analyzed with RNA-seq, this study establishes a gene signature to use for less-invasive endometrial receptivity tests. This report is indeed the first to show that the transcriptome of UF-EVs correlates with the endometrial tissue transcriptome, that RNA signatures in UF-EVs change with endometrial status, and that UF-EVs could serve as a reservoir for potential less-invasive collection of receptivity markers. This article thus represents a step forward in the design of less-invasive approaches for real-time monitoring of endometrial status, necessary for advancing the field of reproductive medicine.
The study was funded by a competitive grant from European Society of Human Reproduction and Embryology (ESHRE Research Grant 2016-1). The authors have no financial or non-financial competing interests to disclose.
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This paper investigates the applicability of two computational intelligence methods for the plasma cutting process modelling: artificial neural networks and adaptive neuro-fuzzy inference systems. ...After exploring the possibilities of neural networks learning from the experimental data for the prediction of the plasma cutting parameter, the adaptive neuro-fuzzy inference system approach was used in order to compare the prediction properties of neural networks and adaptive neuro-fuzzy models. These two methods resulted in the kerf width prediction models for different combinations of input process parameters: cutting current, cutting speed and material thickness, whose significance was assessed by multi-criteria analysis. Descriptive statistics and a visual exploration of two sets of neural-network and adaptive neuro-fuzzy modelled data showed good agreement with the experimental results and their trends. This was additionally confirmed by the t-test and Analysis of Variance, which also provided the selection of the most favourable method for plasma cutting modelling. Accordingly, the adaptive neuro-fuzzy inference systems showed superior modelling capabilities over artificial neural networks for this particular problem setting.
A number of microRNAs have been shown to regulate skeletal muscle development and differentiation. MicroRNA-222 is downregulated during myogenic differentiation and its overexpression leads to ...alteration of muscle differentiation process and specialized structures. By using RNA-induced silencing complex (RISC) pulldown followed by RNA sequencing, combined with in silico microRNA target prediction, we have identified two new targets of microRNA-222 involved in the regulation of myogenic differentiation, Ahnak and Rbm24. Specifically, the RNA-binding protein Rbm24 is a major regulator of muscle-specific alternative splicing and its downregulation by microRNA-222 results in defective exon inclusion impairing the production of muscle-specific isoforms of Coro6, Fxr1 and NACA transcripts. Reconstitution of normal levels of Rbm24 in cells overexpressing microRNA-222 rescues muscle-specific splicing. In conclusion, we have identified a new function of microRNA-222 leading to alteration of myogenic differentiation at the level of alternative splicing, and we provide evidence that this effect is mediated by Rbm24 protein.
Organophosphate pesticides are used in large quantities. Once accumulated in the environment, they exhibit toxic effects on non-target organisms. Dimethoate, a frequently used insecticide, and its ...oxo-analog omethoate inhibit acetylcholinesterase and are toxic for mammals. However, under environmental conditions, they also undergo chemical transformations and decomposition. Nevertheless, the systematic data about dimethoate and omethoate decomposition are missing. We performed a systematic analysis of dimethoate and omethoate decomposition under different pH conditions and estimated their long-term eco-neurotoxic effects. Dimethoate and omethoate decompose rapidly in alkaline aqueous solutions (half-lives 5.7 ± 1.4 and 0.89 ± 0.21 days) but are highly stable in acidic solutions (half-lives 124 ± 18 and 104 ± 9 days). These differences are explained using quantum chemical calculations, indicating that a weaker P–S bond in omethoate is more susceptible to hydrolysis, particularly at a high pH. The toxicity of these pesticide solutions decreases over time, indicating that no or very little of highly more toxic omethoate is formed during hydrolysis of dimethoate, pointing to the advantage of alkaline hydrolysis over other techniques for dimethoate removal. Presented data are used to benchmark dimethoate and omethoate concentrations and toxicity in contaminated water in the pH range 3 to 9 for up to 70 days upon the release in the environment.
Graphical abstract
Cornelia de Lange syndrome (CdLS) and KBG syndrome are two distinct developmental pathologies sharing common features such as intellectual disability, psychomotor delay, and some craniofacial and ...limb abnormalities. Mutations in one of the five genes NIPBL, SMC1A, SMC3, HDAC8 or RAD21, were identified in at least 70% of the patients with CdLS. Consequently, additional causative genes, either unknown or responsible of partially merging entities, possibly account for the remaining 30% of the patients. In contrast, KBG has only been associated with mutations in ANKRD11. By exome sequencing we could identify heterozygous loss‐of‐function mutations in ANKRD11 in two patients with the clinical diagnosis of CdLS. Both patients show features reminiscent of CdLS such as characteristic facies as well as a small head circumference which is not described for KBG syndrome. Patient A, who carries the mutation in a mosaic state, is a 4‐year‐old girl with features reminiscent of CdLS. Patient B, a 15‐year‐old boy, shows a complex phenotype which resembled CdLS during infancy, but has developed to a more KBG overlapping phenotype during childhood. These findings point out the importance of screening ANKRD11 in young CdLS patients who were found to be negative for mutations in the five known CdLS genes.
•UV-C irradiation of chlorpyrifos and its formulations enhances their toxic effects.•Irradiated chlorpyrifos and its formulations induce DNA damage and oxidative stress.•Technical chlorpyrifos is a ...more potent genotoxic inducer than its formulations.•Irradiated chlorpyrifos formulations affect acetylcholinesterase differently.
UV-C irradiation is widely used in the food industry. However, the health effects from dietary exposure to the irradiated pesticide residues retained in foodstuffs are underestimated. In this study, technical chlorpyrifos (TCPF) and its oil in water (EW) and emulsifiable concentrate (EC) formulations were irradiated by UV-C, and their photodegradation products were subjected to toxicity assessment, including determination of acetylcholinesterase (AChE) activity, genotoxicity and oxidative stress using human blood cells as a model system. Toxicity studies were performed using the chlorpyrifos concentrations in the range of those proposed as the maximum residue levels in plant commodities. TCPF, EW and EC photodegradation products induced DNA damage and oxidative stress, and their genotoxicity did not decrease as a function of irradiation time. Irradiated TCPF and EC are more potent AChE inhibitors than irradiated EW. Accordingly, the application of UV-C irradiation must be considered when processing the plants previously treated with chlorpyrifos formulations.
(1) Background: We aimed to analyze the oxidative status of patients with unstable angina pectoris (UA), as well as to determine the correlation of these parameters between coronary arterial and ...peripheral venous blood samples. (2) Methods: The study included 47 human subjects with UA and 45 control subjects. We performed clinical examinations, hemodynamic and coronary angiography measures. Also, in the blood samples, we measured routine laboratory markers and the concentration of pro-oxidants: index of lipid peroxidation (TBARS), superoxide anion radical (O
), hydrogen peroxide (H
O
) and nitrites (NO
), while antioxidant parameters were determined from red blood cells: reduced glutathione (GSH), catalase (CAT) and superoxide dismutase (SOD). All parameters were determined spectrophotometrically. (3) Results: Significantly higher values of TBARS and all measured antioxidants SOD, CAT and GSH were observed in the coronary arterial blood of the UA group relative to coronary arterial blood of the control subjects. On the other hand, in the peripheral venous blood samples, a significantly lower GSH value was found in the UA group compared to the control. (4) Conclusions: This study has shown that the majority of changes in all measured redox markers are found in coronary blood, especially related to the activity of antioxidant components. In patients with an unstable form of angina, prooxidants (superoxide anion radical and index of lipid peroxidation) and endogenous antioxidants (catalase, superoxide dismutase and reduced glutathione) are in direct correlation with the course of ischemic disease. Future studies, where participants would be randomized depending on symptom duration, are necessary to confirm these conclusions.
Multiple Sclerosis (MS) is caused by a still unknown interplay between genetic and environmental factors. Epigenetics, including DNA methylation, represents a model for environmental factors to ...influence MS risk.
Twenty-six affected and 26 unaffected relatives from 8 MS multiplex families were analysed in a multicentric Italian study using MeDIP-Seq, followed by technical validation and biological replication in two additional families of differentially methylated regions (DMRs) using SeqCap Epi Choice Enrichment kit (Roche®).
Associations from MeDIP-Seq across families were combined with aggregation statistics, yielding 162 DMRs at FDR ≤ 0.1. Technical validation and biological replication led to 2 hypo-methylated regions, which point to NTM and BAI3 genes, and to 2 hyper-methylated regions in PIK3R1 and CAPN13.
These 4 novel regions contain genes of potential interest that need to be tested in larger cohorts of patients.
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•The 233 genetic loci associated with MS explain less than 40% of disease heritability, leaving a role to epigenetics in disease risk•We compared whole-genome methylation profiles in whole blood of affected and unaffected relatives of 8 multiplex MS families•We used MeDIP-seq and technical and biological replication in 2 additional families using a custom panel•Due to the heterogeneity of results in families, we adopted a method which leveraged consistency of signal across families•Filtering criteria lead to 2 hypo- and 2 hyper-methylated DMRs which relate to NTM, BAI3, PIK3R1 and CAPN13 genes•Replication of these signals is needed in additional cohort of MS patients
The aim of this work was to investigate the different types of radiation detectors commonly used for radiation protection purposes as survey meters. The study was performed on survey meters that use ...different detectors as ionisation chamber, Geiger Mueller (GM) counter and scintillation detector. For each survey meter, energy dependence and angular response in X- and gamma-radiation fields was tested. The following commercially available survey meters were investigated: ionisation chambers Victoreen 451P, Babyline 31 and VA-J-15A, Geiger counter MRK-M87, 6150 AD6 and FAG FH 40F2 and scintillation counter 6150 ADB. As a source of gamma radiation, (137)Cs and (60)Co were used whereas X-ray radiation fields were generated using an X-ray unit. The radiation characteristics of the survey meters were mostly in compliance with references estimated by standard IEC 1017-2. However, some of them showed larger deviation at lower energies. GM counters exhibit strong energy dependence for low-energy photons.
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