Effects of chlorpyrifos and chlorpyrifos-methyl on the outgrowth of axon-like processes, tubulin, and GAP-43 in N2a cells

The aim of this work was to study the neurodegenerative effects of the organophosphate (OP) pesticides chlorpyrifos (CPF) and chlorpyrifos-methyl (CHM) on cultured mouse N2a neuroblastoma cells. CPF or CHM, at a subcytotoxic concentration of 3 μM, were added to the cells either at the time of the induction of cell differentiation (codifferentiation) or 16 h after the induction of differentiation (postdifferentiation). CPF and CHM were similar in inhibiting significantly the outgrowth of axon-like processes from N2a cells after only 4 h exposure under both co- and postdifferentiation exposure conditions. Densitometric scanning of Western blots of extracts of cells treated with CPF or CHM for 4 h revealed significantly decreased cross-reactivity with a monoclonal antibody recognizing the protein GAP-43 under post- but not under codifferentiation exposure conditions. Exposure to CPF or CHM for 4 h under postdifferentiation conditions also resulted in reduced fluorescence of N2a cell body staining with anti-GAP-43. Cross-reactivity of Western blots with a monoclonal antibody recognizing -tubulin was not significantly affected by OP treatment. These data indicate that a disturbance in GAP-43 may be important in the retraction of axons in predifferentiated N2a cells and support the notion that the mechanisms involved in CPF-and CHM-induced inhibition of axonal outgrowth may be different under co- and postdifferentiation exposure conditions

Diazoxon disrupts the expression and distribution of βIII-tubulin and MAP 1B in differentiating N2a cells

This study aimed at assessing the effects of diazoxon (DZO), a major metabolite of the insecticide diazinon (DZ), on key cytoskeletal proteins in differentiating N2a neuroblastoma cells. Initial experiments established that sub-lethal concentrations of 1, 5 and 10 μM DZO produced profound inhibition of neurite outgrowth. Densitometric scanning of probed immunoblots of N2a cell lysates demonstrated that DZO had no effect on total β-tubulin levels. However, probing with a monoclonal antibody that recognised specifically the βIII-tubulin isotype revealed that 10 μM DZO induced a significant reduction in the levels of this particular form. Levels of polyglutamylated tubulin were not altered. Exposure to 10 μM DZO also decreased the expression of microtubule associated protein 1B (MAP 1B). However, DZO had no effect on the expression of MAP tau. DZO also failed to affect the levels neurofilament light (NFL) and neurofilament medium (NFM) chain levels. Indirect immunofluorescence demonstrated that the staining of neurites in treated cells was weaker than in the controls for βIII-tubulin. In conclusion, DZO disrupts the microtubule (MT) network affecting the expression and distribution of two specific MT proteins known to be important in neuritogenesis. DZO may contribute to the developmental neurotoxicity seen following exposure to DZ

Inhibition of neurite outgrowth in differentiating mouse N2a neuroblastoma cells by phenyl saligenin phosphate: Effects on MAP kinase (ERK 1/2) activation, neurofilament heavy chain phosphorylation and neuropathy target esterase activity

Sub-lethal concentrations of the organophosphate phenyl saligenin phosphate (PSP) inhibited the outgrowth of axon-like processes in differentiating mouse N2a neuroblastoma cells (IC50 2.5 μM). A transient rise in the phosphorylation state of neurofilament heavy chain (NFH) was detected on Western blots of cell extracts treated with 2.5 μM PSP for 4 h compared to untreated controls, as determined by a relative increase in reactivity with monoclonal antibody Ta51 (anti-phosphorylated NFH) compared to N52 (anti-total NFH). However, cross-reactivity of PSP-treated cell extracts was lower than that of untreated controls after 24 h exposure, as indicated by decreased reactivity with both antibodies. Indirect immunofluorescence analysis with these antibodies revealed the appearance of neurofilament aggregates in the cell bodies of treated cells and reduced axonal staining compared to controls. By contrast, there was no significant change in reactivity with anti-a tubulin antibody B512 at either time point. The activation state of the MAP kinase ERK 1/2 increased significantly after PSP treatment compared to controls, particularly at 4 h, as indicated by increased reactivity with monoclonal antibody E-4 (anti-phosphorylated MAP kinase) but not with polyclonal antibody K-23 (anti-total MAP kinase). The observed early changes were concomitant with almost complete inhibition of the activity of neuropathy target esterase (NTE), one of the proposed early molecular targets in organophosphate-induced delayed neuropathy (OPIDN)

Designing Solar Still and Study it\u27s Operating Parameters for Avoiding Chemical Health Risk: A Review

Life depends on water, and in recent years, the importance of  having a sufficient supply of drinkable water cannot be overstated. The traditional methods of desalination necessitate for toxic salt removal expenditure in order to transform chemical toxic brackish water into potable pure water for human health and industrial use with avoiding chemical health risk . After decades of intensive research on a variety of desalination methods, solar desalination is one of the most promising approaches to harnessing renewable sources to provide the human health world with high-quality pure water from chemical toxic water . The necessity of the hour is a small-scale, self-contingent desalination equipment .A cutting-edge tool called a solar still uses solar energy to distil chemical toxic brackish water into pure water for avoiding chemical health risk. Approximately 97% of the water on Earth is chemical toxic brackish or saline salts, making it unfit for human health issues. There are several distillation processes available to turn salty or chemical toxic brackish water into drinkable water using various approaches. Nevertheless, the performance of basin-type solar stills is limited by certain factors, and various research, both experimental and numerical, have explored ways to improve both design and operational techniques. As a result, there has been little use of the most current theoretical initiatives and concepts addressing this issue. This article aims to give a thorough overview of the thermal models created for different kinds of solar stills and the adjustments made to enhance their functionality over time. Our research suggests that a few more factors and design considerations should be made while creating a new solar still. This study is effective because it gives energy researchers ideas into designing solar stills for the production of clean water from chemical toxic brackish or saline salts , which in turn encourages the commercialization of this product for rural human health development by avoiding chemical health risk . Lastly, a broad plan of action is provided for choosing a solar still with a versatile, reliable design. There are also suggestions for additional research included. DOI: https://doi.org/10.52783/jchr.v14.i3.519

Chandrayaan-3 Alternate Landing Site: Pre-Landing Characterisation

India's third Moon mission Chandrayaan 3 will deploy a lander and a rover at a high latitude location of the Moon enabling us to carry out first ever in-situ science investigations of such a pristine location that will potentially improve our understanding on primary crust formation and subsequent modification processes. The primary landing site (PLS), is situated at 69.367621 degS, 32.348126 degE. As a contingency, an alternate landing site (ALS) was also selected at nearly the same latitude but nearly 450 km west to PLS. In this work, a detailed study of the geomorphology, composition, and temperature characteristics of ALS has been carried out using the best-ever high resolution Chandrayaan 2 OHRC DEMs and Ortho images, datasets obtained from Chandrayaan 1 and on-going Lunar Reconnaissance Orbiter. For understanding the thermophysical behaviour, we used a well-established thermophysical model. We found that the Chandrayaan 3 ALS is characterised by a smooth topography with an elevated central part. The ALS is a scientifically interesting site with a high possibility of sampling ejecta materials from Tycho and Moretus. Based on the spectral and elemental analysis of the site, Fe is found to be near approx. 4.8 wt.%, with Mg approx. 5 wt.%, and Ca approx. 11 wt.%. Compositionally, ALS is similar to PLS with a highland soil composition. Spatial and diurnal variability of around 40 K and 175 K has been observed in the surface temperatures at ALS. Although belonging to similar location like PLS, ALS showed reduced daytime temperatures and enhanced night-time temperatures compared to PLS, indicating a terrain of distinctive thermophysical characteristics. Like PLS, ALS is also seems to be an interesting site for science investigations and Chandrayaan 3 is expected to provide new insights into the understanding of lunar science even if it happens to land in the alternate landing site.Comment: 13 pages, 7 figure

Beyond chemicals: opportunities and challenges of integrating non-chemical stressors in adverse outcome pathways

The adverse outcome pathways (AOPs) were developed to accelerate evidence-based chemical risk assessment by leveraging data from new approach methodologies. Thanks to their stressor-agnostic approach, AOPs were seen as instrumental in other fields. Here, we present AOPs that report non-chemical stressors along with the challenges encountered for their development. Challenges regarding AOPs linked to nanomaterials include non-specific molecular initiating events, limited understanding of nanomaterial biodistribution, and needs for adaptations of the in silico modeling and testing systems. Development of AOPs for radiation face challenges in how to incorporate ionizing events type, dose rate, energy deposition, and how to account for targeting multiple macromolecules. AOPs for COVID-19 required the inclusion of SARS-CoV-2-specific replicative steps to capture the essential events driving the disease. Developing AOPs to evaluate efficacy and toxicity of cell therapies necessitates addressing the cellular nature and the therapeutic function of the stressor. Finally, addressing toxicity of emerging biological stressors like microbial pesticides can learn from COVID-19 AOPs. We further discuss that the adaptations needed to expand AOP applicability beyond chemicals are mainly at the molecular and cellular levels while downstream key events at tissue or organ level, such as inflammation, are shared by many AOPs initiated by various stressors. In conclusion, although it is challenging to integrate non-chemical stressors within AOPs, this expands opportunities to account for real-world scenarios, to identify vulnerable individuals, and to bridge knowledge on mechanisms of adversity.info:eu-repo/semantics/publishedVersio

Building an adverse outcome pathway network for COVID-19

The COVID-19 pandemic generated large amounts of data on the disease pathogenesis leading to a need for organizing the vast knowledge in a succinct manner. Between April 2020 and February 2023, the CIAO consortium exploited the Adverse Outcome Pathway (AOP) framework to comprehensively gather and systematically organize published scientific literature on COVID-19 pathology. The project considered 24 pathways relevant for COVID-19 by identifying essential key events (KEs) leading to 19 adverse outcomes observed in patients. While an individual AOP defines causally linked perturbed KEs towards an outcome, building an AOP network visually reflect the interrelatedness of the various pathways and outcomes. In this study, 17 of those COVID-19 AOPs were selected based on quality criteria to computationally derive an AOP network. This primary network highlighted the need to consider tissue specificity and helped to identify missing or redundant elements which were then manually implemented in the final network. Such a network enabled visualization of the complex interactions of the KEs leading to the various outcomes of the multifaceted COVID-19 and confirmed the central role of the inflammatory response in the disease. In addition, this study disclosed the importance of terminology harmonization and of tissue/organ specificity for network building. Furthermore the unequal completeness and quality of information contained in the AOPs highlighted the need for tighter implementation of the FAIR principles to improve AOP findability, accessibility, interoperability and re-usability. Finally, the study underlined that describing KEs specific to SARS-CoV-2 replication and discriminating physiological from pathological inflammation is necessary but requires adaptations to the framework. Hence, based on the challenges encountered, we proposed recommendations relevant for ongoing and future AOP-aligned consortia aiming to build computationally biologically meaningful AOP networks in the context of, but not limited to, viral diseases.info:eu-repo/semantics/publishedVersio

Workgroup Report: Incorporating In Vitro Alternative Methods for Developmental Neurotoxicity into International Hazard and Risk Assessment Strategies

This is the report of the first workshop on Incorporating In Vitro Alternative Methods for Developmental Neurotoxicity (DNT) Testing into International Hazard and Risk Assessment Strategies, held in Ispra, Italy, on 19–21 April 2005. The workshop was hosted by the European Centre for the Validation of Alternative Methods (ECVAM) and jointly organized by ECVAM, the European Chemical Industry Council, and the Johns Hopkins University Center for Alternatives to Animal Testing. The primary aim of the workshop was to identify and catalog potential methods that could be used to assess how data from in vitro alternative methods could help to predict and identify DNT hazards. Working groups focused on two different aspects: a) details on the science available in the field of DNT, including discussions on the models available to capture the critical DNT mechanisms and processes, and b) policy and strategy aspects to assess the integration of alternative methods in a regulatory framework. This report summarizes these discussions and details the recommendations and priorities for future work