Advances in CRISPR-Cas systems for epigenetics.

The CRISPR-Cas9 method has revolutionized the gene editing. Epigenetic changes, including DNA methylation, RNA modification, and changes in histone proteins, have been intensively studied and found to play a key role in the pathogenesis of human diseases. CRISPR-While the utility of DNA and chromatin modifications, known as epigenetics, is well understood, the functional significance of various alterations of RNA nucleotides has recently gained attention. Recent advancements in improving CRISPR-based epigenetic modifications has resulted in the availability of a powerful source that can selectively modify DNA, allowing for the maintenance of epigenetic memory over several cell divisions. Accurate identification of DNA methylation at specific locations is crucial for the prompt detection of cancer and other diseases, as DNA methylation is strongly correlated to the onset as well as the advancement of such conditions. Genetic or epigenetic perturbations can disrupt the regulation of imprinted genes, resulting in the development of diseases. When histone code editors and DNA de-/ methyltransferases are coupled with catalytically inactive Cas9 (dCas9), and CRISPRa and CRISPRi, they demonstrate excellent efficacy in editing the epigenome of eukaryotic cells. Advancing and optimizing the extracellular delivery platform can, hence, further facilitate the manipulation of CRISPR-Cas9 gene editing technique in upcoming clinical studies. The current chapter focuses on how the CRISP/ Cas9 system provides an avenue for the epigenetic modifications and its employability for human benefit.

A Meta Modeling-Based Interoperability and Integration Testing Platform for IoT Systems.

The rapid growth of the Internet of Things (IoT) and its integration into various industries has made it extremely challenging to guarantee IoT systems' dependability and quality, including scalability, dynamicity, and integration with existing IoT frameworks. However, the essential principles, approaches, and advantages of model-driven IoT testing indicate a promising strategy for overcoming these. This paper proposes a metamodeling-based interoperability and integration testing approach for IoT systems that automates the creation of test cases and the assessment of system performance by utilizing formal models to reflect the behavior and interactions of IoT systems. The proposed model-based testing enables the systematic verification and validation of complex IoT systems by capturing the essential characteristics of IoT devices, networks, and interactions. This study describes the key elements of model-driven IoT testing, including the development of formal models, methods for generating test cases, and the execution and assessment of models. In addition, it examines various modeling formalisms and their use in IoT testing, including state-based, event-driven, and hybrid models. This study examines several methods for creating test cases to ensure thorough and effective testing, such as constraint-based strategies and model coverage requirements. Model-driven IoT testing improves defect detection, expands test coverage, decreases testing effort, and increases system reliability. It also offers an organized and automated method to confirm the efficiency and dependability of IoT systems.

Synthesis and biological evaluation of carboxamide and quinoline derivatives as P2X7R antagonists.

P2X7 receptor (P2X7R) has a key role in different pathological conditions, importantly overexpressed and activated in cancers. We explored the structure activity relationship (SAR) of three novel pyrazines, quinoline-carboxamide and oxadiazole series. Their selective inhibitory potency in Ca2+ mobilization assay using h-P2X7R-MCF-7 cells improved with phenyl ring substitutions (-OCF3, -CF3, and -CH3) in carboxamide and oxadiazole derivatives, respectively. However, highly electronegative fluoro, chloro, and iodo substitutions enhanced affinity. 1e, 2f, 2e, 1d, 2 g and 3e were most potent and selective toward h-P2X7R (IC50 values 0.457, 0.566, 0.624, 0.682, 0.813 and 0.890 µM, respectively) and were inactive at h-P2X4R, h-P2X2R, r-P2Y6R, h-P2Y2R, t-P2Y1R expressed in MCF-7 and 1321N1 astrocytoma cells. Cell viability (MTT assay at 100 µM, cell line) for 3e was 62% (HEK-293T), 70% (1321N1 astrocytoma) and 85% (MCF-7). >75% cell viability was noted for 2 g and >80% for 2e and 1d in all non-transfected cell lines. Anti-proliferative effects, compared to control (Bz-ATP), of selective antagonists (10 µM) were 3e (11%) 1d, (19%) 1e, (70%, P = 0.005) and 2f, (24%), indicating involvement of P2X7R. Apoptotic cell death by flow cytometry showed 1e to be most promising, with 35% cell death (PI positive cells), followed by 2e (25%), 2f (20%), and 1d (19%), compared to control. Fluorescence microscopic analysis of apoptotic changes in P2X7R-transfected cell lines was established. 1e and 2f at 1X and 2X IC50 increased cellular shrinkage, nuclear condensation and PI/DAPI fluorescence. In-silico antagonist modeling predicted ligand receptor interactions, and all compounds obeyed Lipinski rules. These results suggest that pyrazine, quinoline-carboxamide and oxadiazole derivatives could be moderately potent P2X7R antagonists for in vivo studies and anti-cancer drug development.

Influence of access to clean fuels and technology, food production index, consumer price index, and income on greenhouse gas emissions from food system: evidence from developed countries.

Sustainable development objectives heavily promote the advancement of cleaner production technologies to reduce emissions and conserve the average world temperature. For the years 1990-2020, the USA, China, Japan, Russia, Germany, and Australia are studied by using the panel fully modified ordinary least square (FMOLS). The results show that clean fuels and technologies and a consumer price index are helpful to reduce greenhouse gas emissions from food system which reduce environmental degradation. Contrarily, increased income and food production contribute to environmental deterioration. There are bidirectional Dumitrescu-Hurlin causal relationships between access to clean fuels and technology and greenhouse gas emissions from food system; real income and greenhouse gas emissions from food system; income and access to clean fuels and technology; income and consumer price index; and income and food production index. This research also revealed a unidirectional causation between the consumer price index and greenhouse gas emissions from food system; food production index and greenhouse gas emissions from food system; access to clean fuels and technology and the consumer price index; and access to clean fuels and technology and the food production index. These findings provide policymakers with relevant content: to promote the goal of green growth, the government should implement consistent measures to subsidize the food industry. Incorporating carbon pricing into food system emissions models would serve to lower production of polluting foods, which would enhance air quality indicators. Finally, a consumer price index should be controlled by controlling prices of green technologies in environmental modeling to improve sustainable development globally and reduce environmental pollution.

Determination of possible contraceptive potential of methanolic leaf extract of Mentha longifolia L. in adult male rats: a biochemical and histological study.

Mentha longifolia L (ML) is locally used as an herbal contraceptive. We aim to assess the toxicity profile and validate the antifertility effects of its leaves in male rats. Adult male Sprague Dawley rats (n = 10 animals/group) were divided into four groups. The Control group received .9% saline, while groups II, III, and IV received 50, 75, and 100 mg/kg/day ML doses for 28 days. ML induced a dose-dependent decrease in percent fertility as well as the number of pups born in ML preexposed pairs, with maximum change observed in the ML3 group (60%, 3.75 ± 1.25) as compared to control (100%, 8.75 ± .48) group. A significant (P < .01) decrease in the rate of daily sperm production was observed in the ML3 group (.86 ± .16) in contrast to the control (2.93 ± .05). The levels of catalase and superoxide dismutase declined significantly in treatment groups (ML1-P < .001, ML2-P < .05, and ML3-P < .01). A significant increase in the production of reactive oxygen species (P < .001) was found in all treatment groups. The concentrations of LH (P < .001), FSH (P < .001), and testosterone (P < .001) were significantly reduced in a dose-dependent manner among ML-treated groups. Decreases in spermatogonial populations, mature spermatids, seminiferous tubule diameter, lumen diameter, and epithelial height were noticed at higher doses. The reversibility study depicted a reduction in both pregnancy outcomes and litter size. Besides contraceptive effects, M. longifolia L usage is associated with oxidative stress; therefore, it is important to consume this herb appropriately and its excessive usage should be prohibited.

Synthesis and Evaluation of Amide and Thiourea Derivatives as Carbonic Anhydrase (CA) Inhibitors.

Series of sulfonamide-substituted amide (9-11), benzamide (12-15), and 1,3-disubstituted thiourea (17-26) derivatives were synthesized from a common precursor, i.e., substituted benzoyl chlorides. Structures of all of the synthesized compounds were characterized by spectroscopic techniques (1H nuclear magnetic resonance (NMR),13C NMR, and Fourier transform infrared spectroscopy (FTIR)). All of the amide (9-15) and thiourea (17-26) derivatives were screened against human carbonic anhydrases, hCA-II, hCA IX, and hCA-XII. Sulfonamide-substituted amides 9, 11, and 12 were found to be excellent selective inhibitors with IC50 values of 0.18 ± 0.05, 0.17 ± 0.05, and 0.58 ± 0.05 µM against hCA II, hCA IX, and hCA XII, respectively. Compound 9 was found to be highly selective for hCA II and about 6-fold more potent as compared to the standard antagonist, acetazolamide. Safe toxicity profiling of the most potent and selective compounds was determined against normal BHK-21 and HEK-293 T cells. Molecular docking studies were performed, which described the type of interactions between the synthesized compounds and enzyme proteins. In addition, in silico absorption, distribution, metabolism, and excretion (ADME) studies were performed, which showed that all of the synthesized molecules fulfilled the druggability criteria.

Influence of green technology, green energy consumption, energy efficiency, trade, economic development and FDI on climate change in South Asia.

Climate change policy has several potential risks. The purpose of this study is to investigate the impact of green technology development, green energy consumption, energy efficiency, foreign direct investment, economic growth, and trade (imports and exports) on greenhouse gas (GHG) emissions in South Asia from 1981 to 2018. We employed Breusch Pagan LM, bias-corrected scaled LM, and Pesaran CD as part of a series of techniques that can assist in resolving the problem of cross-sectional dependence. First and second generation unit root tests are used to assess the stationarity of the series, Pedroni and Kao tests are used to test co-integration. The long-term associations are examined using fully modified ordinary least square (FMOLS) and panel dynamic ordinary least square (DOLS) for robustness. The results revealed that trade, growth rate, and exports significantly increase GHG emissions. This accepted the leakage phenomenon. The results also demonstrated that green technology development, green energy consumption, energy efficiency, and imports all have a significant negative correlation with GHG emissions. Imports, advanced technical processes, a transition from non-green energy to green energy consumption, and energy efficiency are thus critical components in executing climate change legislation. These findings highlight the profound importance of green technology development and green energy for ecologically sustainable development in the South Asian countries and act as a crucial resource for other nations throughout the world when it comes to ecological security. This research recommends the consumption of environmentally friendly and energy-efficient technologies in order to mitigate climate change and the government's implementation of the most recent policies to neutralize GHG emissions in order to achieve sustainable development.

Comparison of the Efficacy and Safety of Apixaban and Warfarin in the Prevention of Stroke in Patients With Non-valvular Atrial Fibrillation: A Meta-Analysis.

Atrial fibrillation is an irregular heart rhythm, and it is one of the most common cardiac arrhythmias. It is associated with a five times increase in the risk of stroke. Anti-coagulants are prescribed routinely to prevent strokes, especially in patients with atrial fibrillation for many years decreasing the risk of stroke among patients with atrial fibrillation. Non-vitamin K oral anticoagulants especially apixaban and rivaroxaban are frequently used and they are considered to be safe and more effective than warfarin. The aim of this meta-analysis is to compare the efficacy and safety of apixaban and warfarin in preventing stroke among patients with non-valvular arterial fibrillation. The current meta-analysis was conducted using the guidelines established by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). A systematic search was done using databases, including PubMed, EMBASE, and Cochrane Library, with no restrictions on language and year of publication. The current meta-analysis included randomized control trials and non-randomized control trials (prospective and retrospective cohort studies) comparing the efficacy and safety of apixaban and warfarin in preventing stroke in patients with non-valvular atrial fibrillation. The primary efficacy outcome was stroke or systemic embolism while the primary safety outcome was major bleeding events. Overall, nine articles were included in the current meta-analysis with a pooled sample size of 267998 patients with non-valvular atrial fibrillation. The administration of apixaban was associated with a significant decrease in stroke or systemic embolism (RR: 0.77, 95% CI: 0.67-0.90) and major bleeding events (RR=0.63, 95% CI: 0.58-0.68) as compared to warfarin. However, no significant difference was reported in all-cause mortality (RR=0.80, 95% CI: 0.30-2.14) between the two groups. The current meta-analysis concluded that apixaban, compared to warfarin in patients with non-valvular atrial fibrillation showed a reduction in stroke and systemic embolism. Apixaban has also a better safety profile in terms of reduction in overall major bleeding events.

Synthesis and pharmacological characterization of multiply substituted 2H-chromene derivatives as P2Y6 receptor antagonists.

P2Y6 receptor (P2Y6R) antagonists represent potential drugs for treating cancer, pain, neurodegeneration, asthma, diabetes, colitis and other disorders. However, there are few chemical classes of known competitive antagonists. We recently explored the structure activity relationship (SAR) of 2H-chromene derivatives as P2Y6R antagonists of moderate affinity. New analogues in this series modified at five positions were synthesized and shown to antagonize Ca2+ transients induced by the native agonist UDP in human (h) P2Y6R-expressing (but not turkey P2Y1R-, hP2Y2R- or hP2Y4R-expressing) astrocytoma cells. Alternatives to the reported 2-(trifluoromethyl)- and 3-nitro- substitutions of this scaffold were not identified. However, 6­fluoro 11 and 6­chloro 12 analogues displayed enhanced potency compared to other halogens, although still in the 1 - 2 µM range. Similar halogen substitution at 5, 7 or 8 positions reduced affinity. 5- or 8­Triethylsilylethynyl extension maintained hP2Y6R affinity, with IC50 0.46 µM for 26 (MRS4853). The 6,8­difluoro analogue 27 (IC50 2.99 µM) lacked off-target activities among 45 sites examined, unlike earlier analogues that bound to biogenic amine receptors. 11 displayed only one weak off-target activity (σ2). Mouse P2Y6R IC50s of 5, 25, 26 and 27 were 4.94, 17.6, 6.15 and 17.8 µM, respectively, but most other analogues had reduced affinity (>20 µM) compared to the hP2Y6R. These analogues are suitable for evaluation in in vivo inflammation and cancer models, which will be performed in the future studies.

Efficacy and Safety of Colchicine in Prevention of Secondary Cardiovascular Outcomes Among Patients With Coronary Vessel Disease: A Meta-Analysis.

Coronary vessel disease (CVD) is a class of diseases that impacts the blood vessels and heart and is one of the leading causes of disability and death. CVD includes cerebrovascular disease and coronary heart disease, both illnesses of the vessels transporting the oxygenated blood to the brain or heart. Colchicine is an inexpensive and old drug with strong anti-inflammatory effects. Numerous randomized control trials (RCTs) have demonstrated the effectiveness of low-dose colchicine for the prevention of severe cardiovascular events without showing any signs of serious adverse effects within the regime of treatment. In the current meta-analysis, we aim to assess the efficacy and safety of colchicine for secondary cardiovascular outcome prevention among patients with clinically proven CVD. The current meta-analysis was carried out using the preferred reporting items for systematic reviews and meta-analysis (PRISMA) guidelines. PUBMED, Cochrane, and EMBASE databases were used to search for RCTs comparing colchicine and placebos for the prevention of secondary cardiovascular outcomes. The primary efficacy endpoint was mortality due to cardiovascular disease, stroke, urgent coronary revascularization, and myocardial infarction. Secondary efficacy outcomes included death due to all-cause mortality. Seven RCTs were reviewed, with a pooled sample size of 12114, out of which 6099 were randomized to the colchicine group, and 6015 were randomized to the control group. The decrease in cardiovascular events, including myocardial infarction, stroke, urgent coronary revascularization, and cardiac-related death, was significantly lower in patients randomized to colchicine (p-value<0.05). The incidence of safety outcomes did not vary significantly different between groups (p>0.05). In patients with CVD, compared to standard medical therapy, colchicine significantly decreases the risk of cardiovascular events such as cardiovascular-related death, myocardial infarction, stroke, and urgent coronary revascularizations.

Bridged Piperidine Analogues of a High Affinity Naphthalene-Based P2Y14R Antagonist.

High affinity phenyl-piperidine P2Y14R antagonist 1 (PPTN) was modified with piperidine bridging moieties to probe receptor affinity and hydrophobicity. Various 2-azanorbornane, nortropane, isonortropane, isoquinuclidine, and ring-opened cyclopentylamino derivatives preserved human P2Y14R affinity (fluorescence binding assay), and their pharmacophoric overlay was compared. Enantiomeric 2-azabicyclo[2.2.1]hept-5-en-3-one precursors assured stereochemically unambiguous, diverse products. Pure (S,S,S) 2-azanorbornane enantiomer 15 (MRS4738) displayed higher affinity than 1 (3-fold higher affinity than enantiomer 16) and in vivo antihyperallodynic and antiasthmatic activity. Its double prodrug 143 (MRS4815) dramatically reduced lung inflammation in a mouse asthma model. Related lactams 21-24 and dicarboxylate 42 displayed intermediate affinity and enhanced aqueous solubility. Isoquinuclidine 34 (IC50 15.6 nM) and isonortropanol 30 (IC50 21.3 nM) had lower lipophilicity than 1. In general, rigidified piperidine derivatives did not lower lipophilicity dramatically, except those rings with multiple polar groups. P2Y14R molecular modeling based on a P2Y12R structure showed stable and persistent key interactions for compound 15.

Antifertility effect of methanolic leaf extract of Chenopodium ambrosioides Hook. in male Sprague Dawley rats.

This study evaluated the antifertility activity of methanolic extract of Chenopodium ambrosioides leaf on male rats. During the experiment, different doses of extract (0, 50, 100 and 150 mg/kg) were orally administered to rats for 28 days. Analysis of sperm parameters revealed a dose-dependent decrease in sperm motility, viability and daily sperm production (DSP). While, increased oxidative stress in reproductive organs; and impaired testicular and epididymal histology was also evident in high dose regimen. Furthermore, a reduction in concentrations of plasma testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) was also recorded. Reduced pregnancy outcome and small litter size in the females paired with treated male rats after 30 days of treatment withdrawn was noted in higher doses. From these findings, it is concluded that the methanolic leaf extract of C. ambrosioides is quite effective in reversible suppression of male fertility.

The burden and high prevalence of hypertension in Pakistani adolescents: a meta-analysis of the published studies.

BACKGROUND: Hypertension has been recognized as a global health concern for developing countries and is scarcely described in many of these countries. In Pakistan, few population-based surveys evaluated the prevalence of hypertension and there is no current nationally representative study (the latest nationwide survey was conducted more than two decades ago). Objective: The goal of the current study was to estimate the pooled prevalence of hypertension in Pakistani population using meta-analysis approach. METHODS: We searched the published literature using PubMed, Google and Scopus supplemented by a manual search of bibliographies of retrieved articles for population studies providing estimates on the prevalence of hypertension between 1990 and 2017. Studies were included if they defined hypertension as ≥140/90 mmHg and conducted in adults (≥15 years). From the extracted results, the heterogeneity index of the studies was determined using Chi-squared I2 tests and on the basis of heterogeneity, a fixed or random effect model was used to estimates the pooled prevalence of hypertension. Meta-regression was performed to determine those factor of generating heterogeneity. RESULTS: Of a total of 1240 articles, 18 studies comprising 42,618 participants met the eligibility criteria. The overall pooled prevalence of hypertension was 26.34% (25.93%, 26.75%). Subgroup analysis showed higher urban prevalence 26.61% (21.80%, 31.42%) than the rural dwellers 21.03% (10.18%, 31.87%). The prevalence by decade in 1990s was 19.55% (18.07%, 21.05%), in 2000s 23.95% (16.60%, 31.30%) and in 2010s 29.95% (24.13%, 35.77%). Similarly, the pooled prevalence was 24.99% (19.70%, 30.28%) in males and 24.76% (16.76%, 32.76%) in females. We recorded high burden of hypertension among the adult Pakistanis when compared to the data published in local and international journals 23.32% (18.9%, 27.74%) and 27.44% (20.97%, 33.91%). We also found differences in the prevalence of hypertension among small, medium and large studies. CONCLUSION: Comparing data from previous studies in Pakistan, we found a higher prevalence in urban areas and among males. The prevalence over time is likely to increase faster, further our results underscore the importance of good quality long-term studies that will help to understand hypertension better and implement effective prevention and management programs.