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This book is composed of four papers prepared to illuminate the problem areas which might arise if the policies of the 1925 Geneva Protocol and other measures to limit chemical and biological weapons are ratified by the United States Senate. The papers included are: Legal Aspects of the Geneva Protocol of 1925; The Use of Herbicides in War: A Political/Military Analysis; The Military Value and Political Implications of the Use of Riot Control Agents in Warfare; and Limitations on Chemical and Biological Warfare Going Beyond Those of the Geneva Protocol. A list of references is included with each paper. Comments on the papers by an advisory group are included, and the appendix contains a copy of the Geneva Protocol of 1925, a copy of the United Kingdom Draft Convention on biological warfare, and statements by President Nixon. (PR)

Carbohydrazides and their Schiff bases are important classes of heterocycles that are not only employed in the area of organic chemistry but also have tremendous applications in physical and inorganic chemistry. A series of potentially bioactive compounds containing carbohydrazide functionality and their hydrazone derivatives have been synthesized and screened for antibacterial, anticancer, antifungal and anti-inflammatory, etc. This brief review discloses some synthetic routes to so many reported carbohydrazides, their Schiff bases, their biological activities, and their structure-activity relationship.

Background Celiac disease is an autoimmune disorder that causes an intolerance to gluten. Owing to hidden sources, lack of clear labeling, and cross-contamination, it is not uncommon for individuals with celiac disease to inadvertently ingest gluten. A strict gluten-free diet is the only treatment. Objectives The purpose of this study was to identify the frequency of activated charcoal use in celiac disease as a purported remedy despite the lack of literature on the safety and efficacy of the practice and to elucidate the manner in which using activated charcoal as a treatment for acute gluten ingestion is being communicated. Methods Using a descriptive study design, a Web-based survey was sent to members of the celiac disease community via social media pages for voluntary participants 18 years and older diagnosed with celiac disease. Participants were stratified into 3 major groups: (1) those who had heard of using activated charcoal as a remedy to counteract gluten ingestion and used it, (2) those who had heard of using activated charcoal but had not used it, and (3) those who had not heard of using activated charcoal. Results In subjects with celiac disease, 424 of 1613 respondents (26%) had heard of using activated charcoal as a potential remedy for gluten ingestion, and 12% had used it. The top source of information regarding any general information including remedies for maladies related to celiac disease was social media networks 72.3% (879/1613). Of those who had used activated charcoal, 122 participants (61.3%) reported subjectively feeling a reduced severity of symptoms. Conclusion Pharmacists should be aware of the off-label use of activated charcoal for accidental gluten ingestion in celiac disease. Pharmacists should educate patients with celiac disease that there is not sufficient evidence to support the use of activated charcoal.

To evaluate the noninferiority of intraocular pressure (IOP)-lowering latanoprost without benzalkonium chloride (BAK) versus latanoprost with BAK (for treatment of open-angle glaucoma or ocular hypertension).Overall, 578 patients were randomized 1:1 to latanoprost without BAK or latanoprost with BAK once daily in the affected eye(s) for 12 weeks. The primary efficacy endpoint was IOP, measured on days 0, 7, 28, 56, and 84 (8 am, 10 am, and 4 pm). Noninferiority was established if the following criteria were met: 95% confidence interval (CI) of the mean difference between treatments included 0 mm Hg for all time points (N1), 95% CI upper limit less than 1.5 mm Hg (N2), and less than 1 mm Hg for≥7 of 12 time points (N3). Primary efficacy analysis was performed on the intent-to-treat population. Safety measurements included ocular and systemic adverse event (AE).The 95% CI included 0 mm Hg for 7/12 time points (N1), 95% CI upper limit was less than 1.5 mm Hg for 12/12 time points (N2), and less than 1.0 mm Hg for 4/7 time points (N3). AEs were mild and similarly distributed between groups.Latanoprost without BAK did not meet two of three criteria for noninferiority and showed a similar safety profile relative to latanoprost with BAK.

Low-expression levels remain a challenge in the quest to use the small laccase (rSLAC) as a viable catalyst. In this study, a recombinant Pichia pastoris strain (rSLAC-GAP-AOX) producing rSLAC under both AOX and GAP promoters (located in two different plasmids) was generated and cultivated in the presence of methanol and mixed feed (methanol:glycerol). Induction with methanol resulted in a maximum laccase activity of 1200 U/L for rSLAC-GAP-AOX which was approximately 2.4-fold higher than rSLAC-AOX and 5.1-fold higher than rSLAC-GAP. The addition of methanol:glycerol in a stoichiometric ratio of 9:1 consistently improved biomass and led to a 1.5-fold increase in rSLAC production as compared to induction with methanol alone. The rSLAC removed 95% of 5 mg/L ciprofloxacin (CIP) and 99% of 100 mg/L tetracycline (TC) in the presence of a mediator. Removal of TC resulted in complete elimination of antibacterial activity while up to 48% reduction in antibacterial activity was observed when CIP was removed. Overall, the present study highlights the effectiveness of a double promoter system in enhancing SLAC production.

Gamma-butyrolactone (GBL), a commonly used industrial solvent, is used recreationally as a central nervous system (CNS) depressant and, therefore, is a United States Drug Enforcement Agency List 1 chemical of the Controlled Substances Act. GBL was identified presumptively in the liquid from JUUL Virginia Tobacco flavored pods during routine untargeted screening analysis of e-cigarette products by gas chromatography–mass spectrometry (GC–MS). Methods for the analysis of GBL were developed for GC–MS and liquid chromatography–tandem mass spectrometry (LC–MS-MS) in the liquids and the aerosol generated from the liquid. Three flavors of JUUL pods available at the time of analysis were obtained by direct purchase from the manufacturer, purchase from a local vape shop and submission from a third party. The only liquid flavor to contain GBL was Virginia Tobacco, with an average of 0.37 mg/mL of GBL, and it was detected in the aerosol. Studies evaluating the pharmacological effects of inhaling GBL do not exist; however, a case report of chronic oral GBL ingestion indicates acute lung injury. The identification of GBL in an e-cigarette product purportedly compliant with federal regulation continues to demonstrate public health and public safety concerns.

Mercury pollution of water bodies constitutes a hazard to both the aquatic life and the populations consuming fishery products. To anticipate the probable eco-toxicological risks that may arise from the largest lake in southern Benin, the present work aims at assessing the mercury status of the sediments of Lake Nokoué, while identifying the areas exposed to anthropogenic mercury contamination. 45 sites were sampled according to a grid plan covering the entire ecosystem of the lake. Employing a Van Veen grab, surface sediments are collected at a depth from 0 to 5 cm in three catches at each site. The composite samples formed at equal mass on the three portions were frozen, lyophilized at - 40 °C, crushed, sieved and homogenized. Fraction less than 63 µm was collected for the mercury content determination by Direct Mercury Analyzer (DMA) operating by atomic absorption according to the US EPA 7473 method. The mercury concentration varies from 1.56 µg/kg to 164.88 µg/kg with an average value of 35.18 µg per kg of sediment. The south-eastern zone of the lake is more concentrated in mercury with a summation of the mercury content of 530.3 µg/kg of sediment. However, the lake has moderate contamination overall (1 ≤ FC ≤ 3). Keywords: Pollution, contamination, mercury, sediment, Lake Nokoué.

Vitex doniana leaves are used in traditional medicine systems for the treatment of inflammatory-related disorders and breast cancer in West Africa. However, there is a dearth of scientific evidence on the effectiveness of the plant leaves on cancer cells or on the safety of the plant if used continuously. This study examined the effects of V. doniana extract on the growth of MCF-7 breast cancer cells, B16-F10 melanoma cells and sEnd.2 endothelial cells. As well, the study investigated the effects of the extract on breast cancer cell morphology, cell cycle progression, migration and invasion, and levels of apoptosis regulatory proteins (Bad, PARP and p53). In vivo, the sub-acute toxicity of the extract in female Wistar rats was examined. The effects of V. doniana extract on MCF-7 cell growth, cell morphology and cell cycle were studied using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, light microscopy and flow cytometry respectively. The multi-target enzyme-linked immunosorbent assay (ELISA) was used to study apoptosis proteins, and the XCELLigence system was used to measure cell invasion and migration. In the sub-acute toxicity study, female Wistar rats were administered 0, 50, 100 and 200 mg/kg of the extract daily for 28 days. Histopathology techniques were used to determine possible toxic effects of the extract on liver, kidney and mammary tissues. The MTT assay showed a concentration-dependent reduction in MCF-7 cell growth in the treated cells compared to the untreated control, and also inhibited the growth of melanoma cells. Furthermore, the results showed that the extract inhibited growth in cancer cells with more potency compared to the non-cancerous endothelial cells. Light microscopy revealed features of apoptosis in V. doniana–treated cells, while flow cytometry showed accumulation of MCF-7 cells in the G1 and sub-G1 phases of the cell cycle following treatment with the plant extract at 25 and 50 μg/ml respectively. In vivo sub-acute toxicity studies showed no significant (P>0.05) difference in serum alanine aminotransferase, gamma glutamyltransferase, urea and creatinine levels between the treatment group and the control, while histopathological examination showed normal architecture of liver, kidney and mammary tissues across the groups. This study provides evidence that V. doniana has anti-cancer effects against MCF-7 breast cancer cells and exhibits no toxic effects in rats at the tested doses. Therefore, V. doniana may have potential in the treatment of breast cancer, thereby necessitating further in vivo anti-cancer studies.

Chinese herbal medicines (CHMs) are classified as dietary supplements. Interactions with western medications, the presence of contaminants or adulterants, or a mis-labeled or mis-used CHM may lead to toxicological emergencies that can be undetected in death investigations. Laboratories must be able to efficiently analyze cases in which CHMs are suspected. Five extractions were evaluated for their ability to extract pharmacologically active compounds from herbal matrices: water, ethanol, microwave-assisted (MAE), ethanol : chloroform, and acid-wash. Anticonvulsive and other pharmacologically active compounds in Gou Teng, Tian Ma, and Jiang Can purchased from Beijing, China and New York were compared in the powder and the extracts using Direct Analysis in Real Time-Mass Spectrometry (DART-MS). Approximately 0.25 g of macerated herb was used per extraction. The water and ethanol extractions were simple liquid extractions. For the MAE, powdered herb was soaked in 65% ethanol, microwaved, and concentrated. The ethanol : chloroform extraction involved soaking in 1 : 1 ethanol : chloroform, sonication, and concentration. In the acid-wash extraction, powdered herb was soaked in acetic acid, followed by addition of sodium hydroxide, hexane extraction, and reconstitution in ethyl acetate. The powdered herbs and extracts were analyzed using a Jeol JMS T100LC AccuTOF DART-MS in positive and negative mode. Of the evaluated methods, no single extraction worked for all active compounds from the three CHMs. The MAE extract contained the most pharmacologically active compounds, while the acid-wash contained the least for the three products. Gou Teng purchased from different sources did exhibit a difference in pharmacologically active compounds, potentially from different species., With the popularity of natural products, rapid analysis of pharmacologically active analytes from natural matrices is critical for public health. Microwave-assisted extraction with DART-MS provides efficient analysis of analytes from herbal matrices.

Since many years ago, the substitution of cement by other cementitious supplementary elements has being a purpose for many researchers. This is to reduce the impact of producing cement on our environment. In this article, we are interested in the possibility of substituting cement with titanium dioxide and titanium dioxide + fly ash. To achieve this purpose, we have manufactured mortars and cement pastes specimens with different rates of replacement of cement by titanium dioxide (0%, 0.1%, 1%) on the one hand and titanium dioxide + fly ash on the overhand. The flexural and compressive strength of each specimen has been determined.

Humans are in–separately linked to the existence of vegetables, as they are the source of several bio–products essential for the survival of the animal kingdom. The importance of vegetables from the point of view of the food industry is determined by their complex chemical content that is important to the human body and this includes organic substances (lipids, proteins, carbohydrates and organic acids). This study examines comparatively the levels of lipid compositions in the samples of dried carrot (Daucus carota L) and cucumber (Cucumis sativus L). The fatty acid, phospholipid and sterol compositions were determined from the samples using Gas Chromatography method. The most concentrated fatty acid (%) was linoleic acid (C18:2) (54.04 and 57.62) and the least was arachidic acid (C20:0) (0.01 and 0.01) for Daucus carota and Cucumis sativus, respectively. The result showed the quality parameters of fatty acids investigated in the Daucus carota and Cucumis sativus samples as: SFA (23.36 and 20.15 %); MUFA (15.27 and 15.5 %); PUFA (60.37 and 64.58 %); DUFA (54.04 and 57.62 %); TUFA (75.64 and 79.83 %); MUFA/SFA (0.63 and 0.76 %); PUFA/SFA (2.87 and 3.47 %); O/L (0.23 and 0.23 %). The total phospholipid contents present in the Daucus carota and Cucumis sativus were 546.11 and 594.51 mg/100 g while that of phytosterols were 366.16 and 376.69 mg/100 g, respectively. Phosphatidycholine has the highest content in both samples (265.80 and 283.64 mg/100 g). The concentrations of phytosterols were very low except in sitosterol (198.71 and 200.53 mg/100 g), stig–masterol (118.42 and 120.39) and campesterol (34.48 and 34.44 mg/100 g) for the Daucus carota and Cucumis sativus samples, respectively. This study revealed that Daucus carota and Cucumis sativus have high values of UFA that make them a special kind of vegetables for nutritional and health applications, and may be a good source of phytosterols.

In this manuscript, the authors explore the antimicrobial and anti-inflammatory properties of extracts from two commonly found and used leaves from species Murraya koenigii and Ocimum gratissimum L., Ethanolic extracts were obtained by maceration and were then concentrated. Serial dilutions of the extracts were done for testing. The extracts were tested for growth inhibition of four bacterial and two fungal strains. Visual examination of growth inhibition, compared to standard antibiotic controls, provided minimum inhibitory concentrations for the two extracts against the six microbial strains. In another experiment, the authors test the anti-inflammatory properties of the two leaf extracts in the standard rat-paw model of inflammation. Results from growth inhibition studies suggest that the two leaf extracts inhibit the growth of all six microbes, with varying levels of sensitivity. Results from anti-inflammatory studies are more impressive, with leaf extracts inhibiting inflammation at levels competitive with the standard drug, indomethacin.

Aim and Objectives: The present study aimed to examine the antiproliferative activity of ethanol extract derived from the Boswellia dalzielii stem bark in breast carcinoma. Methodology: Ethanolic extract of stem bark of Boswellia dalzielii were prepared. Antiproliferative activity was assessed in estrogen receptor (ER)-negative breast carcinoma (MDA-MB-231) cells by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. Results: Obtained results in 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay indicated that ethanol extract of Boswellia dalzielii stem bark showed significant antiproliferative activity in MDA-MB-231 cells in a dose-dependent manner. IC50 of Boswellia dalzielii stem bark ethanol extract in MDA-MB231 was 98. 12 μg/mL. Conclusion: The results obtained suggest that Boswellia dalzielii stem bark extract possesses significant antiproliferative potential which could be mediated by the chemical constituents present in the plant. However, further research needs to be carried out to determine the effect of the extract on the cell cycle and also to determine the type of cell death produced by the extract.

BACKGROUND Diabetes is a serious public health problem which poses serious socio-economic burden at the national and global level. Most synthetic agents for treating diabetes are expensive and not devoid of adverse effect hence, the need for continuous effort in the search for affordable and natural effective treatment of diabetes. This study investigated the anti-diabetic, anti-hyperlipidemic, and hepatoprotective effects of Citrus maxima peel extract on alloxan-induced diabetic rats. METHODS Twenty-eight adult male Wistar rats were categorized into four groups of seven rats with similar body weights. Diabetes was induced using alloxan and baseline data obtained. Rats in groups A, B, and C were administered with 200 mg, 400 mg, and 600 mg of Citrus maxima peel extract per kg body weight/day, respectively for 14 days. Group D (control) received 200 mg of plain water per kg body weight/day. Biochemical analyses of blood glucose, lipid profile, and liver enzyme activities were performed afterthe feeding trial. Data were analyzed using SPSS, version 21. RESULTS Citrus maxima peel extract significantly decreased (p < 0.05) the blood glucose level by 70.17%. The extract (600 mg) resulted in a percentage increase in high-density lipoprotein cholesterol (4.43%) and decrease in total cholesterol (30.86%), triglyceride (10.58%) and low-density lipoprotein cholesterol (10.20%). Liver enzyme activities of the control groups significantly increased while the treated groups showed no significant change. CONCLUSIONS Citrus maxima peel extract has proven to be effective in diabetes management. However, use of standardized extract and proper laboratory tests are necessary.

PurposeWheat flour (whole grain or white wheat flour) is a major ingredient for production of baked goods and confectionery products in Nigeria. However, due to unfavourable climatic and soil conditions, there is over-dependence on importation of wheat having a negative impact on the nation's gross domestic product (GDP). This has therefore spurred increasing popularity of partial or full replacement of wheat flour with other flour types for economic or nutritional reasons. Therefore, the purpose of this study is to evaluate the effect of partial substitution of whole wheat flour with indigenous, underutilized crops.Design/methodology/approachWhole wheat flour was partially (0–40%) replaced with acha and/or pigeon pea flours in ratios of 100:0:0 (WAPK), 90:10:0 (WAPL), 80:20:0 (WAPM), 70:30:0 (WAPN), 80:10:10 (WAPO), 70:20:10 (WAPP), 70:15:15 (WAPQ) and 60:20:20 (WAPR). The study evaluated the effects of supplementation on rheological, functional properties of composite flours and nutritional composition of composite bread.FindingsResults showed that dough development and stability time, values of most pasting properties (peak viscosity, final viscosity, setback and pasting temperature) were significantly higher in the composite flours than in WAPK. However, incorporation of acha and pigeon pea flours resulted in significant decrease in breakdown value. Composite flours produced protein-enriched breads with improved essential amino acids exceeding WHO/FAO reference for adults. Composite flours from blends of whole wheat, acha and pigeon pea flours may serve as a potential raw material suitable for production of nutritious and functional baked products.Originality/valueThe present study confirms effective supplementation of whole grain wheat flour with either acha or both acha and pigeon pea flours. Composite flours showed improved functional and pasting properties; thus, it may be suitable for production of baked products such as bread and biscuits.

Increasing demand for fresh okra (Abelmoschus esculentus L.) has necessitated that shelf-life be extended. Natural edible coatings do not have human safety issues associated with chemical preservat...

Various anthropogenic activities simultaneously alter essential mineral nutrients and contaminant content in the environment. Depending on essential nutrient conditions, the uptake and effects of contaminants in exposed organisms may be altered. The addressing of ecological risk assessment (ERA) of contaminant mixtures has proven difficult. Furthermore, most assessments involving single contaminant exposures do not consider the interaction of essential nutrients on toxicological end points. Hypotheses for toxicological effects of cadmium (Cd), arsenic (As), and their binary mixture (Cd/Asmix) include alteration under varying dietary and media phosphorus (P) conditions. However, interactive effects and effect size (η2) are largely unknown. Here, we investigated the toxicities of Cd-, As-, and Cd/Asmix-treated media and diets on Scenedesmus acutus (a primary producer) and Daphnia pulex (a primary consumer), under varied media and dietary P conditions [low (LP), median (MP), and optimum (COMBO)]. Our results showed significant (p < 0.05) interactive effects and concentration dependent growth inhibition of S. acutus. The toxicity (at day 7) of Cd against S. acutus was 2×, 11×, and 4× that of As in LP, MP, and COMBO conditions, respectively, while the joint toxicity effects of Cd/Asmix were partially additive in LP and COMBO, and synergistic in MP media. Furthermore, acute lethal toxicity (96 h) of Cd in D. pulex was ∼60× that of As, while Cd/Asmix joint toxicity was synergistic. Chronic toxicity (14 d) in D. pulex showed significant (p < 0.05) interaction of As and P-availability on survival, reproduction, and behavior (distance moved, velocity, acceleration and mobility), while Cd and P availability showed significant interactive effect on rotational behavior. Dose response effects of Cd, As, and Cd/Asmix in S. acutus and D. pulex were either monophasic or biphasic under varying nutrient conditions. This study provides empirical evidence of the interactive effects of media/dietary P and toxic metals (Cd, As, and Cd/Asmix) at environmentally relevant concentrations, emphasizing the need for consideration of such interactions during ERA.

Glioblastoma is the most malignant and fatal of all primary brain tumors. Each year in the United States, about 22,000 cases of glioma are diagnosed and about 70% of these patients die. The current treatment modality options available for glioma patients are limited to temozolomide, radiation, and surgery. The poor outcome with these treatment options has necessitated the search for a better chemotherapeutic strategy to improve patient outcomes. In a previous study, a genome-scale clustered regularly interspaced short palindromic repeats (CRISPR) screen in glioblastoma was performed under human type II topoisomerase (TOP2) poison selection and found that various aminoacyl tRNA synthetases (AARS) were the most enriched in our screen. This high expression of AARS was validated by western blot and the gliomas were treated with sub-nanomolar doses of AARS inhibitors. Our results showed that AARS inhibitor treatment effectively killed 75 to 90% of the tumors within 72 h and that this killing is independent of DNA damage repair machinery. Taken together, the findings suggest that application of AARS inhibitors might be curative for glioma, but more experimental in-vivo tests will be needed to validate this. Key words: Borrelidin, mupirocin.

Studies in ecological stoichiometry highlight that grazer dynamics are affected by insufficient food nutrient content (low phosphorus (P)/carbon (C) ratio) as well as excess food nutrient content (high P:C). Contaminant stressors affect all levels of the biological hierarchy, from cells to organs to organisms to populations to entire ecosystems. Eco-toxicological modeling under the framework of ecological stoichiometry predicts the risk of bio-accumulation of a toxicant under stoichiometric constraints. In this paper, we developed and analyzed a Lotka-Volterra type predator- prey model which explicitly tracks the environmental toxicant as well as the toxicant in the populations under stoichiometric constraints. Analytic, numerical, slow-fast steady state and bifurcation theory are employed to predict the risk of toxicant bio-accumulation under varying food conditions. In some cases, our model predicts different population dynamics, including wide amplitude limit cycles where producer densities exhibit very low values and may be in danger of stochastic extinction.

A tool to study SARS-CoV-2 Work with infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) requires high-level biocontainment facilities, making it important to develop safer molecular tools that can potentially be used under less stringent conditions. Self-replicating RNAs known as replicons have long been used to study pathogenic RNA viruses; however, developing replicons to study SARS-SoV-2 has been challenging because of its large genome. Ricardo-Lax et al . used a yeast-based system to construct SARS-CoV-2 replicons that cannot assemble infectious virus because they lack the spike protein required for host cell entry. Transfecting cells with a spike-expressing plasmid and separately with the replicon generates replicon delivery particles (RDPs) that are only capable of one cycle of infection. The replicons and the RDPs can be used in different contexts for drug screening, and viral assays. —VV

This review article seeks to provide relevant information about the applicability of cell-based assays in assessing cytotoxicity of phytochemicals in light of several traditional methods available. Phenolic compounds and other phytochemicals are significant resources for drug discovery and development, thus underlining the enormous potentials of plant-derived natural products for the prevention and management of oxidative stress associated with cancer and other diseases. These effects have been linked to the content of phytochemicals such as phenolic compounds and their antioxidant properties. The abundance and complexity of these bio-constituents highlight the need for well-defined methods for the in vitro characterization and quantification of extracts and/or preparations that can translate to in vivo effects. Cell culture systems provide a useful model for basic research and a wide range of clinical in vitro studies and physiological processes as well as assessing the toxicity and therapeutic potential of compounds including plant extracts with potential medicinal benefits. Their use in cancer research provide a useful insight into possible therapeutic properties of phytochemicals at the cellular level. This approach has been instrumental in the initial stages of developing chemotherapeutic agents where human cancer cell lines are used as experimental models. These models have continued to contribute to elucidating specific requirements for certain biochemical events associated with proliferation, metabolism, loss of cell viability/apoptosis. Cell culture systems remain a promising tool in natural product development.

The P2X4 purinergic receptor is targeted to endolysosomes, where it mediates an inward current dependent on luminal ATP and pH. Activation of P2X4 receptors was previously shown to trigger lysosome fusion, but the regulation of P2X4 receptors and their role in lysosomal Ca2+ signaling are poorly understood. We show that lysosomal P2X4 receptors are activated downstream of plasma membrane P2X7 and H1 histamine receptor stimulation. When P2X4 receptors are expressed, the increase in near-lysosome cytosolic [Ca2+] is exaggerated, as detected with a low-affinity targeted Ca2+ sensor. P2X4-dependent changes in lysosome properties were triggered downstream of P2X7 receptor activation, including an enlargement of lysosomes indicative of homotypic fusion and a redistribution of lysosomes towards the periphery of the cell. Lysosomal P2X4 receptors, therefore, have a role in regulating lysosomal Ca2+ release and the regulation of lysosomal membrane trafficking.

The price of palm oil in the market is rising because of its industrial and domestic applications. Different types of wastes are generated from palm oil processing, ranging from palm oil sludge, palm shell, and waste palm mesocarp fiber. Pressed palm mesocarp fiber contains 5–8% palm oil, which can be recovered using the appropriate method. In this study, an attempt has been made to recover palm oil from pressed oil palm mesocarp, with Soxhlet extraction method using different organic solvents (n-hexane, acetone, dichloromethane, and ethanol). The percentage oil extraction yield (%OEY) of the oil recovered was four time higher with ethanol, and acetone compared with dichloromethane and n-hexane. The %OEY of the oil recovered using n-hexane and dichloromethane was comparable and those recovered using ethanol and acetone were also comparable. Physicochemical analyses, such as viscosity, density, moisture content, flash point, fire point, cloud point, pour point, and acid value, showed that the oils recovered using different solvents were different from each other. Acetone and ethanol performed better because of their properties such as high solubility, high dipole moment, low octanol water partitioning coefficient, and their reactivity. Gas chromatography coupled with mass spectrometry (GC–MS) analyses similarly showed different fragmentation patterns and peaks, which confirmed the various fatty acid components of the recovered oil that were mainly palmitic acid (n-hexadecanoic acid, C16:0) and oleic acid (oleic acid, cis-vaccenic acid, trans-13-octadecenoic acid, cis-13-octadecenoic acid, and 9-octadecenoic acid, C18:1). The ratios of C16:0 to C18:1 were 1:9, 5.3:4.7, 3.5:6:5, and 3.8:6.2 for oil recovered with acetone, dichloromethane, n-hexane, and ethanol, respectively. Acetone and ethanol performed better in oil recovery from pressed mesocarp fiber compared with n-hexane and dichloromethane. The study therefore confirms that more oil could be recovered from pressed oil palm mesocarp for industrial purposes with the use of appropriate solvents.

Crude caffeine was isolated from kola nut, by treating the filtrate obtained from powdered kola nut with lead (II) acetate to precipitate acids and albumins, along with HCl to precipitate lead ions, and to make some insoluble tannins to be soluble, then Na2CO3 solution was added to neutralise the acidic HCl solution. Dichloromethane is added to the filtrate to extract the caffeine from the aqueous layer and then distilled to obtain the crude caffeine. The extracted crude caffeine is recrystallized with activated charcoal and tested for its antioxidant capacity, using the DPPH assay, which screens the antioxidant activity of plant extracts. The DPPH (2, 2-diphenyl-1-picrylhydrazyl) in methanol helps to facilitate the extraction of the antioxidant compounds from the sample. Ascorbic acid (Vitamin C) is used as the standard for this assay, due to its appreciable antioxidant activity. The IC50 value was calculated from the graph of % inhibition against concentration, and the extracted caffeine from kola nut was found out to have a high antioxidant activity comparable to that of the standard, ascorbic acid.

Osteoarthritis (OA), the most common arthritis worldwide, is a degenerative joint disease characterized by progressive cartilage breakdown, subchondral remodeling, and synovial inflammation. Although conventional pharmaceutical therapies aimed to prevent further cartilage loss and joint dysfunction, there are no ideal strategies that target the pathogenesis of OA. Melatonin exhibits a variety of regulatory properties by binding to specific receptors and downstream molecules and exerts a myriad of receptor-independent actions via intracellular targets as a chondrocyte protector, an anti-inflammation modulator, and a free radical scavenger. Melatonin also modulates cartilage regeneration and degradation by directly/indirectly regulating the expression of main circadian clock genes, such as transcriptional activators [brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein (Bmal) and circadian locomotor output cycles kaput (Clock)], transcriptional repressors [period circadian regulator (Per)1/2, cryptochrome (Cry)1/2, and Dec2], and nuclear hormone receptors [Rev-Erbs and retinoid acid-related orphan receptors (Rors)]. Owing to its effects on cartilage homeostasis, we propose a potential role for melatonin in the prevention and therapy of OA via the modulation of circadian clock genes, mitigation of chondrocyte apoptosis, anti-inflammatory activity, and scavenging of free radicals.

Regulation of IP3 receptors (IP3Rs) by IP3 and Ca2+ allows regenerative Ca2+ signals, the smallest being Ca2+ puffs, which arise from coordinated openings of a few clustered IP3Rs. Cells express thousands of mostly mobile IP3Rs, yet Ca2+ puffs occur at a few immobile IP3R clusters. By imaging cells with endogenous IP3Rs tagged with EGFP, we show that KRas-induced actin-interacting protein (KRAP) tethers IP3Rs to actin beneath the plasma membrane. Loss of KRAP abolishes Ca2+ puffs and the global increases in cytosolic Ca2+ concentration evoked by more intense stimulation. Over-expressing KRAP immobilizes additional IP3R clusters and results in more Ca2+ puffs and larger global Ca2+ signals. Endogenous KRAP determines which IP3Rs will respond: it tethers IP3R clusters to actin alongside sites where store-operated Ca2+ entry occurs, licenses IP3Rs to evoke Ca2+ puffs and global cytosolic Ca2+ signals, implicates the actin cytoskeleton in IP3R regulation and may allow local activation of Ca2+ entry., Calcium signals initiated by IP3 receptors in ER membranes regulate most cellular activities. Here, the authors show that KRas-induced actininteracting protein (KRAP) tethers a small subset of IP3 receptors to actin and licenses them to evoke cytosolic calcium signals.

A tripodal Schiff base ligand, 2,4,6-Tris(4-carboxybenzimino)-1,3,5-triazine (MT) and its trinuclear Dy(III), Er(III), and Gd(III) complexes were synthesized. These were characterized using UV-visible, IR, 1H, and 13C NMR spectroscopies, elemental analysis, and molar conductivity measurements. The spectral studies indicate that the ligand is hexadentate and coordinates to the Ln(III) ions through the oxygen atoms of the carboxylic group. The trinuclear complexes were characterized as being bridged by carboxylate anions to the Dy(III), Er(III), and Gd(III) salen centers and displaying a coordination number of six. Biological studies revealed that MT is more active against the test micro-organisms relative to the trinuclear complexes. Acute toxicity studies revealed that MT is safe and has a wide range of effective doses (ED50). In vivo antimalarial studies indicate that MT could serve as an effective antimalarial agent since it has parasitemia inhibition of 84.02% at 50 mg/kg and 65.81% at 25 mg/kg, close to the value (87.22%) of the standard drug—Artesunate. Molecular docking simulation studies on the compounds against SARS-CoV-2 (6Y84) and E. coli DNA gyrase (5MMN) revealed effective binding interactions through multiple bonding modes. The binding energy calculated for Er(III)MT-6Y84 and Er(III)MT-5MMN complexes showed active molecules with the ability to inhibit SARS-CoV-2 and E. coli DNA gyrase.

To augment orthodox anti-inflammatory drugs, which are scarce in some areas, local dwellers in some parts of Nigeria and Asia rely on herbal products such as Combretum zenkeri Engl. & Diels leaves for the treatment of inflammatory diseases. This study investigated the anti-inflammatory activities of crude ethanol extract of C. zenkeri leaves (EECZ). EECZ was prepared and subjected to anti-inflammatory activity tests. The ability of EECZ to inhibit the release of inflammatory mediators in rats was used as a measure of its anti-inflammatory property. The inhibitory effects of EECZ on phospholipase A2 (PLA2) activity, platelet aggregation and haemolysis of human red blood cells (HRBCs) were determined using standard methods. The phytochemical constituents and acute toxicity profile were also evaluated. EECZ at oral doses of 50 and 100 mg/kg b.w. and at 6 h inhibited egg albumin-induced rat paw oedema by 67.36% and 86.19% respectively. The extract similarly inhibited PLA2 activity causing 58.23% and 60.24% inhibition at concentrations of 400 and 800 μg/ml, respectively. At 100–800 μg/ml, EECZ also significantly inhibited haemolysis of HRBCs. In a concentration-dependent manner, EECZ significantly inhibited CaCl2-induced platelet aggregation. The presence of flavonoids, terpenoids, alkaloids, phenol, saponins, steroids, tannins and glycosides were detected in EECZ. The acute toxicity test showed that EECZ is safe at doses less than 5,000 mg/kg b.w. This study shows that EECZ possess anti-inflammatory activities that could be through the inhibition of PLA2 activity and platelet aggregation, and membrane stabilization.

Background: Protein glutaminase specifically deamidates glutamine residue in protein and therefore significantly improves protein solubility and colloidal stability of protein solution. In order to improve its preparation efficiency, we exploited the possibility for its secretory expression mediated by twin-arginine translocation (Tat) pathway in Bacillus licheniformis. Results: The B. licheniformis genome-wide twin-arginine signal peptides were analyzed. Of which, eleven candidates were cloned for construction of expression vectors to mediate the expression of Chryseobacterium proteolyticum protein glutaminase (PGA). The signal peptide of GlmU was confirmed that it significantly mediated PGA secretion into media with the maximum activity of 0.16 U/ml in Bacillus subtilis WB600. A mutant GlmU-R, being replaced the third residue aspartic acid of GlmU twin-arginine signal peptide with arginine by site-directed mutagenesis, mediated the improved secretion of PGA with about 40% increased (0.23 U/ml). In B. licheniformis CBBD302, GlmU-R mediated PGA expression in active form with the maximum yield of 6.8 U/ml in a 25-l bioreactor. Conclusions: PGA can be produced and secreted efficiently in active form via Tat pathway of B. licheniformis, an alternative expression system for the industrial-scale production of PGA.How to cite: Niu D, Li C, Wang P, et al. Twin-arginine signal peptide of Bacillus licheniformis GlmU efficiently mediated secretory expression of protein glutaminase. Electron J Biotechnol 2019;42. https://doi.org/10.1016/j.ejbt.2019.10.006 Keywords: Arginine, Aspartic acid, Bacillus licheniformis, Bacillus subtilis, Escherichia coli, Prostaglandins A, Protein glutaminase, Protein sorting signals, Secretion expression, Signal peptide, Tat pathway

The use of electronic cigarettes (e-cigs) has expanded from a nicotine delivery system to a general drug delivery system. The internet is rife with websites, blogs and forums informing users how to modify e-cigs to deliver illicit drugs while maintaining optimal drug delivery of their device. The goal of this study was to qualitatively identify the presence of methamphetamine in the aerosol produced by an e-cig and to quantitatively assess the effect voltage on the concentration of aerosolized methamphetamine. A KangerTech AeroTank electronic cigarette containing a 30, 60 or 120 mg/mL of methamphetamine in 50:50 propylene glycol: vegetable glycerin formulation was used to produce the aerosol. To qualitatively identify aerosolized methamphetamine, the aerosol was generated at 4.3 V, trapped in a simple glass trapping system, extracted using solid-phase microextraction (SPME), and analyzed by high-resolution Direct Analysis in Real Time AccuTOF™ Mass Spectrometry (DART-MS). To assess the effect of voltage on the concentration of aerosolized methamphetamine, the aerosol was generated at 3.9, 4.3 and 4.7 V, trapped and quantified using gas chromatography mass spectrometry (GC/MS). SPME-DART-MS and SPME-GC-MS demonstrated the aerosolization of methamphetamine. The concentration of aerosolized methamphetamine at 3.9, 4.3 and 4.7 V was not statistically different at 800 ± 600 ng/mL, 800 ± 600 ng/mL and 1,000 ± 800 ng/mL, respectively. The characterization of the vapors produced from e-liquids containing methamphetamine provides an understanding of the dose delivery dynamics of e-cigarettes.

Introduction: Citrus aurantifolia (Christm) is a plant used for the treatment of various ailments including malaria. This study aimed to evaluate the in vivo antiplasmodial efficacy of methanol leaf extract of C. aurantifolia in Swiss albino mice. Methods: The median lethal dose (LD50) was determined by intraperitoneal administration of different doses of the extract (100–4000 mg/kg) to 6 groups of 3 mice each and the animals were observed for 24 hours for physical signs of toxicity. To evaluate the antiplasmodial activity of the extract, three models were used: suppressive, curative and repository. Doses of the extract used were 320, 640 and 960 mg/kg/d in mice, with Chloroquine (5 mg/kg/d) as standard drug. Pyrimethamine (1.2 mg/kg/d) was used as the standard drug for the repository test and distilled water (10 mL/kg/d) as control in all models. Results: In all models, the low dose (320 mg/kg) of the extract produced the highest chemosuppressive effects in all models (P < 0.001). Mice treated with extract lived longer than those in the control group (P < 0.001). Phytochemical screening revealed the presence of alkaloids, flavonoids, saponins, tannins and cardiac glycosides and the LD50 of 3280 mg/kg ± 0.01 shows that the extract has low toxicity. Conclusion: The result of this study shows that C. aurantifolia has antiplasmodial properties which support its use in ethnomedicine in the treatment of malaria.

Synthesis of angular polycyclic phenoxazine derivatives incorporating different phenols is reported in 30-99% yields. O-arylation of 6-chlorodibenzo[a,j] phenoxazin-5-one with a variety of electron-deficient, electron-neutral and electron-rich phenols under the catalytic palladium (II) acetate/t-BuXphos system furnished the compounds of interest. The highest yields were obtained when the intermediate was coupled with electron-rich phenols. IR, 1H-NMR, 13C-NMR and Mass spectra data, confirmed the structures of all the synthesized compounds. Study on the in vitro biological evaluation of the compounds against microorganisms revealed that they are potent antibacterial and antifungal agents, as they showed significant biological activity against Staphylococcus aureus, Streptococcus pneumonia, Escherichia coli, Salmonella typhi, klebsiella pneumonia, Pseudomonas aeruginasa, Basillus substilis, Candida albicans and Aspergillus niger.

4-(Pyrimidin-4-yl)morpholines are privileged pharmacophores for PI3K and PIKKs inhibition by virtue of the morpholine oxygen, both forming the key hydrogen bonding interaction and conveying selectivity over the broader kinome. Key to the morpholine utility as a kinase hinge binder is its ability to adopt a coplanar conformation with an adjacent aromatic core favored by the morpholine nitrogen nonbonding pair of electrons interacting with the electron deficient pyrimidine π-system. Few selective morpholine replacements have been identified to date. Herein we describe the discovery of a potent non-nitrogen containing morpholine isostere with the ability to mimic this conformation and its application in a potent selective dual inhibitor of mTORC1 and mTORC2 (29b).

The goal of this work was to explore the action of ethylenediaminetetraacetic (EDTA) in pursuit of escalating the optical transmittance of the Fricke gel dosimeter prepared with low bloom strength gelatin. The Fricke-xylenol-gelatin (FXGConv) dosimeter recipe used in this study contained a mixture of 5% gelatin, 50 mM sulphuric acid (H2SO4), 0.3 mM ammonium ferrous sulphate (FAS), 0.05 mM xylenol orange (XO) and triple distilled water. Several batches of gels were prepared with varying concentrations of EDTA incorporated in the FXGConv gel. The gel cuvettes were stored at 4 °C for 12 h after preparation and irradiated using gamma rays from a telecobalt unit for doses ranging from 0 to 15 Gy. Dosimetric parameters including the optical background absorbance of the unirradiated gel, stability of dose after irradiation, self-oxygenation (auto-oxidation) and dose response were studied using a spectrophotometer at 585 nm. Based on the response of the EDTA infused FXGConv gels, further investigations were carried out on FXG-Glycine gels incorporated with EDTA. Diffusion coefficient measurements were performed on large volume phantoms irradiated in a linear accelerator. Optical transmittance of unirradiated FXG-Glycine gels improved approximately 40% with EDTA while maintaining acceptable control over Fe2+ auto-oxidation as well as Fe3+ diffusion. An optimal concentration of EDTA in the glycine-based FXG gel dosimeter can provide an overall benefit with low background absorbance and acceptable diffusion control while using lower bloom gelatin for preparation of gel dosimeters.

Introduction: Ricinus communis is used by the people of Niger-Delta region of Nigeria, for the treatment of various ailments, especially malaria. This study evaluated the antiplasmodial potentials of the aqueous seed extract of R. communis, using Plasmodium berghei berghei. Methods: Acute toxicity study was carried out to determine the median lethal dose (LD50) of the extract. Antiplasmodial effect of the extract was assessed in suppressive, repository/ prophylactic and curative models, using Swiss albino mice (15-29 g). Mice were infected intraperitoneally with 0.2 mL of parasitized blood. Extract doses administered were 54.77, 109.54 and 164.32 mg/kg/d of the seed extract and each dose had 6 replicates. Artesunate (5 mg/kg/d) and pyrimethamine (1.2 mg/kg/d) were used as standard drugs, while distilled water (10 mL/kg/d) served as control. Results: Acute toxicity study produced LD50 of 547.72 mg/kg. The extract demonstrated a dosedependent reduction in parasitaemia in all tests. At the end of 4-day test, suppressive effect of 20.80, 49.00, 75.00 and 88.40% were obtained for doses 54.77, 109.54 and 164.32 mg/kg/d of the seed extract and artesunate, respectively. In the repository test pyrimethamine was more potent (72.26%) than the seed extract (9.47%–51.42%). The extract also exhibited appreciable curative effect. The activity of the seed extract was significant when compared with the control (P < 0.05). Mice treated with the seed extract and drugs survived for longer duration than the control group. Conclusion: The aqueous seed extract of R. communis has antiplasmodial potential and its active principle should be elucidated and further investigated to help in the ongoing fight against malaria.

Individual mitochondria undergo an intermittent, all-or-none electrochemical excitation termed “mitoflash.” Feng et al. show that mitoflash occurs following build-up of mitochondrial electrochemical potential and may serve to autoregulate mitochondrial proton electrochemical potential., Respiring mitochondria undergo an intermittent electrical and chemical excitation called mitochondrial flash (mitoflash), which transiently uncouples mitochondrial respiration from ATP production. How a mitoflash is generated and what specific role it plays in bioenergetics remain incompletely understood. Here, we investigate mitoflash biogenesis in isolated cardiac mitochondria by varying the respiratory states and substrate supply and by dissecting the involvement of different electron transfer chain (ETC) complexes. We find that robust mitoflash activity occurs once mitochondria are electrochemically charged by state II/IV respiration (i.e., no ATP synthesis at Complex V), regardless of the substrate entry site (Complex I, Complex II, or Complex IV). Inhibiting forward electron transfer abolishes, while blocking reverse electron transfer generally augments, mitoflash production. Switching from state II/IV to state III respiration, to allow for ATP synthesis at Complex V, markedly diminishes mitoflash activity. Intriguingly, when mitochondria are electrochemically charged by the ATPase activity of Complex V, mitoflashes are generated independently of ETC activity. These findings suggest that mitoflash biogenesis is mechanistically linked to the build up of mitochondrial electrochemical potential rather than ETC activity alone, and may functionally counteract overcharging of the mitochondria and hence serve as an autoregulator of mitochondrial proton electrochemical potential.