Receptor for Advanced Glycation End Product (RAGE) Modulates Inflammation During Feeding of the Hard Tick, Haemaphysalis longicornis in Mice.

Ticks are notorious blood-sucking ectoparasites that affect both humans and animals. They serve as a unique vector of various deadly diseases. Here, we have shown the roles of the receptor for advanced glycation end products (RAGE) during repeated infestations by the tick Haemaphysalis longicornis using RAGE-/- mice. In primary infestation, a large blood pool developed, which was flooded with numerous RBCs, especially during the rapid feeding phase of the tick both in wild-type (wt) and RAGE-/- mice. Very few inflammatory cells were detected around the zones of haemorrhage in the primary infestations. However, the number of inflammatory cells gradually increased in the subsequent tick infestations, and during the third infestations, the number of inflammatory cells reached to the highest level (350.3 ± 16.8 cells/focus). The site of attachment was totally occupied by the inflammatory cells in wt mice, whereas very few cells were detected at the ticks' biting sites in RAGE-/- mice. RAGE was highly expressed during the third infestation in wt mice. In the third infestation, infiltration of CD44+ lymphocytes, eosinophils and expression of S100A8 and S100B significantly increased at the biting sites of ticks in wt, but not in RAGE-/- mice. In addition, peripheral eosinophil counts significantly increased in wt but not in RAGE-/- mice. Taken together, our study revealed that RAGE-mediated inflammation and eosinophils played crucial roles in the tick-induced inflammatory reactions.

Lipidation alters the phase-separation of resilin-like polypeptides.

Biology exploits biomacromolecular phase separation to form condensates, known as membraneless organelles. Despite significant advancements in deciphering sequence determinants for phase separation, modulating these features in vivo remains challenging. A promising approach inspired by biology is to use post-translational modifications (PTMs)-to modulate the amino acid physicochemistry instead of altering protein sequences-to control the formation and characteristics of condensates. However, despite the identification of more than 300 types of PTMs, the detailed understanding of how they influence the formation and material properties of protein condensates remains incomplete. In this study, we investigated how modification with myristoyl lipid alters the formation and characteristics of the resilin-like polypeptide (RLP) condensates, a prototypical disordered protein with upper critical solution temperature (UCST) phase behaviour. Using turbidimetry, dynamic light scattering, confocal and electron microscopy, we demonstrated that lipidation-in synergy with the sequence of the lipidation site-significantly influences RLPs' thermodynamic propensity for phase separation and their condensate properties. Molecular simulations suggested these effects result from an expanded hydrophobic region created by the interaction between the lipid and lipidation site rather than changes in peptide rigidity. These findings emphasize the role of "sequence context" in modifying the properties of PTMs, suggesting that variations in lipidation sequences could be strategically used to fine-tune the effect of these motifs. Our study advances understanding of lipidation's impact on UCST phase behaviour, relevant to proteins critical in biological processes and diseases, and opens avenues for designing lipidated resilins for biomedical applications like heat-mediated drug elution.

Electronic aggregated data collection on cervical cancer screening in Bangladesh since 2014: what the data tells us?

INTRODUCTION: To reduce the high prevalence of cervical cancers among the Bangladeshi women, the Government of Bangladesh established a national cervical cancer screening programme in 2005 for women aged 30 to 60 years. The District Health Information System Version 2 (DHIS2) based electronic aggregated data collection system is used since the year 2013. This study summarises data from the year 2014 to 2022 to assess the effectiveness of the electronic data collection system in understanding the outcome of the screening programme. METHODS: This is a descriptive study based on secondary data extracted in MS Excel from the DHIS2-based electronic repository of the national cervical cancer screening programme of Bangladesh. The respondents were women aged 30-60 years, screened for cervical cancer using VIA (Visual Inspection of cervix with Acetic acid) method in 465 government health facilities. The data were collected on the participants' residential location, month and year of screening, name and type of health facilities performing VIA, and VIA screening results. RESULTS: The national screening programme reported a total 3.36 million VIA tests from 465 government hospitals in 8 years (2014 to 2022). The national average VIA-positivity rate was 3.6%, which varied from 1.4 to 9.5% among the districts. This national screening programme witnessed an exponential growth, year after year, with 83.3% increase in VIA test from 2014 to 2022. The primary and the secondary care hospitals were the highest collective contributors of VIA tests (86.2%) and positive cases (77.8%). The VIA-positivity rates in different hospital types varied widely, 7.0% in the medical university hospital, 5.7% in the medical college hospitals, 3.9% in the district/general hospitals, and 3.0% in the upazila health complexes. CONCLUSIONS: A national cervical cancer screening programme using VIA method and a DHIS2-based electronic data collection backbone, is effective, sustainable, and useful to understand the screening coverage, VIA positivity rate and geographic distribution of the participants and case load to initiate policy recommendations and actions. Decentralization of the screening programme and more efforts at the primary and secondary care level is required to increase screening performances.

Lipidation Alters the Structure and Hydration of Myristoylated Intrinsically Disordered Proteins.

Lipidated proteins are an emerging class of hybrid biomaterials that can integrate the functional capabilities of proteins into precisely engineered nano-biomaterials with potential applications in biotechnology, nanoscience, and biomedical engineering. For instance, fatty-acid-modified elastin-like polypeptides (FAMEs) combine the hierarchical assembly of lipids with the thermoresponsive character of elastin-like polypeptides (ELPs) to form nanocarriers with emergent temperature-dependent structural (shape or size) characteristics. Here, we report the biophysical underpinnings of thermoresponsive behavior of FAMEs using computational nanoscopy, spectroscopy, scattering, and microscopy. This integrated approach revealed that temperature and molecular syntax alter the structure, contact, and hydration of lipid, lipidation site, and protein, aligning with the changes in the nanomorphology of FAMEs. These findings enable a better understanding of the biophysical consequence of lipidation in biology and the rational design of the biomaterials and therapeutics that rival the exquisite hierarchy and capabilities of biological systems.

Microplastic contamination in commercial fish feeds: A major concern for sustainable aquaculture from a developing country.

Plastic pollution has become a global issue nowadays. Due to the increased population in developing countries, we largely depend on fish from our aquaculture industry to meet the required protein demand. Though several studies documented plastic ingestion in freshwater and marine organisms, very limited studies have been conducted to elucidate microplastic (MP) contamination in commercial fish feed. Therefore, this study was designed to identify, quantify, and characterize microplastics (MPs) in commercial fish feeds in Bangladesh and assess possible health risks in fish consuming different commercial fish feeds. All fish feed samples were 100 % contaminated with MPs, where the mean abundance of MPs ranged between 500 and 2200 MPs/kg. No significant differences among different types of feeds (e.g., starter, grower, and finisher) were observed in terms of MPs abundance (F = 0.999, p = 0.385). This study revealed that fiber was the most dominant shape of MPs (90 %), while the most dominant color of MPs was red (34 %), followed by black (31 %) and blue (19 %). The 100-1500 µm size class covers 88 % of the total MPs in the collected fish feed samples. Identified polymers in the samples were polyethylene (PE, 37.71 %), polyvinyl chloride (PVC, 27.14 %), polypropylene (PP, 22.08 %), and polyethylene terephthalate (PET, 13.07 %), respectively, where PE and PVC fall under the risk category IV to V. The Pollution load index (PLI) values of all fish feed samples were <10, indicating the risk category of I (low risk). Therefore, this study highly recommended avoiding plastic materials in the packaging and storing purposes of feed ingredients in the feed mills to ensure contamination-free fish feed for sustainable aquaculture.

COVID-19: Modeling Out-of-Home and In-Home Activity Participation during the Pandemic.

Understanding the interaction between in-home and out-of-home activity participation decisions is important, particularly at a time when opportunities for out-of-home activities such as shopping, entertainment, and so forth are limited because of the COVID-19 pandemic. The travel restrictions imposed as a result of the pandemic have had a massive impact on out-of-home activities and have changed in-home activities as well. This study investigates in-home and out-of-home activity participation during the COVID-19 pandemic. Data comes from the COVID-19 Survey for assessing Travel impact (COST), conducted from March to May in 2020. This study uses data for the Okanagan region of British Columbia, Canada to develop the following two models: a random parameter multinomial logit (RPMNL) model for out-of-home activity participation and a hazard-based random parameter duration (HRPD) model for in-home activity participation. The model results suggest that significant interactions exist between out-of-home and in-home activities. For example, a higher frequency of out-of-home work-related travel is more likely to result in a shorter duration of in-home work activities. Similarly, a longer duration of in-home leisure activities might yield a lower likelihood for recreational travel. Health care workers are more likely to engage in work-related travel and less likely to participate in personal and household maintenance activities at home. The model confirms heterogeneity among the individuals. For instance, a shorter duration of in-home online shopping yields a higher probability for participation in out-of-home shopping activity. This variable shows significant heterogeneity with a large standard deviation, which reveals that sizable variation exists for this variable.

Adaptive Recombinant Nanoworms from Genetically Encodable Star Amphiphiles.

Recombinant nanoworms are promising candidates for materials and biomedical applications ranging from the templated synthesis of nanomaterials to multivalent display of bioactive peptides and targeted delivery of theranostic agents. However, molecular design principles to synthesize these assemblies (which are thermodynamically favorable only in a narrow region of the phase diagram) remain unclear. To advance the identification of design principles for the programmable assembly of proteins into well-defined nanoworms and to broaden their stability regimes, we were inspired by the ability of topologically engineered synthetic macromolecules to acess rare mesophases. To test this design principle in biomacromolecular assemblies, we used post-translational modifications (PTMs) to generate lipidated proteins with precise topological and compositional asymmetry. Using an integrated experimental and computational approach, we show that the material properties (thermoresponse and nanoscale assembly) of these hybrid amphiphiles are modulated by their amphiphilic architecture. Importantly, we demonstrate that the judicious choice of amphiphilic architecture can be used to program the assembly of proteins into adaptive nanoworms, which undergo a morphological transition (sphere-to-nanoworms) in response to temperature stimuli.

Temporal landscape of mutational frequencies in SARS-CoV-2 genomes of Bangladesh: possible implications from the ongoing outbreak in Bangladesh.

Along with intrinsic evolution, adaptation to selective pressure in new environments might have resulted in the circulatory SARS-CoV-2 strains in response to the geoenvironmental conditions of a country and the demographic profile of its population. With this target, the current study traced the evolutionary route and mutational frequency of 198 Bangladesh-originated SARS-CoV-2 genomic sequences available in the GISAID platform over a period of 13 weeks as of 14 July 2020. The analyses were performed using MEGA X, Swiss Model Repository, Virus Pathogen Resource and Jalview visualization. Our analysis identified that majority of the circulating strains strikingly differ from both the reference genome and the first sequenced genome from Bangladesh. Mutations in nonspecific proteins (NSP2-3, NSP-12(RdRp), NSP-13(Helicase)), S-Spike, ORF3a, and N-Nucleocapsid protein were common in the circulating strains with varying degrees and the most unique mutations (UM) were found in NSP3 (UM-18). But no or limited changes were observed in NSP9, NSP11, Envelope protein (E) and accessory factors (NSP7a, ORF 6, ORF7b) suggesting the possible conserved functions of those proteins in SARS-CoV-2 propagation. However, along with D614G mutation, more than 20 different mutations in the Spike protein were detected basically in the S2 domain. Besides, mutations in SR-rich region of N protein and P323L in RDRP were also present. However, the mutation accumulation showed a significant association (p = 0.003) with sex and age of the COVID-19-positive cases. So, identification of these mutational accumulation patterns may greatly facilitate vaccine development deciphering the age and the sex-dependent differential susceptibility to COVID-19.

Genetically Encoded Inverse Bolaamphiphiles.

Developing new protein-based materials with a programmable assembly is of great scientific interest and technological importance. Inspired by nature's use of post-translational modifications (PTMs) to control the function and location of proteins, we have leveraged lipidation-the PTM of proteins with lipids-to synthesize genetically encoded lipidated proteins with controllable hierarchical assembly. Specifically, we envisioned the combination of two orthogonal lipidation pathways with different regioselectivity and substrate preferences inside Escherichia coli to produce recombinant nanomaterials with distinct lipidation domains at each terminus of proteins. In this study, we demonstrate the orthogonality of N-myristoylation and C-cholesterylation pathways for recombinant production of lipidated proteins with a unique triblock architecture, which is a hydrophilic protein block flanked by two lipid tails, i.e., inverse bolaamphiphiles. Our study indicates that the architecture of lipidated protein and the sequence of the polypeptide can be used to control the hierarchical self-assembly of these materials. We envision this bio-enabled approach yielding unexplored recombinant hybrid biomaterials with tunable nanoscale structure and morphology with applications in nanobiotechnology.

Tebuconazole and trifloxystrobin regulate the physiology, antioxidant defense and methylglyoxal detoxification systems in conferring salt stress tolerance in Triticum aestivum L.

Fungicides are widely used for controlling fungi in crop plants. However, their roles in conferring abiotic stress tolerance are still elusive. In this study, the effect of tebuconazole (TEB) and trifloxystrobin (TRI) on wheat seedlings (Triticum aestivum L. cv. Norin 61) was investigated under salt stress. Seedlings were pre-treated for 48 h with fungicide (1.375 µM TEB + 0.5 µM TRI) and then subjected to salt stress (250 mM NaCl) for 5 days. Salt treatment alone resulted in oxidative damage and increased lipid peroxidation as evident by higher malondialdehyde (MDA) and hydrogen peroxide (H2O2) content. Salt stress also decreased the chlorophyll and relative water content and increased the proline (Pro) content. Furthermore, salt stress increased the dehydroascorbate (DHA) and glutathione disulfide (GSSG) content while ascorbate (AsA), the AsA/DHA ratio, reduced glutathione (GSH) and the GSH/GSSG ratio decreased. However, a combined application of TEB and TRI significantly alleviated growth inhibition, photosynthetic pigments and leaf water status improved under salt stress. Application of TEB and TRI also decreased MDA, electrolyte leakage, and H2O2 content by modulating the contents of AsA and GSH, and enzymatic antioxidant activities. In addition, TEB and TRI regulated K+/Na+ homeostasis by improving the K+/Na+ ratio under salt stress. These results suggested that exogenous application of TEB and TRI rendered the wheat seedling more tolerant to salinity stress by controlling ROS and methylglyoxal (MG) production through the regulation of the antioxidant defense and MG detoxification systems.

Explicating physiological and biochemical responses of wheat cultivars under acidity stress: insight into the antioxidant defense and glyoxalase systems.

Soil acidity causes proton (H+) rhizotoxicity, inhibits plant growth and development, and is a major yield-limiting factor for wheat production worldwide. Therefore, we investigated the physiological and biochemical responses of wheat (Triticum aestivum L.) to acidity stress in vitro. Five popular wheat cultivars developed by Bangladesh Agricultural Research Institute (BARI), namely, BARI Gom-21, BARI Gom-24, BARI Gom-25, BARI Gom-26, and BARI Gom-30, were studied in growing media under four different pH levels (3.5, 4.5, 5.5, and 6.5). We evaluated the cultivars based on their relative water content, proline (Pro) content, growth, biomass accumulation, oxidative damage, membrane stability, and mineral composition, as well as the performance of the antioxidant defense and glyoxalase systems. Although decrements of pH significantly reduced the tested morphophysiological and biochemical attributes in all the cultivars, there was high variability among the cultivars in response to the varying pH of the growing media. Acidity stress reduced growth, biomass, water content, and chlorophyll content in all the cultivars. However, BARI Gom-26 showed the least damage, with the lowest H2O2 generation, lipid peroxidation (MDA), and greater membrane stability, which indicate better tolerance against oxidative damage. In addition, the antioxidant defense components, ascorbate (AsA) and glutathione (GSH), and their redox balance were higher in this cultivar. Maximum H2O2 scavenging due to upregulation of the antioxidant enzymes [AsA peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), GSH reductase (GR), GSH peroxidase (GPX), and GSH-S-transferase (GST)] was observed in BARI Gom-26, which also illustrated significant enhancement of methylglyoxal (MG) detoxification by upregulating glyoxalase I (Gly I) and glyoxalase II (Gly II). This study also showed that balanced essential nutrient content as well as lower toxic micronutrient content was found in BARI Gom-26. Therefore, considering the physiological and biochemical attributes and growth, we conclude that BARI Gom-26 can withstand acidity stress during the early seedling stage, by regulating the coordinated action of the antioxidant defense and glyoxalase systems as well as maintaining nutrient balance.

Acetate-induced modulation of ascorbate: glutathione cycle and restriction of sodium accumulation in shoot confer salt tolerance in Lens culinaris Medik.

Physiological and biochemical changes in six-day-old hydroponically grown lentil seedlings exposed to 100 mM salinity stress with or without 5 and 10 mM Na-acetate were studied. Results showed that salt stress reduced recovery percentage, fresh weight (FW), chlorophyll (chl) content, disturbed water balance, disrupted antioxidant defense pathway by decreasing reduced ascorbate content, and caused ion toxicity resulting from increased Na+ accumulation, severe K+ loss from roots in hydroponic culture. However, exogenous application of Na-acetate improved the seedling growth by maintaining water balance and increasing chl content. Furthermore, Na-acetate application reduced oxidative damage by modulating antioxidant defense pathway, and sustained ion homeostasis by reducing Na+ uptake and K+ loss. In the second experiment in glass house, we investigated the role of Na-acetate on lentil for long-term salinity. Acetate application increased FW and dry weight, reduced oxidative and membrane damage, and lowered the accumulation of Na+ in shoot compared with salt stressed seedlings alone. From the results of both experiments, it is clear that the exogenous application of Na-acetate enhanced salt tolerance in lentil seedlings.

Maleic acid assisted improvement of metal chelation and antioxidant metabolism confers chromium tolerance in Brassica juncea L.

Chromium (Cr) is a highly toxic environmental pollutant that negatively affects plant growth and development. Thus, remediating Cr from soil or increasing plant tolerance against Cr stress is urgent. Organic acids are recognized as agents of phytoremediation and as exogenous protectants, but using maleic acid (MA) to attain these results has not yet been studied. Therefore, our study investigated the effects of MA on Cr uptake and mitigation of Cr toxicity. We treated 8-d-old Indian mustard (Brassica juncea L.) seedlings with Cr (0.15mM and 0.3mM K2CrO4, 5 days) alone and in combination with MA (0.25mM) in a semi-hydroponic medium. Under Cr stress, plants accumulated Cr in both the roots and shoots in a dose-dependent manner, where the roots showed higher accumulation. Chromium stress reduced the growth and biomass of the Indian mustard plants by reducing water status and photosynthetic pigments, and increased oxidative damage due to generation of toxic reactive oxygen species (ROS) and methylglyoxal (MG). Chromium stress also interfered with the function of the antioxidant defense and glyoxalase systems. However, using MA in the Cr-stressed plants further increased Cr uptake in the roots, but it slightly reduced the translocation of Cr from the roots to the shoots at a lower dose of Cr and significantly at a higher dose. Moreover, MA also increased the other non-protein thiols (NPTs) containing phytochelatin (PC) content of the seedlings, which reduced Cr toxicity. Supplementing the stressed plants with MA upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH); the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT); and the enzymes of the glyoxalase system including glyoxalase I (Gly I) and glyoxalase II (Gly II); and finally reduced oxidative damage and increased the chlorophyll content and water status as well the growth and biomass of the plants. Our findings suggested two potential uses of MA: first, enhancing phytoremediation, principally phytostabilization and second, working as an exogenous protectant to enhance Cr tolerance.

γ-aminobutyric acid (GABA) confers chromium stress tolerance in Brassica juncea L. by modulating the antioxidant defense and glyoxalase systems.

Chromium (Cr) toxicity is hazardous to the seed germination, growth, and development of plants. γ-aminobutyric acid (GABA) is a non-protein amino acid and is involved in stress tolerance in plants. To investigate the effects of GABA in alleviating Cr toxicity, we treated eight-d-old mustard (Brassica juncea L.) seedlings with Cr (0.15 and 0.3 mM K2CrO4, 5 days) alone and in combination with GABA (125 µM) in a semi-hydroponic medium. The roots and shoots of the seedlings accumulated Cr in a dose-dependent manner, which led to an increase in oxidative damage [lipid peroxidation; hydrogen peroxide (H2O2) content; superoxide (O2•-) generation; lipoxygenase (LOX) activity], methylglyoxal (MG) content, and disrupted antioxidant defense and glyoxalase systems. Chromium stress also reduced growth, leaf relative water content (RWC), and chlorophyll (chl) content but increased phytochelatin (PC) and proline (Pro) content. Furthermore, supplementing the Cr-treated seedlings with GABA reduced Cr uptake and upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH) and the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II), and finally reduced oxidative damage. Adding GABA also increased leaf RWC and chl content, decreased Pro and PC content, and restored plant growth. These findings shed light on the effect of GABA in improving the physiological mechanisms of mustard seedlings in response to Cr stress.

Coordinated Actions of Glyoxalase and Antioxidant Defense Systems in Conferring Abiotic Stress Tolerance in Plants.

Being sessile organisms, plants are frequently exposed to various environmental stresses that cause several physiological disorders and even death. Oxidative stress is one of the common consequences of abiotic stress in plants, which is caused by excess generation of reactive oxygen species (ROS). Sometimes ROS production exceeds the capacity of antioxidant defense systems, which leads to oxidative stress. In line with ROS, plants also produce a high amount of methylglyoxal (MG), which is an α-oxoaldehyde compound, highly reactive, cytotoxic, and produced via different enzymatic and non-enzymatic reactions. This MG can impair cells or cell components and can even destroy DNA or cause mutation. Under stress conditions, MG concentration in plants can be increased 2- to 6-fold compared with normal conditions depending on the plant species. However, plants have a system developed to detoxify this MG consisting of two major enzymes: glyoxalase I (Gly I) and glyoxalase II (Gly II), and hence known as the glyoxalase system. Recently, a novel glyoxalase enzyme, named glyoxalase III (Gly III), has been detected in plants, providing a shorter pathway for MG detoxification, which is also a signpost in the research of abiotic stress tolerance. Glutathione (GSH) acts as a co-factor for this system. Therefore, this system not only detoxifies MG but also plays a role in maintaining GSH homeostasis and subsequent ROS detoxification. Upregulation of both Gly I and Gly II as well as their overexpression in plant species showed enhanced tolerance to various abiotic stresses including salinity, drought, metal toxicity, and extreme temperature. In the past few decades, a considerable amount of reports have indicated that both antioxidant defense and glyoxalase systems have strong interactions in conferring abiotic stress tolerance in plants through the detoxification of ROS and MG. In this review, we will focus on the mechanisms of these interactions and the coordinated action of these systems towards stress tolerance.

Manganese-induced salt stress tolerance in rice seedlings: regulation of ion homeostasis, antioxidant defense and glyoxalase systems.

Hydroponically grown 12-day-old rice (Oryza sativa L. cv. BRRI dhan47) seedlings were exposed to 150 mM NaCl alone and combined with 0.5 mM MnSO4. Salt stress resulted in disruption of ion homeostasis by Na+ influx and K+ efflux. Higher accumulation of Na+ and water imbalance under salinity caused osmotic stress, chlorosis, and growth inhibition. Salt-induced ionic toxicity and osmotic stress consequently resulted in oxidative stress by disrupting the antioxidant defense and glyoxalase systems through overproduction of reactive oxygen species (ROS) and methylglyoxal (MG), respectively. The salt-induced damage increased with the increasing duration of stress. However, exogenous application of manganese (Mn) helped the plants to partially recover from the inhibited growth and chlorosis by improving ionic and osmotic homeostasis through decreasing Na+ influx and increasing water status, respectively. Exogenous application of Mn increased ROS detoxification by increasing the content of the phenolic compounds, flavonoids, and ascorbate (AsA), and increasing the activities of monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), superoxide dismutase (SOD), and catalase (CAT) in the salt-treated seedlings. Supplemental Mn also reinforced MG detoxification by increasing the activities of glyoxalase I (Gly I) and glyoxalase II (Gly II) in the salt-affected seedlings. Thus, exogenous application of Mn conferred salt-stress tolerance through the coordinated action of ion homeostasis and the antioxidant defense and glyoxalase systems in the salt-affected seedlings.

The association of CYP2C19 LoF alleles with adverse clinical outcomes in stroke patients taking clopidogrel: An updated meta-analysis.

The aggregated risk of recurrent stroke in stroke/transient ischemic attack (TIA) patients carrying CYP2C19 LoF alleles who take clopidogrel has not been investigated recently, and the available research is limited. This study aimed to perform an updated meta-analysis to assess the association between CYP2C19 LoF alleles and the risk of recurrent stroke in patients taking clopidogrel. Databases were searched for the literature on eligible studies. The end points were recurrent stroke, composite vascular events, and bleeding events. Odds ratios (ORs) were calculated using RevMan software, where p < 0.05 was considered statistically significant. Patients carrying CYP2C19 LoF alleles who were treated with clopidogrel had a significantly increased risk of recurrent ischemic stroke compared with non-carriers (OR 2.18, 96% CI 1.80-2.63; p < 0.00001). The risk of recurrent stroke was only significantly different in Asian patients (OR 2.29, 96% CI 1.88-2.80; p < 0.00001) but not in patients of other ethnicities; however, there were a limited number of studies in other ethnic groups. Both observational studies (OR 2.83, 96% CI 2.20-3.65; p < 0.00001) and RCTs (OR 1.48, 96% CI 1.10-1.98; p = 0.009) found associations with a significantly increased risk of recurrent ischemic stroke. Asian stroke patients or TIA patients carrying CYP2C19 LoF alleles and taking clopidogrel were at a significantly higher risk of recurrent ischemic stroke than non-carriers. Significantly increased risk of recurrent ischemic stroke was found in both observational studies and RCTs.

Cholelithiasis in a goat associated with chronic fascioliosis in Bangladesh: A case report and review of literatures.

The objective of the present study was to describe a very rare case of gallstone (cholelithiasis) in a goat associated with chronic fasciolosis. During a routine slaughterhouse-based survey, a two-and-half-year-old female Black Bengal Goat was found to be affected with severe chronic fascioliosis characterized by the massive damage in the liver. Through systemic dissection of liver, we isolated 94 adult Fasciola spp., and by PCR, we confirmed the fluke as Fasciola gigantica. The gallbladder of the goat was oedematous. On opening the gallbladder, we recovered 255 stones of variable sizes. Stones were whitish in colour and friable, and some of the fragile stones were attached to the wall of the gallbladder. To the authors' knowledge, this is the first report of the cholelithiasis in a goat associated with F. gigantica.

Ascaridia galli, a common nematode in semiscavenging indigenous chickens in Bangladesh: epidemiology, genetic diversity, pathobiology, ex vivo culture, and anthelmintic efficacy.

Ascaridia galli is the most common nematode in chickens. Ascaridia galli is highly prevalent in chickens reared in scavenging or semiscavenging systems. Here, we studied the epidemiology, pathology, genetic diversity, ex vivo culture protocol and anthelmintic sensitivity of A. galli prevalent in indigenous chickens in Bangladesh. Through morphological study and molecular analyses, the isolated worms were confirmed as A. galli. Of the chickens examined, 45.6% (178 out of 390) were found infected. The male and young chickens were significantly (P < 0.05) more prone to A. galli infection. Prevalence of the infection was significantly (P < 0.05) lower in the summer season. In heavy infections, A. galli blocked the small intestine. Marked inflammation, increased mucus production and petechial hemorrhages were evident in the small intestine, particularly in the duodenum. Also, there were desquamation and adhesion of the mucosal villi; degeneration, necrosis of the epithelial cells and goblet cell hyperplasia. The mucosal layer was infiltrated mainly with eosinophils and heterophils. We developed a hen egg white-based long-term ex vivo culture protocol which supported the survival and reproduction of A. galli for more than a week. Levamisole (LEV) and ivermectin (IVM) efficiently killed A. galli. However, albendazole (ABZ), mebendazole (MBZ), and piperazine (PPZ) did not kill the worms even at 120 µg/mL and 1mg/mL concentrations, respectively. Taken together, our results suggest that A. galli is highly prevalent in semiscavenging chickens in Bangladesh. Ascaridia galli can be easily maintained ex vivo in egg white supplemented M199 medium. LEV and IVM, but not ABZ, MBZ and PPZ, can be used for treating and controlling A. galli infections in chickens.

A computational approach for structural and functional analyses of disease-associated mutations in the human CYLD gene.

Tumor suppressor cylindromatosis protein (CYLD) regulates NF-κB and JNK signaling pathways by cleaving K63-linked poly-ubiquitin chain from its substrate molecules and thus preventing the progression of tumorigenesis and metastasis of the cancer cells. Mutations in CYLD can cause aberrant structure and abnormal functionality leading to tumor formation. In this study, we utilized several computational tools such as PANTHER, PROVEAN, PredictSNP, PolyPhen-2, PhD-SNP, PON-P2, and SIFT to find out deleterious nsSNPs. We also highlighted the damaging impact of those deleterious nsSNPs on the structure and function of the CYLD utilizing ConSurf, I-Mutant, SDM, Phyre2, HOPE, Swiss-PdbViewer, and Mutation 3D. We shortlisted 18 high-risk nsSNPs from a total of 446 nsSNPs recorded in the NCBI database. Based on the conservation profile, stability status, and structural impact analysis, we finalized 13 nsSNPs. Molecular docking analysis and molecular dynamic simulation concluded the study with the findings of two significant nsSNPs (R830K, H827R) which have a remarkable impact on binding affinity, RMSD, RMSF, radius of gyration, and hydrogen bond formation during CYLD-ubiquitin interaction. The principal component analysis compared native and two mutants R830K and H827R of CYLD that signify structural and energy profile fluctuations during molecular dynamic (MD) simulation. Finally, the protein-protein interaction network showed CYLD interacts with 20 proteins involved in several biological pathways that mutations can impair. Considering all these in silico analyses, our study recommended conducting large-scale association studies of nsSNPs of CYLD with cancer as well as designing precise medications against diseases associated with these polymorphisms.