Discovery of a novel natural compound, vitekwangin B, with ANO1 protein reduction properties and anticancer potential.

Background: Prostate cancer and non-small cell lung cancer (NSCLC) present significant challenges in the development of effective therapeutic strategies. Hormone therapies for prostate cancer target androgen receptors and prostate-specific antigen markers. However, treatment options for prostatic small-cell neuroendocrine carcinoma are limited. NSCLC, on the other hand, is primarily treated with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors but exhibits resistance. This study explored a novel therapeutic approach by investigating the potential anticancer properties of vitekwangin B, a natural compound derived from Vitex trifolia. Methods: Vitekwangin B was chromatographically isolated from the fruits of V. trifolia. ANO1 protein levels in prostate cancer and NSCLC cells were verified and evaluated again after vitekwangin B treatment. Results: Vitekwangin B did not inhibit anoctamin1 (ANO1) channel function but significantly reduced ANO1 protein levels. These results demonstrate that vitekwangin B effectively inhibited cancer cell viability and induced apoptosis in prostate cancer and NSCLC cells. Moreover, it exhibited minimal toxicity to liver cells and did not affect hERG channel activity, making it a promising candidate for further development as an anticancer drug. Conclusion: Vitekwangin B may offer a new direction for cancer therapy by targeting ANO1 protein, potentially improving treatment outcomes in patients with prostate cancer and NSCLC. Further research is needed to explore its full potential and overcome existing drug resistance challenges.

Disease spectrum and its molecular characterisation in the lentil production system of lower-Indo Gangetic plains.

Lentil is a food legume grown in the Indo-Gangetic plains including lower Gangetic Bengal (LGB). Lentil productivity in this zone is severely impeded because of the prevalence of several biotic cues. Plausible reports regarding the status of disease scenario and the associated risk factors are missing. Therefore, judicious crop management strategies are lacking. An intensive survey of 267 farmers' fields was conducted over 3 years in major lentil-growing districts of LGB to evaluate the disease incidence and prevalence. Additional insights were generated, apprehending isolation and characterisation of associated pathogens through spore morphology and molecular markers as well as elucidating the role of biophysical factors in influencing disease development. Climate change has shifted the disease dimension of lentil and precipitated new disease complexes of great risk, which was reflected through geospatial mapping results in the present study. The prevalence of three major diseases, namely collar rot (Sclerotium rolfsii), lentil blight complex (LBC) incited by both Alternaria and Stemphylium, and lentil rust (Uromyces viciae-fabae), was ascertained through cultural and molecular studies and contextualised through pathogenicity appraisal. This study is the first to investigate the complex mixed infection of Alternaria alternata and Stemphylium botryosum, successfully isolating S. botyrosum in India, and confirming the pathogens through sequencing by using internal transcribed spacer (ITS) primers and Stemphylium-specific Glycerol-3-phosphate dehydrogenase 1 (gpd1) and gpd2 primers. Unlike late planting, early planting promoted collar rot infestation. LBC and rust incidence were magnified in late planting. Soil texture resulted in the spatial distribution of collar rot disease. The surveyed data also highlighted the potential role of resistant cultivars and cropping pattern intervention to ensure associational resistance towards addressing the disease bottleneck in lentil.

ANO1-downregulation induced by schisandrathera D: a novel therapeutic target for the treatment of prostate and oral cancers.

Anoctamin 1 (ANO1), a drug target for various cancers, including prostate and oral cancers, is an intracellular calcium-activated chloride ion channel that plays various physiopathological roles, especially in the induction of cancer growth and metastasis. In this study, we tested a novel compound isolated from Schisandra sphenanthera, known as schisandrathera D, for its inhibitory effect on ANO1. Schisandrathera D dose-dependently suppressed the ANO1 activation-mediated decrease in fluorescence of yellow fluorescent protein; however, it did not affect the adenosine triphosphate-induced increase in the intracellular calcium concentration or forskolin-induced cystic fibrosis transmembrane conductance regulator activity. Specifically, schisandrathera D gradually decreased the levels of ANO1 protein and significantly reduced the cell viability in ANO1-expressing cells when compared to those in ANO1-knockout cells. These effects could be attributed to the fact that schisandrathera D displayed better binding capacity to ANO1 protein than the previously known ANO1 inhibitor, Ani9. Finally, schisandrathera D increased the levels of caspase-3 and cleaved poly (ADP-ribose) polymerase 1, thereby indicating that its anticancer effect is mediated through apoptosis. Thus, this study highlights that schisandrathera D, which reduces ANO1 protein levels, has apoptosis-mediated anticancer effects in prostate and oral cancers, and thus, can be further developed into an anticancer agent.

Unveiling the Potentiality of Shikonin Derivatives Inhibiting SARS-CoV-2 Main Protease by Molecular Dynamic Simulation Studies.

Shikonin, a phytochemical present in the roots of Lithospermum erythrorhizon, is well-known for its broad-spectrum activity against cancer, oxidative stress, inflammation, viruses, and anti-COVID-19 agents. A recent report based on a crystallographic study revealed a distinct conformation of shikonin binding to the SARS-CoV-2 main protease (Mpro), suggesting the possibility of designing potential inhibitors based on shikonin derivatives. The present study aimed to identify potential shikonin derivatives targeting the Mpro of COVID-19 by using molecular docking and molecular dynamics simulations. A total of 20 shikonin derivatives were screened, of which few derivatives showed higher binding affinity than shikonin. Following the MM-GBSA binding energy calculations using the docked structures, four derivatives were retained with the highest binding energy and subjected to molecular dynamics simulation. Molecular dynamics simulation studies suggested that alpha-methyl-n-butyl shikonin, beta-hydroxyisovaleryl shikonin, and lithospermidin-B interacted with two conserved residues, His41 and Cys145, through multiple bonding in the catalytic sites. This suggests that these residues may effectively suppress SARS-CoV-2 progression by inhibiting Mpro. Taken together, the present in silico study concluded that shikonin derivatives may play an influential role in Mpro inhibition.

In silico design of epitope-based peptide vaccine against non-typhoidal Salmonella through immunoinformatic approaches.

Non-typhoidal Salmonella (NTS) is one of the leading bacterial causes of many invasive human infections with a high antibiotic resistance profile. With this concern, the current study aimed to design an effective epitope-based peptide vaccine against NTS species as a successive and substitutive protective measure of invasive NTS disease. To design rationally, the current study considered a comprehensive in silico workflow combination of both immunoinformatics and molecular modeling approaches, including molecular docking and molecular dynamics (MD) simulation. We identified the two most promising T cell epitopes KVLYGIFAI and YGIFAITAL, and three B cell epitopes AAPVQVGEAAGS, TGGGDGSNT, and TGGGDGSNTGTTTT, in the outer membrane of NTS. Using these epitopes, a multiepitope vaccine was subsequently constructed along with appropriate adjuvant and linkers, which showed a good binding affinity and stability with toll-like receptor 2 (TLR2) in both molecular docking and MD simulation. Furthermore, in silico immune simulation described a strong immune response with a high number of antibodies, interferon-γ, and activated B and T cells. This study collectively suggests that predicted vaccine constructs could be considered potential vaccine candidates against common NTS species.Communicated by Ramaswamy H. Sarma.

Anticancer effect of verteporfin on non-small cell lung cancer via downregulation of ANO1.

Anoctamin 1 (ANO1) is a calcium-activated chloride channel found in various cell types and is overexpressed in non-small cell lung cancer (NSCLC), a major cause of cancer-related mortality. With the rising interest in development of druggable compounds for NSCLC, there has been a corresponding rise in interest in ANO1, a novel drug target for NSCLC. However, as ANO1 inhibitors that have been discovered simultaneously exhibit both the functions of an inhibition of ANO1 channel as well as a reduction of ANO1 protein levels, it is unclear which of the two functions directly causes the anticancer effect. In this study, verteporfin, a chemical compound that reduces ANO1 protein levels was identified through high-throughput screening. Verteporfin did not inhibit ANO1-induced chloride secretion but reduced ANO1 protein levels in a dose-dependent manner with an IC50 value of ~300 nM. Moreover, verteporfin inhibited neither P2Y receptor-induced intracellular Ca2+ mobilization nor cystic fibrosis transmembrane conductance regulator (CFTR) channel activity, and molecular docking studies revealed that verteporfin bound to specific sites of ANO1 protein. Confirming that verteporfin reduces ANO1 protein levels, we then investigated the molecular mechanisms involved in its effect on NSCLC cells. Interestingly, verteporfin decreased ANO1 protein levels, the EGFR-STAT3 pathway as well as ANO1 mRNA expression. Verteporfin reduced the viability of ANO1-expressing cells (PC9, and gefitinib-resistant PC9) and induced apoptosis by increasing caspase-3 activity and PARP-1 cleavage. However, it did not affect hERG channel activity. These results show that the anticancer mechanism of verteporfin is caused via the down-regulation of ANO1.

Evaluation of occupational health management status and safety issues of the small-scale fisheries sector in Bangladesh.

BACKGROUND: Small-scale fishing is one of the most precarious occupations, with high rates of threats and hazards. The present study was undertaken to evaluate the health hazards and safety issues of fishers involved in small-scale fisheries (SSF). MATERIALS AND METHODS: Fifty SSF fishers (n = 50) were surveyed by using a pre-tested questionnaire between October 2019 and March 2020 at the lower Meghna River in the northern tip of the Bay of Bengal, Bangladesh. RESULTS: Results revealed that 56% of SSF fishermen belong to a nuclear family, and 42% completed primary education. Forty per cent had an annual income of between 1,000 and 1,500 USD. Seventy-six per cent of fishermen were found to suffer from fever, and 72%, and 60% from diarrhoea and skin diseases over the last 5 years (2015-2020), respectively. During fishing, 78% of fishermen also suffered from red-eye problems, dizziness, and headache, and 68% struggled with musculoskeletal complaints during the last 5 years. Extreme cyclonic occurrences and sudden storms were experienced by 66% and 32% of fishermen, respectively, during the last 5 years. Local pharmacies were visited by 46% of fishermen for treatment due to ease of access. Sixty-four per cent of participants applied their local indigenous knowledge to treat health-related problems. Twenty-eight per cent and 32% of fishermen used a first aid box and stored medicine on board, respectively. CONCLUSIONS: Most of the fishers are in great risk of medium- to high-range danger while fishing in the SSF sector in Bangladesh. Many countries have developed protocols for safe and responsible fishing. In Bangladesh, adequate attention is needed for the sustainable development of the SSF sector.

Nematicidal Activity of Holigarna caustica (Dennst.) Oken Fruit Is Due to Linoleic Acid.

Holigarna caustica (Dennst.) Oken is used by the tribes of Northeast India for the treatment of intestinal problems. Therefore, the present study was undertaken to investigate the active principles of this plant responsible for its anthelmintic activity, using bioassay-guided fractionation. An ethanol extract of H. caustica fruit was fractionated on a silica gel column, followed by HPLC, while nematicidal activity was followed throughout on Caenorhabditis (C.) elegans as a model organism. Our study constitutes the first nematicidal report for this plant. Bioassay-guided purification led to the isolation of one compound (IC50 = 0.4 µM) as the only active constituent in the most active fraction. The compound was identified as linoleic acid based on spectroscopic data (1H and 13C NMR and ESI-MS). No cytotoxicity was observed in the crude extract or in linoleic acid (up to 356 µM). The results support the use of H. caustica for the treatment of intestinal problems by traditional healers in India.

In silico analysis of nonsynonymous single-nucleotide polymorphisms (nsSNPs) of the SMPX gene.

Mutations in the SMPX gene can disrupt the regular activity of the SMPX protein, which is involved in the hearing process. Recent reports showing a link between nonsynonymous single-nucleotide polymorphisms (nsSNPs) in SMPX and hearing loss, thus classifying deleterious SNPs in SMPX will be an uphill task before designing a more extensive population study. In this study, damaging nsSNPs of SMPX from the dbSNP database were identified by using 13 bioinformatics tools. Initially, the impact of nsSNPs in the SMPX gene were evaluated through different in silico predictors; and the deleterious convergent changes were analyzed by energy-minimization-guided residual network analysis. In addition, the pathogenic effects of mutations in SMPX-mediated protein-protein interactions were also characterized by structural modeling and binding energy calculations. A total of four mutations (N19D, A29T, K54N, and S71L) were found to be highly deleterious by all the tools, which are located at highly conserved regions. Furthermore, all four mutants showed structural alterations, and the communities of amino acids for mutant proteins were readily changed, compared to the wild-type. Among them, A29T (rs772775896) was revealed as the most damaging nsSNP, which caused significant structural deviation of the SMPX protein, as a result reducing the binding affinity to other functional partners. These findings reflect the computational insights into the deleterious role of nsSNPs in SMPX, which might be helpful for subjecting wet-lab confirmatory analysis.

Current Knowledge on Pathogenicity and Management of Stemphylium botryosum in Lentils (Lens culinaris ssp. culinaris Medik).

Stemphylium blight (SB) caused by Ascomycete, Stemphylium botryosum Wallr. has been a serious threat to lentil cultivation, mainly in Bangladesh, Nepal, India, and Canada since its first outbreak in Bangladesh in 1986. The genus Stemphylium Wallr., a dematiaceous hyphomycete, comprises up to 150 species, and is pathogenic on a wide range of plants infecting leguminous as well as nonleguminous crops. In recent years, studies indicated overlapping in morphological characters among the different species under the genus Stemphylium, making the identification and description of species difficult. This necessitates different molecular phylogenetic analysis in species delimitation. Therefore, a detailed understanding of spatial diversity and population structure of the pathogen is pertinent for producing source material for resistance breeding. The role of different weather variables as predisposing factors for the rapid spread of the pathogen necessitates devising a disease predictive model for the judicial application of fungicides. A dearth of information regarding spore biology, epidemiology, race diversity, host-pathogen interaction, and holistic disease management approach necessitates immediate attention towards more intensive research efforts. This is the first comprehensive review on the current state of knowledge and research efforts being made for a better understanding of the SB resistance through cognizing biology, ecology, and epidemiology of S. botryosum and effective disease management strategies to prevent widespread outbreaks of SB. The information regarding the biology and epidemiology of S. botryosum is also crucial for strengthening the "Integrated Disease Management" (IDM) programme. The need for a regional research network is advocated where the disease is becoming endemic.

Antibacterial, anthelmintic, and analgesic activities of Piper sylvaticum (Roxb.) leaves and in silico molecular docking and PASS prediction studies of its isolated compounds.

Background In the present study, we investigated the antibacterial, anthelmintic, and analgesic activities of methanol extract of P. sylvaticum leaves (MEPSL) in experimental models. Then, computational analysis (in silico molecular docking and PASS prediction) was performed to determine the potent phytoconstituents of total six isolated compounds of this plant for antibacterial and anthelmintic activities. Methods Qualitative and quantitative phytochemical studies were carried out by established methods. In vitro antibacterial activity was determined by disc diffusion technique and anthelmintic activity was tested against Tubifex tubifex worm whereas analgesic activity was determined by the acetic acid-induced writhing test in mice. Molecular docking study was performed using Schrödinger Maestro 10.1 and an online tool used for PASS prediction. Results Our phytochemical study revealed the presence of alkaloids, flavonoids, saponins, and also indicated a substantial amount of phenols (65.83 mg), flavonoids (102.56 mg), and condensed tannins (89.32 mg). MEPSL showed good antibacterial activity against both gram-positive and gram-negative bacteria. Our result exhibited that MEPSL has strong anthelmintic action compared to standard levamisole. In addition, the extract also showed a dose-dependent and statistically significant analgesic activity at the doses of 200 and 400 mg/kg, body weight. Docking studies showed that piperine and piperlonguminine have the best scores for the tested enzymes. PASS predicted the antibacterial and anthelmintic activity of both phytoconstituents. Conclusions This study suggests that MEPSL possess significant antibacterial, anthelmintic, and analgesic activities which could be related to the presence of several phytochemicals. The phytoconstituents, i.e. piperine and piperlonguminine were found to be most effective in computational studies.

Assessing medicinal plants traditionally used in the Chirang Reserve Forest, Northeast India for antimicrobial activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Knowledge on the use of plants for different ailments by the tribals of the Chirang Reserve Forest (CRF, Northeast India) was used to assess the potential of these plants for treating viral, bacterial and fungal infections. MATERIALS AND METHODS: Fieldwork in the CRF documented the use of plants for the treatment of various human ailments. The ethnobotanical data were analysed using different quantitative indices viz. Informant Consensus Factor (ICF), Relative Frequency Citation (RFC), Fidelity Level (FL), Relative Popularity Level (RPL) and Rank Order Priority (ROP). For each plant, four extracts were prepared with different solvents (water, ethanol, acetone and hexane) and tested for bioactivity such as antiviral (Enterovirus 71) and antimicrobial (E. coli, S. aureus, C. albicans) effects. To develop fingerprints, thin layer chromatography (TLC) was carried out with at least one extract of each active plant. RESULTS: Thirty-seven plant species belonging to 26 families used frequently in the CRF were categorised into twenty disease(s) and markedly high ICF values (0.5-1) were found. The FL of the 19 most important plant species ranged from 10% to 100%. Aglaia spectabilis, Actinodaphne obovata, Bischofia javanica, Gmelina arborea, Hodgsonia macrocarpa and Mesua ferrea were the most popular plant species with RPL values > 0.4. The most commonly treated diseases were skin infections, worm infections, diarrhoea, dysentery, common cold and throat infections, cough and respiratory problems, fever, malaria, rheumatism, pain, stomach ache and gastric problems. Among the taxonomic families, the most represented are Lauraceae (4 species); Meliaceae (3 species), Euphorbiaceae, Elaecarpoceae, Magnoliaceae, Malvaceae, Moraceae and Myrtaceae (2 species each). Of these, 35 plants (95%) with 91 of their extracts were found to be active against S. aureus (inhibition > 50%). Similarly, over half of the tested plants inhibited growth of E. coli (19 plants with 31 extracts) while 9 plants with 14 extracts were active against C. albicans. Moreover, one-third (12) of the plants showed anti-enteroviral activity. TLC analysis of the extracts indicates the presence of different phytochemical classes such as alkaloids, flavonoids, glycosides, terpenoids and saponins based on observing characteristic spots under visible or UV light (254 or 360 nm), with or without derivatisation. CONCLUSIONS: Thirty-seven plants commonly used by the tribes of the CRF as anti-infective agents offer perspectives for further research, as the phytochemistry and phytopharmacology for most of these plants have not been published to date.

Biodiversity of epilithic cyanobacteria from freshwater streams of Kakoijana reserve forest, Assam, India.

The biodiversity of epilithic cyanobacteria from one of the unexplored habitats of freshwater streams of Kakoijana reserve forest of Assam, India was estimated. This paper lists a total of 29 species representing 18 genera of 12 families and 4 orders as per recent system of classification. Morphological descriptions, common habitats and distribution pattern were described for each species identified that were represented systematically. Of these 29 species, 11 were unicellular, 9 non-heterocytous filamentous and 9 heterocytous filamentous forms. All the unicellular (Aphanocapsa crassa, A. muscicola, Aphanothece nidulans, A. saxicola, Chlorogloea purpurea, Chroococcus cohaerens, C. minimus, C. minor, Cyanobacterium cedrorum, Cyanocystis versicolor and Gloeocapsopsis crepidinum) and 13 (Calothrix epiphytica, C. scopulorum, Leptolyngbya boryana, L. calotrichoides, L. fragilis, L. notata, Lyngbya arboricola, Nostoc humifusum, N. oryzae, N. punctiforme, Parthasarathiella prolifica, Porphyrosiphon ceylanicus and Scytonema millei) of the remaining 18 species were recorded for the first time as freshwater epiliths. While, 5 species (Hapalosiphon welwitschii, Leptolyngbya tenuis, Oscillatoria pseudogeminata, Phormidium laetevirens, Tolypothrix fragilis) and 8 species (Aphanothece saxicola, Calothrix scopulorum Chlorogloea purpurea, Chroococcus minor, Gloeocapsopsis crepidinum, Leptolyngbya calotrichoides, L. fragilis and L. tenuis) were reported earlier as freshwater-and marine-epilithic forms respectively. All are new records for Assam except 6 species (A. nidulans, H. welwitschii, N. punctiforme, N. oryzae, O. pseudogeminata and P. ceylanicus), while 3 species (C. purpurea, L. boryana and L. calotrichoides) are new records for India. Six nitrogen fixing heterocytous forms such as, C. epiphytica, C. scopulorum, N. humifusum, N. punctiforme, N. oryzae and S. millei, were common to the neighboring paddy fields.