VDR downregulation and promoter hypermethylation as one of the causes for triggering type 2 diabetes mellitus: Clinical and molecular studies.

Background: Our present study was to investigate the methylation and Gene expression of the vitamin D receptor (VDR) gene in the causing T2DM and to determine the inflammatory biomarkers in exaggerating T2DM in Kashmiri population. Methods: In this study, T2DM cases (n = 100) and controls (n = 100) of Kashmiri population were designed. Blood samples were taken from both groups, and serum vitamin D levels, inflammatory biomarkers (TNF-α, IL-6, IL-10, CRP, Leptin and adiponectin) were estimated by ELISA. By using methylation-specific PCR (MS-PCR) and RT-PCR, respectively, the levels of methylation and expression were measured after the extraction of DNA and RNA. Results: Studies using RT-PCR demonstrated that patients with diabetes had a lower degree of VDR expression than control subjects (P > 0.05). The T2DM was shown to be strongly correlated with hypermethylation (p-value < 0.001, OR 2.9; 95%CI 1.6-5.54). When compared to control groups, T2DM patients' levels of vitamin D in their serum were considerably lower (p < 0.01). Pro-inflammatory mediators like TNF-α, CRP, IL-6, and leptin levels were discovered to be higher, and concentrations of anti-inflammatory mediators like IL-10 and adiponectin were observed to be lower in people with T2DM than in people without the condition (p < 0.05). Conclusions: This study suggests the hypermethylation and down expression of VDR as one of the basis for causing T2DM in kashmiri individuals, exaggerated by enhanced degree of TNF-α, CRP, IL-6 and leptin and diminished concentration of IL-10 and adiponectin in T2DM. Supplementary Information: The online version contains supplementary material available at 10.1007/s40200-023-01266-6.

Molecular characterization of fungal endophyte diversity isolated from Aconitum heterophyllum: a critically endangered medicinal plant of Kashmir Himalaya.

Aconitum heterophyllum is a rare perennial herb from Kashmir Himalayas. Due to its threatened status and dependence on its environment, the plant was examined for any potential endophytes, which is of utmost importance for bioprospection. In the current study, endophytic fungal diversity associated with A. heterophyllum was examined, and 328 fungal isolates were found in the plant's leaf, stem, and root tissues. Twelve (12) endophytic fungal species were identified utilizing, molecular analysis of the nuclear ribosomal DNA Internal Transcribes Spacer (ITS), rLSU, and rSSU sequences. Maximum likelihood analysis was used to determine the phylogenetic connection between each isolate. The genera Arthrinium, Chaetomium, Purpureocillium, Alternaria, Penicillium, Aspergillus, Cladosporium, and Bjerkandera species dominated the ascomycete and basidiomycete fungal endophytes.

Phytochemical screening and antibacterial activity of Skimmia anquetilia N.P. Taylor and Airy Shaw: A first study from Kashmir Himalaya.

The present study aimed to explore the antibacterial activity of various organic root extracts of Skimmia anquetilia N.P. Taylor and Airy Shaw and the identification of major functional groups and phytoconstituents through fourier transform infrared spectrometer (FTIR) and gas chromatography-mass spectrometer (GC-MS). The extracts were evaluated for antibacterial activity against multidrug-resistant (MDR) strains viz., Pseudomonas aeruginosa (MTCC424), Escherichia coli (MTCC739), Klebsiella pneumoniae (MTCC139), Salmonella typhi (MTCC3224), and Staphylococcus aureus (MTCC96). ESKAPE pathogens such as S. aureus, K. pneumoniae, and P. aeruginosa are responsible for a majority of all healthcare acquired infections. The ethyl acetate extract showed the highest zone of inhibition against P. aeruginosa (18 mm) followed by S. aureus (17 mm). The minimum inhibitory concentration (MIC) of ethyl acetate extract against strain of S. aureus (4 mg mL-1) demonstrated therapeutically significant antibacterial activity. The FTIR spectra of root extracts revealed the occurrence of functional characteristic peaks of alcohols, carboxylic acids, aromatic compounds, alkanes, alkenes, and amines that indicates the presence of various metabolites in the extracts. The GC-MS investigation led to the identification of diverse phytoconstituents in each of the extracts with varying concentrations and molecular masses. The highest number of compounds were identified from the methanol extract (112), followed by n-hexane extract (88) and ethyl acetate extract (74). The most predominant compounds were 5, 10-pentadecadien-1-ol, (Z,Z)-(33.94%), n-hexadecanoic acid (13.41%) in n-hexane extract, 5,10-pentadecadien-1-ol, (Z,Z)-(10.48%), 1-hexyl-2-nitrocyclohexane (7.94%) in ethyl acetate extract, and 1-hexyl-2-nitrocyclohexane (15.43%), cis,cis,cis-7,10,13-hexadecatrienal (13.29%) in methanol extract. The results of the present study will create a way for the invention of plant-based medicines for various life-threatening microbial infections using S. anquetilia, which may lead to the development of novel drugs against drug-resistant microbial infections.

Skimmia anquetilia N.P. Taylor and Airy Shaw (Rutaceae): A Critical Appriasal of its Ethnobotanical and Pharmacological Activities.

Skimmia anquetilia N.P. Taylor and Airy Shaw (Rutaceae) is a perennial, aromatic, gregarious wild ornamental shrub native to the Western Himalaya. The plant is used in the traditional medicinal system to treat copious health conditions like rheumatism, fever, inflammation, headache, influenza, body-ache, clearing of the nose, diabetes, lowering the body temperature, smallpox, wounds, burns, snake, and scorpion bites. Phytochemical and gas chromatography-mass spectrometer (GC-MS) analysis of S. anquetilia showed the presence of alkanes, alkenes, carboxylic acids, fatty acids, and their esters, simple coumarins, terpenes, phenylpropanoid, and so on. These active principles exhibit a wide array of pharmacological effects, including anti-inflammatory, antioxidant, anti-cancerous, anti-feedant, and antibacterial properties. Most pharmacological studies were based on the essential oil and the crude extracts of the plant and the bioactive compounds responsible for the bioefficacy have not been well-identified. Further investigations are required to transform the experience-based claims on the use of S. anquetilia in traditional medicine practices into evidence-based information. Detailed in-vitro and in-vivo studies on the mechanisms of action of pure bioactive compounds and more elaborate toxicity studies to ensure plant safety for human use should be conducted. This review recapitulates the current status of its use in the ethnobotany, phytochemistry, and pharmacological activities. It also offers a critical assessment of the plant's existing information which would help to recuperate its potential as a source for drug development of lead molecules.

Marine bacteria and omic approaches: A novel and potential repository for bioremediation assessment.

Marine environments accommodating diverse assortments of life constitute a great pool of differentiated natural resources. The cumulative need to remedy unpropitious effects of anthropogenic activities on estuaries and coastal marine ecosystems has propelled the development of effective bioremediation strategies. Marine bacteria producing biosurfactants are promising agents for bio-remediating oil pollution in marine environments, making them prospective candidates for enhancing oil recovery. Molecular omics technologies are considered an emerging field of research in ecological and diversity assessment owing to their utility in environmental surveillance and bioremediation of polluted sites. A thorough literature review was undertaken to understand the applicability of different omic techniques used for bioremediation assessment using marine bacteria. This review further establishes that for bioremediation of environmental pollutants (i.e. heavy metals, hydrocarbons, xenobiotic and numerous recalcitrant compounds), organisms isolated from marine environments can be better used for their removal. The literature survey shows that omics approaches can provide exemplary knowledge about microbial communities and their role in the bioremediation of environmental pollutants. This review centres on applications of marine bacteria in enhanced bioremediation, using the omics approaches that can be a vital biological contrivance in environmental monitoring to tackle environmental degradation. The paper aims to identify the gaps in investigations involving marine bacteria to help researchers, ecologists and decision-makers to develop a holistic understanding regarding their utility in bioremediation assessment.

Relevance of vitamin D3 in COVID-19 infection.

SARS-CoV-2 virus, the main culprit for COVID-19 disaster, has triggered a gust of curiosity both in the mechanism of action of this infection as well as potential risk factors for disease generation and regimentation. The prime focus of the present review, which is basically a narrative one, is in utilizing the current concepts of vitamin D3 as an agent with myriad functions, one of them being immunocompetence and a promising weapon for both innate and adaptive immunity against COVID-19 infection. Some of the manifestations of SARS-CoV-2 virus such as Acute Respiratory Distress Syndrome (ARDS) overlap with the pathophysiological effects that are overcome due to already established role of vitamin D3 e.g., amelioration of cytokine outburst. Additionally, the cardiovascular complications due to COVID-19 infection may also be connected to vitamin D3 levels and the activity of its active forms. Eventually, we summarise the clinical, observational and epidemiological data of the respiratory diseases including COVID-19 disease and try to bring its association with the potential role of vitamin D3, in particular, the activity of its active forms, circulating levels and its supplementation, against dissemination of this disease.

Artemisia amygdalina Upregulates Nrf2 and Protects Neurons Against Oxidative Stress in Alzheimer Disease.

Alzheimer disease is a complex neurodegenerative disorder. It is the common form of dementia in elderly people. The etiology of this disease is multifactorial, pathologically it is accompanied with accumulation of amyloid beta and neurofibrillary tangles. Accumulation of amyloid beta and mitochondrial dysfunction leads to oxidative stress. In this study, neuroprotective effect of Artemisiaamygdalina against H2O2-induced death was studied in differentiated N2a and SH-SY5Ycells. Cells were treated with H2O2 to induce toxicity which was attenuated by Artemisia amygdalina. The nuclear factor erythroid 2-related factor 2 (Nrf2) is an emerging regulator of cellular resistance to oxidants. It controls the basal and induced expression of antioxidant response element-dependent genes. Further, we demonstrated that Artemisia amygdalina protects neurons through upregulation of Nrf2 pathway. Moreover, reactive oxygen species and mitochondrial membrane potential loss formed by H2O2 was attenuated by Artemisia amygdalina. Thus, Artemisia amygdalina may have the possibility to be a therapeutic agent for Alzheimer disease.

Scientific Study of Gentiana kurroo Royle.

The present investigation was carried out to review and highlight the potential phytochemicals and medicinal phenomena of the critically endangered medicinal plant, Gentiana kurroo Royle of the western and north-western Himalayas. The medicinal plant is heavily exploited for root and rhizome. Due to its endemic nature and the high rate of exploitation from its natural habitat, this species had become critically endangered. The phytochemical screening of the plant revealed that the plant contains some vital phyto-constituents (iridoids, xanthones, C-glucoxanthone mangiferin, and C-glucoflavones) that have a medicinal value for various acute and chronic diseases. Several researchers have carried out experimental work to validate the folkloric use of the medicinal plant for different ailments like antibacterial, antioxidant, anti-arthritic, anti-inflammatory, analgesic activities and anti-diabetic activity. However, it is yet to be confirmed the antifungal activity of the same plant. Because of endemic nature and high rate of exploitation there is need for alternative method called bio-prospecting of Endophytes from the plant, to carry out the production and characterization of bioactive metabolites for pharmacological uses and can become a conservative tool for the medicinal plant.

Podophyllum hexandrum ameliorates endosulfan-induced genotoxicity and mutagenicity in freshwater cyprinid fish crucian carp.

CONTEXT: Medicinal plants continue to act as a repository for novel drug leads with novel mechanisms of action. Podophyllum hexandrum Royale (Berberideceae) treats diverse conditions in folk medicine. OBJECTIVE: The antimutagenic potential of P. hexandrum was evaluated against endosulfan-induced clastogenicity in a piscine model by cytogenetic endpoints. MATERIALS AND METHODS: Podophyllum hexandrum rhizomes were subjected to successive solvent extraction. Fish were exposed to hexane, chloroform, ethyl acetate, methanol and aqueous extracts (15 mg/L each) of plant and endosulfan (0.05 mg/L) alone followed by their combination for antimutagenicity estimates. Chromosomal aberrations (CA) were made from kidney cells and micronuclei (MN) slides from peripheral blood erythrocytes at 48, 72 and 96 h. Antioxidant activity was analyzed by the DPPH assay. Phytochemical analyses were carried out using chromatographic and spectroscopic techniques. RESULTS: Endosulfan induced significant (p < .05) MN, authenticated by scanning electron microscopy, and CA in a time-dependent manner. However, methanol and ethyl acetate extracts revealed ameliorating effects. The column eluted methanolic fraction-2 (ME-F2) showed highest reduction profile of 83 and 84% in CA and MN, followed in its extent (73 and 72%) by ethyl acetate fraction-4 (EE-F4). ME-F2 and EE-F4 showed three and six major peaks when analyzed by GC-MS. To explore possible mechanism of action, ME-F2 showed potent antioxidant potential and strong correlation (R2 = .900) with antimutagenic activity, whereas EE-F4 seemed to act through a different mechanism. DISCUSSION AND CONCLUSION: This study confirms the antimutagenic potential of the subject plant with the identification of some novel compounds, justifying their use in folk medicine, and their corresponding benefit to mankind.

Isolation and antiproliferative activity of Lotus corniculatus lectin towards human tumour cell lines.

The objective of the study was to investigate the anti cancer activity of a lectin isolated from Lotus corniculatus seeds. A tetrameric 70kDa galactose specific lectin was purified using two step simple purification protocol which involved affinity chromatography on AF-BlueHC650M and gel filtration on Sephadex G-100. The lectin was adsorbed on AF-BlueHC650M and desorbed using 1M NaCl in the starting buffer. Gel filtration on Sephadex G-100 yielded a major peak absorbance that gave two bands of 15kDa and 20kDa in SDS PAGE. Hemagglutination activity was completely preserved, when the temperature was in the range of 20-60°C. However, drastic reduction in activity occurred at temperatures above 60°C. Full hemagglutination activity was retained at ambient pH 4-12. Thereafter no activity was observed above pH 13. Hemaglutination of the lectin was inhibited by d-galactose. The lectin showed a strong antiproliferative activity towards human leukemic (THP-1) cancer cells followed by lung cancer (HOP62) cells and HCT116 with an IC50 of 39µg/ml and 50µg/ml and 60µg/ml respectively. Flow cytometry analysis showed an increase in the percentage of cells in sub G0G1 phase confirming that Lotus corniculatus lectin induced apoptosis. Morphological observations showed that Lotus corniculatus lectin (LCL) treated THP-1 cells displayed apparent apoptosis characteristics such as nuclear fragmentation, appearance of membrane enclosed apoptotic bodies and DNA fragmentation. Lotus corniculatus lectin (LCL) effectively inhibits the cell migration in a dose dependent manner as indicated by the wound healing assay.

Variation in bioactive principles of Artemisia amygdalina Decne. in wild and tissue culture regenerants.

Wild and tissue culture raised regenerants of Artemisia amygdalina, a critically endangered and endemic plant of Kashmir and North West Frontier Provinces of Pakistan were screened for the amount of bioactive principles and in particular antimalarial compound artemesinin. Phytochemical screening of extracts revealed the presence of terpenes, alkaloids, phenolics, tannins (polyphenolics), cardiac glycosides and steroids in wild (aerial, inflorescence) and tissue culture regenerants (in vitro grown plant, callus and green house acclimatized plants). HPLC of Artemisia amygdalina revealed the presence of artemesinin in petroleum ether extracts of wild aerial part, tissue culture raised plant and green house acclimatized plants. Acetonitrile and water in 70:30 ratios at flow rate of 1ml/min was standardised as mobile phase. Retention time for standard chromatogram was 6.7. Wild inflorescences and callus does not produce artemesinin. This is the first report of phytochemical screening and artemesinin estimation of wild and tissue culture raised regenerants of Artemisia amygdalina.

Free radical scavenging activity of Elsholtzia densa.

Medicinal plants have been used traditionally to cure a variety of diseases since ancient times. Elsholtzia densa, a rare annual herb of the Kashmir valley, was assessed for its antioxidant efficacy. Antioxidant activity of the crude extracts was evaluated using 1, 1- diphenyl-2-picryl hydrazyl free radical (DPPH), DNA sugar damage, lipid peroxidation, ferric thiocyanate (FTC) and hydrogen peroxide scavenging assays. The maximum percentage decrease of 1,1-diphenyl-2-picryl hydrazyl radical (DPPH) standard solution was recorded for the 50% ethanolic extract (90.48%). The extracts were further evaluated using the thiobarbituric acid reactive substances assay. The methanolic extract showed the highest activity (32.02%) in reducing oxidative damage to DNA. The antioxidant activity of the extracts was also determined using the linoleic acid system and the highest antioxidant activity (49.64%) was found in the 50% ethanolic extract. In the case of the FTC assay, the 50% ethanolic extract showed the highest activity (70.14%) which was comparable to that of α-tocopherol. Moreover, total phenolics concentration was found to be 62.5mg% and 77.5mg% in the cases of absolute ethanolic and 50% ethanolic extracts, respectively. These findings indicate promising antioxidant activity of crude extracts of the plant and the need for further exploration of their effective use in both modern and traditional systems of medicine.