In-vitro cytotoxicity in relation to chemotypic diversity in diploid and tetraploid populations of Gentiana kurroo Royle.

ETHNOPHARMACOLOGICAL RELEVANCE: Gentiana kurroo is a multipurpose critically endangered medicinal herb prescribed as medicine in Ayurveda in India and exhibits various pharmacological properties including anti-cancer activity. The species is rich repository of pharmacologically active secondary metabolites together with secoiridoidal glycosides. AIM OF THE STUDY: The study aimed to investigate the chemical diversity in different populations/cytotypes prevailing in G. kurroo to identify elite genetic stocks in terms of optimum accumulation/biosynthesis of desired metabolites and having higher in-vitro cytotoxicity potential in relation to chemotypic diversity. MATERIAL AND METHODS: The wild plants of the species were collected from different ranges of altitudes from the Kashmir Himalayas. For cytological evaluation, the standard meiotic analysis was performed. The standard LC-MS/MS technique was employed for phytochemical analysis based on different marker compounds viz. sweroside, swertiamarin, and gentiopicroside. Different tissues such as root-stock, aerial parts, and flowers were used for chemo-profiling. Further, the methanolic extracts of diploid and tetraploid cytotypes were assessed for cytotoxic activity by using MTT assay against four different human cancer cell lines. RESULTS: The quantification of major bioactive compounds based on tissue- and location-specific comparison, as well as in-vitro cytotoxic potential among extant cytotypes, was evaluated. The comprehensive cytomorphological studies of the populations from NW Himalayas revealed the occurrence of different chromosomal races viz. n = 13, 26. The tetraploid cytotype was hitherto unreported. The tissue-specific chemo-profiling revealed relative dominance of different phytoconstituents in root-stock. There was a noticeable increase in the quantity of the analyzed compounds in relation to increasing ploidy status along the increasing altitudes. The MTT assay of methanolic extracts of diploid and tetraploid cytotypes displayed significant cytotoxicity potential in tetraploids. The root-stock extracts of tetraploids were highly active extracts with IC50 value ranges from 5.65 to 8.53 µg/mL against HCT-116 colon cancer. CONCLUSION: The chemical evaluation of major bioactive compounds in diverse cytotypes from different plant parts along different altitudes presented an appreciable variability in sweroside, swertiamarin, and gentiopicroside contents. Additionally, the concentrations of these phytoconstituents varied for cytotoxicity potential among different screened cytotypes. This quantitative difference of active bio-constituents was in correspondence with the growth inhibition percentage of different tested cancer cell lines. Thus, the present investigation strongly alludes towards a prognostic approach for the identification of elite cytotypes/chemotypes with significant pharmacological potential.

Thyroid Function, Urinary Iodine, and Thyroid Antibody Status Among the Tribal Population of Kashmir Valley: Data From Endemic Zone of a Sub-Himalayan Region.

Background: There are scarce data on the prevalence of thyroid disorders and urinary iodine status among tribal populations of India, with no reported data from Kashmir valley. Objectives: To estimate the prevalence of thyroid disorders and evaluate urinary iodine concentration (UIC) and thyroid autoantibody status among Gujjar and Bakerwal tribes of Kashmir valley. Methods: This cross-sectional study recruited tribal subjects using multistage cluster sampling from 5 out of 22 districts of Jammu and Kashmir (J&K). Using a predesigned questionnaire, the details of past or current medical history and drug intake, including thyroid hormone medication etc. were recorded after obtaining an informed consent. Examination included anthropometry (height, weight, waist circumference), blood pressure measurement, and relevant general physical examination focusing on goiter palpation, while as laboratory assessment included estimation of serum thyroid hormone levels, antithyroid peroxidase antibody (anti-TPO Ab), and urinary iodine concentration. Results: A total of 763 subjects (56.4% women and 43.6% men) with a mean(±SD) age of 39.46 (±17.51) ranging from 10 to 85 years and mean(±SD) body mass index (BMI) of 21.28 (±4.16) kg/m2 were studied. Goiter was detected in 6.8%, while 33.2% subjects had some form of thyroid dysfunction (including 24.1% subclinical and 6.8% overt hypothyroidism). Subclinical and overt hyperthyroidism were observed in 1.3 and 0.9% of cases, respectively. Anti-TPO Ab was elevated in 13.6%, while the median [interquartile range (IQR)] for UIC was 154.50 (135) µg/L [156.13 (134) µg/L in men and 147.26 (136) µg/L in women]. A negative correlation was observed between UIC and anti-TPO Ab (r = -0.087, P = <0.05). Conclusion: These novel data on iodine and thyroid status among a tribal population of India generally inhabiting in remote sub-Himalayan belts demonstrate high prevalence of subclinical hypothyroidism (SCH) with persistent iodine deficiency. These preliminary data may warrant large well-designed studies to carry out comprehensive assessment of the problem in this high-risk and marginalized population.

Cross sectional study on Kashmiri tribal population: Their demo-economic status and behavioural risk factors.

BACKGROUND: The demographic particulars of Gujar-Bakarwals are mostly dominant on mountainous regions of Kashmir Valley. Their housing, sanitation, health care facilities are very low sub-standard than other sections of population. Behavioral risk factors including tobacco use and skipping meals are prevalent. OBJECTIVE: The present study focuses on the socio-economic and demographic profile among the tribal population of Kashmir; their major risk factors of some non-communicable diseases. METHODS: The study is community based cross sectional survey undertaken in selected districts of Jammu and Kashmir. RESULTS: Around 94.3% of the tribal population fell under low income groups with an annual income of Rs. <25000 per year. Only 37.1% subjects were educated. 61.0% of tribal subjects lack access to pure drinking water and proper sanitation. Interestingly, 63-66% of the population was younger with a high prevalence of smoking among both males and females (33.3% males and 7.3%, respectively). Among non-communicable diseases, diabetes was less prevalent whereas a comparatively higher prevalence of hypertension, dyslipidemia, thyroid dysfunction, and vitamin D deficiency was present with significant associations with the risk factors. CONCLUSIONS: There is widespread poverty, illiteracy, and lack of basic amenities among the tribal people which makes it imperative to address these concerns to improve the socioeconomic disparities and health indices of the marginalized population. Smoking and inadequate consumption of meals was prevalent. There is an urgent need to address behavioral risk factors such as smoking and skipping meals through primary prevention.

Simultaneous quantification of triterpenoic acids by high performance liquid chromatography method in the extracts of gum resin of Boswellia serrata obtained by different extraction techniques.

BACKGROUND: Boswellia serrata, also known as Indian frankincense is a commercially important medicinal plant which has been used for hundreds of years as an Ayurvedic medicine for the attempted treatment of arthritis. It contains naturally occurring triterpenoic acids, called as boswellic acids (BA's). RESULTS: A highly reproducible High performance liquid chromatography-ultraviolet diode array detection (HPLC-UV-DAD) method was developed for the simultaneous determination and quantitative analysis of eight major triterpenoic acids in Boswellia serrata gum resin obtained by different extraction techniques. All the calibration curves exhibited good linear regression (R(2) > 0.997) within the test ranges. The established method showed good precision and overall recoveries of the boswellic acids. CONCLUSIONS: The eight triterpenoic acids coded as BS-1 (11-keto-beta-boswellic acid), BS-2 (3-O-acetyl-11-keto-beta-boswellic acid), BS-3 (3-keto tirucallic acid), BS-4 (3-O-acetyl-alpha-tirucallic acid), BS-5 (3-O-acetyl-beta-tirucallic acid), BS-6 (alpha-boswellic acid), BS-7 (beta-boswellic acid) and BS-8 (3-O-acetyl-beta-boswellic acid) were isolated from the processed gum resin of Boswellia serrata by column chromatography. The proposed HPLC method is simple, reliable and has been very useful for the qualitative as well as quantitative analysis of boswellic acids in the gum resin of Boswellia serrata. The proposed method allows to quantify boswellic acids in appreciable amounts by HPLC-UV (DAD) method in the extracts and the available marketed formulations.Graphical abstractIsolation & separation of eight Triterpenoic acids from Boswellia serrata.

TNF-α inhibitory effect of Euphorbia hirta in rats.

CONTEXT: Euphorbia hirta L. (Euphorbiaceae) (E. hirta) is a tree locally used as a traditional medicine in Africa and Australia to treat numerous diseases such as hypertension, respiratory ailments, tumors, and wounds, and it has reported antiallergic, antipyretic, anti-inflammatory activities, etc. OBJECTIVE: This study evaluated the ability of fresh leaves of E. hirta ethanol extract to inhibit the intracellular tumor necrosis factor α (TNF-α) level in the synovial fluid and neutrophils in lipopolysaccharide (LPS)-induced inflamed rat knees. MATERIALS AND METHODS: Female Wister albino rats 140-160 g were used. E. hirta ethanol extract was given orally at 25, 50, 100, and 200 mg/kg, 2 h before an intra-articular (i.a.) injection of LPS. Two and three hours later, synovial fluid and neutrophils levels of intracellular TNF-α production were measured. RESULTS: In the time course of the experiment, E. hirta maximum inhibition at 100 and 200 mg/kg (p.o.) dose showed 16.5 ± 1.34 and 14.4 ± 1.30% of synovial fluid, 4.26 ± 0.36 and 3.78 ± 0.29% of neutrophils levels of intracellular TNF-α productions at 2 h after LPS injection. LPS control displayed 22.97 ± 1.61 and 6.78 ± 0.34% of synovial fluid and neutrophils levels of intracellular TNF-α at 2 h after LPS injection. Intracellular TNF-α was also estimated at 3 h after LPS injection. DISCUSSION AND CONCLUSION: The LPS-injected rat knee model gives a comparative study of acute anti-inflammatory responses. E. hirta inhibition of proinflammatory intracellular cytokine TNF-α production with LPS-induced inflamed rat knee is of great importance in defining the anti-arthritic potential of E. hirta.

Immunosuppressive effects of Euphorbia hirta in experimental animals.

Euphorbia hirta L. (Euphorbiaceae) (E. hirta) is a tree locally used as a traditional medicine in Africa and Australia to treat numerous diseases such as hypertension, respiratory ailments, tumors, wounds, antipyretic, anti-inflammatory activities, etc. Therefore, we undertook to investigate their immunomodulatory effect on T lymphocytes (CD3+, CD4+ and CD8+ receptors) and Th1 cytokines (IL-2, TNF-α, IFN-γ) in a dose-dependent manner. E. hirta ethanol extract at 25, 50, 100 and 200 mg/kg doses was given orally for 7 days from the day of immunization. E. hirta maximum inhibition at 100 and 200 mg/kg p.o. was found to significantly block the production of the cell-mediated immune response, (CD3+, CD4+ and CD8+ receptors) and (IL-2, TNF-α, IFN-γ) and also prolongs graft rejection. E. hirta also showed a decrease of delayed hypersensitivity (DTH) response and dose-related decrease in the primary antibody response, respectively. Based on the data, it can be suggested that E. hirta is a potent and non-toxic immunosuppressor, which can be further explored for the development of potent immunosuppressor.