Om Prakash Chaurasia

Traditional medicinal plants of cold desert Ladakh-used against kidney and urinary disorders.

AIM OF THE STUDY Traditional medicine of clod desert Ladakh has large potential to treat various ailments among tribal communities inhabited in the remotest region of Indian subcontinent. This study was conducted to document the new ethno-medico-botanical information and traditional use of medicinal plants against kidney and urinary disorders, and thus to conserve the rapidly disappearing traditional knowledge system of Amchis of Ladakh. MATERIALS AND METHODS The information was collected from 105 villages of Leh and Kargil districts of Ladakh region by involving 47 Amchis (the herbalists), village heads and old aged persons including women population through on spot interview and repeated queries among other interviewees over a period of 3 years from 2004-2006. RESULTS The use of 68 medicinal plants belonging to 29 families and 58 genera of clod desert was documented against the treatment of kidney and urinary disorders in the tribal communities of Ladakh region in India. These species were used in combination of some exotic species such as Bergenia ligulata, Cinnamomum zeylanicum, Crocus sativus, Elettaria cardamomum, Emblica officinalis, Ficus religiosa, Mangifera indica, Punica granatum, Santalum album, Spondiax axillaris, Terminalia belerica, Terminalia chebula, Zingiber officinale and some medicinal stones, minerals and salts etc. Problem in urine discharge, burning sensation and painful urination, inflammation and bleeding in the kidney, irritable condition of bladder, haemorrhage of kidney and removal of blocked urine and kidney stone were the frequently reported disorders in the study area. CONCLUSION The effectiveness of traditional system of medicine, role of Amchis in preparation of remedies according to age, sex and severity of ailment, method of preparation, doses and its administrations among tribal communities of Ladakh provides certain new information. Though the system is extensively used among the tribal communities in the remotest regions but still it has a great scope of proper phytochemical and pharmacological validation of the medicinal plants used in different remedies for conservation and development of traditional system of medicine according to modernization.

Acetyl-L-carnitine-mediated neuroprotection during hypoxia is attributed to ERK1/2-Nrf2-regulated mitochondrial biosynthesis

Neuronal damage in hypoxia and several neurodegenerative disorders is invariably associated with oxidative damage and mitochondrial dysfunction. Administration of acetyl‐L‐carnitine (ALCAR) on the other hand attenuates neuronal damage, prevents apoptosis, and improves energy status in hypoxic stress through less understood mechanisms. Becasue mitochondrial biogenesis could be a possible mechanism for ALCAR‐induced improvement in bioenergetics in neurons, the present study aimed at exploring signaling pathways of ALCAR‐induced neuroprotection in hypoxia and possible occurrence of mitochondrial biogenesis. To create global hypoxia, adult Sprague–Dawley rats were exposed to a simulated altitude of 7,620 m at standard temperature and humidity conditions. We here demonstrate that administration of ALCAR to hypoxic rats for a period of 2 weeks effectively protected hippocampal neurons from mitochondrial dysfunction, excitotoxicity, and neurodegeneration. ALCAR administration resulted in peroxisome proliferator‐activated receptor γ coactivator‐1α and nuclear respiratory factor‐1‐induced mitochondrial biogenesis, the expression of which was regulated by an extracellular‐related kinase‐nuclear factor erythroid 2‐related factor 2 (ERK‐Nrf2)‐mediated mechanism. Most notably, calcium buffering into nonfunctional mitochondria ameliorated excitotoxicity and improved bioenergetic status of the hippocampal neurons. Together, the data reveal the immense therapeutic potential of ALCAR for the treatment of ischemia, stroke, and other neurodegenerative disorders associated with hypoxic stress and excitotoxicity.

Sequential determination of fat- and water-soluble vitamins in Rhodiola imbricata root from trans-Himalaya with rapid resolution liquid chromatography/tandem mass spectrometry.

A rapid method was developed to determine both types of vitamins in Rhodiola imbricata root for the accurate quantification of free vitamin forms. Rapid resolution liquid chromatography/tandem mass spectrometry (RRLC-MS/MS) with electrospray ionization (ESI) source operating in multiple reactions monitoring (MRM) mode was optimized for the sequential analysis of nine water-soluble vitamins (B1, B2, two B3 vitamins, B5, B6, B7, B9, and B12) and six fat-soluble vitamins (A, E, D2, D3, K1, and K2). Both types of vitamins were separated by ion-suppression reversed-phase liquid chromatography with gradient elution within 30 min and detected in positive ion mode. Deviations in the intra- and inter-day precision were always below 0.6% and 0.3% for recoveries and retention time. Intra- and inter-day relative standard deviation (RSD) values of retention time for water- and fat-soluble vitamin were ranged between 0.02-0.20% and 0.01-0.15%, respectively. The mean recoveries were ranged between 88.95 and 107.07%. Sensitivity and specificity of this method allowed the limits of detection (LOD) and limits of quantitation (LOQ) of the analytes at ppb levels. The linear range was achieved for fat- and water-soluble vitamins at 100-1000 ppb and 10-100 ppb. Vitamin B-complex and vitamin E were detected as the principle vitamins in the root of this adaptogen which would be of great interest to develop novel foods from the Indian trans-Himalaya.

Antioxidant capacities, phenolic profile and cytotoxic effects of saxicolous lichens from trans-Himalayan cold desert of Ladakh.

Fourteen saxicolous lichens from trans-Himalayan Ladakh region were identified by morpho-anatomical and chemical characteristics. The n-hexane, methanol and water extracts of the lichens were evaluated for their antioxidant capacities. The lichen extracts showing high antioxidant capacities and rich phenolic content were further investigated to determine their cytotoxic activity on human HepG2 and RKO carcinoma cell lines. The ferric reducing antioxidant power (FRAP), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging capacities and β-carotene-linoleic acid bleaching property exhibited analogous results where the lichen extracts showed high antioxidant action. The lichen extracts were also found to possess good amount of total proanthocyanidin, flavonoid and polyphenol. The methanolic extract of Lobothallia alphoplaca exhibited highest FRAP value. Methanolic extract of Xanthoparmelia stenophylla showed the highest ABTS radical scavenging capacity. The n-hexane extract of Rhizoplaca chrysoleuca exhibited highest DPPH radical scavenging capacity. Highest antioxidant capacity in terms of β-carotene linoleic acid bleaching property was observed in the water extract of Xanthoria elegans. Similarly, Melanelia disjuncta water extract showed highest NO scavenging capacity. Among n-hexane, methanol and water extracts of all lichens, the methanolic extract of Xanthoparmelia mexicana showed highest total proanthocyanidin, flavonoid and polyphenol content. From cytotoxic assay, it was observed that the methanolic extracts of L. alphoplaca and M. disjuncta were exhibiting high cytotoxic effects against cancer cell growth. Similarly, the water extract of Dermatocarpon vellereum, Umbilicaria vellea, X. elegans and M. disjuncta and the methanolic extract of M. disjuncta and X. stenophylla were found to possess high antioxidant capacities and were non-toxic and may be used as natural antioxidants for stress related problems. Our studies go on to prove that the unique trans-Himalayan lichens are a hitherto untapped bioresource with immense potential for discovery of new chemical entities, and this biodiversity needs to be tapped sustainably.

Evaluation of anti-oxidant activities and total phenol and flavonoid content of the hydro- alcoholic extracts of Rhodiola sp.

Abstract Rhodiola sp. ( R. heterodanta and R. imbricata ), belonging to the stone crop family Crassulaceae , is an important medicinal plant and food crop in Trans-Himalayan cold desert. It is a well known medicinal plant in the Amchi system of medicine (Tibetan system of medicine) being used in various ailments like anti-stress, radio-protective, anticancer, anti-inflammatory agent, adaptogen etc. Hydro- alcoholic extracts of Rhodiola sp. root and shoot were investigated in the present study to quantify total Phenolic as well as anti-oxidant activity. Total phenolic contents were determined as total flavonoids (TF), and total polyphenols (TPP). Rhodiola heterodanta root extract had highest amount of total phenol and flavonoids are found to be 79.21±.26 mg GAE/g and 269.3±.82mg Qc/g respectively. In vitro antioxidant activity was investigated by DPPH radical scavenging activity, reducing power assay. R. imbricata hydro-alcoholic root extract showed the highest reducing power and DPPH. radical scavenging. A significant correlation existed between concentrations of the extract and percentage inhibition of free radicals and reducing power. These results clearly indicate that Rhodiola sp. are effective against free radical mediated diseases and also helpful to draw special attention for further studies.

Acute and sub acute toxicity and efficacy studies of Hippophae rhamnoides based herbal antioxidant supplement

Objectives: Present study was carried out to evaluate acute and subacute toxicity and efficacy of Seabuckthorn (Hippophae rhamnoides) based herbal antioxidant supplement (HAOS). Materials and Methods: In vivo toxicity studies were performed in male balb ‘C’ mice by oral administration. Acute toxicity study was done at doses ranging from 2000 to 10 000 mg/ kg while in subacute studies, HAOS was given at doses of 2000, 4000, and 8000 mg/kg body weight. Animals were observed for any toxic sign and symptoms periodically. At completion of study animals were sacrificed; their hematological, biochemical parameters were analyzed and histopathology of vital organs was done. In vivo efficacy studies in human volunteers were done and the levels of vitamin A and Vitamin C in blood samples were analyzed in comparison to a similar commercially available formulation. Results: No mortality and any clinical signs of toxicity were found in HAOS administered group of animals. There were no significant alterations in hematological and biochemical parameters. Histopathological analysis of vital organs showed normal architecture in all the HAOS administered groups. Human studies showed an increase of 32% and 172% in Vitamin A and Vitamin C levels respectively in term of bioavailability. Conclusion: The data obtained indicate no toxicity of this antioxidant supplement up to the highest dose studied. Efficacy in terms of increased bioavailability of vitamin A and C in human volunteers indicates the clinical usefulness of the supplement.

In Vitro Antioxidant Activity Of Methanolic Extract Of Rhodiola Imbricata Edgew.

Abstract Rhodiola imbricata (Roseroot) is a perennial plant distributed in South Asia viz., Pakistan, Nepal, China and India. In India it is endemic to Trans Himalayan cold desert of Ladakh. It is a well known medicinal plant in the Amchi system of medicine (Tibetan system of medicine) being used in various ailments like anti-stress, radio-protective, anticancer, anti-inflammatory agent, adaptogen etc. The aim of this study was to assess the in vitro potential of methanol extract of Rhodiola imbricata roots. The DPPH activity of the extract (0.1–1.2 mg/ml) was increased in a dose dependent manner, which was found in the range of (39.55–70.76%) as compared to ascorbic acid (46.78–81.47%). The IC50 values of methanol extract in DPPH radical, nitric oxide, hydroxyl radical were obtained to be 0.33, 0.47, 0.58 mg/ml, respectively. However, the IC50 values for the standard ascorbic acid were noted to be 0.42, 0.43, 0.51 mg/ml, respectively. Measurement of total phenolic content of the methanol extract of R. imbricata was achieved using Folin-Ciocalteau reagent containing 185.7 mg/g of phenolic content, which was found significantly higher when compared to reference standard gallic acid. The results obtained in this study clearly indicate that R. imbricata has a significant potential to use as a natural anti-oxidant agent.

Determination of total phenols, free radical scavenging and antibacterial activities of Mentha longifolia Linn. Hudson from the cold desert, Ladakh, India.

Abstract The aim of the study was to evaluate the antioxidant and free radical scavenging properties, total phenolic contents and antibacterial activities of the methanol extract of Mentha longifolia Linn. Hudson. growing in the high altitude cold desert region of Ladakh, Jammu and Kashmir, India. The antioxidant activities of the extracts were investigated with four different methods such as 2,2′-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS), Reducing power assay and Ferric reducing antioxidant property (FRAP). M. longifolia exhibited the strongest activity as ABTS scavenger compared to DPPH and FRAP. All the assays carried out found that the plant has antioxidant potential. The total polyphenol content (TPC) and total flavonoid content of the extract was determined as 107.208 ± 34.2 mg GAE/g dry wt. and 42.47 ± 7.97 mg QE/g dry wt. respectively where 39.61% of TPC were flavonoids. A high correlation was found between the DPPH and ABTS free radical scavenging potency (R 2 = 0.869). The study has validated the medicinal potential of Mentha longifolia Linn. Hudson and revealed that the consumption of the plant could be beneficial as it possesses the desirable quality of antioxidant activity.

Nutritional Profile of Phytococktail from Trans-Himalayan Plants

We estimated the nutritive value, vitamin content, amino acid composition, fatty acid content, and mineral profile of a phytococktail comprising sea buckthorn (Hippophae rhamnoides), apricot (Prunus armeniaca), and roseroot (Rhodiola imbricata) from trans-Himalaya. The free vitamin forms in the phytococktail were determined by rapid resolution liquid chromatography/tandem mass spectrometry (RRLC-MS/MS). Vitamin E and B-complex vitamins were detected as the principle vitamins. Reversed-phase high performance liquid chromatography (RP-HPLC) with pre-column derivatization was used for identification and quantification of amino acids. Eight essential and eleven non-essential amino acids were quantified, and the content ranged between 76.33 and 9485.67 µg/g. Among the essential amino acids, L-methionine, L-phenylalanine, L-lysine, L-leucine, and L-histidine were found to be the dominant contributors. We also quantified the fatty acids in the phytococktail by using gas chromatography coupled with a flame ionization detector (GC-FID) with fatty acid methyl esters (FAMEs) derivatization. The analysis revealed the presence of 4 major fatty acids contributing to the total lipid content. Palmitic acid was found to be the rich source of saturated fatty acid (SFA) and constituted ∼31% of the total lipid content. Among the unsaturated fatty acids (UFAs), palmitoleic acid (43.47%), oleic acid (20.89%), and linoleic acid (4.31%) were prominent. The mineral profiling was carried out by inductively coupled plasma optical emission spectrometer (ICP-OES), and it was found to contain a number of important dietary mineral elements. The harsh climatic conditions, difficult terrain, and logistic constraints at high altitude regions of Indian trans-Himalayan cold desert lead to the scarcity of fresh fruits and vegetables. Therefore, the source of multiple vitamins, essential amino acids, fatty acids, and dietary minerals from the phytococktail would provide great health benefit in the stressful environment and could be used as a high value nutritional supplement.

One pot green preparation of Seabuckthorn silver nanoparticles (SBT@AgNPs) featuring high stability and longevity, antibacterial, antioxidant potential: a nano disinfectant future perspective

In recent years antimicrobial resistance (AMR) threatens the effective prevention and treatment of an ever-increasing range of infections caused by bacteria, parasites, viruses, fungi and development of effective therapeutics is a major challenge to the scientific community. Silver Nanoparticles (AgNPs) well know antimicrobials, but limited information available on the effect of ‘Phytosynthesis’ and there clinical applications. Phytochemicals occluded in Seabuckthorn (SBT) have been extensively used as dietary supplements and as natural pharmaceuticals in the treatment of various diseases including human cancer. In this context, we present a kind of eco-friendly, economically viable, possible to commercialization, free from chemical agents and rapid preparation of SBT@AgNPs employing the leaves extracts of Seabuckthorn (SBT). The SBT leaf extracts the rich source of organic acids and inorganic compounds may be envisaged for the formation of the SBT@AgNPs at room temperature (25 °C ± 1.5 for 60 minutes). Physicochemical properties of synthesized nanoparticles were characterized using various spectroscopic techniques. The results depicted that SBT leaf mediated SBT@AgNPs are spherical with an average particles size of ∼10–40 nm. The present SBT@AgNPs has shown long time stability (one year), indicated by their unchanged SPR band, TEM image, and zeta-potential value. The evaluation of antimicrobial properties of SBT@AgNPs indicated significant inhibition of Gram-positive and Gram-negative bacterial strains even at much lower than the minimum inhibitory concentration (2 μg mL−1). Live and dead bacterial assay with confocal microscopy image demonstrated that the SBT@AgNPs disrupt the bacterial cell membrane which subsequently results in cellular material leakage and cell death. Antioxidant assay results suggest that DPPH radical scavengers increased more than 10 fold efficacy as compared to SBT leaves extract. Interestingly, this study also demonstrated that SBT@AgNPs impregnated paper preparation and there antibacterial activity against bacterial cocktail, the results shown that higher than the commercially available medicated dressing materials and deterioration and longevity of SBT@AgNPs impregnated paper also studied. Overall, the present study has disclosed a sustainable and green preparation of SBT@AgNPs and depicted their antibacterial, antioxidant agents and routine infection-control in day to day life. We strongly believe that SBT@AgNPs has great potential as an effective antibacterial tool for controlling and prevention of currently emerging bacterial infections in everyday life.