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Ophiocordyceps sinensis, a high-altitude medicinal mushroom, is widely revered in traditional medicine for its antiproliferative, antihypercholesterolemic, energy enhancement, etc. properties. These properties are attributed to the presence of steroids, terpenoids, polyphenolics, glycosides, and glycoproteins in it. The current study presents characterization of three phenolic rich fractions (PRFs) separated from aqueous extract of O. sinensis using diethyl ether and ethyl acetate. These fractions possessed considerable antioxidant potential in terms of total phenolic content, total flavonoid content, ferric ion reducing power, and free radical scavenging efficiency. Chemical characterization of the PRFs was carried out using HPTLC and GC-MS. The diethyl ether and ethyl acetate fractions elucidated appreciable antibacterial activity against Salmonella typhi. The aqueous extract and residual phenolic fraction displayed protective effect on HEK 293 cell lines against hypoxia stress and also bestowed appreciative adaptogenic role against cold, hypoxia, and restraint stress in vivo, by decreasing levels of malondialdehyde and superoxide dismutase and incrementing reduced glutathione concentrations.

Hippophae rhamnoides L. commonly known as Seabuckthorn (SBT), a wild shrub of family Elaegnacea, has extensively used for treating various ailments like skin diseases, jaundice, asthma, lung troubles. SBT leaves have been reported to possess several pharmacological properties including immunomodulatory, antioxidant, anti-inflammatory, antimicrobial and tissue regeneration etc. The present study was undertaken to evaluate the adjuvant property of supercritical carbon dioxide extracts (SCEs 300ET and 350ET) of SBT leaves in balb/c mice immunized with Tetanus and Diphtheria toxoids. The dynamic changes in the immune response were measured in terms of humoral and cell-mediated immune responses. We have seen the effect of SCEs on immunoglobulin subtypes and secondary immune response generation. In addition, the effect of SCEs on antigen specific cellular immunity was evaluated. Our results show that SCEs 300ET and 350ET significantly enhanced antibody titers in response to both TT and DT antigens. The secondary immune response generated was significantly increased in case of TT immunized animals. SCEs also enhanced cytokine levels (IFN-γ, IL-4, TNF-α and IL-1β) and increased lymphoproliferation. Besides, both SCEs did not show any toxic effects. Therefore, the study suggests that SCEs are safe and have potent immunostimulatory activity and hence, seems to be a promising balanced Th1 and Th2 directing immunological adjuvant for various veterinary as well as human vaccines., (Copyright © 2017. Published by Elsevier B.V.)

The toxicity and disposal concerns of organic solvents used in conventional extraction purposes has entailed the need for greener alternatives. Among such techniques, supercritical fluid extraction (SFE) has gained popularity by yielding extracts of high purity in a much faster manner. Carbon dioxide (CO2) is generally preferred as a supercritical solvent because of its lower temperature requirements, better diffusivity and easy removal. The present study describes the characterization of supercritical CO2 extracts of Indian variety of Cordyceps sinensis (CS)- a high-altitude medicinal mushroom widely revered in traditional medicine for its extensive anti-hypercholesterolemic, anti-inflammatory, anti-proliferative and energy-enhancing properties. Experimental parameters viz. 300 and 350 bar of extraction pressure, 60°C of temperature, 0.4°L/h CO2 of flow rate and use of 1% (v/v) of ethanol as entrainer were optimized to prepare three different extracts namely, CSF1, CSF2 and CSF3. High-performance thin-layer chromatography (HPTLC) was used for assessing the quality of all the extracts in terms of cordycepin, the pivot biomarker compound in CS. Characterization by HPTLC and GC-MS confirmed the presence of flavonoids and nucleobases and, volatile organic compounds (VOCs), respectively. The chromatographic data acquired from metabolite profiling were subjected to chemometric analysis in an open source R studio which illustrated interrelatedness between CSF1 and CSF2 in terms of two major principal components. i.e. Dim 1 and Dim 2 whose values were 40.33 and 30.52% in variables factor map plotted using the HPTLC-generated retardation factor values. The factor maps based on retention times of the VOCs exhibited a variance of Dim 1 = 43.95% and Dim 2 = 24.85%. Furthermore, the extracts demonstrated appreciable antibacterial activity against Escherichia coli and Salmonella typhi by generation of reactive oxygen species (ROS), protein leakage and efflux pump inhibition within bacterial pathogens. CSFs were elucidated to be significantly cytoprotective (p < 0.05) in a simulated hypobaric hypoxia milieu (0.5% oxygen). CSF2 showed the best results by effectively improving the viability of human embryonic kidney (HEK 293) cells to 82.36 ± 1.76% at an optimum dose of 100 µg/ml. Levels of hypoxia inducible factor-1 alpha (HIF-1α) were modulated four-fold upon supplementation with CSF2. The results collectively evinced that the CSF extracts are substantially bioactive and could be effectively utilized as mycotherapeutics for multiple bioeffects. [ABSTRACT FROM AUTHOR]

The present study aims to establish a high-performance thin layer chromatography (HPTLC)-based comparative analysis, directed toward characterization of nucleobases in aqueous and alcoholic extracts of sea buckthorn leaves from three different varieties:Hippophae salicifolia, Hippophae rhamnoides mongolica, andHippophae rhamnoides turkestanica. The alcoholic and aqueous leaf extracts from these sea buckthorn varieties were prepared using accelerated solvent extraction technique. A novel HPTLC method for separating and identifying six nucleobases, namely, guanosine, guanine, cytosine, adenine, uracil, and thymine were adopted. HPTLC analysis indicated the presence of one or more of these nucleobases in a total of six leaf extracts evaluated, their quantities varying from 0.23 to 7.76 µg nucleobase per mg of extract. Though a typical trend could not be observed in the values obtained, the extracts were found to be considerably rich with respect to nucleobase contents. The results acquired from HPTLC were subsequently validated by hyphenation with mass spectrometry and also by applying chemometric tools in form of heat maps, hierarchical cluster dendrograms, and principal component analysis. The presence of nucleobases in the leaf extracts was confirmed by HPLC as well but HPTLC proved to be a better approach for characterization of nucleobases in plant extracts, than high performance liquid chromatography (HPLC). [ABSTRACT FROM AUTHOR]