Endophytic fungi from Himalayan silver birch as potential source of plant growth enhancement and secondary metabolite production.

Mountain biodiversity is under unparalleled pressure due to climate change, necessitating in-depth research on high-altitude plant's microbial associations which are crucial for plant survival under stress conditions. Realizing that high-altitude tree line species of Himalaya are completely unexplored with respect to the microbial association, the present study aimed to elucidate plant growth promoting and secondary metabolite producing potential of culturable endophytic fungi of Himalayan silver birch (Betula utilis D. Don). ITS region sequencing revealed that the fungal isolates belong to Penicillium species, Pezicula radicicola, and Paraconiothyrium archidendri. These endophytes were psychrotolerant in nature with the potential to produce extracellular lytic activities. The endophytes showed plant growth promoting (PGP) traits like phosphorus solubilization and production of siderophore, indole acetic acid (IAA), and ACC deaminase. The fungal extracts also exhibited antagonistic potential against bacterial pathogens. Furthermore, the fungal extracts were found to be a potential source of bioactive compounds including the host-specific compound-betulin. Inoculation with fungal suspension improved seed germination and biomass of soybean and maize crops under net house conditions. In vitro PGP traits of the endophytes, supported by net house experiments, indicated that fungal association may support the growth and survival of the host in extreme cold conditions.

Deciphering the Antimicrobial Potential of Taxus wallichiana Zucc: Identification and Characterization Using Bioassay-Guided Fractionation.

Taxus wallichiana Zucc. is a high valued medicinal plant and has been mainly studied for its anti-cancer properties. However, research on its other important biological activities, such as its antimicrobial potential, still needs attention. The focus of the present study is to investigate the antimicrobial activity of secondary metabolites of T. wallichiana needles against 3 different groups of microorganisms, i. e., bacteria, actinobacteria, and fungi. Bioactive compounds from T. wallichiana needles were separated through column chromatography, and, TLC-bioautography. Mobile phases were optimized using Snyder's selectivity triangle. Antimicrobial spots were fractionated and compounds were identified by gas chromatography-mass spectroscopy (GC/MS) and liquid chromatography-mass spectrometry (LC/MS). Functional groups were characterized using Fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance (NMR) was used to identify the molecular structures. GC/MS and LC/MS data analysis confirm the presence of fatty acids (arachidic acid, behenic acid, palmitic acid, and stearic acid), vitamins (nicotinamide), and alkaloids (cinchonine, timolol), aminobenzamides (procainamide), carbocyclic sugar (myoinositol), and alkane hydrocarbon (hexadecane), having antimicrobial activity in the needles of T. wallichiana. To the best of our knowledge, this is the first report on the isolation and characterization of antimicrobial compounds from the needles of Taxus wallichiana (Himalayan yew). The data obtained from the present study will be supportive to the new drug discoveries in modern medicine with various combinations of medicinal plant's active constituents that can be used for curing many diseases.

Uncommon Presentation of Metastatic Squamous Cell Carcinoma of the Skin and Treatment Challenges.

BACKGROUND Squamous cell carcinoma is one of the most common keratinocytic skin cancers, the other being basal cell carcinoma. It is the second most common skin cancer after melanoma. Cutaneous squamous cell carcinoma is mostly a localized disease. The metastatic presentation is rare even in the presence of invasive disease. The metastatic potential depends on the presence of high-risk features at the time of diagnosis. Lung, liver, and bone are the frequent sites of metastasis. Local and locoregional disease undergoes excision with or without adjuvant radiation. However, we lack proper treatment paradigms for this metastatic disease. CASE REPORT We are reporting a case of an elderly female with a history of high-risk localized cutaneous squamous cell carcinoma treated with complete local excision and radiation presenting 5 years later with extensive disease to the lung and liver, abdominal nodes, and spinal fracture. The patient was not a candidate for chemotherapy due to kidney failure. On the basis of ongoing separate trials on different immunotherapies, she was started on nivolumab. CONCLUSIONS Treating metastatic cutaneous squamous cell carcinoma is a challenge considering the absence of phase III trials due to the rarity of this disease. Historically, platinum with or without 5-FU (fluorouracil), bleomycin, doxorubicin, and retinoic acid were used with variable responses. Data on epidermal growth factor receptor (EGFR) inhibitors on EGFR expressing tumors are available. However, even with the most recent reports on immunotherapy in patients with high programmed death-1 expression or high mutation burden, it is difficult to achieve good response.

Ginkgo biloba flavonoid glycosides in antimicrobial perspective with reference to extraction method.

The present study aims to investigate the effect of extraction method on the recovery of flavonoid glycosides, antimicrobials and antioxidants from Ginkgo leaves collected from six different locations in Uttarakhand, Indian Himalaya. Four extraction methods, namely maceration, reflux, shaker and soxhlet were considered, where reflux extracts showed higher antimicrobial antioxidant activity and higher content of flavonoid glycosides. The reference standards of Ginkgo flavonoid glycosides (quercetin, kaempferol and isorhamnetin) and crude extracts were tested for their antimicrobial activity against gram positive and gram negative bacteria and fungi following disc diffusion method and minimum inhibitory concentration (MIC). All the test microorganisms were observed to be inhibited significantly by Ginkgo flavonoids in plate based assays. Correlation coefficients exhibited the extent of contribution of flavonoid glycosides in antimicrobial activity and confirmed the reflux method as a potential method for extraction. Moreover, antioxidant activity as measured by DPPH assay was also found to be higher in reflux method. Significant variation (p < 0.05) in the flavonoid glycosides among the locations was also observed and sample collected from GB6 location was found to be the best for quercetin and isorhamnetin, while GB5 for kaempferol. Significant correlation (r < 0.05, r < 0.001) was obtained while developing the relationship between total flavonoid glycosides and antimicrobials. The present study, thus suggests the reflux method of extraction to be the best for maximum recovery of flavonoid glycosides with higher antioxidant and antimicrobial activities from Ginkgo extract.

Optimisation and characterisation of the orange pigment produced by a cold adapted strain of Penicillium sp. (GBPI_P155) isolated from mountain ecosystem.

With globalisation and growing health risks of synthetic colourants, search for pigments from natural sources has increased owing to their non-toxic nature. The present study highlights the bioprospection of natural pigment from a cold adapted fungal strain of Penicillium sp. (GBPI_P155), isolated from soil of Indian Himalayan region. The fungus produced insoluble and orange-coloured pigment in liquid medium with maximum production recorded in potato dextrose (PD) broth at 15°C and 3 pH, while maximum biomass was produced at 25°C and pH 3. While examining the effect of different mineral salts, and carbon and nitrogen sources on pigment production, maximum accumulation of pigment was recorded in PD broth supplemented with 2% maltose. Following production, extraction of pigment was performed using chloroform and characterised partially by UV/vis (λmax at 495 nm and a shoulder peak at 530 nm) and Fourier Transform Infrared (FT-IR) spectroscopy. Thin layer chromatography of chloroform extract resulted in separation of pigment in three fractions with Rf values 0.911, 0.852 and 0.808, which were further analysed using Liquid Chromatography Mass Spectrometry (LC/MS). The overall approach resulted in identification of pigment as a mixture of different derivatives of carotenoids. The extracted pigment also possessed antimicrobial activity against Gram-positive and Gram-negative bacteria and actinobacteria.

Antioxidant, antimicrobial activity and bioactive compounds of Bergenia ciliata Sternb.: A valuable medicinal herb of Sikkim Himalaya.

Bergenia ciliata Sternb., commonly known as Paashaanbhed, is a well known herb of Sikkim Himalaya with various pharmaceutical properties. However, scientific exploration of B. ciliata, growing in the Sikkim Himalaya, for phytochemicals and pharmacological properties is in infancy. With this view, the present study was undertaken to investigate B. ciliata leaf extracts for antioxidant, antimicrobial activity and bioactive compounds. Three solvents viz., methanol, ethyl acetate and hexane were used for extraction and the respective leaf extracts were analyzed for total phenolic and flavonoid contents along with the antioxidant and antimicrobial activities. Amongst the tested solvents, methanol was found to be the best solvent for extraction with highest total phenolic contents and the lowest IC50 values for the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. Methanol extract also exhibited effective antimicrobial activity, particularly against bacteria and actinomycetes. Further, high performance liquid chromatography (HPLC) analysis revealed that methanolic extract contains the highest amount of all the three analyzed bioactive compounds viz. bergenin, catechin and gallic acid. The current study suggests that the methanol extract of B. ciliata is a potential source of natural antioxidant and antimicrobial compounds that can be used in food and drug industries.

Antimicrobial and antioxidant phytochemicals in leaf extracts of Bergenia ligulata: a Himalayan herb of medicinal value.

Phenolic and flavonoid contents in leaf extracts of Bergenia ligulata have been analysed for their contribution in antimicrobial and antioxidant activities. The extracts were prepared in three solvents (separately) following maceration and soxhelt methods. The antimicrobial activity was tested using disc diffusion assay against a range of microorganisms along with the determination of minimum inhibitory concentration (MIC), while the antioxidant activity was tested following DPPH assay. Leaf extracts exhibited antimicrobial activity against all the three groups of microorganisms; results coincided with respective MIC. In general, the methanol extracts prepared through maceration favoured the determination of antimicrobial and antioxidant activities. Maximum values for phenolic and flavonoid contents were obtained in macerated methanol and ethyl acetate extracts, respectively. The statistical correlations exhibited the extent of the contribution of phenolic and flavonoid contents in antimicrobial and antioxidant activities and also indicated the involvement of other plant metabolites.

Prolonged Laccase Production by a Cold and pH Tolerant Strain of Penicillium pinophilum (MCC 1049) Isolated from a Low Temperature Environment.

Production of laccase by a cold and pH tolerant strain of Penicillium pinophilum has been investigated under different cultural conditions for up to 35 days of incubation. The fungus was originally isolated from a low temperature environment under mountain ecosystem of Indian Himalaya. The estimations were conducted at 3 temperatures (15, 25, and 35°C), a range of pH (3.5-11.5), and in presence of supplements including carbon and nitrogen sources, vitamins, and antibiotics. Optimum production of laccase was recorded at 25°C (optimum temperature for fungal growth) and 7.5 pH. The production of enzyme was recorded maximum on day 28 (11.6 ± 0.52 U/L) following a slow decline at day 35 of incubation (10.6 ± 0.80 U/L). Fructose and potassium nitrate (0.2%) among nutritional supplements, chloramphenicol (0.1%) among antibiotics, and folic acid (0.1%) among vitamins were found to be the best enhancers for production of laccase. Relatively lower but consistent production of laccase for a longer period is likely to be an ecologically important phenomenon under low temperature environment. Further, enhancement in production of enzyme using various supplements will be useful for its use in specific biotechnological applications.