Cymbidium aloifolium is known for its ornamental and medicinal values. It has been listed as threatened orchid species. In this study, in vitro propagated C. aloifolium plantlets were interacted with the Piriformospora indica. The growth assay was performed for 45 days; the plant growth pattern such as number and length of roots and shoots were measured. Microscopic study of the root section stained by trypan blue was done to detect the peloton formation. The methanol extracts of the fungal colonized plant as well as uncolonized (control) plant were prepared and various metabolites were identified by gas chromatography mass spectroscopy. Acclimatization was done in a substrate composition of coco peat: gravel: charcoal in ratio 2:2:1. P. indica-colonized plantlet showed the highest growth with the formation of clamdospore in the root section. The growth regulator such as auxin, ascorbic acid, andrographolide, hexadecanoic acid, and DL-proline were identified. After three months of field transfer, plantlet colonized by P. indica survived and remained healthy as compared to uncolonized control plantlet.
The immature seeds of Dendrobiumchryseum, asympodial epiphytic orchid with yellow flowers, were cultured in vitro, and the resultant protocorms were used as explants for seedling development. Protocorms were cultured on½ M.S. medium fortified with Kinetin (Kn), 6-Benzylaminopurine (BAP), and Gibberellic Acid (GA3) in three concentrations (0.5mg/l, 1.0mg/land 2.0 mg/l) both alone and supplemented with 5% and 10% coconut water (C.W.). The highest number of shootsofD. chryseum developed on ½ - M.S. medium fortified with 2.0mg/lofKn and10% C.W. and the longest shootsdeveloped on ½ M.S. media fortified with 1.0mg/lGA3, and 10% C.W. The shoot derived from protocorms were placed in ½ M.S. medium fortified with three different rooting hormones, Indole -3- acetic acid (IAA), Indole -3-butyric acid (IBA) and α-Naphthalene acetic acid (NAA) in different concentrations alone as well as with each 1.0mg/l hormone combined with 10% C.W. The most effective of these media was ½ M.S. medium fortified with 1.5 mg/l IAA for rooting as well as for the production of longest roots. The present study could be useful for standardizing the protocol for mass propagation of the endangered orchid Dendrobiumchryseum.
Objectives: The emergence of carbapenem resistant Klebsiella pneumoniae has been problematic in health sectors. It has been increasing widely in both hospital and community settings. The resistance acquired due to presence of Klebsiella pneumoniae carbapenemase (KPC) and Oxacillinase-48 carbapenemase (OXA-48) is of great concern. This cross-sectional study was designed to detect the carbapenem resistant gene namely blaKPC and blaOXA-48 in K. pneumoniae obtained from different clinical specimens.Results: Among the clinical isolates tested, culture positivity was 51.31% with K. pneumoniae as predominant isolates with 28.20%. AST revealed 38 (69.09%) carbapenemase producing K. pneumoniae while MHT con rmed 31 (56.36%) isolates as carbapenemase producer phenotypically. Similarly, highest percentage of carbapenemase production was seen in sputum (45.16%), in male (67.74%), in age group 51-60 (25.80%) and in OPD (45.16%). PCR screening was done for 31 carbapenemase producing isolates to detect blaKPC and blaOXA-48 genes. Two (6.45%) isolates were positive for blaKPC gene and 9 (29.03%) for blaOXA-48 genes. Signi cant numbers of blaKPC and blaOXA-48 genes were detected in carbapenem resistant Klebsiella pneumoniae from clinical isolates.
From the medicinal orchid Dendrobium chryseum Rolfe, which is used in traditional and folk Chinese medicine, the protocorms were raised in Murashige and Skoog (MS) media in three strengths, full strength (FMS), half strength (1/2 MS), and quarter strength (1/4 MS), with or without the phytohormones 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) and coconut water (CW). The comparative cytotoxic activities of the wild and in vitro-raised protocorms were evaluated in human cervical carcinoma (HeLa) and human glioblastoma (U251) cell lines by MTT assay. In in vivo and in vitro, the methanol extracts of D. chryseum showed significant cytotoxic activities. Significant growth inhibition (%) and potent IC50 values were demonstrated in HeLa cell lines (49.79% (210.5 μg/mL) for in vitro-raised Dendrobium chryseum (DCT) versus 46.97% (226.5 μg/mL) for wild Dendrobium chryseum (DCW)). Similarly, activities against U251 cell lines exhibited also significant inhibition (28.76% (612.54 μg/mL) for DCW and 17.15% (1059.92 μg/mL) for DCT). The cytotoxic activities of both, wild and tissue-cultured samples, were superior in HeLa cells. In U251 cells, the wild sample was more active than the tissue-cultured one with a moderate cytotoxic effect. Hence, protocorm culture may therefore be a promising future tool for producing pharmacologically bioactive compounds in medicinal orchids. Such sustainable technology approach will minimize the pressure on the natural population of threatened but commercially important medicinal orchids.
Ex-situ conservation of the ornamental and medicinal orchid, Coelogyne stricta, was performed by mass propagation using seed culture. Propagation stages were optimized using full-and half-strength solidified MS medium with different phytohormones. Maximum seed germination (88 ± 0.5% over 6 weeks of culture) was achieved on half-strength MS medium supplemented with 15% coconut water. Maximum shoot numbers were found on full-strength MS medium supplemented with 1 mg/L BAP, 2 mg/L Kinetin, and 10% coconut water, while the longest root was developed on full-strength MS medium with 1.5 mg/L IBA. A 2:1:1 combination of coco-peat, pine bark, and sphagnum moss was found to be a suitable potting mixture resulting in 80% seedling survivability. The cytotoxic activity of extracts of both wild plants and in vitro-developed protocorms was determined using an MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay on a cervical cancer cell line. The wild plant extract inhibited the growth of 41.99% of cells, showing that this extract has moderate cytotoxic activity toward cervical cancer cells.
Reactive oxygen species (ROS) at high concentrations induce oxidative stress, an imbalanced redox state that is a prevalent cause of neurodegenerative disorders. This study aimed to investigate the protective effect of Capsosiphon fulvescens (CF) extract on oxidative stress-induced impairment of cognitive function in models of neurodegenerative diseases. CF was extracted with subcritical water and several solvents and H 2 O 2 (0.25 mM) or aluminum chloride (AlCl 3 ; 25 µM) as an inducer of ROS was treated in PC12 neuronal cells and zebrafish larvae. All statistical analyses were performed using one-way analysis of variance and Dunnett's test using GraphPad Prism. H 2 O 2 and AlCl 3 were found to significantly induce ROS production in PC12 neuronal cells and zebrafish larvae. In addition, they strongly affected intracellular Ca 2+ levels, antioxidant enzyme activity, brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) signaling, acetylcholinesterase (AChE) activity, and hallmarks of Alzheimer's disease. However, treatment of H 2 O 2 -induced PC12 cells or AlCl 3 -induced zebrafish larvae with CF subcritical water extract at 90℃ and CF water extract effectively regulated excessive ROS production, intracellular Ca 2+ levels, and mRNA expression of superoxide dismutase, glutathione peroxide, glycogen synthase kinase-3 beta, β-amyloid, tau, AChE, BDNF, and TrkB. Our study suggested that CF extracts can be a potential source of nutraceuticals that can improve the impairment of cognitive function and synaptic plasticity by regulating ROS generation in neurodegenerative diseases.
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