EVALUATION OF ANTIFUNGAL EFFICIENCY OF 3,7-DIHYDROXY 3′,4′ ORTHODIHYDROXY FLAVONE STUDIED IN RELATION TO SOME MORPHOLOGICAL PARAMETERS IN PISUM SATIVUM L. PLANTS

Objectives: 3,7-dihydroxy 3′,4′ orthodihydroxy flavone isolated and identified from 50% aqueous ethanolic extract [5] and tested positive in antifungalbioassay against Fusarium oxysporum ciceri [6] was taken up for further detailed analysis of the antifungal property for in vitro study in relation to Pisum sativum L. plants. The study incorporated a comparison of some morphological parameters such as root, shoot, internode, petiole length,leaf and leaflet length, leaf and leaflet number/leaf, the total number of flower/plant, pod/plant, and seeds/pod.Methods: Seeds of P. sativum L. plants were taken up as experimental material for in vitro studies. Six different sets were maintained by presoakingthe seeds with water, 3,7-dihydroxy 3′,4′ orthodihydroxy flavone, and fungicide for 3 hrs. The seeds were grown in pot cultures. At seedling stage,three sets, one from each treatment, were exposed to fungal inoculum following soil drenching method. At the 21st day, 42 day of age,the above-mentioned parameters were studied.Results: Results indicated increased growth of internode, petiole, and shoot length in the plants which were administered with the 3,7-dihydroxy3′,4′ orthodihydroxy flavone. The fungus infested plants exhibited reduced growth of internode, petiole, and shoot length. However, griseofulvin, apopularly used fungicide showed inhibition of growth of all the parameters in healthy as well as in the infected plants. Flowering time was delayed inthe infested plants. Remarkably again, administration of the plant extract quickened the flowering time as similarly as in the healthy plants. Finally, thenumber of seeds per pod in the plant also showed the same promising picture. The 3,7-dihydroxy 3′,4′ orthodihydroxy flavone treated plants showedhigher number of seeds a pod when administered to the healthy and infected plants.Keywords: Fungicide, Inhibition, Inoculums, Infestation

Kidney injury molecule-1: a urinary biomarker for contrast induced acute kidney injury.

Back ground: Urinary kidney injury molecule 1 (KIM-1) is early biomarker for renal damage. A few studies have been published analyzing the potential use of urinary kidney injury molecule-1 (KIM-1) as a biomarker for acute kidney injury. However no study has been done related to Acute Kidney Injury associated with contrast administration. Aim: To search for new markers to identify Acute Kidney Injury (ARF) associated with contrast administration earlier than serum creatinine. Material and Methods: We studied 100 consecutive patients with normal serum creatinine undergoing angiographic procedure. We assessed urine KIM-1, at 4h, 8h, and 24 hours after the angiographic procedure. Serum creatinine was measured at basal, 24h and 48 hours after the procedure. Results: There was a significant rise in urinary KIM-1 levels at 24 hours after the angiographic procedure. The presence of contrast induced nephropathy associated with acute Kidney Injury was 12%. Conclusion: The present study highlighted the importance of urinary KIM-1 in detecting Acute Kidney Injury associated with contrast administration earlier than Serum creatinine. Key words: Neutrophil-gelatinase-associated lipocalin. Contrast-induced nephropathy. Cystatin C. Glomerular Filtration Rate (GFR), Kidney injury molecule -1 (KIM-1)

Differential expression and role of p21cip/waf1 and p27kip1 in TNF-α-induced inhibition of proliferation in human glioma cells

<p>Abstract</p> <p>Background</p> <p>The role of TNF-α in affecting the fate of tumors is controversial, while some studies have reported apoptotic or necrotic effects of TNF-α, others provide evidence that endogenous TNF-α promotes growth and development of tumors. Understanding the mechanism(s) of TNF-α mediated growth arrest will be important in unraveling the contribution of tissue associated macrophages in tumor resistance. The aim of this study was to investigate the role of Cyclin Dependent Kinase Inhibitors (CDKI) – p21^cip/waf1and p27^kip1in TNF-α mediated responses in context with p53 and activation of NF-κB and Akt pathways. The study was done with human glioma cell lines -LN-18 and LN-229 cells, using monolayer cultures and Multicellular Spheroids (MCS) as <it>in vitro </it>models.</p> <p>Results</p> <p>TNF-α induced inhibition of proliferation and enhanced the expression of p21^cip/waf1and p27^kip1in LN-18 cells. p21 was induced on exposure to TNF-α, localized exclusively in the nucleus and functioned as an inhibitor of cell cycle but not as an antiapoptotic protein. In contrast, p27 was constitutively expressed, localized predominantly in the cytoplasm and was not involved in arrest of proliferation. Our data using IκBα mutant LN-18 cells and PI3K/Akt inhibitor-LY294002 revealed that the expression of p21 is regulated by NF-κB. Loss of IκBα function in LN-229 cells (p53 positive) did not influence TNF-α induced accumulation of pp53 (Ser-20 p53) suggesting that p53 was not down stream of NF-κB. Spheroidogenesis enhanced p27 expression and p21 induced by TNF-α was significantly increased in the MCS compared to monolayers.</p> <p>Conclusion</p> <p>This study demarcates the functional roles for CDKIs-p21^cip/waf1and p27^kip1during TNF-α stimulated responses in LN-18 glioma cells. Our findings provide evidence that TNF-α-induced p21 might be regulated by NF-κB or p53 independently. p21 functions as an inhibitor of cell proliferation and does not have a direct role in rendering the cells resistant to TNF-α mediated cytotoxicity.</p

Colistin the last resort drug in 21st century antibiotics to combat Multidrug resistance superbugs

Polymyxin' E' (Colistin) is considered the last resort therapy against Multidrug resistance (MDR) bacteria, mainly Klebsiella peumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli and play a critical role in causing life-threatening infection, and their prevalence is increasing as a big concern globally. Apart from immunological adaptation, chromosomal mutations and plasmid-mediated genes are mostly associated with this resistance at the molecular level. Therefore, the current review extensively focused on Colistin as a drug in 21st-century antibiotics, the activities spectrum with diverse resistance mechanisms of bacteria against Colistin, and emerging approaches of Colistin from discovery to tackling MDR. In the study, we got to know about the challenges and new developments with old weapons like phage therapy as well as new approaches like Phage display and drug repurposing, in addition to the chromosomal and plasmid-mediated genes that play a role in antimicrobial resistance (AMR). The present study would provide insight into the prognostic aspect of combating MDR

Malabaricone-A Induces A Redox Imbalance That Mediates Apoptosis in U937 Cell Line

BACKGROUND: The 'two-faced' character of reactive oxygen species (ROS) plays an important role in cancer biology by acting both as secondary messengers in intracellular signaling cascades and sustaining the oncogenic phenotype of cancer cells, while on the other hand, it triggers an oxidative assault that causes a redox imbalance translating into an apoptotic cell death. PRINCIPAL FINDINGS: Using a tetrazolium [{3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl}-2H-tetrazolium] based cell viability assay, we evaluated the cytotoxicity of a plant derived diarylnonanoid, malabaricone-A on leukemic cell lines U937 and MOLT-3. This cytotoxicity hinged on its ability to cause a redox imbalance via its ability to increase ROS, measured by flow cytometry using 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate and by decreasing glutathione peroxidase activity. This redox imbalance mediated apoptosis was evident by an increase in cytosolic [Ca(2+)], externalization of phosphatidyl serine as also depolarization of the mitochondrial membrane potential as measured by flow cytometry. There was concomitant peroxidation of cardiolipin, release of free cytochrome c to cytosol along with activation of caspases 9, 8 and 3. This led to cleavage of the DNA repair enzyme, poly (ADP-ribose) polymerase that caused DNA damage as proved by labeling with 4',6-diamidino-2-phenylindole (DAPI); furthermore, terminal deoxy ribonucleotide transferase catalysed incorporation of deoxy uridine triphosphate confirmed DNA nicking and was accompanied by arrest of cell cycle progression. CONCLUSIONS: Taken together, compounds like MAL-A having pro-oxidant activity mediate their cytotoxicity in leukemic cells via induction of oxidative stress triggering a caspase dependent apoptosis

Studies on phytochemicals: Part XV-Investigation on the alkaloidal constituents of Indian Mappia foetida

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MENCA experiment aboard India’s Mars Orbiter Mission

The Mars Exospheric Neutral Composition Analyser (MENCA) aboard the Indian Mars Orbiter Mission (MOM) is a quadrupole mass spectrometer-based experiment. Making use of the highly elliptical and low inclination (~150°) orbit of MOM, MENCA will conduct in situ measurements of the composition and radial distribution of the Martian neutral exosphere in the 1–300 amu mass range in the equatorial and low latitudes of Mars. The functionality of MENCA has been tested during the Earth-bound and heliocentric phases of MOM before its operation in the Martian orbit. This article describes the scientific objectives, instrument details, design and development, test and evaluation, and calibration of the MENCA instrument

A potent betulinic acid analogue ascertains an antagonistic mechanism between autophagy and proteasomal degradation pathway in HT-29 cells

Betulinic acid (BA), a member of pentacyclic triterpenes has shown important biological activities like anti-bacterial, anti-malarial, anti-inflammatory and most interestingly anticancer property. To overcome its poor aqueous solubility and low bioavailability, structural modifications of its functional groups are made to generate novel lead(s) having better efficacy and less toxicity than the parent compound. BA analogue, 2c was found most potent inhibitor of colon cancer cell line, HT-29 cells with IC50 value 14.9 μM which is significantly lower than standard drug 5-fluorouracil as well as parent compound, Betulinic acid. We have studied another mode of PCD, autophagy which is one of the important constituent of cellular catabolic system as well as we also studied proteasomal degradation pathway to investigate whole catabolic pathway after exploration of 2c on HT-29 cells. Mechanism of autophagic cell death was studied using fluorescent dye like acridine orange (AO) and monodansylcadaverin (MDC) staining by using fluorescence microscopy. Various autophagic protein expression levels were determined by Western Blotting, qRT-PCR and Immunostaining. Confocal Laser Scanning Microscopy (CLSM) was used to study the colocalization of various autophagic proteins. These were accompanied by formation of autophagic vacuoles as revealed by FACS and transmission electron microscopy (TEM). Proteasomal degradation pathway was studied by proteasome-Glo™ assay systems using luminometer.The formation of autophagic vacuoles in HT-29 cells after 2c treatment was determined by fluorescence staining – confirming the occurrence of autophagy. In addition, 2c was found to alter expression levels of different autophagic proteins like Beclin-1, Atg 5, Atg 7, Atg 5-Atg 12, LC3B and autophagic adapter protein, p62. Furthermore we found the formation of autophagolysosome by colocalization of LAMP-1 with LC3B, LC3B with Lysosome, p62 with lysosome. Finally, as proteasomal degradation pathway downregulated after 2c treatment colocalization of ubiquitin with lysosome and LC3B with p62 was studied to confirm that protein degradation in autophagy induced HT-29 cells follows autolysosomal pathway. In summary, betulinic acid analogue, 2c was able to induce autophagy in HT-29 cells and as proteasomal degradation pathway downregulated after 2c treatment so protein degradation in autophagy induced HT-29 cell