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BACKGROUND: Exploration of microbes isolated from north western Himalayas for bioactive natural products. RESULTS: A strain of Trichoderma lixii (IIIM-B4) was isolated from Bacopa monnieri L. The ITS based rDNA gene sequence of strain IIIM-B4 displayed 99% sequence similarity with different Trichoderma harzianum species complex. The highest score was displayed for Hypocrea lixii strain FJ462763 followed by H. nigricans strain NBRC31285, Trichoderma lixii strain CBS 110080, T. afroharzianum strain CBS124620 and Trichoderma guizhouense BPI:GJS 08135 respectively. Position of T. lixii (IIIM-B4) in phylogenetic tree suggested separate identity of the strain. Microbial dynamics of T. lixii (IIIM-B4) was investigated for small peptides. Medium to long chain length peptaibols of 11 residue (Group A), 14 residue (Group B) and 17 residue (Group C) were identified using Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) mass spectrometer. Optimization is undeniably a desideratum for maximized production of desirable metabolites from microbial strain. Here optimization studies were carried out on T. lixii (IIIM-B4) using different growth media through Intact Cell Mass Spectrometry (ICMS). A multifold increase was obtained in production of 11 residue peptaibols using rose bengal medium. Out of these, one of them named as Tribacopin AV was isolated and sequenced through mass studied. It was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH. It was found to have antifungal activity against Candida albicans (25 µg/mL MIC). CONCLUSION: In this study, we isolated a strain of T. lixii (IIIM-B4) producing medium and long chain peptaibols. One of them named as Tribacopin AV was found novel as having unique sequence Ac-Gly-Leu-Leu-Leu-Ala-Leu-Pro-Leu-Aib-Val-Gln-OH, which had antifungal properties.

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ETHNOPHARMACOLOGICAL RELEVANCE: Tinospora cordifolia is a miraculous ayurvedic herb used in the treatment of innumerable diseases such as diabetes, gonorrhea, secondary syphilis, anaemia, rheumatoid arthritis, dermatological diseases, cancer, gout, jaundice, asthma, leprosy, in the treatment of bone fractures, liver & intestinal disorders, purifies the blood, gives new life to the whole body; (rejuvenating herb) and many more. Recent studies have revealed the anticancer potential of this plant but not much work has been done on the anticancer chemical constituents actually responsible for its amazing anticancer effects. This prompted us to investigate this plant further for new potent anticancer molecules. AIM OF THE STUDY: The present study was designed to isolate and identify new promising anticancer candidates from the aqueous alcoholic extract of T. cordifolia using bioassay-guided fractionation. MATERIALS AND METHODS: The structures of the isolated compounds were determined on the basis of spectroscopic data interpretation and that of new potent anticancer molecule, TC-2 was confirmed by a single-crystal X-ray crystallographic analysis of its corresponding acetate. The in vitro anti-cancer activity of TC-2 was evaluated by SRB assay and the autophagic activity was investigated by immunofluorescence microscopy. Annexin-V FITC and PI dual staining was applied for the detection of apoptosis. The studies on Mitochondrial Membrane potential and ROS (Reactive oxygen species) production were also done. RESULTS: Bioassay guided fractionation and purification of the aqueous alcoholic stem extract of Tinospora cordifolia led to the isolation of a new clerodane furano diterpene glycoside (TC-2) along with five known compounds i.e. cordifolioside A (ß-D-Glucopyranoside,4-(3-hydroxy-1-propenyl)- 2,6-dimethoxyphenyl 3-O-D-apio-ß-D-furanosyl) (TC-1), ß-Sitosterol(TC-3), 2ß,3ß:15,16-Diepoxy- 4α, 6ß-dihydroxy-13(16),14-clerodadiene-17,12:18,1-diolide (TC-4), ecdysterone(TC-5) and tinosporoside(TC-6). TC-2 emerged as a potential candidate for the treatment of colon cancer. CONCLUSION: The overall study on the bioassay guided isolation of T.cordifolia identified and isolated a new clerodane furano diterpenoid that exhibited anticancer activity via induction of mitochondria mediated apoptosis and autophagy in HCT116 cells. We have reported a promising future candidate for treating colon cancer.

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Fruit drinks contain negligible amount of protein as nutritional component. Fortification of fruit drinks with protein is a challenge due to protein stability in acidic and ionic environment. Mango ready-to-serve (RTS) beverage was fortified with modified whey protein and its rheological properties were studied. Whey protein was hydrolysed with papain to improve its stability in acidic medium. The water holding capacity of whey protein increased about two times after hydrolysis. Hydrolysed and native whey protein was used at 2, 3 and 4% levels for fortification of mango based RTS beverage. Addition of hydrolysed whey protein at all the three levels did not significantly change the flow behaviour of the beverage. Native whey protein fortification resulted in precipitation; however, addition of hydrolysed whey protein led to stable beverage formulation at all the three levels. Hydrolysed whey protein imparted slight bitter taste to the RTS beverage, which was masked by ß-cyclodextrin @ 0.15% of total protein. The mango RTS beverage with 3.0% hydrolysed whey protein was found acceptable with good sensory appeal and stability during thermal processing as well storage in glass bottles.

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The marine natural product fascaplysin (1) is a potent Cdk4 (cyclin-dependent kinase 4)-specific inhibitor, but is toxic to all cell types possibly because of its DNA-intercalating properties. Through the design and synthesis of numerous fascaplysin analogues, we intended to identify inhibitors of cancer cell growth with good therapeutic window with respect to normal cells. Among various non-planar tryptoline analogues prepared, N-(biphenyl-2-yl) tryptoline (BPT, 6) was identified as a potent inhibitor of cancer cell growth and free from DNA-binding properties owing to its non-planar structure. This compound was tested in over 60 protein kinase assays. It displayed inhibition of Cdk4-cyclin D1 enzyme in vitro far more potently than many other kinases including Cdk family members. Although it blocks growth of cancer cells deficient in the mitotic-spindle checkpoint at the G0/G1 phase of the cell cycle, the block occurs primarily at the G2/M phase. BPT inhibits tubulin polymerization in vitro and acts as an enhancer of tubulin depolymerization of paclitaxel-stabilized tubulin in live cells. Western blot analyses indicated that, in p53-positive cells, BPT upregulates the expression of p53, p21 and p27 proteins, whereas it downregulates the expression of cyclin B1 and Cdk1. BPT selectively kills SV40-transformed mouse embryonic hepatic cells and human fibroblasts rather than untransformed cells. BPT inhibited the growth of several human cancer cells with an IC50<1 µM. The pharmacokinetic study in BALB/c mice indicated good plasma exposure after intravenous administration. It was found to be efficacious at 1/10th the maximum-tolerated dose (1000 mg/kg) against human tumours derived from HCT-116 (colon) and NCI-H460 (lung) cells in SCID (severe-combined immunodeficient) mice models. BPT is a relatively better anticancer agent than fascaplysin with an unusual ability to block two overlapping yet crucial phases of the cell cycle, mitosis and G0/G1. Its ability to effectively halt tumour growth in human tumour-bearing mice would suggest that BPT has the potential to be a candidate for further clinical development.

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Tumor angiogenesis is a validated target for therapeutic intervention, but agents that are more disease selective are needed. Here, we report the isolation of secalonic acid-D (SAD), a mycotoxin from a novel source that exhibits potent antiangiogenic antitumor activity. SAD inhibited multiple HIF1α/VEGF-arbitrated angiogenesis dynamics as scored in human umbilical vascular endothelial cells and human MCF-7 breast tumor xenografts. Similarly, SAD suppressed VEGF-induced microvessel sprouting from rat aortic ring and blood vessel formation in the Matrigel plug assay in C57/BL6J mice. Under normoxic or hypoxic conditions, SAD inhibited cell survival through the Akt/mTOR/p70S6K pathway, with attendant effects on key proangiogenesis factors, including HIF1α, VEGFR, and MMP-2/MMP-9. These effects were reversed by cotreatment with the Akt inhibitors perifosine and GSK69069 or by the addition of neutralizing VEGF antibodies. The apoptotic properties of SAD were determined to be both extrinsic and intrinsic in nature, whereas the cell-cycle inhibitory effects were mediated by altering the level of key G1-S transition-phase proteins. In experimental mouse models of breast cancer, SAD dosing produced no apparent toxicities (either orally or intraperitoneal) at levels that yielded antitumor effects. Taken together, our findings offered a preclinical validation and mechanistic definition of the antiangiogenic activity of a novel mycotoxin, with potential application as a cancer-selective therapeutic agent.

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The effect of hot water blanching treatment on thin layer drying kinetics of aonla shreds was studied at drying air temperatures of 50, 55 and 60 °C with the air velocity of 1.2 m/s. The drying time decreased with the increase in air temperature and blanching. The drying process was observed in falling rate. Drying after blanching reduced the vitamin C content of aonla shreds by 69.36% whereas it decreased by 27.78% in unblanched shreds. Eight commonly used mathematical models were evaluated to predict the drying behavior of aonla shreds. The Midilli model described the drying behaviour of unblanched aonla shreds at all temperatures better than other models whereas two-term model described the drying kinetics of blanched aonla shreds satisfactorily. The effective diffusivities of the unblanched and blanched aonla shreds were determined using Fick's law of diffusion. The activation energy was found to be 47.21 kJ/mol for unblanched and 43.98 kJ/mol for blanched aonla shreds.

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Beam quality correction kQQo (r), which reflects the absorbed energy dependence of the detector, is calculated for solid state detector materials diamond, LiF, Li2B4O7 and Al2O3 for the 137Cs RTR brachytherapy source using the Monte Carlo-based EGSnrc code system. The study also includes calculation of detector-specific phantom scatter corrections kphant (r) for solid phantoms such as PMMA, polystyrene, RW1, solid water, virtual water and plastic water. Above corrections are calculated as a function of distance r along the transverse axis of the source. kQQo (r) is about unity for the Li2B4O7 detector. LiF detector shows a gradual decrease in kQQo (r) with r (decrease is about 2% over the distance range of 1 - 15 cm). Diamond detector shows a gradual increase in kQQo (r)with r (about 3% larger than unity at 15 cm). In the case of Al2O3 detector, kQQo (r)decreases with r steeply (about 14% over the distance range of 1 - 15 cm). The study shows that some solid state detectors demonstrate distance-dependent kphant (r)values, but the degree deviation from unity depends on the type of solid phantom and the detector.

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Erythrina suberosa is an ornamental tall tree found in India, Pakistan, Nepal, Bhutan, Burma, Thailand and Vietnam. We have isolated four known distinct metabolites designated as α-Hydroxyerysotrine, 4'-Methoxy licoflavanone (MLF), Alpinumisoflavone (AIF) and Wighteone. Among the four isolated metabolites the two flavonoids, MLF and AIF were found to be the most potent cytotoxic agent with IC50 of ∼20µM in human leukemia HL-60 cells. We are reporting first time the anticancer and apoptotic potential of MLF and AIF in HL-60 cells. Both MLF and AIF inhibited HL-60 cell proliferation and induce apoptosis as measured by several biological endpoints. MLF and AIF induce apoptosis bodies formation, enhanced annexinV-FITC binding of the cells, increased sub-G0 cell fraction, loss of mitochondrial membrane potential (Δψm), release of cytochrome c, Bax, activation of caspase-9, caspase-3 and PARP (poly ADP Ribose polymers) cleavage in HL-60 cells. MLF and AIF also increase the expression of apical death receptor, Fas, with inhibition of anti-apoptotic protein Bid. All the above parameters revealed that these two flavonoids induce apoptosis through both extrinsic and intrinsic apoptotic pathways in HL-60 cells. In spite of apoptosis, these two flavonoids significantly inhibit nuclear transcription factor NF-κB and STAT (Signal Transducer and Activator of Transcription) signaling pathway, which are highly expressed in leukemia. The present study provide an insight of molecular mechanism of cell death induced by MLF and AIF in HL-60 cells which may be useful in managing and treating leukemia.

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UNLABELLED: Hertz's theory of contact stresses was applied to predict the splitting of guar seeds during uni-axial compressive loading between 2 rigid parallel plates. The apparent modulus of elasticity of guar seeds varied between 296.18 and 116.19 MPa when force was applied normal to hilum joint (horizontal position), whereas it varied between 171.86 and 54.18 MPa when force was applied in the direction of hilum joint (vertical position) with in moisture content range of 5.16% to 15.28% (d.b.). At higher moisture contents, the seeds yielded after considerable deformation, thus showing ductile nature. Distribution of stresses below the point of contact were plotted to predict the location of critical point, which was found at 0.44 to 0.64 mm and 0.37 to 0.53 mm below the contact point in vertical and horizontal loading, respectively, depending upon moisture content. The separation of cotyledons from each other initiated before yielding of cotyledons and thus splitting of seed took place. The relationships between apparent modulus of elasticity, principal stresses with moisture content were described using second-order polynomial equations and validated experimentally. PRACTICAL APPLICATION: Manufacture of guar gum powder requires dehulling and splitting of guar seeds. This article describes splitting behavior of guar seeds under compressive loading. Results of this study may be used for design of dehulling and splitting systems of guar seeds.

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Marine natural products offer an abundant source of pharmacologically active agents with great diversity and complexity, and the potential to produce valuable therapeutic entities. Indole alkaloids is one of the important class of marine-derived secondary metabolites, with wide occurrence amongst variety of marine sources such as sponges, tunicates, algae, worms and microorganisms and have been extensively studied for their biological activities. Among this chemical family, a sponge-derived bis-indole alkaloid fascaplysin (1) exhibited broad range of bioactivities including antibacterial, antifungal, antiviral, anti-HIV-1-RTase, p56 tyrosine kinase inhibition, antimalarial, anti-angiogenic, antiproliferative activity against numerous cancer cell lines, specific inhibition of cyclin-dependent kinase-4 (IC(50) 350 nM) and action as a DNA intercalator. In the present review, the chemical diversity of natural as well as synthetic analogues of fascaplysin has been reviewed with a detailed account on synthetic reports and pharmacological studies. Our analysis of the structure-activity relationships of this family of compounds highlights the existence of various potential leads for the development of novel anticancer agents.

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Marine invertebrates are a rich source of novel, bioactive secondary metabolites and have attracted a great deal of attention from scientists in the fields of chemistry, pharmacology, ecology, and molecular biology. This profilic natural source has produced several antitumor secondary metabolites and amongst these, indole alkaloids are of wide occurrence. Meridianins A-G (1-7) are indole alkaloids isolated from tunicate Aplidium meridianum and are known to inhibit variety of protein kinases associated with cancer and neurodegenerative diseases. These compounds also exhibited promising antiproliferative activity in several cancer cell lines. Amongst natural meridianins, meridianin E (5) showed potent and selective inhibition of CDK-1 and CDK-5. Several synthetic meridianin analogs exhibited potent and selective inhibition of glycogen synthase-3 (GSK-3) and dual-specificity tyrosine-phosphorylation regulated kinase 1A (Dyrk-1A) which are known to be implicated in progression of Alzheimer's disease. The present review provides the critical account of isolation, medicinal chemistry and pharmacology of meridianins. Our analysis of the structure-activity relationships of this family of compounds highlights the existence of various potential leads for the development of novel anticancer and anti-Alzheimer's agents.

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Chromone alkaloids and flavoalkaloids are an important group of natural products possessing promising medicinal properties. A chromone alkaloid rohitukine is a major bioactive chemical constituent of plant Dysoxylum binectariferum (Meliaceae) Hook. which is phylogenetically related to the Ayurvedic plant, D. malabaricum Bedd. used for treatment of rheumatoid arthritis. This chromone alkaloid led to discovery of two synthetic flavoalkaloids: flavopiridol (Sanofi) and P-276-00 (Piramal) which have reached to advanced stages of clinical development for cancer treatment. Flavopiridol (Alvocidib; L868275; HMR-1275; NSC 649890 of Sanofi-Aventis + NCI) is approved as an orphan drug for treatment of chronic lymphocytic leukemia and is currently undergoing phase II studies as monotherapy and also as in combination regimes with traditional chemotherapy agents. P-276-00 (12) is currently in phase II clinical studies for advanced refractory neoplasms and multiple myeloma. Extensive amount of medicinal chemistry efforts have been reported on these flavoalkaloids. Flavopiridol demonstrated potent and specific in vitro inhibition of variety of cyclindependent kinases with clear block in cell cycle progression at the G1/S and G2/M phases. Preclinical studies demonstrated the capacity of flavopiridol to induce programmed cell death, promote differentiation, inhibit angiogenic processes and modulate transcriptional events. The co-crystallised structure of deschloro-flavopiridol with CDK-2 is available and key interactions in the ATP binding site have been reported. Flavopiridol has also been studied for the treatment of arthritis and atherosclerotic plaque formation. The present review comprises discovery, medicinal chemistry, pharmacology and preclinical/clinical development of flavoalkaloids as CDK inhibitors.

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Ras proteins regulate diverse cellular pathways that are important in the growth and spread of malignancies, including cell proliferation, cell cycle regulation, cell survival, angiogenesis and cell migration. These proteins lack the conventional transmembrane or hydrophobic domain typical of membrane associated proteins. Being small and hydrophilic in nature, these proteins undergo four-stage post-translational lipid modifications viz. prenylation, AAX proteolysis, carboxymethylation and palmitoylation for membrane localization which is important for their function. Therefore, enzymes involved in these modifications viz. farnesyl transferase (FTase), geranylgeranyl transferase-I (GGTase-I), geranylgeranyl transferase-II (GGTase-II), Ras converting enzyme-1 (Rce-1) and isoprenyl cysteine methyl transferase (ICMT) are emerging as potential therapeutic targets for the discovery of newer anticancer therapeutics. Several natural products have shown modulation of these post-translational enzymes. In the present review, natural products isolated from terrestrial as well as marine sources showing ability to modulate these k-Ras post-translational targets and their promise as potential anticancer agents have been discussed. A total of 157 natural products with 141 corresponding references have been covered.

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Bioassay-guided fractionation of the fruits of Piper longum afforded two new minor amides, piperlongimin A (2) [2E-N-isobutyl-hexadecenamide] and piperlongimin B (4) [2E-octadecenoylpiperidine] together with five known compounds with moderate cytotoxic activity. The structures were elucidated on the basis of spectroscopic evidences. All these compounds inhibited cell proliferation of human leukemia, HL-60 cell lines, and displayed major apoptosis-inducing effects.

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The reference medium for brachytherapy dose measurements is water. Accuracy of dose measurements of brachytherapy sources is critically dependent on precise measurement of the source-detector distance. A solid phantom can be precisely machined and hence source-detector distances can be accurately determined. In the present study, four different solid phantom materials such as polymethylmethacrylate (PMMA), polystyrene, Solid Water, and RW1 are modeled using the Monte Carlo methods to investigate the influence of phantom material on dose rate distributions of the new model of BEBIG (60)Co brachytherapy source. The calculated dose rate constant is 1.086 +/- 0.06% cGy h(-1) U(-1) for water, PMMA, polystyrene, Solid Water, and RW1. The investigation suggests that the phantom materials RW1 and Solid Water represent water-equivalent up to 20 cm from the source. PMMA and polystyrene are water-equivalent up to 10 cm and 15 cm from the source, respectively, as the differences in the dose data obtained in these phantom materials are not significantly different from the corresponding data obtained in liquid water phantom. At a radial distance of 20 cm from the source, polystyrene overestimates the dose by 3% and PMMA underestimates it by about 8% when compared to the corresponding data obtained in water phantom.

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PURPOSE: AAPM TG-56 recommends the use of a specific dosimetric dataset for each brachytherapy source model. In this study, a full dosimetric dataset for indigenously developed 137Cs source models, namely, the CSA1 and CSA2, in accordance with the AAPM TG-43U1 formalism is presented. The study includes calculation of dose-to-kerma ratio D/K in water around these sources including stainless steel encapsulated 137Cs sources such as RTR, 3M, and selectron/LDR 137Cs. METHODS: The Monte Carlo-based EGSnrcMP code system is employed for modeling the sources in vacuum and in water. Calculations of air-kerma strength, S(K) for the investigated sources and collision kerma in water along the transverse axis of the RTR source are based on the FLURZnrc code. Simulations of water-kerma and dose in water for the CSA1, CSA2, RTR, 3M, and selectron/ LDR 137Cs sources are carried out using the DOSRZnrc code. In DOSRZnrc calculations, water-kerma and dose are scored in a cylindrical water phantom having dimensions of 80 cm diameter x 80 cm height. RESULTS: The calculated dose-rate constants for the CSA1 and CSA2 sources are 0.945(1) and 1.023(1) cGy/(h U), respectively. The calculated value of S(K) per unit source activity, S(K)/A for the CSA1 and CSA2 sources is 7.393(7) x 10(-8) cGy cm2/(h Bq). The EGSnrcMP-based collision kerma rates for the RTR source along the transverse axis (0.25-10 cm) agree with the corresponding GEANT4-based published values within 0.5%. Anisotropy profiles of the CSA1 and CSA2 sources are significantly different from those of other sources. For the selectron/LDR single pellet 137Cs spherical source (modeled as a cylindrical pellet with dimensions similar to the seed selectron), the values of D/K at 1 and 1.25 mm from the capsule are 1.023(1) and 1.029(1), respectively. The value of D/K at 1 mm from the CSA1, CSA2, RTR, and 3M 137Cs source capsules (all sources have an external radius of 1.5 mm) is 1.017(1) and this ratio is applicable to axial positions z = 0 to z = -L/2. This is in contrast to a published GEANT4-based Monte Carlo dosimetric study on RTR and 3M 137Cs sources wherein the authors have assumed that collision kerma is approximately equal to absorbed dose at 1 mm from the source capsules. Collision kerma is approximately equal to absorbed dose for distances > or = 2 mm from source capsules as opposed to > or = 1 mm reported in published studies. A detailed electron transport is necessary up to 2 mm from source capsules. CONCLUSIONS: The Monte Carlo-calculated dose-rate data for the CSA1 and CSA2 sources can be used as input data for treatment planning or to verify the calculations by radiotherapy treatment planning system.

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The present study is on the growth inhibitory effect of Withania somnifera methanolic leaf extract and its active component, withanolide on HL-60 promyelocytic leukemia cells. The decrease in survival rate of HL-60 cells was noted to be associated with a time dependent decrease in the Bcl-2/Bax ratio, leading to up regulation of Bax. Both the crude leaf extract and the active component activated the apoptotic cascade through the cytochrome c release from mitochondria. The activation of caspase 9, caspase 8 and caspase 3 revealed that caspase was a key mediator in the apoptotic pathway. DNA fragmentation analysis revealed typical ladders as early as 12h indicative of caspase 3 role in the apoptotic pathway. Flow cytometry data demonstrated an increase of sub-G1 peak upon treatment by 51% at 24h, suggesting the induction of apoptotic cell death in HL-60 cells.

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