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        First draft genome sequencing of indole acetic acid producing and plant growth promoting fungus Preussia sp. BSL10

        Khan, A.L.,Asaf, S.,Khan, A.R.,Al-Harrasi, A.,Al-Rawahi, A.,Lee, I.J. Elsevier Science Publishers 2016 Journal of biotechnology Vol.225 No.-

        <P>Preussia sp. BSL10, family Sporormiaceae, was actively producing phytohormone (indole-3-acetic acid) and extra-cellular enzymes (phosphatases and glucosidases). The fungus was also promoting the growth of arid-land tree-Boswellia sacra. Looking at such prospects of this fungus, we sequenced its draft genome for the first time. The Illumina based sequence analysis reveals an approximate genome size of 31.4 Mbp for Preussia sp. BSL10. Based on ab initio gene prediction, total 32,312 coding sequences were annotated consisting of 11,967 coding genes, pseudogenes, and 221 tRNA genes. Furthermore, 321 carbohydrate-active enzymes were predicted and classified into many functional families. (C) 2016 Elsevier B.V. All rights reserved.</P>

      • Endophytes from medicinal plants and their potential for producing indole acetic acid, improving seed germination and mitigating oxidative stress

        Khan, A. L.,Gilani, S. A.,Waqas, M.,Al-Hosni, K.,Al-Khiziri, S.,Kim, Y. H.,Ali, L.,Kang, S. M.,Asaf, S.,Shahzad, R. Springer Science + Business Media 2017 Journal of Zhejiang University. Science B. Biomedi Vol.18 No.2

        <P>Medicinal plants have been used by marginal communities to treat various ailments. However, the potential of endophytes within these bio-prospective medicinal plants remains unknown. The present study elucidates the endophytic diversity of medicinal plants (Caralluma acutangula, Rhazya stricta, and Moringa peregrina) and the endophyte role in seed growth and oxidative stress. Various organs of medicinal plants yielded ten endophytes, which were identified as Phoma sp. (6 isolates), Alternaria sp. (2), Bipolaris sp. (1), and Cladosporium sp. (1) based on 18S rDNA sequencing and phylogenetic analysis. The culture filtrates (CFs; 25%, 50%, and 100% concentrations) from these endophytes were tested against the growth of normal and dwarf mutant rice lines. Endophytic CF exhibited dose-dependent growth stimulation and suppression effects. CF (100%) of Phoma sp. significantly increased rice seed germination and growth compared to controls and other endophytes. This growth-promoting effect was due to the presence of indole acetic acid in endophytic CF. The gas chromatography/mass spectrometry (GC/MS) analysis showed the highest indole acetic acid content ((54.31 +/- 0.21) mu mol/L) in Bipolaris sp. In addition, the isolate of Bipolaris sp. exhibited significantly higher radical scavenging and anti-lipid peroxidation activity than the other isolates. Bipolaris sp. and Phoma sp. also exhibited significantly higher flavonoid and phenolic contents. The medicinal plants exhibited the presence of bio-prospective endophytic strains, which could be used for the improvement of crop growth and the mitigation of oxidative stresses.</P>

      • SCIESCOPUSKCI등재

        Potential Health Benefits of Conjugated Linoleic Acid (CLA): A Review

        Khanal, R.C. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.9

        Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of octadecadienoic acid with two conjugated double bonds. Of more than a dozen isomers of CLA found naturally in dairy and meat products from ruminants, c-9, t-11 and t-10, c-12 are the two isomers with known physiological importance, including anticarcinogenic, antidiabetic, antilipogenic, and antiatherosclerotic effects. Positive effects of CLA on immune function and bone modeling have also been reported. In spite of the compelling findings in tissue cultures and experimental animal models, its effect, dose, and mechanism of action vis-à-vis specific isomers remains speculative. Results obtained from animal models are inconclusive and conflicting at times in humans, where the research data is limited. It appears that there is a long way to go before CLA could be accepted unequivocally as having definite effects in any or all of these physiological states and how such effects actually occur in humans. The objective of this review is to critically examine the available literature on potential health benefits of CLA observed in cell culture, animal models, and human subjects, wherever possible and to a certain extent the mechanism of action associated with these biological activities.

      • Aglycon of Rhizochalin from the <i>Rhizochalina incrustata</i> Induces Apoptosis <i>via</i> Activation of AMP-Activated Protein Kinase in HT-29 Colon Cancer Cells

        Khanal, Prem,Kang, Bong Seok,Yun, Hyo Jeong,Cho, Hae-Guk,Makarieva, Tatyana Nikolaevna,Choi, Hong Seok Pharmaceutical Society of Japan 2011 BIOLOGICAL & PHARMACEUTICAL BULLETIN Vol.34 No.10

        <P>Rhizochalin is a two-headed sphingolipid-like compound isolated from the sponge <I>Rhizochalina incrustata</I>. It has been reported that rhizocalin and its derivates have a chemopreventive and chemotherapeutic effect. However, the molecular mechanism of these effects is not understood. Here, we demonstrate that aglycon of rhizochalin (AglRhz) from the <I>Rhizochalina incrustata</I> induces AMP-activated protein kinase (AMPK) phosphorylation, and thereby inhibits mammalian target of rapamycin (mTOR)-p70S6 kinase-extracellular signal-regulated kinase (ERK) signaling and activator protein 1 (AP-1) activity <I>via</I> phosphorylation of Raptor in HT-29 cells. In addition, AglRhz induced activation of caspase-3 and poly(ADP-ribose) polymerase (PARP), and DNA fragmentation in HT-29 cells, leads to induction of apoptosis as well as suppression of tumorigenicity of HT-29 cells. Notably, AglRhz inhibits insulin-like growth factor (IGF)-1-induced AP-1 activity and cell transformation in JB6 Cl41 cells. Overall, our findings identify AMPK as an important target protein for mediating the anti-tumor properties of AglRhz in HT-29 colon cancer cells and have important implication for sponges, the most important marine source, in colon cancer.</P>

      • SCIESCOPUSKCI등재

        Effect of Feeding Urea Treated Rice and Wheat Straw on Intake and Milk Yield of Lactating Buffaloes under Farmers Conditions

        Khanal, R.C.,Gurung, D.B.,Kadariya, R.K. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.8

        Two experiments were conducted to study the effect of urea treatment of rice and wheat straw on feed intake, dry matter (DM) digestibility and milk yield of lactation buffaloes in their late lactation under farmers' management conditions in the western hills of Nepal during 1995 and 1997. Dry matter intake (DMI) from urea treated rice and wheat straw was not improved significantly (p<0.05) nor the total DMI of the lactating buffaloes was improved significantly. However, feeding urea treated rice straw increased straw DMI by 14.2% and total DMI by 10.63% units over the untreated rice straw. Similarly, the increase in straw and total DMI were 20.18 and 17.40% units over the untreated wheat straw fed animals. Although there was no significant effect of urea treatment of both straw on DM digestibility, it was higher for treated than untreated straw at all locations. An overall increment of 18.1% units for rice straw and 13.3% units for wheat straw was observed. There was a significant effect (p<0.01) of feeding urea treated rice and wheat straw on the milk yield of lactating buffaloes during late lactation under farmers conditions. Post experiment milk yield was also significantly (p<0.05) higher for the animals fed treated straw in both the experiments. Buffalo milk yield was also significantly affected by breed (p<0.01), location (p<0.01) and parity (p<0.01) of the animals. General response of the farmers about the technology and their observed effect on animal performance was also very positive.

      • SCISCIESCOPUS

        Proyl isomerase Pin1 facilitates ubiquitin-mediated degradation of cyclin-dependent kinase 10 to induce tamoxifen resistance in breast cancer cells

        Khanal, P,Yun, H J,Lim, S C,Ahn, S G,Yoon, H E,Kang, K W,Hong, R,Choi, H S Macmillan Publishers Limited 2012 Oncogene Vol.31 No.34

        Endocrine therapies that inhibit estrogen receptor (ER)-α signaling are the most common and effective treatment for ER-α-positive breast cancer. However, the use of these agents is limited by the frequent development of resistance. The aim of this study was to elucidate the mechanisms by which downregulation of CDK10 expression confers resistance to tamoxifen in breast cancer. Here, we show that peptidyl-prolyl isomerase Pin1 downregulates CDK10 protein as a result of its interaction with and ubiquitination of CDK10, thereby affecting CDK10-dependent Raf-1 phosphorylation (S338). Pin1<SUP>–/–</SUP> mouse embryonic fibroblasts (MEFs) show higher CDK10 expression than Pin1<SUP>+/+</SUP> MEFs, whereas CDK10 protein was downregulated in the rescued Pin1<SUP>–/–</SUP> MEFs after reexpression of Pin1. Pin1 silencing in SKBR-3 and MCF7 cells increased the CDK10 expression. In human tamoxifen-resistant breast cancer and tamoxifen-resistant MCF7 cells, immunohistochemical staining and immunoblotting analysis shows an inverse correlation between the expression of CDK10 and the degree of tamoxifen resistance. There was also a positive correlation between the high level of P-Raf-1 (Ser338) and Pin1 in human tamoxifen-resistant breast cancer and tamoxifen-resistant MCF7 (TAMR-MCF7) cells. Importantly, 4-OH tamoxifen (4-OHT), when used in combination with overexpressed CDK10 or Raf-1 inhibitor, increased cleaved PARP and DNA fragmentation to inhibit cologenic growth of MCF7 cells and Tamoxifen-resistant MCF7 cells, respectively. On the basis of these findings, we suggest that the Pin1-mediated CDK10 ubiquitination is a major regulator of tamoxifen-resistant breast cancer cell growth and survival.

      • SCISCIESCOPUS

        Tpl-2 kinase downregulates the activity of p53 and enhances signaling pathways leading to activation of activator protein 1 induced by EGF

        Khanal, P.,Lee, K.-Y.,Kang, K.-W.,Kang, B. S.,Choi, H. S. Oxford University Press 2009 Carcinogenesis Vol.30 No.4

        <P>Tumor progression locus-2 (Tpl-2) kinase is a member of the mitogen-activated protein kinase kinase kinase family that has been implicated in cellular transformation. The enhanced expression of this protein has been shown to activate both the mitogen-activated protein kinase and c-Jun N-terminal kinase pathways. However, the molecular mechanisms responsible for the oncogenic potential of Tpl-2 are still largely unknown. Here, we showed that Tpl-2 interacted with p53 both in vitro and ex vivo. The overexpression of Tpl-2 inhibited the epidermal growth factor (EGF)-induced p53 phosphorylation (Ser15) through upregulating the activity of protein phosphatase 2A, which interacted with p53 stimulated by EGF. Also, the EGF-induced p53 activity was suppressed in the Tpl-2 wild-type (WT)-transfected HEK 293 cells, but had no effect in the Tpl-2-mutant (S413A)-transfected cells. Furthermore, introduction of small interfering RNA-Tpl-2 into HEK 293 cells resulted in decreased cell viability compared with only adenovirus-p53-infected cells. In addition, the Tpl-2 WT, but not Tpl-2 mutant (S413A), showed increased EGF-induced c-fos promoter activity, followed by activator protein 1 (AP-1) transactivation activity, which was associated with the cell transformation prompted by the H-Ras-Tpl-2-AP-1 signaling axis. These results indicated that the Ser413 of Tpl-2 plays an important role in EGF-induced carcinogenesis as well as inactivation of the p53.</P>

      • SCIESCOPUSKCI등재

        Influence of Supplementing Dairy Cows Grazing on Pasture with Feeds Rich in Linoleic Acid on Milk Fat Conjugated Linoleic Acid (CLA) Content

        Khanal, R.C.,Dhiman, T.R.,Boman, R.L.,McMahon, D.J. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.9

        Three experiments were conducted to investigate the hypothesis that cows grazing on pasture produce the highest proportion of c-9 t-11 CLA in milk fat and no further increase can be achieved through supplementation of diets rich in linoleic acid, such as full-fat extruded soybeans or soybean oil. In experiment 1, 18 lactating Holstein cows were used in a randomized complete block design with measurements made from wk 4 to 6 of the experiment. In experiment 2, three cannulated lactating Holstein cows were used in a $3{\times}3$ Latin square design. Each period was 4 wk with measurements made in the final wk of each period. Cows in both experiments were assigned at random to treatments: a, conventional total mixed ration (TMR); b, pasture (PS); or c, PS supplemented with 2.5 kg/cow per day of full-fat extruded soybeans (PES). In both experiments, feed intake, milk yield, milk composition, and fatty acid profile of milk and blood serum were measured, along with fatty acid composition of bacteria harvested from rumen digesta in experiment 2. In experiment 3, 10 cows which had continuously grazed a pasture for six weeks were assigned to two groups, with one group (n = 5) on pasture diet alone (PS) and the other group (n = 5) supplemented with 452 g of soy oil/cow per day for 7 d (OIL). In experiment 1, cows in PS treatment produced 350% more c-9, t-11 CLA compared with cows in TMR treatment (1.70 vs. 0.5% of fat), with no further increase for cows in PES treatment (1.50% of fat). Serum c-9, t-11 CLA increased by 233% in PS treatment compared with TMR treatment (0.21 vs. 0.09% of fat) with no further increase for cows in PES treatment (0.18% of fat). In experiment 2, cows in PS treatment produced 300% more c-9 t-11 CLA in their milk fat compared with cows in TMR treatment (1.77 vs. 0.59% of fat), but no further increase for cows in PES treatment (1.84% of fat) was observed. Serum c-9, t-11 CLA increased by 250% for cows in PS treatment compared with cows in TMR treatment (0.27 vs. 0.11% of fat), with no further increase for cows in PES treatment (0.31% of fat). The c-9, t-11 CLA content of ruminal bacteria for cows in PS treatment was 200% or more of TMR treatment, but no further increase in bacterial c-9, t-11 CLA for cows in PES treatment was observed. Supplementation of soy oil in experiment 3 also did not increase the c-9 t-11 CLA content of milk fat compared with cows fed a full pasture diet (1.60 vs. 1.54% of fat). Based on these findings, it was concluded that supplementing with feeds rich in linoleic acid, such as full-fat extruded soybeans or an equivalent amount of soy oil, to cows grazing perennial ryegrass pasture may not increase milk fat c-9 t-11 CLA contents.

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