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Characterization of a glutathione metabolic mutant of Mycobacterium tuberculosis and its resistance to glutathione and nitrosoglutathione 下载免费PDF全文
Glutathione is a tripeptide and antioxidant, synthesized at high levels by cells during the production of reactive oxygen and nitrogen intermediates. Glutathione also serves as a carrier molecule for nitric oxide in the form of S-nitrosoglutathione. Previous studies from this laboratory have shown that glutathione and S-nitrosoglutathione are directly toxic to mycobacteria. Glutathione is not transported into the cells as a tripeptide. Extracellular glutathione is converted to a dipeptide due to the action of transpeptidase, and the dipeptide is then transported into the bacterial cells. The processing of glutathione and S-nitrosoglutathione is brought about by the action of the enzyme gamma-glutamyl transpeptidase. The function of gamma-glutamyl transpeptidase is to cleave glutathione and S-nitrosoglutathione to the dipeptide (Cys-Gly), which is then transported into the bacterium by the multicomponent ABC transporter dipeptide permease. We have created a mutant strain of Mycobacterium tuberculosis lacking this metabolic enzyme. We investigated the sensitivity of this strain to glutathione and S-nitrosoglutathione compared to that of the wild-type bacteria. In addition, we examined the role of glutathione and/or S-nitrosoglutathione in controlling the growth of intracellular M. tuberculosis inside mouse macrophages. 相似文献
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Mark Richter Tajhal Dayaram Aidan G. Gilmartin Gopinath Ganji Sandhya Kiran Pemmasani Harjeet Van Der Key Jason M. Shohet Lawrence A. Donehower Rakesh Kumar 《PloS one》2015,10(2)
The wild-type p53-induced phosphatase 1 (WIP1) is a serine/threonine phosphatase that negatively regulates multiple proteins involved in DNA damage response including p53, CHK2, Histone H2AX, and ATM, and it has been shown to be overexpressed or amplified in human cancers including breast and ovarian cancers. We examined WIP1 mRNA levels across multiple tumor types and found the highest levels in breast cancer, leukemia, medulloblastoma and neuroblastoma. Neuroblastoma is an exclusively TP53 wild type tumor at diagnosis and inhibition of p53 is required for tumorigenesis. Neuroblastomas in particular have previously been shown to have 17q amplification, harboring the WIP1 (PPM1D) gene and associated with poor clinical outcome. We therefore sought to determine whether inhibiting WIP1 with a selective antagonist, GSK2830371, can attenuate neuroblastoma cell growth through reactivation of p53 mediated tumor suppression. Neuroblastoma cell lines with wild-type TP53 alleles were highly sensitive to GSK2830371 treatment, while cell lines with mutant TP53 were resistant to GSK2830371. The majority of tested neuroblastoma cell lines with copy number gains of the PPM1D locus were also TP53 wild-type and sensitive to GSK2830371A; in contrast cell lines with no copy gain of PPM1D were mixed in their sensitivity to WIP1 inhibition, with the primary determinant being TP53 mutational status. Since WIP1 is involved in the cellular response to DNA damage and drugs used in neuroblastoma treatment induce apoptosis through DNA damage, we sought to determine whether GSK2830371 could act synergistically with standard of care chemotherapeutics. Treatment of wild-type TP53 neuroblastoma cell lines with both GSK2830371 and either doxorubicin or carboplatin resulted in enhanced cell death, mediated through caspase 3/7 induction, as compared to either agent alone. Our data suggests that WIP1 inhibition represents a novel therapeutic approach to neuroblastoma that could be integrated with current chemotherapeutic approaches. 相似文献
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Jitendra Gopichand Patil Mahendra Laxman Ahire Kirti Manik Nitnaware Sayantan Panda Vijay P. Bhatt Polavarapu B. Kavi Kishor Tukaram Dayaram Nikam 《Applied microbiology and biotechnology》2013,97(6):2379-2393
Digitalis purpurea L. (Scrophulariaceae; Foxglove) is a source of cardiotonic glycosides such as digitoxin and digoxin which are commercially applied in the treatment to strengthen cardiac diffusion and to regulate heart rhythm. This investigation deals with in vitro propagation and elicited production of cardiotonic glycosides digitoxin and digoxin in shoot cultures of D. purpurea L. In vitro germinated seedlings were used as a primary source of explants. Multiple shoot formation was achieved for three explant types (nodal, internodal, and leaf) cultured on Murashige and Skoog (MS) medium with several treatments of cytokinins (6-benzyladenine—BA; kinetin—Kin; and thidiazuron—TDZ) and auxins (indole-3-acetic acid—IAA; α-naphthaleneacetic acid—NAA; and 2,4-dichlorophenoxy acetic acid—2,4-D). Maximum multiple shoots (12.7?±?0.6) were produced from nodal explants on MS?+?7.5 μM BA. Shoots were rooted in vitro on MS containing 15 μM IAA. Rooted plantlets were successfully acclimatized. To further maintain the multiple shoot induction, mother tissue was cut into four equal parts and repeatedly sub-cultured on fresh shoot induction liquid medium after each harvest. On adaptation of this strategy, an average of 18 shoots per explant could be produced. This strategy was applied for the production of biomass and glycosides digitoxin and digoxin in shoot cultures on MS medium supplemented with 7.5 μM BA and several treatments with plant growth regulators, incubation period, abiotic (salicylic acid, mannitol, sorbitol, PEG-6000, NaCl, and KCl), biotic (Aspergillus niger, Helminthosporium sp., Alternaria sp., chitin, and yeast extract) elicitors, and precursors (progesterone, cholesterol, and squalene). The treatment of KCl, mycelial mass of Helminthosporium sp., and progesterone were highly effective for the production of cardenolides. In the presence of progesterone (200 to 300 mg/l), digitoxin and digoxin accumulation was enhanced by 9.1- and 11.9-folds respectively. 相似文献
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Vinayak Haribhau Lokhande Tukaram Dayaram Nikam Suprasanna Penna 《Plant Cell, Tissue and Organ Culture》2010,102(1):17-25
In vitro-grown cells of Sesuvium portulacastrum L., an important ‘salt accumulator’ mangrove associate, were incubated on a medium containing different levels of salt, including
0, 100, 200, or 400 mM NaCl, in order to evaluate biochemical, physiological, and growth responses. A significant decrease
in callus growth, water status, and cell membrane damage was observed under salt stress. Osmotic adjustment was revealed by
the accumulation of inorganic ions, such as sodium (Na+), and organic osmolytes (proline, glycine betaine, and total soluble sugars) in NaCl-treated calli compared to control. However,
accretion of osmolytes and inorganic ions did not support growth of calli under NaCl stress conditions. The observed reduced
growth rate in calli subjected to stress, up to 200 mM NaCl, was coupled with lower catalase and ascorbate peroxidase activities
and with a significantly higher superoxide dismutase activity. These findings suggested that S. portulacastrum cell cultures exhibited higher osmotic adjustment to salt stress. 相似文献
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Respiratory failure due to differentiation arrest and expansion of alveolar cells following lung-specific loss of the transcription factor C/EBPalpha in mice 下载免费PDF全文
Bassères DS Levantini E Ji H Monti S Elf S Dayaram T Fenyus M Kocher O Golub T Wong KK Halmos B Tenen DG 《Molecular and cellular biology》2006,26(3):1109-1123
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Mahendra Laxman Ahire Savaliram Goga Ghane Vinayak Haribhau Lokhande Penna Suprasanna Tukaram Dayaram Nikam 《In vitro cellular & developmental biology. Plant》2011,47(4):488-495
Uraria picta is extensively used in the Asian traditional systems of medicine. Overexploitation of the species for preparation of the
drug Dashmula has led to the plant becoming rare and endemic. In the present investigation, an efficient micropropagation
protocol has developed from leaf-derived callus of U. picta. Among the various concentrations of cytokinins (6-benzyladenine—BA; kinetin—Kin; and thidiazuron—TDZ) used, a significantly
higher number of shoots per culture (58.8 ± 0.8) was observed on Murashige and Skoog (MS) medium fortified with 4.44 μM BA.
The shoot regeneration frequency was sustained upon transfer to the same fresh medium at 4-wk intervals over a period of 2 yr.
The medium containing various concentrations of auxins (α-napthalene acetic acid (NAA) or indole-3-acetic acid (IAA)) showed
callus interspersed root formation; however, MS basal medium containing 3% sucrose revealed direct root induction from in vitro raised shoots. The acclimatized in vitro grown plants showed almost 98% survival upon transfer to soil in earthen pots and grown ex vitro. Randomly amplified polymorphic DNA analysis of 25 arbitrarily selected regenerants and mother plants revealed 100% uniformity
and true-to-type nature of the regenerants. Methanolic extracts of callus showed strong antibacterial activity against pathogenic
bacteria as compared to leaf and root extracts of in vitro raised plants and wild plants, suggesting the presence of higher concentrations of active chemical constituents (isoflavanoids)
in callus cultures of U. picta. 相似文献
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