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Basu A.,Saha Institute of Nuclear Physics | Saha S.,Saha Institute of Nuclear Physics | Karmakar S.,Saha Institute of Nuclear Physics | Chakravarty S.,Institute of Genetic Engineering | And 3 more authors.
Proteomics | Year: 2013

Sickle cell disease (SCD) is a hemolytic disorder caused by a mutation in beta-globin gene and affects millions of people worldwide. Though clinical manifestations of the disease are quite heterogeneous, many of them occur due to erythrocyte sickling at reduced oxygen concentration and vascular occlusion mediated via blood cell adhesion to the vessel wall. We have followed proteomic approach to resolve the differentially regulated proteins of erythrocyte cytosol. The deregulated proteins mainly fall in the group of chaperone proteins such as heat shock protein 70, alpha hemoglobin stabilizing protein, and redox regulators such as aldehyde dehydrogenase and peroxiredoxin-2 proteoforms. Proteasomal subunits are found to be upregulated and phospho-catalase level also got altered. Severe oxidative stress inside erythrocyte is evident from the ROS analysis and OxyblotTM experiments. Peroxiredoxin-2 shows significant dimerization in the SCD patients, a hallmark of oxidative stress inside erythrocytes. One interesting fact is that most of the differentially regulated proteins are also common for hemoglobinopathies such as Eβ thalassemia. These could provide important clues in understanding the pathophysiology of SCD and lead us to better patient management in the future.© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source


Lahiri K.,University of Calcutta | Mukhopadhyay M.J.,Laval University | Desjardins Y.,Institute of Genetic Engineering | Mukhopadhyay S.,University of Calcutta
Nucleus (India) | Year: 2012

Mucuna pruriens L. is an important medicinal plant reported to contain L-3,4-dihydroxy phenylalanine (L-DOPA). The present report describes a simple protocol for plant regeneration through callus morphogenesis in four strains belonging to two different varieties of Mucuna pruriens. Friable, soft and nodular callus was induced from nodal and internodal segments of in vitro-grown seedlings on modified Murashige and Skoog's (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP). For regeneration of shoots, the callus tissues were transferred to medium supplemented with various levels of cytokinin. Highest shoot regeneration from the callus tissue was achieved in modified MS medium fortified with BAP at 1.33 μM level. The regenerated shoots were rooted in vitro in half-strength of liquid MS medium supplemented with various levels of NAA. The regenerates were acclimatized for 2-3 weeks and showed about 80 % survival rate after transferring to the field. Cytological analysis revealed chromosome number stability of all the regenerates (2n022) with complete absence of aneuploidy. Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) analyses of total genomic DNA supported stability at the molecular level among all the clones of these strains when compared with their mother plants. © Archana Sharma Foundation of Calcutta 2012. Source


Lahiri K.,University of Calcutta | Mukhopadhyay M.J.,Institute of Genetic Engineering | Mukhopadhyay S.,University of Calcutta
Cytologia | Year: 2016

The present article has reported a detailed karyotype analysis and 4C nuclear DNA estimation in different cultivars and populations of Clitoria ternatea L. of Fabaceae. This plant, widely distributed in tropical regions in India, is a climber and is an economically important medicinal plant. The karyotype study has revealed a constancy in the diploid somatic chromosome number (2n=16) in all these cultivars and populations. However, the nature and number of nucleolar chromosomes varied. The centromeric chromosomes were either metacentric or sub-metacentric in all these cultivars. The total chromosome length and volume differed among these cultivars and populations having the same somatic chromosome number. The differential condensation of chromosomes and the association of variable amounts of proteins in chromosome composition might have been responsible for such differences. In situ cytophotometric studies revealed an intermediate genome size in this species. This is the first report on the detailed karyotype and genome size of cultivars of Clitoria ternatea L. © 2016 The Japan Mendel Society. Source


Ghareeb D.A.,Alexandria University | Hussien H.M.,Alexandria University | Khalil A.A.,Institute of Genetic Engineering | El-Saadani M.A.,Alexandria University | Ali A.N.,Alexandria University
Toxicological and Environmental Chemistry | Year: 2010

The current study was carried out to investigate the effects of low level lead (Pb) exposure on brain tissue antioxidant enzymes activities and acetylcholinesterase (AChE), inflammatory markers (nitrites (NO) and TNF-α), and lipid profile. Furthermore, the possible effects of flaxseed extract to reverse PB-induced toxicity were examined. Female Sprague-Dawley rats were exposed to Pb (200 mg L-1 in drinking water) for three weeks followed by 21 days of orally administrated flaxseed extract (300 mg kg-1). AChE activity increased by 64% and a significant decrease in glutathione (GSH) levels, total antioxidants capacity, glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) activities after Pb exposure. Moreover, NO and α-TNF were increased by 166.5% and 400%, respectively. Finally, Pb exposure increased the brain cholesterol and triglycerides levels. Chronic treatment with flaxseed significantly attenuated cholinergic dysfunction, oxidative stress, and inflammation in the brain after a three week treatment period. Data showed the involvement of factors such as oxidative stress, inflammation, and high expression of AChE activity in Pb-induced neurotoxicity, and showed that flaxseed prevented these adverse effects. © 2010 Taylor & Francis. Source


Lahiri K.,University of Calcutta | Mukhopadhyay M.J.,Institute of Genetic Engineering | Mukhopadhyay S.,University of Calcutta
Plant Tissue Culture and Biotechnology | Year: 2011

Reports on increment in L-DOPA content in micropropagated plants, regenerated through a simple technique following shoot bud multiplication in four strains of two different varieties of Mucuna pruriens are made. Nodal segments from in vitro grown seedlings were cultured on modified MS with various concentrations and combinations of BAP, 2iP, Kn either alone or with NAA. Highest shoot regeneration from the callus was achieved in modified MS fortified with BAP at 1.33 μM level. The regenerated shoots were rooted in vitro in half-strength of liquid MS supplemented with various levels of NAA (0.54 - 10.7 μM) or IBA (0.49 - 9.85 μM). However, roots of superior quality were obtained at 5.4 μM NAA level after two weeks in culture. The regenerants were acclimatized for 2 - 3 weeks and about 85% survival rate was observed after transferring to the field. Both chromosome number stability as well as stability in nuclear DNA contents of the regenerants of all these strains was recorded with complete absence of aneuploidy. The different strains have revealed two to threefolds increase in L-DOPA contents in the cultured plants. The L-DOPA concentration in leaves of the regenerated plants varied from11.85 - 15.42 mg/g dry weight in all these accessions. However, the highest amount was observed in the wild strain of M. pruriens. This is the first report on enhancement of L-DOPA content in differentiated tissue of cultured plants of both the varieties of M. pruriens. Source

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