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PubMed | IPGMER, Institute of Genetic Engineering and Pt J N M Medical College
Type: | Journal: New microbes and new infections | Year: 2015

We studied the roles of vitamin D and its receptor, VDR, in the progression of leprosy. The majority of individuals with leprosy from Kolkata, India, with a type 1 or type 2 reaction have low levels of vitamin D3 in serum samples. Interestingly, individuals with a type 2 reaction associated with neuritis/erythema nodosum leprosum had very low VDR mRNA expression levels, ranging from 5% to 10%, compared to that of healthy control subjects; these patients also had a high bacilli index, ranging from 3+ to 5+. This is the first report to indicate that VDR expression levels may determine the complexity and severity of the progression of leprosy.

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.

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.

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.

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.

De P.,Institute of Genetic Medicine and Genomic science | Chakravarty S.,Institute of Genetic Engineering | Chakravarty A.,Institute of Genetic Engineering
Journal of Human Reproductive Sciences | Year: 2015

Two couples with a history of recurrent pregnancy losses were referred to the Institute of Genetic Medicine and Genomic Science for cytogenetic evaluation. Chromosomal analysis of the phenotypically normal couples was done to investigate whether there are any new chromosomal abnormalities present in either of the couples caused recurrent pregnancy losses. Clinical and hormonal profile of the couples revealed normal parameters. The ultrasound scan of the females showed normal uterine and ovarian structures. Chromosomal analysis of the couples revealed normal 46, XY karyotypes in the both the male partners, and novel balanced reciprocal translocations 46, XX, t (5;8) (q35.3;q24.23) and 46, XX, t (4;13) (q12;q14) chromosomal constitutions in the female partners. Further, corroboration of the chromosome abnormalities was carried out by high resolution banding analysis. Unique and novel balanced reciprocal translocations were reported as an original investigation in two female partners from two different unrelated families both with the history of recurrent pregnancy losses. © 2015 Journal of Human Reproductive Sciences.

Chatterjee T.,Institute of Genetic Medicine and Genomic Science | Chakravarty A.,Institute of Genetic Medicine and Genomic Science | Chakravarty S.,Thalassaemia Foundation | Chakravarty S.,Institute of Genetic Engineering
Hemoglobin | Year: 2015

We evaluated population screening programs (1999-2011), conducted by the Thalassaemia Foundation, Kolkata, India, for the first time in Eastern India in different districts of West Bengal, for prevention of thalassemia comprising screening of heterozygotes and β-thalassemia intermedia (β-TI) cases [β+, β++, β0/β+, βE/βE (codon 26 or HBB: c.79G > A), Hb-E-β-thalassemia (Hb E-β-thal)]. Among 18,166 cases, we found 2092 heterozygotes and 2245 β-TI individuals (who had no information about their disorders). Results were evaluated with standard hematological analyses including erythrocyte indices, hemoglobin (Hb) typing and quantification. Participants were divided into five groups (children, pre-marriage cases, pre-pregnancy cases, affected family members, pregnant women). The objectives of this evaluation were to fix cut-off values of red blood cells (RBCs), mean corpuscular volume (MCV), mean corpuscular Hb (MCH), red blood cell distribution width (RDW) and Hb A2, as the standard World Health Organization (WHO) guidelines were not strictly followed in mass-scale screening programs. We have observed many dilemmas in considering the status of the thalassemia subject, due to presence of some other clinical conditions such as iron deficiency anemia, α-thalassemia (α-thal), δ-thalassemia (δ-thal), clinically silent Hb variants, and some cases of non hemoglobinopathies (such as pregnancy) along with thalassemia. The MCV values varied greatly in different conditions of hemoglobinopathies, whereas MCH provided a more stable measurement. We found an MCH value of <27.0 pg is a suitable cut-off point for screening in this population. Participants with an MCH of <27.0 pg should be investigated further to confirm or exclude a diagnosis of β-thal trait. © 2015 Taylor & Francis.

PubMed | Institute of Genetic Medicine and Genomic Science and Institute of Genetic Engineering
Type: Journal Article | Journal: Journal of pediatric genetics | Year: 2016

Here, we present two thalassemic patients (one male and one female), having unusual clinical phenotypes. Both had mental retardation in which one was associated with microcephaly and other had congenital cataract. They were referred to our institute for clinical evaluation and cytogenetic testing. Both patients were tested for presence of abnormal hemoglobin by high performance liquid chromatography and found to be thalassemic. Their -globin mutation was also determined by amplification refractory mutation system-polymerase chain reaction. The male patient was found to have intervening sequence 1-5 (G-C)/+, indicating -thalassemia trait and the female was found to have Cod 26 (G-A)/IVS 1-5 (G-C), indicating hemoglobin E- thalassemia. Their cytogenetic analysis of blood lymphocytes were studied with high-resolution GTG-banding analysis by using chromosome profiling (Cyto-vision software 3.6) on their chromosomes. Results revealed 46,XY,del(1)(p36.21) in the male and 46,XX,del(1)(p36.3) in the female. Their genotype variation showed (based on genome browser) significant gene loss which probably leads to marked phenotype variation. We believe, thalassemia with mental retardation associated with microcephaly and congenital cataract, both having loss in chromosome 1, p36 position, is reported probably first time from India. This report will definitely enlighten all concerns and add to the information in growing literature.

Bhattacharya D.,Saha Institute of Nuclear Physics | Saha S.,Saha Institute of Nuclear Physics | Basu S.,Saha Institute of Nuclear Physics | Chakravarty S.,Institute of Genetic Engineering | And 4 more authors.
Proteomics - Clinical Applications | Year: 2010

Purpose: In (hemoglobin, Hb) HbEβ-thalassemia, HbE (β-26 Glu→ Lys) interacts with b-thalassemia to produce clinical manifestation of varying severity. This is the first proteomic effort to study changes in protein levels of erythrocytes isolated from HbEβ-thalassemic patients compared to normal. Experimental design: We have used 2-DE and MALDI-MS/MS-based techniques to investigate the differential proteome profiling of membrane and Hb-depleted fraction of cytosolic proteins of erythrocytes isolated from the peripheral blood samples of HbEβ-thalassemia patients and normal volunteers. Results: Our study showed that redox regulators such as peroxiredoxin 2, Cu-Zn superoxide dismutase and thioredoxin and chaperones such as α-hemoglobin stabilizing protein and HSP-70 were upregulated in HbEβ-thalassemia. We have also observed larger amounts of membrane associated globin chains and indications of disruption of spectrin-based junctional complex in the membrane skeleton of HbEβ-thalassemic erythrocytes upon detection of low molecular weight fragments of β-spectrin and decrease in b-actin and dematin content. Conclusion and clinical relevance: We have observed interesting changes in the proteomic levels of redox regulators and chaperons in the thalassemic hemolysates and have observed strong correlation or association of the extent of such proteomic changes with HbE levels. This could be important in understanding the role of HbE in disease progression and pathophysiology.

De P.,Institute of Genetic Engineering | Mukhopadhyay M.J.,Institute of Genetic Engineering | Chakravarty A.,Institute of Genetic Engineering
Nucleus (India) | Year: 2015

Cymbopogon citratus, commonly known as lemongrass - a herb in Asian cuisine, is well known for its medicinal properties since ancient period and have been reported in ancient Indian Ayurvedic Medicine for relieving cough and nasal congestion. Since the early 1980s, an alarming problem of groundwater arsenic (As) contamination has devastated eastern Indian regions. Arsenic is a known human carcinogen and an environmental pollutant known to cause adverse health effects such as liver injury, neurotoxicity, increase the risk of cancers of skin, lung, bladder and liver. DNA damage and genetic instability induced by the inorganic arsenicals present in water are thought to be prerequisites for the initiation of carcinogenesis. Many natural herbs, which are consumed through our daily diet, possess excellent cancer chemopreventive properties. The present study was conducted to examine the anticlastogenic action of lemongrass extract against the As-induced chromosomal aberrations. Our results demonstrate that lemongrass extract particularly Citral (60 mg/kg/b.w) is effective in counteracting the clastogenicity, of the most potent form of As, sodium arsenite in mice. These results suggest that the use of lemongrass in diet may give an effective protection against the health crisis generated by As. © 2015, Archana Sharma Foundation of Calcutta.

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