Manipal Hospital Diagnostic Services

Bangalore, India

Manipal Hospital Diagnostic Services

Bangalore, India
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Bhat V.S.,Manipal Hospital Diagnostic Services | Mandal A.K.,St Johns Research Institute | Mathew B.,St Johns Research Institute
Indian Journal of Clinical Biochemistry | Year: 2012

Hemoglobin J-Rajappen (alpha)90 Lys → Thr is an alpha chain variant found in heterozygous state and presents normal hematological blood picture. Due to the ambiguity in results obtained while analyzing by HPLC and alkaline gel electrophoresis, we report this rare case of HbJ-Rajappen using non denaturing gel electrophoresis and matrix assisted laser desorption ionization mass spectrometry. Though HbJ-Rajappen has earlier been reported using different techniques, this is the first report being validated using mass spectrometry technique. © 2012 Association of Clinical Biochemists of India.


Bhat V.S.,Manipal Hospital Diagnostic Services | Dewan K.K.,Cauvery Medical Center | Krishnaswamy P.R.,Cauvery Medical Center
Indian Journal of Clinical Biochemistry | Year: 2010

We report a case of hemolytic anemia that was subsequently identified to be a case of α-thalassaemia harboring the common rightward 3.7 kb deletion/HbH. The diagnosis was based on sequential analyses using BioRad D10 HPLC, Alkaline gel electrophoresis, GPO α THAL-IC strips and the identification of the specific genetic lesion using an α Globin reverse dot blot hybridization assay. Supravital stain of RBCs helped in identifying classical HbH inclusions. In a background of a variable clinical presentation, lack of definitive hematological markers, and general under-diagnosis of α-thalassaemias we have used this case to highlight the features and sequence of techniques involved in identifying and characterizing an α-globin chain mutation, starting from a diffuse clinical history and presentation up to the identification of a specific genetic lesion involved. © Association of Clinical Biochemists of India 2010.


Bhat V.S.,Manipal Hospital Diagnostic Services | Dewan K.K.,Cauvery Medical Center | Krishnaswamy P.R.,Cauvery Medical Center | Mandal A.K.,St Johns Research Institute | Balaram P.,Indian Institute of Science
Indian Journal of Clinical Biochemistry | Year: 2010

Hemoglobin Q-India (alpha) 64 Asp →His is an alpha chain variant which is generally found in heterozygous state and presents normal hematological blood picture. Here we report a rare case of HbQ-India with a thalassemic phenotype that has been analyzed using a combination of mass spectrometry, gene sequencing and PCR analysis. This combined analyses revealed the HbQ variant to be associated with a beta chain mutation, IVS 1-1 [G>T]. Though HbQ has earlier been reported with thalassemic trait using different techniques, this is the first report of a compound α and β chain Hb heterozygous mutant involving HbQ and IVS1-1 being validated using Mass Spectrometry and Reverse dot blot hybridization.


Bhat V.S.,Manipal Hospital Diagnostic Services | Mandal A.K.,St Johns Research Institute | Mathew B.,St Johns Research Institute
Indian Journal of Clinical Biochemistry | Year: 2012

The present report describes the molecular study of HbD ran (beta) 22 Glu → Gln associated with β-Thalassemia IVS1-5 (G > C) found in India, and the first case in which mutation has been identified using mass spectrometry. Given the apparent ethnic origin and the mobility of the variant hemoglobin at alkaline pH, hemoglobin D-Punjab would be suspected, but HPLC excluded this possibility. Further characterization of hemoglobinopathy was made by using nondenaturing gel electrophoresis and matrix assisted laser desorption ionization mass spectrometry and IVS1-5 being validated by reverse dot blot hybridization followed by sequencing of the β-globin gene. © 2012 Association of Clinical Biochemists of India.


Bhat V.S.,Manipal Hospital Diagnostic Services | Dewan K.K.,Cauvery Medical Center | Krishnaswamy P.R.,Cauvery Medical Center
Indian Journal of Clinical Biochemistry | Year: 2011

We report a case of a diabetic, heterozygote with near normal hematology, marginally low level of hemoglobin A2(HbA2) having an increased level of hemoglobin F(HbF) that was pancellularly distributed among the red cells. BioRad DiaSTAT measurements gave a high glycated hemoglobin A1c(HbA1c) of 31.5% and the BioRad Variant analyzer recorded an HbA1c value which was very low, in discordance with the detected blood glucose levels. Flow cytometry and polymerization chain reaction (PCR) based studies were carried out which revealed the case to be that of the common hereditary persistence of fetal hemoglobin (HPFH)-3, an Asian Indian mutation. Fructosamine estimation and HbA1c by Boronate affinity chromatography were able to resolve the discordant value detected and was able to confirm the diabetes status. The case would have been a diagnostic dilemma, if reported without correlation. © 2010 Association of Clinical Biochemists of India.


PubMed | Manipal Hospital Diagnostic Services
Type: Journal Article | Journal: Indian journal of clinical biochemistry : IJCB | Year: 2012

Hemoglobin Q- India (alpha) 64 Asp His is an alpha chain variant which is generally found in heterozygous state and presents normal hematological blood picture. Here we report a rare case of HbQ-India with a thalassemic phenotype that has been analyzed using a combination of mass spectrometry, gene sequencing and PCR analysis. This combined analyses revealed the HbQ variant to be associated with a beta chain mutation, IVS 1-1 [G>T]. Though HbQ has earlier been reported with thalassemic trait using different techniques, this is the first report of a compound and chain Hb heterozygous mutant involving HbQ and IVS1-1 being validated using Mass Spectrometry and Reverse dot blot hybridization.


PubMed | Manipal Hospital Diagnostic Services
Type: Journal Article | Journal: Indian journal of clinical biochemistry : IJCB | Year: 2013

Hemoglobin J-Rajappen (alpha)90 LysThr is an alpha chain variant found in heterozygous state and presents normal hematological blood picture. Due to the ambiguity in results obtained while analyzing by HPLC and alkaline gel electrophoresis, we report this rare case of HbJ-Rajappen using non denaturing gel electrophoresis and matrix assisted laser desorption ionization mass spectrometry. Though HbJ-Rajappen has earlier been reported using different techniques, this is the first report being validated using mass spectrometry technique.


Bhat V.S.,Manipal Hospital Diagnostic Services | Lokeshwarappa S.,Manipal Hospital Diagnostic Services | Mandal A.K.,St Johns Research Institute
Indian Journal of Clinical Biochemistry | Year: 2015

The present case report describes the molecular and proteomic based study of Hb variant HbE associated with β+ thalassemia IVS 1-1 G>T, in a juvenile diabetic patient. Given the ethnic origin and mobility of the variant hemoglobin at alkaline pH, HbE would be suspected. But hematologically and clinically abnormality being detected, HPLC and Electrophoresis not being able to characterize due to retention time and band being in region of HbA2, respectively, further characterization of hemoglobinopathy was made using MALDI and IVS 1-1 G>T being validated by reverse dot blot hybridization. Capillary electrophoresis was also employed in order to separate HbE and HbA2 bands. This case report being first of its kind, wherein a HbE/β+ thalassemia has been characterized using multiple techniques. © 2014, Association of Clinical Biochemists of India.


Sivasubramaniyan K.,Stempeutics Research Pvt. Ltd. | Pal R.,Stempeutics Research Malaysia Sdn Bhd | Totey S.,Stempeutics Research Malaysia Sdn Bhd | Bhat V.S.,Manipal Hospital Diagnostic Services
Current Stem Cell Research and Therapy | Year: 2010

Human embryonic stem cells (hESC) differentiate spontaneously in culture and develop a complex microenvironment comprising of autologously derived niche that in turn supports their pluripotency. The basic hypothesis that we deal with is that hESCs undergoing differentiation, sequentially generate trophectoderm and endoderm lineages and thereafter influence further events through the production of growth factors. These factors control the fate of hESCs either by promoting or retarding the recruitment of new cells in the differentiation program. This scenario therefore represents an analog of the in vivo situation in which extra-embryonic tissues influence the behavior of the inner cell mass (ICM). The premise of the paper is the Rho kinase inhibitor Y27632 that can spatiotemporally alter this balance between pluripotency and differentiation. To evaluate the composition and inclination of lineage specification during spontaneous differentiation, we have studied the hESC colonies and their surrounding niche as interdependent entities. We show that the population of fibroblastic niche that surrounds hESC colonies co-expresses trophectoderm and niche cell markers including SSEA1, hCG, progesterone, HAND1, pSmad1 and FGFR1 as early as day 4. A sudden increase in the expression of GATA4 and AFP secretion indicated putative endoderm formation on day 6 in both control and Y27632 treated cultures. On day 6, 20 μM of Y27632 supplementation significantly reduced the trophectoderm-like niche population without affecting endoderm formation, enhanced the average size and number of hESC colonies, decreased IGF1 secretion thereby improving the pluripotency. Overall our findings support the afore mentioned hypothesis and demonstrate that closely packed epithelial trophectoderm-like cells bordering the hESC colonies present an initial and imminent localized niche which is spatiotemporally regulated. Such advances in understanding the behavior and modulation of hESC and its surrounding niche would facilitate better differentiation protocols for applications in regenerative medicine and drug screening. © 2010 Bentham Science Publishers Ltd.


PubMed | St Johns Research Institute and Manipal Hospital Diagnostic Services
Type: Journal Article | Journal: Indian journal of clinical biochemistry : IJCB | Year: 2015

The present case report describes the molecular and proteomic based study of Hb variant HbE associated with (+) thalassemia IVS 1-1 G>T, in a juvenile diabetic patient. Given the ethnic origin and mobility of the variant hemoglobin at alkaline pH, HbE would be suspected. But hematologically and clinically abnormality being detected, HPLC and Electrophoresis not being able to characterize due to retention time and band being in region of HbA2, respectively, further characterization of hemoglobinopathy was made using MALDI and IVS 1-1 G>T being validated by reverse dot blot hybridization. Capillary electrophoresis was also employed in order to separate HbE and HbA2 bands. This case report being first of its kind, wherein a HbE/(+) thalassemia has been characterized using multiple techniques.

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