Pratap Mukhopadhyaya - Academia.edu (original) (raw)

Papers by Pratap Mukhopadhyaya

BLAD is an economically important genetic disorder in cattle. The incidence rate in India is curr... more BLAD is an economically important genetic disorder in cattle. The incidence rate in India is currently reported to be 3.23%. This study was therefore aimed to design a new set of improved PCR primers that can be used for convenient detection of the diagnostic SNP at codon 128 of the CD18 gene of cattle responsible for causing the disease, using Taq I enzyme-mediated RFLP analysis. In depth, bioinformatic analysis was performed to confirm the suitability of primers to substitute published versions that generated shorter fragments and required polyacrylamide gel for detection. The new set of primers generated 184 bp am- plicon with an asymmetrically placed diagnostic Taq I recognition site that cleaved it into 132 bp and 52 bp, respectively. It was demonstrated that the 132 bp and 184 bp fragments were sufficient to diagnose carriers excluding the need for identifying the 52 bp fragment in a regular, 3% agarose gel run on 0.5% TBE instead of conventional non-denaturing polyacrylamide gel.

Roumanian archives of microbiology and immunology

A gram negative chemolithotrophic bacterium (RPI) with facultative mode of nutrition was isolated... more A gram negative chemolithotrophic bacterium (RPI) with facultative mode of nutrition was isolated from the soil. Enzymological studies confirmed presence of Thiosulphate oxidase, sulphite oxidase and Rhodanese, all of which play role in sulfur metabolism pathway. A set of degenerate oligonucleotide primer pairs was used for thermal amplification of a major part of the coding region of the Cytochrome c gene locus of this bacterium. Nucleotide and translated amino acid sequence revealed the gene to be a diheme Cytochrome c, different from the monoheme Cytochrome c observed in Chloribium limicola, a photosynthetic green sulfur bacterium. Significant homology at the nucleotide level could be detected only with Pseudoaminobacter salicylatoxidans. On the contrary, significant homology at the amino acid level was observed with Bradyrhizobium japonicum, Silicobacter pomeroyi apart from P. salicylatoxidans. This is possibly because of codon degeneracy observed within the diverse members of c...

Journal of Applied Genetics, 2008

Possibility of perchlorate reduction by microbes raises hope for an eco-friendly mode of degradat... more Possibility of perchlorate reduction by microbes raises hope for an eco-friendly mode of degradation of this toxic rocket fuel. This study reports 3 isolates (A1, A2 and A3) capable of molybdenum-independent degradation of perchlorate under aerobic conditions. The rate of degradation was the highest when perchlorate concentration was 17 mM, and then 3.2 mM, 4.7 mM and 4.1 mM of perchlorate was reduced by isolates A1, A2 and A3 (respectively) after 72 h at 28 o C under aerobic conditions. Presence of perchlorate at a concentration higher than 17 mM resulted in some inhibition of perchlorate reduction. 16S ribosomal RNA gene analysis revealed isolate A1 to be Pseudomonas stutzeri (Proteobacteria) while isolates A2 ad A3 where found to belong to the genus Arthrobacter (Actinobacteria). The study, apart from demonstrating ribotyping as a rapid method of identification of economically important soil microbes, also raised prospects for using artificial consortia for environmental degradation of perchlorate, without apparent domination of Dechloromonas spp. (a group of microbes known for perchlorate remediation in the environment).

Current HIV Research, 2013

A portion of the gag gene cDNA for p24 protein from 30 Indian HIV-1 proviral DNA was amplified by... more A portion of the gag gene cDNA for p24 protein from 30 Indian HIV-1 proviral DNA was amplified by PCR and sequenced. Phylogenetic analysis with reference samples of A1, A2, B, C, D, F1, F2, G, H, J, K, N and O subtypes revealed that 29 test samples aligned with subtype C reference strain while 1 matched with HIV-1 subtype A. Multiple alignment of predicted amino acid sequence of the Indian test samples and reference C subtype of HIV-1 samples from other countries indicated a molecular signature by way of rigid conservation of the amino acid 'S' at position 41 of the gag p24 protein in all Indian HIV-1 samples analyzed in this study as opposed to 'T' in the same position in C subtype sequences from other parts of the world. A phylogenetic analysis and visualization of the resulting tree in radial position showed distinct clubbing of all Indian C subtypes and formation of a cluster when compared to C subtype sequences from other countries with a single Chinese sample as an exception which was found in the Indian cluster. The use of a portion of p24 gene sequence as tool for subtyping as well as phylogenetic grouping with special reference to its geographical location is discussed.

Nucleotide positions 5063534 to 5063554 and 5063973 to 5063952 within the Human Janus Kinase 2 (J... more Nucleotide positions 5063534 to 5063554 and 5063973 to 5063952 within the Human Janus Kinase 2 (JAK2) gene (Gene bank accession number NT_008413.17) were identified as novel, optimized oligonucleotide primer hybridization sites for thermal amplification of a robust 440 bp PCR amplicon which encompassed the V617F mutation linked to occurrence of Polycythemia vera (PV), a well known myeloproliferative disorder (MPD) in humans. The single strand oligonucleotide primer pair was also found suitable to generate high quality fluorescent nucleotide sequence data from both strands of DNA that was sufficient for detection of all possible genotypes of JAK2 gene with regard to V617F mutation. The accuracy and reproducibility of the method was satisfactory ((r2=0.99, p

Genetics and Molecular Research, 2008

Proteases form nematophagous fungi are most important extracellular hydrolytic enzymes paying a c... more Proteases form nematophagous fungi are most important extracellular hydrolytic enzymes paying a centra l role in cuticle degradation. In the present study, prote ase from two nematode-trapping fungi, A. conoides GenBank accession no. JX979095 and D. flagrans JX979096 were studied at biochemical and molecular level. Crude protease of A. conoides showed maximum activity at pH 7 and temperature 50°C while protese of D. flagrans was functioning best at pH 8 and temperature 55°C. 726 and 716 bp serine protease gene fragment from each fungus was amplified and sequences were submitted to GenBank under the accession No. KC769585 and KC862257. Phylogenetic relationships showed homology with serine proteases of other nematode-trapping fungi. Moreover protease activity in culture broth of fungi inoculated with nematodes was increased. Biochemical assay demonstrated 2.14 and 6.4 fold increase in protease activity in induced culture of A. conoides and D. flagrans respectively. Real Time-PCR assa...

Oncology Research and Treatment, 2014

Introduction: The aim of this study was to investigate the relation between A1 allele of the DRD2... more Introduction: The aim of this study was to investigate the relation between A1 allele of the DRD2 gene encoding the D2 sub type of the dopamine receptor with obesity and depression in obese, female Indian population. Methods: A resource population of 270 obese female individuals with mean body weight of 96.3 ± 9.4 Kg and body mass index of 33.1 ± 2.7 Kg/m were genotyped for the DRD2 A1 allele using the PCRRFLP method. Multivariate analysis was performed using a logistic regression model to test the association between phenotype variables and genotypes. Results: Out of 270 obese subjects, 35.55% were heterozygous for the DRD2 A1 allele. No subjects homozygous for this pathogenic allele were encountered. Out of the sub population bearing one copy of the DRD2 A1 allele, 65.62% were suffering from depression. Conclusion: The pathogenic DRD2 allele was found to be significantly associated with conditions of obesity and depression in a section of obese, female Indian population.

Roumanian archives of microbiology and immunology, 2012

Human MECP2 gene located at q28 arm of X chromosome was identified as target for thermal co-ampli... more Human MECP2 gene located at q28 arm of X chromosome was identified as target for thermal co-amplification with HIV-1 proviral DNA of infected individuals. The selected MECP2 gene-specific primers functioned at a wide range of annealing temperature, extension time and exhibited no significant interaction with pathogen specific primers. A 466 bp PCR amplicon originating from human MECP2 gene was found to be diagnostic for inhibition-free PCR reaction when co-amplified with the HIV-1 target gene in a multiplexed, nested PCR reaction. The 5' end of the MECP2 primers were engineered to position an EcoRI restriction endonuclease site to facilitate rapid cloning in various DNA vector molecules at the corresponding EcoRI sites. Cell mass of Escherichia coli (XL1Blue) harboring the recombinant plasmid when added to pleural fluid of HIV-1 infected individuals co-infected with Mycobacterium tuberculosis, generated the diagnostic 466 bp MECP2 PCR amplicon as well as the 194 bp PCR amplicon ...

for generating viable protoplasts from isolate of the nematophagous fungus Arthobotrys oviformis ... more for generating viable protoplasts from isolate of the nematophagous fungus Arthobotrys oviformis is developed. The lytic enzyme Novozyme 234 is used in the study. Maximum protoplasts are released with 24 hours old A. oviformis mycelia after 35 minutes of treatment with the lytic enzyme Novozyme 234 at a concentration of 10 mg ml -1 . The nematophagous fungus Dactylaria parvispora however is found to be refractive to the process and protoplasts could not be generated under the range of conditions used in this study. The yield of protoplasts and the frequency of regeneration of A. oviformis is found to be highest when 0.6 molar Sodium Chloride is used as the osmotic stabilizer. This is the first report of an optimized method for generating viable protoplast from young mycelia of the nematophagous fungus A. oviformis.

Aims: Development of a JAK2 Allelic burden estimation kit using Taqman probes and determine trend... more Aims: Development of a JAK2 Allelic burden estimation kit using Taqman probes and determine trend in shifting of JAK2 V617F allelic burden in and its use in analysis of patients with essential thrombocythemia (ET) and polycythemia vera (PV) to predict thrombotic complications. Original Research Article Syed et al.; IRJO, 2(4): 1-16, 2019; Article no.IRJO.53378 2 Methodology: Through a retrospect study, a total of 412 ET and PV patient, divided into 3 groups (0-2, 2-5 and 5-10 years) based on time of detection of the disease, were tested in retrospect for presence of the JAK2 V617F mutant allele burden using an in-house developed Taqman probe-based kit, trend in shifting of the mutant allele burden was studied and segregated into ET (n=167) and PV (n=126) group based on their disease profile. It was then categorized into 3 time periods (0-2, 2-5 and 5-10 years) based on time of detection of the disease. Results: Around 293 (71%) were positive for JAK2 V617F while 59 (14.2%) and 8 (1.9%) positive for CALR exon 9 frame shift & MPL mutations (W515L/W515K) respectively. The 1-25% allelic burden group size gradually fell in ET population over time and this trend continued in the PV population also. In the former the fall was 7% & 11% for 2-5 & 5-10 years category while in the later, it was 1% and 15% respectively for the same time period-category. Conclusions: There is a distinct molecular continuum in the JAK2 V617F allelic burden in the ET & PV patients which followed a predictable trend and was associated with increasingly complicated vascular events.

In recent times, the demand for prescribed medicines has increased substantially. The Royal Pharm... more In recent times, the demand for prescribed medicines has increased substantially. The Royal Pharmaceutical Society of Great Britain (RPSGB) felt the burden and introduced supplementary prescribing in their system in the year 2003. The UK government authorities and other related bodies designed special courses which was required to be attended by pharmacists as well as nurses who intended to prescribe medicines. However, after the introduction of this legislation there was a need to assess whether the above legislation was beneficial or not. Limited studies exist that explore the current scenario of supplementary prescribing in the UK. This book is aimed at analyzing view of nurses and pharmacists on supplementary prescribing. Different sets of questionnaire were used to collect response from both pharmacists as well as nurses. Analysis of the data indicated unambiguous significance and increasing importance of the role of supplementary prescribing in England.

Cell free DNA (cfDNA) is now emerging as potent biomarker in cancer diagnostics as well as an use... more Cell free DNA (cfDNA) is now emerging as potent biomarker in cancer diagnostics as well as an useful tool for non-invasive diagnostic methods related to a wide range of clinical conditions. Its extraction from blood is crucial to exploitation as a biomarker. Link of cfDNA has been established with a wide range of health conditions including autoimmune diseases, sepsis and trauma, heart diseases and dialytic processes. However, its utility in prenatal diagnosis has been one of the most widely investigated areas of research. Association of cfDNA with cancer and its methylation status as a potent prognostic marker is also assuming increasing significance in recent times. Its association with different forms of cancer with special reference to breast cancer is notable. A doubtful mammographic findings does indicate other testing that involve imaging techniques but the final diagnosis is established by a biopsy in particular to differentiate malignant from benign tumors. Although this is...

The book discusses the medicinal properties of plants such as, Bacopa monnieri, Centella asiatica... more The book discusses the medicinal properties of plants such as, Bacopa monnieri, Centella asiatica, Rubia cordifolia, Hemidesmus indicus and Phyllanthus amarus. These plants have several therapeutic uses and are indigenous to India. They have immense value since ancient times and are still used in Ayurvedic preparations and hence in demand in the herbal market. The book describes in-vitro cultures which were established using various explants. It further discusses various analytic techniques such as Thin Layer Chromatography (TLC), High Performance Thin Layer Chromatography (HPTLC) & High Performance Liquid Chromatography (HPLC) which were exploited during the course of study of these plant extracts. Description of animal study techniques, acute toxicity on Swiss mice and efficacy studies of Phyllanthus amarus and Andrographis paniculata extracts on Albino Wistar rats with damaged liver as well as screening of hepatoprotective activity of pure phyllanthin are part of this book. The s...

International Journal of Agricultural Technology, 2014

Insufficient nutrient and pest infection are two major predicaments for low crop yield in agricul... more Insufficient nutrient and pest infection are two major predicaments for low crop yield in agriculture. Nematodes, a most infectious parasite pest in plants responsible for major yield loss. Chemical phosphate fertilizer made large reserves of it in soil, but part of it is made available to the plants because of irreversible fixation of it with divalent cation. In the present study, Arthrobotrys conoides (JX979095) and Duddingtonia flagrans (JX979096) a nematode trapping fungi were isolated from the arable soil of Anand district and studied for its phosphate solubilization potential. Among the different P sources, bioleaching of Tricalcium Phosphate (TCP) was found to be more efficient compared to other P salts. Phosphate solubilizing ability was enhanced in presence of D-glucose as compared to other C sources. In case of nitrogen source ammonium sulfate at 0.5 % showed best P solubilization followed by casein, urea and sodium nitrate respectively. Duddingtonia flagrans was showing m...

Possibility of reduction of perchlorate using microbes raises hope for an eco friendly mode of de... more Possibility of reduction of perchlorate using microbes raises hope for an eco friendly mode of degradation of this toxic rocket fuel. This study reports three isolates, viz., Al, A2 and A3 capable of molybdenum-independent degradation of perchlorate under aerobic conditions. The rate of degradation was found to be highest at substrate concentration of 17 mM of perchlorate, where isolate Al, A2 and A3 reduced 3.2mM, 4.7mM and 4.1 mM of perchlorate respectively under aerobic condition when incubated for 72 hours at 28 C. Presence of perchlorate at a concentration higher than 17mM exhibited inhibition in reduction of the compound. 16S ribosomal RNA gene analysis revealed isolate Al to be Pseudomonas stutzeri (Proteobacteria) while isolates A2 ad A3 where found to be different species of the genus AnhTobacter (Actinobacteria). The study apart from demonstrating ribotyping as a rapid method of identification of economically important soil microbes also raised prospect of using artificial...

A gram negative chemolithotrophic bacterium (RP1) with facultative mode of nutrition was isolated... more A gram negative chemolithotrophic bacterium (RP1) with facultative mode of nutrition was isolated from the soil. Enzymological studies confirmed presence of Thiosulphate oxidase, sulphite oxidase and Rhodanese, all of which play role in sulphur metabolism pathway. A set of degenerate oligonucleotide primer pairs was used for thermal amplification of a major part of the coding region of the Cytochrome c gene locus of this bacterium. Nucleotide and translated amino acid sequence revealed the gene to be a diheme Cytochrome c, different from the monoheme Cytochrome c observed in Chloribium limicola, a photosynthetic green sulphur bacterium. Significant homology at the nucleotide level could be detected only with Pseudoaminobacter salicylatoxidans. On the contrary, significant homology at the amino acid level was observed with Bradyrhizobium japonicum, Silicobacter pomeroyi apart from P. salicylatoxidans. This is possibly because of codon degeneracy observed within the diverse members of...

An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle.... more An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle. This has opened the prospect for large-scale screening of cattle using the polymerase chain reaction (PCR) technique for the rapid identification of heterozygous animals. To facilitate such a screening process, the mutant and normal alleles of factor XI gene, represented by 244- and 320-bp PCR amplified fragments, were individually cloned in Escherichia coli using a multicopy plasmid cloning vehicle to generate pFXI-N and pFXI-M, respectively. The authenticity of the inserts was confirmed by nucleotide sequencing. A nested PCR method was developed, by which PCR amplicons generated from primers with annealing sites on the recombinant plasmids and by flanking the insert were used as templates for amplification of the diagnostic products using factor XI gene-specific primers. An equimolar mixture of both PCR amplicons, originating from pFXI-N and pFXI-M, constituted the carrier control whi...

BLAD is an economically important genetic disorder in cattle. The incidence rate in India is curr... more BLAD is an economically important genetic disorder in cattle. The incidence rate in India is currently reported to be 3.23%. This study was therefore aimed to design a new set of improved PCR primers that can be used for convenient detection of the diagnostic SNP at codon 128 of the CD18 gene of cattle responsible for causing the disease, using Taq I enzyme-mediated RFLP analysis. In depth, bioinformatic analysis was performed to confirm the suitability of primers to substitute published versions that generated shorter fragments and required polyacrylamide gel for detection. The new set of primers generated 184 bp am- plicon with an asymmetrically placed diagnostic Taq I recognition site that cleaved it into 132 bp and 52 bp, respectively. It was demonstrated that the 132 bp and 184 bp fragments were sufficient to diagnose carriers excluding the need for identifying the 52 bp fragment in a regular, 3% agarose gel run on 0.5% TBE instead of conventional non-denaturing polyacrylamide gel.

Roumanian archives of microbiology and immunology

A gram negative chemolithotrophic bacterium (RPI) with facultative mode of nutrition was isolated... more A gram negative chemolithotrophic bacterium (RPI) with facultative mode of nutrition was isolated from the soil. Enzymological studies confirmed presence of Thiosulphate oxidase, sulphite oxidase and Rhodanese, all of which play role in sulfur metabolism pathway. A set of degenerate oligonucleotide primer pairs was used for thermal amplification of a major part of the coding region of the Cytochrome c gene locus of this bacterium. Nucleotide and translated amino acid sequence revealed the gene to be a diheme Cytochrome c, different from the monoheme Cytochrome c observed in Chloribium limicola, a photosynthetic green sulfur bacterium. Significant homology at the nucleotide level could be detected only with Pseudoaminobacter salicylatoxidans. On the contrary, significant homology at the amino acid level was observed with Bradyrhizobium japonicum, Silicobacter pomeroyi apart from P. salicylatoxidans. This is possibly because of codon degeneracy observed within the diverse members of c...

Journal of Applied Genetics, 2008

Possibility of perchlorate reduction by microbes raises hope for an eco-friendly mode of degradat... more Possibility of perchlorate reduction by microbes raises hope for an eco-friendly mode of degradation of this toxic rocket fuel. This study reports 3 isolates (A1, A2 and A3) capable of molybdenum-independent degradation of perchlorate under aerobic conditions. The rate of degradation was the highest when perchlorate concentration was 17 mM, and then 3.2 mM, 4.7 mM and 4.1 mM of perchlorate was reduced by isolates A1, A2 and A3 (respectively) after 72 h at 28 o C under aerobic conditions. Presence of perchlorate at a concentration higher than 17 mM resulted in some inhibition of perchlorate reduction. 16S ribosomal RNA gene analysis revealed isolate A1 to be Pseudomonas stutzeri (Proteobacteria) while isolates A2 ad A3 where found to belong to the genus Arthrobacter (Actinobacteria). The study, apart from demonstrating ribotyping as a rapid method of identification of economically important soil microbes, also raised prospects for using artificial consortia for environmental degradation of perchlorate, without apparent domination of Dechloromonas spp. (a group of microbes known for perchlorate remediation in the environment).

Current HIV Research, 2013

A portion of the gag gene cDNA for p24 protein from 30 Indian HIV-1 proviral DNA was amplified by... more A portion of the gag gene cDNA for p24 protein from 30 Indian HIV-1 proviral DNA was amplified by PCR and sequenced. Phylogenetic analysis with reference samples of A1, A2, B, C, D, F1, F2, G, H, J, K, N and O subtypes revealed that 29 test samples aligned with subtype C reference strain while 1 matched with HIV-1 subtype A. Multiple alignment of predicted amino acid sequence of the Indian test samples and reference C subtype of HIV-1 samples from other countries indicated a molecular signature by way of rigid conservation of the amino acid 'S' at position 41 of the gag p24 protein in all Indian HIV-1 samples analyzed in this study as opposed to 'T' in the same position in C subtype sequences from other parts of the world. A phylogenetic analysis and visualization of the resulting tree in radial position showed distinct clubbing of all Indian C subtypes and formation of a cluster when compared to C subtype sequences from other countries with a single Chinese sample as an exception which was found in the Indian cluster. The use of a portion of p24 gene sequence as tool for subtyping as well as phylogenetic grouping with special reference to its geographical location is discussed.

Nucleotide positions 5063534 to 5063554 and 5063973 to 5063952 within the Human Janus Kinase 2 (J... more Nucleotide positions 5063534 to 5063554 and 5063973 to 5063952 within the Human Janus Kinase 2 (JAK2) gene (Gene bank accession number NT_008413.17) were identified as novel, optimized oligonucleotide primer hybridization sites for thermal amplification of a robust 440 bp PCR amplicon which encompassed the V617F mutation linked to occurrence of Polycythemia vera (PV), a well known myeloproliferative disorder (MPD) in humans. The single strand oligonucleotide primer pair was also found suitable to generate high quality fluorescent nucleotide sequence data from both strands of DNA that was sufficient for detection of all possible genotypes of JAK2 gene with regard to V617F mutation. The accuracy and reproducibility of the method was satisfactory ((r2=0.99, p

Genetics and Molecular Research, 2008

Proteases form nematophagous fungi are most important extracellular hydrolytic enzymes paying a c... more Proteases form nematophagous fungi are most important extracellular hydrolytic enzymes paying a centra l role in cuticle degradation. In the present study, prote ase from two nematode-trapping fungi, A. conoides GenBank accession no. JX979095 and D. flagrans JX979096 were studied at biochemical and molecular level. Crude protease of A. conoides showed maximum activity at pH 7 and temperature 50°C while protese of D. flagrans was functioning best at pH 8 and temperature 55°C. 726 and 716 bp serine protease gene fragment from each fungus was amplified and sequences were submitted to GenBank under the accession No. KC769585 and KC862257. Phylogenetic relationships showed homology with serine proteases of other nematode-trapping fungi. Moreover protease activity in culture broth of fungi inoculated with nematodes was increased. Biochemical assay demonstrated 2.14 and 6.4 fold increase in protease activity in induced culture of A. conoides and D. flagrans respectively. Real Time-PCR assa...

Oncology Research and Treatment, 2014

Introduction: The aim of this study was to investigate the relation between A1 allele of the DRD2... more Introduction: The aim of this study was to investigate the relation between A1 allele of the DRD2 gene encoding the D2 sub type of the dopamine receptor with obesity and depression in obese, female Indian population. Methods: A resource population of 270 obese female individuals with mean body weight of 96.3 ± 9.4 Kg and body mass index of 33.1 ± 2.7 Kg/m were genotyped for the DRD2 A1 allele using the PCRRFLP method. Multivariate analysis was performed using a logistic regression model to test the association between phenotype variables and genotypes. Results: Out of 270 obese subjects, 35.55% were heterozygous for the DRD2 A1 allele. No subjects homozygous for this pathogenic allele were encountered. Out of the sub population bearing one copy of the DRD2 A1 allele, 65.62% were suffering from depression. Conclusion: The pathogenic DRD2 allele was found to be significantly associated with conditions of obesity and depression in a section of obese, female Indian population.

Roumanian archives of microbiology and immunology, 2012

Human MECP2 gene located at q28 arm of X chromosome was identified as target for thermal co-ampli... more Human MECP2 gene located at q28 arm of X chromosome was identified as target for thermal co-amplification with HIV-1 proviral DNA of infected individuals. The selected MECP2 gene-specific primers functioned at a wide range of annealing temperature, extension time and exhibited no significant interaction with pathogen specific primers. A 466 bp PCR amplicon originating from human MECP2 gene was found to be diagnostic for inhibition-free PCR reaction when co-amplified with the HIV-1 target gene in a multiplexed, nested PCR reaction. The 5' end of the MECP2 primers were engineered to position an EcoRI restriction endonuclease site to facilitate rapid cloning in various DNA vector molecules at the corresponding EcoRI sites. Cell mass of Escherichia coli (XL1Blue) harboring the recombinant plasmid when added to pleural fluid of HIV-1 infected individuals co-infected with Mycobacterium tuberculosis, generated the diagnostic 466 bp MECP2 PCR amplicon as well as the 194 bp PCR amplicon ...

for generating viable protoplasts from isolate of the nematophagous fungus Arthobotrys oviformis ... more for generating viable protoplasts from isolate of the nematophagous fungus Arthobotrys oviformis is developed. The lytic enzyme Novozyme 234 is used in the study. Maximum protoplasts are released with 24 hours old A. oviformis mycelia after 35 minutes of treatment with the lytic enzyme Novozyme 234 at a concentration of 10 mg ml -1 . The nematophagous fungus Dactylaria parvispora however is found to be refractive to the process and protoplasts could not be generated under the range of conditions used in this study. The yield of protoplasts and the frequency of regeneration of A. oviformis is found to be highest when 0.6 molar Sodium Chloride is used as the osmotic stabilizer. This is the first report of an optimized method for generating viable protoplast from young mycelia of the nematophagous fungus A. oviformis.

Aims: Development of a JAK2 Allelic burden estimation kit using Taqman probes and determine trend... more Aims: Development of a JAK2 Allelic burden estimation kit using Taqman probes and determine trend in shifting of JAK2 V617F allelic burden in and its use in analysis of patients with essential thrombocythemia (ET) and polycythemia vera (PV) to predict thrombotic complications. Original Research Article Syed et al.; IRJO, 2(4): 1-16, 2019; Article no.IRJO.53378 2 Methodology: Through a retrospect study, a total of 412 ET and PV patient, divided into 3 groups (0-2, 2-5 and 5-10 years) based on time of detection of the disease, were tested in retrospect for presence of the JAK2 V617F mutant allele burden using an in-house developed Taqman probe-based kit, trend in shifting of the mutant allele burden was studied and segregated into ET (n=167) and PV (n=126) group based on their disease profile. It was then categorized into 3 time periods (0-2, 2-5 and 5-10 years) based on time of detection of the disease. Results: Around 293 (71%) were positive for JAK2 V617F while 59 (14.2%) and 8 (1.9%) positive for CALR exon 9 frame shift & MPL mutations (W515L/W515K) respectively. The 1-25% allelic burden group size gradually fell in ET population over time and this trend continued in the PV population also. In the former the fall was 7% & 11% for 2-5 & 5-10 years category while in the later, it was 1% and 15% respectively for the same time period-category. Conclusions: There is a distinct molecular continuum in the JAK2 V617F allelic burden in the ET & PV patients which followed a predictable trend and was associated with increasingly complicated vascular events.

In recent times, the demand for prescribed medicines has increased substantially. The Royal Pharm... more In recent times, the demand for prescribed medicines has increased substantially. The Royal Pharmaceutical Society of Great Britain (RPSGB) felt the burden and introduced supplementary prescribing in their system in the year 2003. The UK government authorities and other related bodies designed special courses which was required to be attended by pharmacists as well as nurses who intended to prescribe medicines. However, after the introduction of this legislation there was a need to assess whether the above legislation was beneficial or not. Limited studies exist that explore the current scenario of supplementary prescribing in the UK. This book is aimed at analyzing view of nurses and pharmacists on supplementary prescribing. Different sets of questionnaire were used to collect response from both pharmacists as well as nurses. Analysis of the data indicated unambiguous significance and increasing importance of the role of supplementary prescribing in England.

Cell free DNA (cfDNA) is now emerging as potent biomarker in cancer diagnostics as well as an use... more Cell free DNA (cfDNA) is now emerging as potent biomarker in cancer diagnostics as well as an useful tool for non-invasive diagnostic methods related to a wide range of clinical conditions. Its extraction from blood is crucial to exploitation as a biomarker. Link of cfDNA has been established with a wide range of health conditions including autoimmune diseases, sepsis and trauma, heart diseases and dialytic processes. However, its utility in prenatal diagnosis has been one of the most widely investigated areas of research. Association of cfDNA with cancer and its methylation status as a potent prognostic marker is also assuming increasing significance in recent times. Its association with different forms of cancer with special reference to breast cancer is notable. A doubtful mammographic findings does indicate other testing that involve imaging techniques but the final diagnosis is established by a biopsy in particular to differentiate malignant from benign tumors. Although this is...

The book discusses the medicinal properties of plants such as, Bacopa monnieri, Centella asiatica... more The book discusses the medicinal properties of plants such as, Bacopa monnieri, Centella asiatica, Rubia cordifolia, Hemidesmus indicus and Phyllanthus amarus. These plants have several therapeutic uses and are indigenous to India. They have immense value since ancient times and are still used in Ayurvedic preparations and hence in demand in the herbal market. The book describes in-vitro cultures which were established using various explants. It further discusses various analytic techniques such as Thin Layer Chromatography (TLC), High Performance Thin Layer Chromatography (HPTLC) & High Performance Liquid Chromatography (HPLC) which were exploited during the course of study of these plant extracts. Description of animal study techniques, acute toxicity on Swiss mice and efficacy studies of Phyllanthus amarus and Andrographis paniculata extracts on Albino Wistar rats with damaged liver as well as screening of hepatoprotective activity of pure phyllanthin are part of this book. The s...

International Journal of Agricultural Technology, 2014

Insufficient nutrient and pest infection are two major predicaments for low crop yield in agricul... more Insufficient nutrient and pest infection are two major predicaments for low crop yield in agriculture. Nematodes, a most infectious parasite pest in plants responsible for major yield loss. Chemical phosphate fertilizer made large reserves of it in soil, but part of it is made available to the plants because of irreversible fixation of it with divalent cation. In the present study, Arthrobotrys conoides (JX979095) and Duddingtonia flagrans (JX979096) a nematode trapping fungi were isolated from the arable soil of Anand district and studied for its phosphate solubilization potential. Among the different P sources, bioleaching of Tricalcium Phosphate (TCP) was found to be more efficient compared to other P salts. Phosphate solubilizing ability was enhanced in presence of D-glucose as compared to other C sources. In case of nitrogen source ammonium sulfate at 0.5 % showed best P solubilization followed by casein, urea and sodium nitrate respectively. Duddingtonia flagrans was showing m...

Possibility of reduction of perchlorate using microbes raises hope for an eco friendly mode of de... more Possibility of reduction of perchlorate using microbes raises hope for an eco friendly mode of degradation of this toxic rocket fuel. This study reports three isolates, viz., Al, A2 and A3 capable of molybdenum-independent degradation of perchlorate under aerobic conditions. The rate of degradation was found to be highest at substrate concentration of 17 mM of perchlorate, where isolate Al, A2 and A3 reduced 3.2mM, 4.7mM and 4.1 mM of perchlorate respectively under aerobic condition when incubated for 72 hours at 28 C. Presence of perchlorate at a concentration higher than 17mM exhibited inhibition in reduction of the compound. 16S ribosomal RNA gene analysis revealed isolate Al to be Pseudomonas stutzeri (Proteobacteria) while isolates A2 ad A3 where found to be different species of the genus AnhTobacter (Actinobacteria). The study apart from demonstrating ribotyping as a rapid method of identification of economically important soil microbes also raised prospect of using artificial...

A gram negative chemolithotrophic bacterium (RP1) with facultative mode of nutrition was isolated... more A gram negative chemolithotrophic bacterium (RP1) with facultative mode of nutrition was isolated from the soil. Enzymological studies confirmed presence of Thiosulphate oxidase, sulphite oxidase and Rhodanese, all of which play role in sulphur metabolism pathway. A set of degenerate oligonucleotide primer pairs was used for thermal amplification of a major part of the coding region of the Cytochrome c gene locus of this bacterium. Nucleotide and translated amino acid sequence revealed the gene to be a diheme Cytochrome c, different from the monoheme Cytochrome c observed in Chloribium limicola, a photosynthetic green sulphur bacterium. Significant homology at the nucleotide level could be detected only with Pseudoaminobacter salicylatoxidans. On the contrary, significant homology at the amino acid level was observed with Bradyrhizobium japonicum, Silicobacter pomeroyi apart from P. salicylatoxidans. This is possibly because of codon degeneracy observed within the diverse members of...

An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle.... more An insertion mutation within exon 12 of the factor XI gene has been described in Holstein cattle. This has opened the prospect for large-scale screening of cattle using the polymerase chain reaction (PCR) technique for the rapid identification of heterozygous animals. To facilitate such a screening process, the mutant and normal alleles of factor XI gene, represented by 244- and 320-bp PCR amplified fragments, were individually cloned in Escherichia coli using a multicopy plasmid cloning vehicle to generate pFXI-N and pFXI-M, respectively. The authenticity of the inserts was confirmed by nucleotide sequencing. A nested PCR method was developed, by which PCR amplicons generated from primers with annealing sites on the recombinant plasmids and by flanking the insert were used as templates for amplification of the diagnostic products using factor XI gene-specific primers. An equimolar mixture of both PCR amplicons, originating from pFXI-N and pFXI-M, constituted the carrier control whi...