Molecular Characterization of Rathi and Tharparkar Indigenous Cattle (Bos indicus) Breeds by RAPD-PCR

Sharma, Amit Kumar,Bhushan, Bharat,Kumar, Sanjeev,Kumar, Pushpendra,Sharma, Arjava,Kumar, Satish Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.9

Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

Magnetic Properties and Electronic Structure of Pt₃Ni (001), (110) and (111) Surfaces

Sharma Bharat Kumar,Kwon Oryong,Dorj Odkhuu,Hong Soon Cheol 한국자기학회 2011 한국자기학회 학술연구발표회 논문개요집 Vol.2011 No.6

Magnetism during adsorption of oxygen in Pt segregated Pt<SUB>3</SUB>Ni (111)

Sharma Bharat Kumar,Kwon Oryong,Dorj Odkhuu,Hong Soon Cheol 한국자기학회 2011 한국자기학회 학술연구발표회 논문개요집 Vol.2011 No.12

Electronic structure and magnetism of catalytic material Pt<SUB>3</SUB>Ni surfaces

Bharat Kumar Sharma,Oryong Kwon,Dorj Odkhuu,Soon Cheol Hong 한국자기학회 2012 한국자기학회 학술연구발표회 논문개요집 Vol.2012 No.11

Elevated N1-Acetylspermidine Levels in Doxorubicintreated MCF-7 Cancer Cells: Histone Deacetylase 10 Inhibition with an N1-Acetylspermidine Mimetic

Ajay Kumar Raj,Kiran Bharat Lokhande,Kratika Khunteta,Sachin Chakradhar Sarode,Nilesh Kumar Sharma 대한암예방학회 2024 Journal of cancer prevention Vol.29 No.2

Cancer drug resistance is associated with metabolic adaptation. Cancer cells have been shown to implicate acetylated polyamines in adaptations during cell death. However, exploring the mimetic of acetylated polyamines as a potential anticancer drug is lacking. We performed intracellular metabolite profiling of human breast cancer MCF-7 cells treated with doxorubicin (DOX), a well known anticancer drug. A novel and in-house vertical tube gel electrophoresis assisted procedure followed by LC-HRMS analysis was employed to detect acetylated polyamines such as N1-acetylspermidine. We designed a mimetic N1-acetylspermidine (MINAS) which is a known substrate of histone deacetylase 10 (HDAC10). Molecular docking and molecular dynamics (MDs) simulations were used to evaluate the inhibitory potential of MINAS against HDAC10. The inhibitory potential and the ADMET profile of MINAS were compared to a known HDAC10 inhibitor Tubastatin A. N1-acetylspermidine, an acetylated form of polyamine, was detected intracellularly in MCF-7 cells treated with DOX over DMSO-treated MCF-7 cells. We designed and curated MINAS (PubChem CID 162679241). Molecular docking and MD simulations suggested the strong and comparable inhibitory potential of MINAS (–8.2 kcal/ mol) to Tubastatin A (–8.4 kcal/mol). MINAS and Tubastatin A share similar binding sites on HDAC10, including Ser138, Ser140, Tyr183, and Cys184. Additionally, MINAS has a better ADMET profile compared to Tubastatin A, with a high MRTD value and lower toxicity. In conclusion, the data show that N1-acetylspermidine levels rise during DOX-induced breast cancer cell death. Additionally, MINAS, an N1-acetylspermidine mimetic compound, could be investigated as a potential anticancer drug when combined with chemotherapy like DOX.

Effect of Leptin and IGFBP-3 Gene Polymorphisms on Serum IgG Level of Cattle Calves

Choudhary, Vivek,Kumar, Pushpendra,Saxena, V.K.,Bhattacharya, T.K.,Bhushan, Bharat,Sharma, Arjava,Ahmed, K.A. Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.8

Leptin and IGFBP-3 are two proteins that play an important role in growth and metabolism of the animals. They are also involved in the immune function of animals and, thus, are candidate genes for the study of association with immune functions. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of these two genes was done to screen 64 crossbred (Holstein Friesian${\times}$Hariana) female calves of one year of age. From each RFLPs (fragments) three genotypes were observed. In all the RFLPs the mutant homozygotes were very less in numbers and, hence, were excluded from the least squares analysis. The serum IgG level was estimated using SRID assay. The mean level of serum IgG was $28.83{\pm}2.73mg/ml$. The effect of these identified genotypes on serum IgG level of calves at one year of age was analysed using least squares analysis. The HaeIII RFLP-AB genotype had significantly (p<0.05) higher serum IgG level ($31.86{\pm}3.05$) than the HaeIII RFLP-AA ($25.62{\pm}2.96$) genotype. There was no significant effect of leptin genotypes on the IgG level. The present results indicated a role of the IGFBP-3 gene on serum IgG level of cattle calves.

Characterization of MHC DRB3.2 Alleles of Crossbred Cattle by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

Paswan, Chandan,Bhushan, Bharat,Patra, B.N.,Kumar, Pushpendra,Sharma, Arjava,Dandapat, S.,Tomar, A.K.S.,Dutt, Triveni Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.9

The present investigation was undertaken to study the genetic polymorphism of the DRB3 exon 2 in 75 crossbred cattle by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Five genotypes i.e. HaeIII-a, HaeIII-b, HaeIII-e, HaeIII-ab and HaeIII-ae were observed when the 284 bp PCR products were digested with HaeIII restriction enzyme. The corresponding frequencies of these patterns were 0.53, 0.04, 0.01, 0.38 and 0.04, respectively. Digestion with RsaI restriction enzyme resolved 24 different restriction patterns. The frequencies of these patterns ranged from 0.013 (RsaI-f, RsaI-k and RsaI-c/n) to 0.120 (RsaI-n). The results revealed that the crossbred cows belonged to the RsaI patterns namely b, k, l, a/l, d/s, l/n, l/o and m/n, whose corresponding frequencies were 0.027, 0.013, 0.040, 0.027, 0.040, 0.067, 0.027 and 0.067, respectively. Digestion of the 284 bp PCR product of DRB3.2 gene with PstI in the crossbred cattle did not reveal any restriction site. These results suggested the absence of the recognition site in some of the animals. These results also revealed that the crossbred cows studied were in homozygous as well as heterozygous condition. On the basis of the above results it can be concluded that the DRB3.2 gene was found to be highly polymorphic in the crossbred cattle population.

PCR-SSCP of Serum Lysozyme Gene (Exon-III) in Riverine Buffalo and Its Association with Lysozyme Activity and Somatic Cell Count

Sahoo, Nihar Ranjan,Kumar, Pushpendra,Bhushan, Bharat,Bhattacharya, T.K.,Sharma, Arjava,Dayal, Sanker,Pankaj, Prabhat Kumar,Sahoo, Monalisa Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.8

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

Genetic Identity between Bhadawari and Murrah Breeds of Indian Buffaloes (Bubalus bubalis) Using RAPD-PCR

Saifi, H.W.,Bhushan, Bharat,Kumar, Sanjeev,Kumar, Pushpendra,Patra, B.N.,Sharma, Arjava Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.5

Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out with a battery of 11 random decamer primers to study band frequency (BF), genetic identity index (I) and mean average percentage difference (MAPD) between Bhadawari and Murrah breeds of buffalo. The primers OPA04 and BG15 resolved a band of 460 bp, which was present only in animals of Bhadawari breed. Whereas, the primers OPA14, BG27 and BG28 produced Murrah specific fragments of sizes 730 bp and 1,230 bp, respectively. The estimate of genetic identity index was highest (0.845) with the primer OPA01 and the lowest (0.479) with the primer BG27. The genetic identity index pooled over the primers was 0.596${\pm}$0.037 between these two breeds. The highest MAPD estimate (53.9) between the two breeds was obtained with the primer BG27 and the lowest (14.3) with the primer OPA01. It might be concluded that the genetic identity index between these two breeds calculated on the basis of BF showed moderate level of genetic identity with the primers employed. MAPD calculated on the basis of uncommon bands also demonstrated lower to medium level of genetic difference between Bhadawari and Murrah breeds of buffalo.

Pattern of Liver Enzymes in Alcohol Dependence Syndrome Patients

( Mithileshwer Raut ),( Binod Kumar Yadav ),( Vijay Kumar Sharma ),( Eans Tara Tuladhar ),( Aseem Bhattarai ),( Bharat Jha ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Alcohol dependence syndrome (ADS) has become a global public health challenge because of its high prevalence and the concomitant increase in risk of liver disease, cardiovascular disease and premature death. Influence of alcohol use on liver metabolism is well recognized. This study was aimed at examining the association of liver enzymes like γ-glutamyltransferase (GGT) and aminotransferase, with alcohol dependence syndrome patients. Methods: This cross-sectional study was conducted in Tribhuvan University Teaching Hospital. ADS patients were screened by the consultant psychiatrist using the Alcohol Use Disorder Identification Test (AUDIT) questionnaire. A total of 89 patients scored positive on the AUDIT as having alcohol-related problems and were included in the study. Blood Pressure and other anthropometric parameters were measured while blood samples were analyzed for liver enzymes and serum protein. Results: Mean age of cases and controls was 35.42 ± 5.6 & 34.53 ± 3.5 years respectively. The mean values of Gamma GT, SGOT and SGPT were largely elevated in cases as compared to the controls with a statically significance (P<0.001). Among the ADS cases serum GGT level was elevated in 97% patients. The SGOT/SGPT ratio was also significantly higher in cases (2.02 ± 0.39) and control (1.45±0.62). It was found that 15.1 % cases had low serum protein level and 32.9% cases were low serum albumin level. Albumin to globulin ratio was also significantly decreased in cases (1.16 ±0.29). Conclusions: These findings support the hypothesis that, alcohol may affect the pattern of liver enzymes and also damage the liver cells. Decrease in serum albumin and elevation of SGOT to SGPT ratio more than two is suggestive of development of liver cirrhosis in alcohol dependence patients.