Evaluation of candidate genotype of immune gene MBL1 associated with udder health and performance traits in dairy cattle and buffalo of India.

An investigation was conducted to identify polymorphism in mannose-binding lectin 1 (MBL1) gene and its effect on udder health and performance traits in dairy cattle and buffalo of India. Candidate single-nucleotide polymorphism (SNP) c.2534G > A of MBL1 gene was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). All the possible genotypes for SNP c.2534G > A (GG, AG, and AA) were observed in the studied population. However, Sahiwal cows revealed dimorphic pattern (AG and GG). The effect of targeted SNP on incidence of mastitis was evaluated and found to be significant. Animals with GG genotype were less susceptible to clinical mastitis and had comparatively lower somatic cell score (SCS) in Hardhenu cattle (P < 0.01) and Murrah buffalo (P < 0.05). Animals having GG genotype also exhibited significantly (P < 0.05) lower age at first calving (AFC). AG genotyped Murrah buffalo animals revealed significantly higher second lactation milk yield (P < 0.01). GG genotype with SCS and AFC could therefore be exploited as a promising candidate marker for the genetic improvement of udder health and AFC in dairy animals.

Evaluation of genetic and non-genetic factors on foot and mouth disease (FMD) virus vaccine-elicited immune response in Hardhenu (Bos taurus x Bos indicus) cattle.

Foot and mouth disease (FMD) is the most contagious disease of mammals and a major threat to animal husbandry sector. In India, vaccination with the inactivated trivalent (O, A and Asia1) vaccine is one proven way for protecting the livestock from FMD. However, many outbreaks have been reported in different parts of the country. Therefore, present study was aimed at elucidating the effects of genetic and non-genetic factors on FMD viral vaccine-elicited immune response in Hardhenu cattle. The effect of season of vaccination was not consistent. The effect of status of animal was significant for all the pre and post AB titres except for pre AB titre of serotype O and post AB titre of Asia1.The estimates of heritability for response to vaccination were low to high ranging from 0.11 to 0.45. The highest heritability estimate was obtained for serotype O and the lowest for Asia1. The heritability estimates for pre and post AB titres ranged from 0.15 to 0.33. All the pre and post AB titres and responses to vaccination had genetic correlations ranged from high negative to high positive among them. Results of this study highlight the variation in vaccine response which needs to be further exploited on large-scale animal data for better immunization and protection against highly contagious viral vesicular disease of cloven-hoofed animals.

Antigen levels of the urokinase-type plasminogen activator and its gene polymorphisms in colorectal cancer.

We analysed the distribution of genotypes of two polymorphisms in the urokinase-type plasminogen activator (uPA) gene: C-->T substitution in exon 6 and T-->C substitution in intron 7 in 52 subjects with colorectal cancer. Genotypes were determined in tumour tissue and distant mucosa samples by allele-specific polymerase chain reaction. The antigen levels of uPA in cancer tissue were higher than in distant mucosa as measured by enzyme-linked immunosorbent assay. The level of uPA antigens in cancer samples with the C/C genotype of C-->T polymorphism in exon 6 was higher than in samples with C/T and T/T genotypes. No differences in the level of uPA antigens between the alleles of the intron 7 T-->C polymorphism were found. As uPA can be involved in cancer invasion and metastasis, C/C genotype in exon 6 of uPA gene can be further considered as being related to colorectal cancer progression.

Plasminogen activator inhibitor 1 (PAI-1) levels and gene promoter polymorphisms in subjects with colorectal cancer.

The high level of plasminogen activator inhibitor 1 (PAI-1) in colorectal cancer predicts poor prognosis for patients. The insertion (5G)/deletion (4G) polymorphism (the 4G/5G polymorphism) and G-->A single base substitution (the G/A polymorphism) located at promoter of PAI-1 gene may have functional significance in regulation of its expression. In the present work the level of PAI-1, distribution of genotypes and frequency of alleles of the 4G/5G and G/A polymorphisms in samples of cancer tissue and normal mucosa as well as in blood were investigated. Blood, tumor and normal tissues were obtained from 40 patients with colorectal cancer. The 4G/5G and G/A polymorphism were determined by PCR amplification using the allele specific primers. The PAI-1 level was measured by enzyme linked immunosorbent assay (ELISA). The distribution of the genotypes of both polymorphisms did not differ significantly (p > 0.05) from those predicted by the Hardy-Weinberg distribution. There were no differences in the genotype distributions and allele frequencies between blood, normal mucosa samples and cancer tissue. The 4G/5G and G/A polymorphisms were in linkage disequilibrium. The average level of PAI-1 in tumor samples was significantly (p < 0.05) higher than in normal tissue. The results obtained indicate that a higher level of PAI-1 can be associated with colorectal cancer. On the other hand, in colon cancer, the 4G/5G and G/A polymorphisms are not linked with elevated levels of PAI-1 and therefore may not be used to predict colon cancer prognosis.

A C/T polymorphism in the urokinase-type plasminogen activator gene in colorectal cancer.

Urokinase plasminogen activation system can play an important role in the appearance and progression of many cancers. Urokinase-type plasminogen activator (uPA) is implicated in the control of cell adhesion and invasion, and is regarded as a strong prognostic marker in colorectal cancer. A C-->T substitution (the C/T polymorphism) in the nucleotide sequence encoding the kringle structure of uPA results in an alteration from proline to leucine at position 121. This substitution may be directly or indirectly involved in the decreased affinity for uPA substrates. In the present work the distribution of genotypes and frequencies of alleles of the C/T polymorphism were investigated. Tumour tissues and distal mucosa samples were obtained from 40 patients with colorectal cancer. Blood samples from sex and age matched healthy individuals served as control. The C/T polymorphism was determined by PCR amplification using the allele specific primers. No differences between genotypes of the C/T polymorphism in cancer tissue and distant mucosa of each patient were found. The distributions of the genotypes in both patients and control differed significantly (p < 0.05) from that predicted by the Hardy-Weinberg distribution. A distinct preference of heterozygotes (70% - patients, 65% - controls) was observed in both patients and controls. Additionally, there were no differences in the frequencies of the C and T alleles in both groups. The C/T polymorphism of the uPA gene may not be linked with colorectal cancer.

Synergistic effect of vitamin C on DNA damage induced by cadmium.

Salts of divalent cadmium are well-known human mutagens and carcinogens. In the present work, the ability of vitamin C to modulate genotoxic effects of cadmium chloride on human lymphocytes was assessed using single cell gel electrophoresis (comet assay). Vitamin C at 20 and 100 micromol/l and cadmium at 5, 30 and 150 micromol/l significantly increased the tail moment of lymphocytes. Vitamin C also increased the tail moment of cells exposed to cadmium. This effect was concentration-dependent: the higher the vitamin C concentration the greater the tail moment. The combined effects of cadmium and vitamin C were more pronounced at all concentrations tested than the sum of the effects of the compounds applied separately (p < 0.05), so cadmium and vitamin C can be considered to have synergistic effects. The results obtained can be partly explained by the participation of cadmium in the Fenton reaction and reduction of its oxidized form by vitamin C.