A study on IL8RB gene polymorphism as a potential immuno-compromised adherent in exaggeration of parenteral and mammo-crine oxidative stress during mastitis in buffalo.

The genetic markers in inflammatory responses during mastitis afford a reasonable way for improving milk production in the Egyptian buffalo breed. Among them is the interleukin 8 Receptor Gene (IL8RB) (CXCR2); a chemokine receptor gene augments the neutrophil migration during infection. To understand its role better during mastitis in Egyptian buffalos, twenty-five dairy animals representing the normal, sub-clinically, clinically and chronically affected buffalos were randomly selected from different districts. Screening criteria for mastitis were based on somatic cell count and California mastitis test assays on their milk samples. Biochemically, mastitis induced an increase in milk lactate dehydrogenase, alkaline phosphatase and catalase activities and serum malanoaldehyde concentration. The total antioxidant concentrations, however, decreased in serum and milk during mammary inflammation. The protein profiling of milk whey proved an accelerated mammary inflammatory influx of blood-borne proteins during mastitis. The genomic DNAs were extracted from blood samples and the CXCR2 sequence of 1246 bp covering a part of intron 1, exon 2 and a part of 3'UTR were submitted to Genbank (accession # KY399457.1). The study clearly defined the presence of four SNPs. Three were detected as synonymous substitutions in coding region and one in the 3'UTR region. Only SNP C/A at c.127 was found to be highly associated with mastitis. In conclusion, the results warrant the potential correlation between the genetic SNP variance for certain genes and the incidence of mastitis in buffalo breed.

Current status of the river buffalo (Bubalus bubalis L.) gene map.

Ninety-nine loci have been assigned to river buffalo chromosomes, 67 of which are coding genes and 32 of which are anonymous DNA segments (microsatellites). Sixty-seven assignments were based on cosegregation of cellular markers in somatic cell hybrids (synteny), whereas 39 were based on in situ hybridization of fixed metaphase chromosomes with labeled DNA probes. Seven loci were assigned by both methods. Of the 67 assignments in somatic cell hybrids, 38 were based on polymerase chain reaction (PCR), 11 on isozyme electrophoresis, 10 on restriction endonuclease digestion of DNA, 4 on immunofluorescence, and 4 on chromosomal identification. A genetic marker or syntenic group has been assigned to each arm of the five submetacentric buffalo chromosomes as well as to the 19 acrocentric autosomes, and the X and Y chromosomes. These same markers map to the 29 cattle autosomes and the X and Y chromosomes, and without exception, cattle markers map to the buffalo chromosome or chromosomal region predicted from chromosome banding similarity.

Specific numerical chromosomal aberrations induced by adriamycin.

Fluorescence in situ hybridization with 7, 17, X, and Y chromosome-specific DNA probe was used to investigate the ability of Adriamycin (AM) to induce aneuploidy in interphase human lymphocytes. The reliability of the probes was tested by hybridization to metaphases and interphase nuclei of untreated normal lymphocytes. Two signals were scored in over 87% of the analyzed nuclei with chromosome 7 and 17 probes, whereas one signal was recorded in over 86% of the nuclei with chromosomes X and Y. The same conditions and probe concentrations were used for hybridizing the four probes to interphase nuclei of AM-treated and untreated lymphocytes, cultured from healthy individuals and cancer patients. AM was found to induce significant increases of trisomy 7 and 17 in lymphocytes cultured from healthy individuals and cancer patients, where the interphase nuclei showed three spots in over 70% and 72% of the cells, respectively. Only 6% of interphase nuclei of untreated cells cultured from healthy individuals and cancer patients showed three spots. No significant increase in X or Y aneuploidy was induced by exposure to AM.

Mutagenicity of cidial (phenthoate). I: Effect on maternal and fetal somatic cells.

Cidial, an organophosphorous insecticide (also known as phenthoate), was tested for its genotoxic effect on both maternal and fetal cells. Cidial was administered at three different dose levels (53.5, 106.9, and 171 mg/kg) to pregnant mice on day 16 of gestation. Maternal bone marrow and embryonic liver cells were examined for chromosomal aberrations and cellular proliferation. Cidial was found to increase the percentage of cells with chromosomal aberrations in both mothers and fetuses. It also significantly inhibited the rate of mitotic activity of both maternal and fetal cells, with the inhibitory effect being more appreciable in fetal cells than in maternal cells. The data indicate that cidial, which is widely used in rural areas, is hazardous to both mothers and their transplacentally exposed babies.

Synteny mapping in river buffalo.

The cosegregation of ten coding loci has been investigated, in a panel of 37 somatic cell hybrids resulting from the fusion of a hamster cell line and river buffalo lymphocytes, by use of Southern hybridization technique. Five syntenic groups, TCRB-PGY3, ASS-ABL, FUCA1P-CRYG, MBP-YES1, and CGN1-ACTA1, previously assigned to cattle as U13, U16, U17, U28, and U29 respectively, were also found to be syntenic in buffalo. Based on the extensive syntenic conservation and banding homology between cattle and river buffalo, comparative mapping predicts the localization of these syntenic groups on river buffalo Chromosomes (Chrs) :BBU7, BBU12, BBU2q, BBU22, and BBU4q respectively as they have been previously localized on cattle Chrs BTA4, BTA11, BTA2, BTA24 & BTA28.

Assignment of genes coding for leukocyte surface molecules to river buffalo chromosomes.

A panel of 35 somatic cell hybrids between river buffalo (Bubalus bubalis L.) and Chinese hamster was used in combination with 40 monoclonal antibodies to find synteny among leukocyte antigens and between already established synteny groups and leukocyte antigens. Six synteny groups were identified and three sets of markers were related to already existing synteny groups. Antibody IL-A165 detected CD71, which is syntenic with synteny groups U10 and U25, confirming that U10 and U25 are both are on a biarmed river buffalo chromosome. This would imply that U25 could be assigned to the short arm of the river buffalo chromosome 1 (BBU 1p), as U10 has previously been assigned to this chromosome. Antibodies Buf 32, Buf 24 and BAGB27 detected an antigen syntenic with U1 and U7. This finding suggests that U1, which is assigned to cattle chromosome 16, which corresponds to river buffalo chromosome 5q, is syntenic with U7, and that U7 is located on the short arm of buffalo chromosome 5. Furthermore, the gene coding for CD14 is assigned to U16, which is on cattle chromosome 11 and expected to be on river buffalo chromosome 12.

Radioprotective effect of vitamins C and E.

Albino rats were treated with aqueous vitamin C solution and vitamin E solution dissolved in olive oil at two concentrations, 100 and 300 mg/kg/day, for 6 months. Some of the animals were then subjected to whole-body irradiation. Chromosomal aberrations and mitotic activity in non-irradiated and irradiated groups were recorded. Both vitamins were found to be non-mutagenic. Vitamin C exerted a radioprotective effect but vitamin E was not radioprotective and it suppressed the radioprotection otherwise produced by olive oil.

Chromosome aberrations in spermatogonia and sperm abnormalities in Curacron-treated mice.

Curacron is an organophosphorus pesticide widely used in cotton fields. In order to assay its mutagenic potential in mammalian germ cells chromosomal aberrations in spermatogonial cells and sperm abnormalities were examined in mice after Curacron treatment. For studying chromosomal aberrations mice were treated both acutely (single treatment) and subacutely (for 5 consecutive days) with 3 dose levels of Curacron, 12, 36 and 72 mg/kg. Curacron was found to produce a significant increase in structural chromosomal aberrations after acute and subacute treatments. This increase was dose-dependent. A dose-dependent inhibition in mitotic activity in spermatogonia was also found. For studying sperm abnormalities mice were treated for 5 consecutive days with 20, 40 and 60 mg/kg. Morphological sperm abnormalities increased significantly after treatment with Curacron. The increase was dose-dependent. An inhibition of 40.2% in sperm count and of 74.5% in sperm motility occurred after treatment with 60 mg/kg Curacron. These results show that Curacron has a damaging effect on spermatogonial cells as well as on sperm morphology.

Micronuclei formation in somatic cells of mice exposed to 50-Hz electric fields.

Male Swiss mice are tested under uniform 50-Hz electric field intensities of 100, 170, 220, and 290 kV/m. These values on the basis of equal induced current density are equivalent to the case of a human exposed to field intensities of nearly 8, 14, 18, and 24 kV/m, respectively. The latter values may be found beneath or in the vicinity of extra-high-voltage power lines whose voltages range from 220 to 765 kV. The cytogenetic effect of extremely low-frequency (ELF) electric fields, as expressed by micronuclei formation, is assessed. Mice are exposed for 24 hr, and samples are taken 48, 72, and 96 hr from the beginning of exposure. Sham-exposed mice served as controls. The number of micronucleated polychromatic erythrocytes (PCE) in exposed animals are significantly higher than those of the control. The increase in micronucleated PCE was significantly dose dependent at all times. Samples taken 96 hr after exposure show a decrease in percentages of micronucleated PCE, which may be taken as an indication of recovery.

In vivo evaluation of the genotoxic potential of curacron in somatic cells of mice.

Cytogenetic effects produced in somatic cells by Curacron, a phosphoric acid ester insecticide, were assessed in mice by three criteria: chromosome aberrations, sister chromatid exchange (SCE), and micronucleus induction. Curacron significantly increased the frequency of all three in an apparent dose-dependent manner. Curacron also inhibited the mitotic activity. The increased number of micronuclei is comparable to the increase in chromosome aberrations; the latter were mainly breaks. The frequency of SCE was considerably less than the frequencies of micronuclei and chromosome aberrations.

Chromosomal aberrations induced by sodium nitrite in bone marrow of adult rats and liver cells of transplacentally exposed embryos.

A commonly used food preservative, sodium nitrite, was administered in the drinking water to pregnant (d 5-18 of gestation) and nonpregnant albino rats. Sodium nitrite induced chromosomal aberrations in bone marrow of both pregnant and nonpregnant adults and liver of transplacently exposed embryos. The magnitude of the effect was greater in embryonic liver cells than in adult bone marrow cells.