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In this investigation, the role of autophagy and apoptosis in Solanum nigrum fruit extract-induced cytotoxicity in MDA-MB-468, a triple negative breast cancer cell line was studied. Breast cancer cells were treated with varying concentrations (1 to 5 mg/ml) of Solanum nigrum extract for 24 and 48 h. Autophagy was detected using acridine orange staining and real-time PCR for Lc3 and Beclin1 genes expression. In addition, cell apoptosis was measured by double staining, DNA fragmentation and annexin V-fluorescein isothiocyanate binding assays. The expression levels of apoptosis-related genes Bcl2 and Bax were determined by real-time PCR. Different concentrations of Solanum nigrum extract showed cytotoxic effect on MDA-MB-468 cells. The expression levels of autophagy-related genes, Lc3 and Beclin1 increased in cells treated with 1.5 mg/ml of Solanum nigrum extract. Upregulation of Bax and downregulation of Bcl2 was detected after treatment with 5 mg/ml of Solanum nigrum extract. These findings indicated that Solanum nigrum extract induced cell death in MDA-MB-468 cells by two distinct mechanisms, apoptosis and autophagy, and these findings further suggest a possibility that this extract could be used to treat triple negative breast cancer.

A cattle genetic linkage map was constructed which covers more than 95 percent of the bovine genome at medium density. Seven hundred and forty six DNA polymorphisms were genotyped in cattle families which comprise 347 individuals in full sibling pedigrees. Seven hundred and three of the loci are linked to at least one other locus. All linkage groups are assigned to chromosomes, and all are orientated with regards to the centromere. There is little overall difference in the lengths of the bull and cow linkage maps although there are individual differences between maps of chromosomes. One hundred and sixty polymorphisms are in or near genes, and the resultant genome-wide comparative analyses indicate that while there is greater conservation of synteny between cattle and humans compared with mice, the conservation of gene order between cattle and humans is much less than would be expected from the conservation of synteny. This map provides a basis for high-resolution mapping of the bovine genome with physical resources such as Yeast and Bacterial Artificial Chromosomes as well as providing the underpinning for the interpolation of information from the Human Genome Project.