Desvio para a esquerda associado a compartimento medular de reserva dos neutrófilos não esgotado em cadelas portadoras de piometra / Nuclear left deviation associated with medullary reserve compartment of non exhausted neutrophils in female dogs with pyometra

From fifteen female dogs with clinical diagnoses of pyometra, hematological exams were performed at three times: M0 (prior to the surgery), M24 (24h after ovarysalpingohisterectomy-OSH) and M48 (48h after OSH). Anemia was seen in 80% of the cases, characterized as mild normocytic normochromic type. The means of total leukocyte counts were 27.043, 57.940 and 40.139 céls/µL in M0, M24 and M48. A total of80% of the animals presented neutrophilic left shift in all moments. During medullar exams, the cellular, iron reserve and megakaryocytic concentration were raised as well as the ME ratio, showing a value of 26,3:1,0, probably due to the elevation of granular proliferation and maturation compartment, as the mean of the reserve compartment was within normal range. As 83% of the animals with neutrophilic left shift showed a melullary reserve compartment raised, it can be concluded that female dogs with piometra had left shift of neutrophils with disproportionally between compartments, without segmented medullar saturation.(AU)

Behavior of mesenchymal stem cells stained with 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) in osteogenic and non osteogenic cultures

4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) is a DNA dye widely used to mark and trace stem cells in therapy. We here studied the effect of DAPI staining on the behavior of mesenchymal stem cells cultured in either a control, non-osteogenic medium or in an osteogenic differentiation medium. In the control medium, the number of stem cells/field, as well as the number of fluorescent cells/field increased up to the sixth day in both control and DAPI-treated cultures. Afterwards, both the number of fluorescent cells and their fluorescence intensity decreased. Control cells were fusiform and with some long extensions that apparently linked them to neighboring cells, while DAPI-treated cells were mostly round cells with fine and short extensions. The trypan-blue exclusion method showed 99% cell viability in both groups, however, both alkaline phosphatase activity and the thiazolyl blue formazan assay (indicative of mitochondrial metabolism) gave significantly lower values in DAPI-marked cells The mitochondrial mass, as indicated by specific staining and flow cytometry, showed no differences between groups. Mesenchymal stem cells gave origin to mineralized nodules in the osteogenic differentiation medium and there were not DAPI-marked cells on the ninth day of culture. Alkaline phosphatase activity, viability assay and number of cells/field and of mineralized nodules/field were similar in both groups. So, DAPI treatment did not change cell viability and proliferation during osteogenic differentiation of mesenchymal stem cells. However, since these cells loose DAPI marking after 9 days in osteogenic cultures suggests that DAPI may not be an effective marker for mesenchymal stem cells implanted in bone tissue for long periods.