Procena in vitro aktivacije proizvodnje azot oksida kao odgovor bovinih epitelnih ćelija endometrijuma i vagine i mononuklearnih krvnih ćelija na Mycoplasma bovis, Mycoplasma bovigenitalium i Ureaplasma diversum

Genital mycoplasmosis is a condition present in bovine production systems, and the most important agents involved are Mycoplasma bovis, Mycoplasma bovigenitalium and Ureaplasma diversum. Some aspects of their pathogenesis remain unclear. This study was designed in order to evaluate their ability to stimulate mononuclear cells from the endometrium, vagina and peripheral blood of cycling and healthy cows to produce nitric oxide (NO). Cellular cultures of endometrial, vaginal and peripheral blood cells from 33 healthy cows were cultivated with Mycoplasma bovis, Mycoplasma bovigenitalium and Ureaplasma diversum originated from the 4th passage in culture broth and the NO production was measured by the Greiss reaction. Confi rmation of the presence of mononuclear cells and of the agents during and after the NO assay was done by Giemsa stained smears and further cultivation and detection by PCR reaction. Mononuclear cells from all samples produced NO. Mycoplasma bovigenitalium stimulated higher NO production than the others (p<0.05). Endometrial cells produced less NO than vaginal or blood cultured cells. In conclusion, it seems that Mycoplasma bovis, Mycoplasma bovigenitalium, and Ureaplasma diversum are able to activate mononuclear cells and induce the production of NO, thus suggesting that this pathway is elicited in response to the primary infection by these agents. More studies are necessary to verify why these agents remain in the bovine reproductive tract for long periods and how they reassume deleterious effects.Genitalna mikoplazmoza je stanje koje može biti prisutno u reproduktivnom sistemu goveda, a najznačajniji agensi koji je izazivaju su Mycoplasma bovis, Mycoplasma bovigenitalium i Ureaplasma diversum. Neki od aspekata patogeneze nisu još uvek u potpunosti razjašnjeni. Ova studija je kreirana kako bi se procenila njihova sposobnost stimulisanja mononuklearnih ć elija iz endometrijuma, vagine i periferne krvi krava u ciklusu i zdravih krava da proizvode azotni oksid (NO). Ć elijske kulture ć elija endometrijuma, vagine i periferne krvi 33 zdrave krave uzgajane su sa Mycoplasma bovis, Mycoplasma bovigenitalium i Ureaplasma diversum proisteklim iz 4 pasaže u bujonu, a proizvodnja NO merena je Greiss reakcijom. Potvrda prisustva mononuklearnih ć elija, kao i infektivnih agenasa tokom i posle određivanja NO, urađena je Giemsa bojenjem na razmazima, kao i daljom kultivacijom i detekcijom PCR reakcijom. Mononuklearne ć elije iz svih uzoraka proizvodile su NO. Mycoplasma bovigenitalium stimulisala je već u proizvodnju NO od ostalih (

Increased Levels of NOTCH1, NF-kappa B, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

As previously shown, higher levels of NOTCH1 and increased NF-kappa B signaling is a distinctive feature of the more primitive umbilical cord blood (UCB) CD34+ hematopoietic stem cells (HSCs), as compared to bone marrow ( BM). Differences between BM and UCB cell composition also account for this finding. The CD133 marker defines a more primitive cell subset among CD34+ HSC with a proposed hemangioblast potential. To further evaluate the molecular basis related to the more primitive characteristics of UCB and CD133+ HSC, immunomagnetically purified human CD34+ and CD133+ cells from BM and UCB were used on gene expression microarrays studies. UCB CD34+ cells contained a significantly higher proportion of CD133+ cells than BM (70% and 40%, respectively). Cluster analysis showed that BM CD133+ cells grouped with the UCB cells ( CD133+ and CD34+) rather than to BM CD34+ cells. Compared with CD34+ cells, CD133+ had a higher expression of many transcription factors (TFs). Promoter analysis on all these TF genes revealed a significantly higher frequency ( than expected by chance) of NF-kappa B-binding sites (BS), including potentially novel NF-kappa B targets such as RUNX1, GATA3, and USF1. Selected transcripts of TF related to primitive hematopoiesis and self-renewal, such as RUNX1, GATA3, USF1, TAL1, HOXA9, HOXB4, NOTCH1, RELB, and NFKB2 were evaluated by real-time PCR and were all significantly positively correlated. Taken together, our data indicate the existence of an interconnected transcriptional network characterized by higher levels of NOTCH1, NF-kappa B, and other important TFs on more primitive HSC sets.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Financiadora de Estudos e Projetos (FINEP), Brazi