RESUMO
The immunosuppressor effects of the widely distributed neuropeptide somatostatin were examined on purified peripheral blood human monocytes. Somatostatin, at concentrations thought to be physiologic (10(-10)-10(-7) M), regulated monocyte/macrophage responses to (LPS) stimulation, as reflected by interleukin production. In particular, somatostatin had direct inhibitory effects on TNF-alpha, IL-1 beta, and IL-6 secretion by LPS-activated monocytes, while the decrease on IL-8 synthesis was modulated mainly by the action of somatostatin on TNF-alpha and IL-1 beta. In fact, the addition of these two inflammatory cytokines to the monocyte culture medium was able to induce IL-8 expression, as demonstrated by mRNA analysis, also in presence of the neuropeptide. Although somatostatin affected IL-8 production in an indirect way, it suppressed directly the chemotactic response of neutrophils to IL-8. Finally, somatostatin downregulation of monocyte activation was confirmed by the decrease of HLA-DR expression on cell plasma membranes (52% versus 33%). Our results confirm that somatostatin exerts preferential effects on the suppression of immunoreactions by modulating cytokine production and activity.
Assuntos
Citocinas/biossíntese , Regulação da Expressão Gênica/efeitos dos fármacos , Leucócitos Mononucleares/efeitos dos fármacos , Somatostatina/farmacologia , Células Cultivadas , Antígenos HLA-DR/biossíntese , Humanos , Terapia de Imunossupressão , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Interleucina-8/biossíntese , Leucócitos Mononucleares/imunologia , Lipopolissacarídeos/farmacologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Distraction osteogenesis by the method of Ilizarov has permitted the study of bone formation. This is an analysis of 64 human biopsies in patients who were undergoing tibial lengthening by the method of Ilizarov. After analysis by conventional and polarized light microscopy, four stages of bone formation were identified. Bone formation is of the direct type with no cartilaginous phase.
Assuntos
Matriz Extracelular/patologia , Fixação de Fratura , Osteogênese , Biópsia , Alongamento Ósseo , Matriz Extracelular/fisiologia , Feminino , Humanos , Masculino , Microscopia de Polarização , Tíbia/patologia , Tíbia/fisiologia , Tíbia/cirurgiaRESUMO
A procedure is described for preparing and maintaining in culture isolated lobules of bovine seminal vesicles, consisting of glandular acini, surrounded by little connective tissue and with free access to the external medium, in which secreted material can be collected. After 48 h in culture, the isolated lobules appeared indistinguishable, by morphological and biochemical criteria, from freshly isolated lobules. After much longer culture times about one third of the glandular cells were still capable of effective protein synthesis. Studying the biosynthesis of seminal ribonuclease with preparations of isolated lobules we found that the enzyme was synthesized and secreted; only the fully amidated isoenzyme was synthesized and secreted, indicating that production of the selectively deamidated isoenzymic forms occurred after secretion, newly synthesized protein was rapidly exported, indicating that the high levels of enzyme previously reported for the seminal vesicle tissue were essentially due to its content of stored secretion.
Assuntos
Ribonucleases/biossíntese , Glândulas Seminais/enzimologia , Animais , Bovinos , Cromatografia , Técnicas de Cultura/métodos , Masculino , Ribonucleases/análise , Glândulas Seminais/citologiaRESUMO
Primary cell cultures were established from explants of rat seminal vesicle. The establishment of primary cell cultures required, among other factors, the presence of testosterone. Two cell populations were detected in such primary cultures: fibroblast-like cells and epithelial-like cells; the latter encompassed a subtype of small cells and a subtype of large squamous cells (most likely the result of a degenerative process acting upon the former). Histochemical, as well as electron-microscopical observations, indicated the presence of a persistent secretory activity in the small epithelial cells; fibroblast and large squamous epithelial cells were inactive in this respect. Staining of the cells with a peroxidase-conjugated antibody and analysis of the proteins produced in the presence of labelled methionine, showed that one of the major rat seminal vesicle secretory proteins, namely RSV-IV, was also produced. Conditions which favoured the growth of epithelial cells, rather than of fibroblasts, were determined. The use of nearly homogeneous cell populations and the use of collagen-coated Petri dishes, allowed the cloning of two independently obtained permanent cell lines, namely SVC-1 and SVC-2. The in vitro growth rate of both cell lines was modulated by the amount of testosterone in the medium. Both cell lines were able to synthesize a significant amount of RSV-IV protein under testosterone control.
Assuntos
Proteínas Secretadas pela Próstata , Biossíntese de Proteínas , Glândulas Seminais/citologia , Animais , Células Cultivadas , Histocitoquímica , Masculino , Peso Molecular , Ratos , Proteínas de Plasma Seminal , Glândulas Seminais/metabolismo , Testosterona/farmacologiaRESUMO
The mRNA populations of control and 8-days-denervated adult rat gastrocnemius have been analysed by the translation assay and the cDNA-mRNA molecular hybridization technique. This analysis demonstrates the appearance of marked changes in many of the mRNA sequences present in muscle fibres following denervation and thus gives strong support to the hypothesis that the motoneurons are able to control the gene expression of muscle fibres.
Assuntos
Regulação da Expressão Gênica , Neurônios Motores/fisiologia , Músculos/metabolismo , RNA Mensageiro/metabolismo , Animais , Denervação , Genes Sintéticos , Masculino , Proteínas Musculares/análise , Músculos/inervação , Hibridização de Ácido Nucleico , Biossíntese de Proteínas , RatosRESUMO
Experiments are reported showing that following 8 days of denervation the function of the protein-synthesizing machinery, operating in the rat gastrocnemius fibres, is altered, probably as a consequence of decreased amounts of ribosomes and actively translated mRNA. In addition, the data obtained show that the amount per muscle and the availability per ribosome of the soluble factors involved in the process of protein synthesis are markedly decreased, thus suggesting that the amounts of ribosomes, mRNA and soluble factors are regulated in a concerted fashion when muscular protein synthesis is decreased after denervation.