RESUMO
This study evaluated effects of bone morphogenetic protein II (BMP) on glial fibrillary acidic protein (GFAP) in the brain of female Fischer 344 rats during 14 days of spaceflight. GFAP mRNA decreased in vehicle-implanted rats flown on the space shuttle by 53 and 48% in the stratum moleculare and stratum lacunosum moleculare hippocampal subregions, respectively. GFAP mRNA was not significantly affected by BMP implantation during spaceflight. Rats returning from space exhibited a 56% increase in serum corticosterone. BMP treatment did not additively increase corticosterone elevations in microgravity but appeared to increase serum corticosterone and reduce GFAP mRNA in the stratum moleculare in control rats. These data suggest that exposure to microgravity reduces GFAP expression in hippocampal astrocytes.
Assuntos
Proteínas Morfogenéticas Ósseas/farmacologia , Química Encefálica/efeitos dos fármacos , Química Encefálica/fisiologia , Proteína Glial Fibrilar Ácida/metabolismo , Voo Espacial , Ausência de Peso/efeitos adversos , Animais , Astrócitos/efeitos dos fármacos , Tamanho Celular/efeitos dos fármacos , Corticosterona/sangue , Feminino , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Degeneração Neural/fisiopatologia , Proteínas do Tecido Nervoso/biossíntese , Neurônios/efeitos dos fármacos , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos F344RESUMO
The maturation of dental enamel succeeds the degradation of organic matrix. Inhibition studies have shown that this degradation is accomplished by a serine-type proteinase. To isolate and characterize cDNA clones encoding this proteinase, we used two degenerate primer approaches to amplify part of the coding region using polymerase chain-reaction (PCR). First, we purified the proteinase from porcine transition-stage enamel matrix and characterized it by partial protein sequencing. The enzyme was isolated from the neutral soluble enamel extract by successive ammonium sulfate precipitations, hydroxyapatite HPLC, reverse-phase HPLC, DEAE ion exchange, and affinity chromatography with a Benzamidine Sepharose 6B column. The intact protein and lysylendopeptidase-generated cleavage products were characterized by amino acid sequence analyses. Degenerate oligonucleotide primers encoding two of the polypeptide sequences were synthesized. In a complementary strategy, degenerate oligonucleotide primers were designed against highly conserved active-site regions of chymotrypsin-like proteinases. Both approaches yielded PCR amplification products that served as probes for screening a porcine enamel organ epithelia-specific cDNA library. The longest full-length clone is 1133 nucleotides and encodes a preproprotein of 254 amino acids. We designate this protein enamel matrix serine proteinase 1 or EMSP1. The active protein has 224 amino acids, an isotope-averaged molecular mass of 24.1 kDa, and an isoelectric point of 6.0. Multiple-tissue Northern analysis indicates that EMSP1 is a tooth-specific protein. Gelatin enzymography shows a dramatic increase in EMSP1 activity in the transition-stage enamel matrix. EMSP1 is most homologous to kallikriens and trypsins.
Assuntos
Amelogênese , Proteínas do Esmalte Dentário/química , Esmalte Dentário/enzimologia , Calicreínas , Serina Endopeptidases/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Proteínas do Esmalte Dentário/genética , Proteínas do Esmalte Dentário/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Peso Molecular , Especificidade de Órgãos , Reação em Cadeia da Polimerase , Processamento de Proteína Pós-Traducional , Análise de Sequência , Serina Endopeptidases/genética , Serina Endopeptidases/isolamento & purificação , SuínosRESUMO
The goal of this study was to evaluate the effects of microwave irradiation on immunocytochemical staining in freshly frozen rat brain tissue using an antibody previously shown to successfully stain only formalin-fixed materials. GFAP immunoreactivity was markedly increased in sections exposed to microwave heating when compared with control tissue. Uniform staining enhancement was mainly a result of increased numbers of immunoreactive soma and large astrocytic processes. The integrity of the tissue was not affected by this treatment. Other antibodies need to be tested with this method, but our results suggest that microwave irradiation is useful for rapid antigen retrieval in freshly frozen central nervous system tissue.
Assuntos
Encéfalo/efeitos da radiação , Córtex Cerebral/efeitos da radiação , Hipocampo/efeitos da radiação , Micro-Ondas , Animais , Imuno-Histoquímica , Masculino , Ratos , Ratos Endogâmicos F344RESUMO
Alcohol and other substance abuse treatment programs are focusing more on the wellbeing of the total patient rather than just targeting functional sobriety. Various explanations are offered in this article for the inclusion of a fitness and health promotion component in a chemical abuse treatment program. The treatment program at Charter Hospital of Fort Worth, Texas, is discussed in detail, including a comprehensive medical examination, fitness and nutritional screenings, and appropriate exercise activities. Educational sessions are offered on health fitness, risk reduction, stress management, smoking cessation, proper nutrition and the positive psychological aspects of exercise. The implications of a health/fitness component in the recovery process are emphasized.
