RESUMO
OBJECTIVE: Our purpose was to develop a specific polymerase chain reaction detection method for 16 micro-organisms in amniotic fluid and to correlate its performance with bacterial cultures and preterm delivery occurrence. STUDY DESIGN: The study group was made up of 50 patients with preterm labor and intact membranes. The control group consisted of 23 patients not in labor and undergoing amniocentesis for either karyotype or lung maturity studies. Polymerase chain reaction and bacterial cultures were assayed in amniotic fluid of all patients. Results were correlated with pregnancy outcome. RESULTS: Polymerase chain reaction identified micro-organisms in 23 cases in the study group (46%), whereas cultures identified only 6 (12%). All control samples were negative for polymerase chain reaction and cultures. The sensitivity of polymerase chain reaction and cultures for the identification of patients delivering before 34 weeks' gestation was 64% and 18%, respectively. CONCLUSION: A polymerase chain reaction gene amplification method was developed to identify 16 micro-organisms in amniotic fluid. Compared with bacterial cultures, polymerase chain reaction amplification in amniotic fluid appears to be more sensitive in identifying patients delivering prematurely.
Assuntos
Líquido Amniótico/microbiologia , Parto Obstétrico , Trabalho de Parto Prematuro/epidemiologia , Reação em Cadeia da Polimerase , Adolescente , Adulto , Infecções Bacterianas/epidemiologia , Feminino , Humanos , Incidência , Recém-Nascido , Doenças do Recém-Nascido/epidemiologia , Morbidade , GravidezRESUMO
Recent demonstrations of no changes in hypothalamic gonadotropin releasing hormone (GnRH) gene expression and GnRH levels detected at the pituitary gland in diestrous and lactating rats, indicate that lactational hypogonadotropism in this species is not associated with inhibition of hypothalamic GnRH synthesis and secretion. Hypothalamic galanin potentiates GnRH effects on luteinizing hormone (LH) secretion in male and cycling rats. To explore the interaction between GnRH and galanin during lactation, we studied in vitro the effects of pulsatile stimulation with those peptides upon LH synthesis and secretion from rat pituitaries on diestrous 1 or day 10 of lactation. Hemipituitaries were separately incubated in 1 ml Dulbecco's Minimal Essential Medium supplemented with 1% penicillin-streptomycin and fetal calf serum, at 37 degrees C in 5% CO2-air. The hemipituitaries were stimulated during 12 h with hourly pulses, 6 min each, of (a) gonadotropin releasing hormone (GnRH 25 ng/pulse), (b) rat galanin (600 ng/pulse), (c) GnRH plus galanin, or (d) saline. Medium was collected before each pulse to determine LH by radioimmunoassay. After the 12 h pulsatile regime total RNA was extracted and both actin and beta-LH mRNA were determined by reverse transcriptase polymerase chain reaction. There was a significant stimulation of LH secretion by GnRH (ANOVA, p < 0.001) without significant differences between diestrous and lactation pituitaries. Galanin alone did not modify LH secretion but it potentiated the effect of GnRH upon pituitaries from diestrous (p = 0.036) but not lactating rats. Neither peptide alone or its combination modified pituitary beta-LH mRNA levels. Results show that galanin regulates differently the secretion and synthesis of LH at the pituitary level. The disappearance of galanin-induced potentiation of GnRH effects upon LH secretion during lactation might contribute to the hypogonadotropism of lactation in the rat.
Assuntos
Animais Lactentes/fisiologia , Galanina/fisiologia , Hormônio Liberador de Gonadotropina/fisiologia , Hormônio Luteinizante/metabolismo , Animais , Diestro , Feminino , Técnicas In Vitro , Hormônio Luteinizante/biossíntese , Hipófise , Ratos , Ratos Sprague-DawleyRESUMO
The L5 ribosomal protein gene (rpl5) and a S14 ribosomal protein pseudogene were identified by sequence analysis in the potato mitochondrial genome. The two genes are separated by one nucleotide and are found upstream of the apocytochrome b gene (cob), an arrangement conserved also in Arabidopsis and Brassica. The rpl5 gene has an intact open reading frame while the rps14 locus is disrupted by a five nucleotide duplication that introduces a frameshift in the reading frame. Editing of rpl5 and pseudorps14 cotranscripts has been studied by cDNA sequence analysis. Eight C residues are edited into U in the rpl5 coding region, resulting in eight amino acid changes that increase the homology between potato and other RPL5 polypeptides. Interestingly, the rps14 pseudogene sequence is not edited at any nucleotide position.
Assuntos
Genes de Plantas/genética , Pseudogenes/genética , Proteínas Ribossômicas/genética , Solanum tuberosum/genética , Sequência de Aminoácidos , Apoproteínas/genética , Sequência de Bases , Grupo dos Citocromos b/genética , Citocromos b , DNA Mitocondrial/genética , DNA de Plantas/genética , Mitocôndrias , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , RNA/análise , Edição de RNA/fisiologia , RNA Mitocondrial , RNA de Plantas/análise , Mapeamento por Restrição , Análise de Sequência de DNARESUMO
A reverse transcriptase activity has been detected in potato mitochondria using special RNAs as templates: a bacterial RNA coding for neomycin phosphotransferase (neo pa RNA) and a Neurospora crassa mitochondrial RNA (184 nt RNA). Surprisingly, no exogenous primer addition was required. These RNA templates share a primary and secondary structure similar to the T psi CG loop of tRNAs that could constitute the recognition site for the enzyme. Reverse transcriptase activity was inhibited by ddTTP, ethidium bromide and aphidicolin, while potato mitochondrial DNA polymerase was not inhibited by aphidicolin indicating that these activities correspond to distinct enzymes. A conserved sequence of reverse transcriptases was detected in potato mitochondrial DNA suggesting that this enzyme could be mitochondrially encoded.
Assuntos
Mitocôndrias/enzimologia , DNA Polimerase Dirigida por RNA/metabolismo , Solanum tuberosum/enzimologia , Sequência de Bases , DNA Complementar/genética , DNA Mitocondrial/genética , Genes de Plantas , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Oligonucleotídeos/química , RNA/ultraestrutura , RNA Mitocondrial , DNA Polimerase Dirigida por RNA/genética , Inibidores da Transcriptase Reversa/farmacologiaRESUMO
PIP: The extent and consequences of exclusion of low income strata from maternal and child health programs in Chile are analyzed using available data. Infant mortality has been shown by several studies to be closely associated with socioeconomic status in Chile. Babies of illiterate mothers showed the highest rate of mortality and the least improvement in rate between 1972-78. The effect of socioeconomic status on the mortality rate of infants in greatly influenced by birth weight; low birth weight infants of low income groups suffer significantly higher mortality than among higher income groups. Several national studies in Chile demonstrated a relationship between infant malnutrition and health program coverage. Infant malnutrition is greatest in groups benefiting least from health care. Based on the fact that 90.5% of births in 1980 were professionally attended, it is estimated that 9.5% of the low income population lacks access to health care. A recent survey showed that 9.9% of the population under 6 years, some 105,848 children, was not covered by the National Complementary Feeding Program. Another study showed that 12.3% of mothers had no prenatal medical attention prior to their most recent birth; mothers with little or no education, living in rural areas, and of high parity were most likely not to have received medical attention. Factors responsible for lack of access to health and nutrition programs appeared to include unsatisfactory relationships with the health workers, poor acceptability of foods offered, excessive distance and waiting times, and lack of interest or motivation on the part of the mothers.^ieng