RESUMO
Escherichia coli RNA polymerase was assayed with 4 mM Mg2+ and 1 mM Mn2+ using native DNA, heat-denatured DNA, histone-nucleate and isolated rat liver nuclei as the template source. With purified DNA and either or both divalent metal ions, 0.1--5 mM amine stimulated enzyme activity. Spermidine resulted in the greatest stimulation (1.7-fold at 5 mM); whereas, spermine or methylglyoxal bis(guanylhydrazone) first stimulated, then above 3 mM inhibited, the reaction. The addition of unfractionated histone to purified DNA inhibited the reaction by 90%. The subsequent addition of amines resulted in a slight stimulation in incorporation (1.5-fold) in the range of 1--3 mM amine. Alternatively, when enzyme was combined with DNA before histone, only a 20% inhibition was observed and this could be completely prevented by 3 mM spermidine. The addition of amines to isolated nuclei resulted in marked alterations in ultrastructure and Mg2+ content; however, relatively small effects on RNA polymerase activity were observed. With the E. coli enzyme, 0.1--1.0 mM amine stimulated RNA synthesis (1.5-fold) whereas, none of the amines stimulated endogeneous activity in the absence or presence of 300 mM (NH4)2SO4.
Assuntos
RNA Polimerases Dirigidas por DNA/metabolismo , Guanidinas/farmacologia , Mitoguazona/farmacologia , Poliaminas/farmacologia , Animais , Núcleo Celular/metabolismo , Ativação Enzimática , Histonas/farmacologia , Técnicas In Vitro , Fígado/metabolismo , Magnésio/farmacologia , Masculino , Manganês/farmacologia , Ratos , Moldes GenéticosRESUMO
We have identified and characterized 12 mutations in seven genes that affect the development of the Caenorhabditis elegans hindgut. We find that the mutations can disrupt the postembryonic development of the male-specific blast cells within the hindgut, the hindgut morphology in both males and hermaphrodites, and in some cases, the expression of a hindgut marker in hermaphrodite animals. Mutations in several of the genes also affect viability. On the basis of their mutant phenotypes, we propose that the genes fall into four distinct classes: (1) egl-5 is required for regional identity of the tail; (2) sem-4 is required for a variety of ectodermal and mesodermal cell types, including cells in the hindgut; (3) two genes, lin-49 and lin-59, affect development of many cells, including hindgut; and (4) three genes, mab-9, egl-38, and lin-48, are required for patterning fates within the hindgut, making certain hindgut cells different from others. We also describe a new allele of the Pax gene egl-38 that is temperature sensitive and affects the conserved beta-hairpin of the EGL-38 paired domain. Our results suggest that a combination of different factors contribute to normal C. elegans hindgut development.
Assuntos
Caenorhabditis elegans/fisiologia , Sistema Digestório/crescimento & desenvolvimento , Genes de Helmintos , Mutação , Envelhecimento , Sequência de Aminoácidos , Animais , Caenorhabditis elegans/crescimento & desenvolvimento , Transtornos do Desenvolvimento Sexual , Teste de Complementação Genética , Ligação Genética , Proteínas de Helminto/química , Proteínas de Helminto/genética , Heterozigoto , Masculino , Dados de Sequência Molecular , Mutagênese , Mutação de Sentido Incorreto , Estrutura Secundária de Proteína , Recombinação Genética , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Surgical alteration of the pancreas can result in several anatomic alterations which may affect insulin release. We evaluated the effects of resection, systemic drainage, and autotransplantation of the canine pancreas on peripheral insulin levels and glucose disposal as measured by iv glucose tolerance tests (IVGTT) and a steady state hyperglycemic challenge (clamp). Proximal pancreatectomy (PPx) with reduced beta-cell mass and intact portal drainage resulted in a modestly elevated fasting glucose level and increased integrated glucose response to IVGTT. Compared to preoperative normals, basal insulin was unchanged from preoperative controls; however, peak insulin and integrated insulin response to IVGTT were decreased in PPx animals. Splenocaval drainage or autotransplantation of the distal pancreas resulted in normalization of the severely altered insulin response and fasting glucose levels. K values were significantly reduced after all three procedures. Clamp studies confirmed the basal glucose and insulin findings of the IVGTT. During the clamp, PPx animals had peripheral insulin values approximately 50% of normal controls, while autotransplantation and splenocaval drainage animals had insulin values that approximate normal controls. All three postsurgical groups had blunted insulin levels during stable hyperglycemia. Glucose utilization rates were severely decreased in all three groups. Reduction of beta-cell mass with intact portal drainage resulted in reduced insulin response to glucose challenge by either IVGTT or clamp. Systemic drainage of this same reduced beta-cell mass resulted in peripheral insulin levels comparable to normal controls. Denervation (autotransplantation) had little additive effect. All three groups demonstrated severely decreased rates of glucose disappearance as measured by both IVGTT and clamp studies. Therefore, reduction in beta-cell mass, drained systemically or portally, results in altered glucose disposal regardless of the peripheral insulin levels.
Assuntos
Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Pancreatectomia , Animais , Glicemia/metabolismo , Cães , Feminino , Teste de Tolerância a Glucose , Insulina/sangue , Secreção de Insulina , Sistema Porta , Valores de ReferênciaRESUMO
Anatomic alterations of the pancreas result in physiologic alterations that have not been completely analyzed. Insulin plays a major role in carbohydrate metabolism; nevertheless, as much as 50% of a hyperglycemic load may be metabolized independent of insulin. We analyzed the effects of surgical alterations of the pancreas on postoperative glucose metabolism, including insulin-independent effects. Mongrel female dogs underwent one of three procedures: proximal partial pancreatectomy (PPx), PPx plus diversion of pancreatic venous effluent to the systemic circulation (SC), or PPx plus segmental pancreatic autotransplantation (PAT). Intravenous glucose tolerance tests, with or without a background infusion of somatostatin (SST; 400 ng/kg/min) were performed on all animals preoperatively and postoperatively. SST completely suppressed secretion of assayable peripheral insulin. The rate of glucose disposal during SST suppression approximates the rate of insulin-independent glucose disposal (IIGD). Although there was a significant decrease in the rate of glucose disposal during SST infusion when compared with the rate without SST, no differences in IIGD were found between postoperative groups. IIGD was calculated at 50% to 55% for control, PPx, and SC groups and at 67% for PAT. Peripheral sensitivity to an exogenous insulin infusion (euglycemic clamp) was unchanged by any of the procedures. We conclude that surgical alteration of the pancreas, including pancreas transplantation, results in altered glucose handling in the face of "normal" peripheral levels of insulin. Changes in IIGD and analysis of peripheral sensitivity to insulin do not explain these alterations completely.
Assuntos
Insulina/metabolismo , Pancreatectomia , Animais , Glicemia/metabolismo , Cães , Feminino , Técnica Clamp de Glucose , Teste de Tolerância a Glucose , Insulina/sangue , Secreção de Insulina , Cinética , Transplante de Pâncreas , Transplante AutólogoRESUMO
Isoproterenol (0.3 micronmole/gm body weight) was injected intraperitoneally every 24 h for three days. The synthesis of deoxyribonucleic acid, the concentration of putrescine, spermidine and spermine and the activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase were measured in parotid and submandibular glands at 4 to 8 h after each injection. The parotid glands responded with peaks of DNA synthesis at 24 and 72 h and peaks of putrescine content and decarboxylase activities 8 to 12 h after each injection. Spermidine increased steadily in the parotid, whereas there was little change in the spermine concentration throughout the 72 h. Polyamine metabolism showed much less response in the submandibular gland, and little or no increase in spermidine or spermine levels or in DNA synthesis was observed.
Assuntos
Isoproterenol/farmacologia , Glândula Parótida/metabolismo , Poliaminas/metabolismo , Glândula Submandibular/metabolismo , Adenosilmetionina Descarboxilase/metabolismo , Animais , DNA/biossíntese , Masculino , Camundongos , Tamanho do Órgão/efeitos dos fármacos , Ornitina Descarboxilase , Glândula Parótida/efeitos dos fármacos , Putrescina/metabolismo , Espermidina/metabolismo , Estimulação Química , Glândula Submandibular/efeitos dos fármacosRESUMO
Timed-gestation Wister rats were given free access to isocaloric diets containing 8% protein (malnourished) or 24% protein (controls). The dietary regimen was initiated on the 12th day of gestation and continued to the dams throughout the remainder of gestation and lactation. Postweaning rats were continued on the respective diets until studies were performed. Whole brain norepinephrine concentration was slightly increased on Day 21 after birth and was significantly increased on Day 35 in malnourished rats as compared with controls.
Assuntos
Encéfalo/metabolismo , Norepinefrina/metabolismo , Deficiência de Proteína/metabolismo , Envelhecimento , Animais , Feminino , Masculino , RatosRESUMO
The marginal adaptation and microleakage of the glass-ionomer-lined composite resin restoration in simulated erosion and conventional class 5 preparations were evaluated. The most common site of leakage for all restorations was the gingival margin. In erosion lesion restorations, the etched thick liners were superior to the etched thin liners with respect to marginal leakage. Acid-etched and unetched liners in both the erosion lesion and the conventional class 5 restorations were comparable. Under the SEM a gap was often found in the liner rather than at the liner-resin interface in restorations with etched liners.
Assuntos
Condicionamento Ácido do Dente , Resinas Compostas , Forramento da Cavidade Dentária , Infiltração Dentária , Restauração Dentária Permanente/métodos , Cimentos de Ionômeros de Vidro , Bis-Fenol A-Glicidil Metacrilato , Humanos , Microscopia Eletrônica de Varredura , Propriedades de Superfície , Fatores de Tempo , Erosão Dentária/terapiaAssuntos
Transplante de Rim/fisiologia , Emprego , Feminino , Seguimentos , Nível de Saúde , Humanos , Transplante de Rim/mortalidade , Transplante de Rim/reabilitação , Masculino , Prontuários Médicos , Morbidade , Complicações Pós-Operatórias/epidemiologia , Gravidez , Estudos Retrospectivos , Inquéritos e Questionários , Fatores de TempoAssuntos
Suscetibilidade à Cárie Dentária , Cárie Dentária/terapia , Educação em Odontologia , Medição de Risco , Agendamento de Consultas , Currículo , Cárie Dentária/diagnóstico , Dentística Operatória/educação , Diagnóstico Bucal/educação , Odontologia Geral/educação , Humanos , Odontopediatria/educação , Faculdades de Odontologia , Estados UnidosRESUMO
1. The interaction of polyamines and methylglyoxal bis(guanythydrazone) (1, 1'-[(methylethanediylidene)-dinitrilo]diguanidine) with isolated rat liver nuclei was investigated by electron microscopy. 2. At 4mM, putrescine was without effect; however, spermidine, spermine or methylglyoxal bis(guanythydrazone) resulted in dispersed chromatin and alterations in nucleolar structure. In addition, spermidine or methylglyoxal bis(guanylhydrazone) caused marked aggregation of interchromatin granules. 3. The DNA template property of calf thymus DNA was examined by using DNA polymerases from Escherichia coli, Micrococcus lysodeikticus and calf thymus in the presence of 0-5 mM-amine. 4. In the presence of DNA polymerase, spermine or methylglyoxal bis(guanylhydrazone) inhibited activity, whereas putrescine or spermidine had much less effect or in some cases stimulated [3H]dTMP incorporation. 5. Template activity which was inhibited by spermine or methylglyoxal bis(guanylhydrazone) could be partially restored by additional DNA or enzyme. 6. When mixed with calf thymus DNA, calf thymus histone inhibited template activity as measured with E. coli DNA polymerase. The template activity of such a 'histone-nucleate' could not be restored by putrescine, spermidine, spermine or methylglyoxal bis(guanylhydrazone). 7. DNA template activity of isolated rat liver nuclei was tested by using E. coli DNA polymerase. None of the amines was able to increase the template activity of the nuclear DNA in vitro.
Assuntos
Núcleo Celular/efeitos dos fármacos , DNA/metabolismo , Guanidinas/farmacologia , Mitoguazona/farmacologia , Poliaminas/farmacologia , Animais , Bovinos , Núcleo Celular/metabolismo , Núcleo Celular/ultraestrutura , DNA Nucleotidiltransferases/metabolismo , Histonas/isolamento & purificação , Técnicas In Vitro , Fígado/ultraestrutura , Masculino , Microscopia Eletrônica , Desnaturação de Ácido Nucleico , Putrescina/farmacologia , Ratos , Espermidina/farmacologia , Espermina/farmacologia , Moldes Genéticos , Timo/análiseRESUMO
The nonrecirculating isolated perfused rat liver was used to study biliary antibiotic excretion by the liver in a steady-state, controlled environment in which bile flow, bile salt output, and antibiotic delivery were maintained under constant conditions. The effects of piperacillin, ampicillin, and gentamicin on bile flow and bile salt output were analyzed; none altered bile salt output, and only high concentrations of piperacillin (100 micrograms/mL) increased bile flow. The ratio of antibiotic concentration in bile and perfusate depended on the type of antibiotic and perfusate concentration. Piperacillin infusions at perfusate concentrations of 50 or 100 micrograms/mL (in the presence of 60 microM taurocholate) yielded bile to perfusate ratios of 112 +/- 10 versus 49 +/- 3, respectively. Using similar perfusate, the concentration ratios for ampicillin (20 micrograms/mL) and gentamicin (10 micrograms/mL) were only 3.4 +/- 0.5 and 0.5 +/- 0.1, respectively. By altering the perfusate to contain either 60 microM or 240 microM taurocholate, we found variance in bile salt output from 27 +/- 1 to 115 +/- 2 mumol/h, yet this alteration had little effect on the output of ampicillin (perfusate concentration of 20 micrograms/mL), 73 +/- 7 versus 74 +/- 12 micrograms/h, or piperacillin (perfusate concentration 100 micrograms/mL), 10 +/- 1 versus 11 +/- 2 mg/h. Thus, it appears ampicillin and piperacillin are excreted into bile at high concentrations by bile salt-independent pathways. Partial biliary obstruction (6 cm H2O) results in significant decreases in bile volume. Infusion of 50 micrograms/mL of piperacillin resulted in increased biliary flow that approached nonobstructed values. Obstruction resulted in significant decreases in bile piperacillin concentration. Whether the choleretic effect of high concentrations of piperacillin has any clinical significance in nonobstructed or obstructed conditions remains to be established.