RESUMO
Gastroesophageal reflux disease-related diseases, such as Barrett's esophagus and adenocarcinoma of the esophagogastric junction (AEGJ), are believed to occur less frequently in Asia than in Western countries. However, the number of reported cases is increasing, yet little is known regarding the epidemiology of AEGJ in Japan. The primary study aim is to investigate the clinicoepidemiological characteristics of AEGJ. The secondary aim is to identify factors associated with it. In the 6.5 years between January 2008 and June 2014, we reviewed 88,199 esophagogastroduodenoscopy (EGD) reports and associated medical records (Study 1). We conducted a case-control study to identify factors associated with AEGJ (Study 2). Control subjects were randomly selected and age and sex matched from among subjects who underwent EGD during medical evaluations. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated using an unconditional logistic regression method. During the study period, 83 patients with AEGJ (72 men and 11 women; mean age 68 years) were diagnosed. Six cases were Siewert type I and 77 were type II. The incidence rate of AEGJ was 0.6-1.7/100,000 person-years. Compared with the 101 control subjects, obesity (body mass index ⧠25 kg/m2; [OR = 2.82; 95% CI: 1.13-7.01]) was associated with AEGJ. The incidence rate of AEGJ is lower in Japan than in Western countries, but associated factors similar to those in Western patients were detected, including obesity, a hiatal hernia, smoking, and the male sex.
Assuntos
Adenocarcinoma/epidemiologia , Povo Asiático/estatística & dados numéricos , Neoplasias Esofágicas/epidemiologia , Junção Esofagogástrica/patologia , Adenocarcinoma/etiologia , Adenocarcinoma/patologia , Idoso , Índice de Massa Corporal , Estudos de Casos e Controles , Neoplasias Esofágicas/etiologia , Neoplasias Esofágicas/patologia , Feminino , Hérnia Hiatal/complicações , Humanos , Incidência , Japão/epidemiologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Razão de Chances , Fatores de Risco , Fatores Sexuais , Fumar/efeitos adversosRESUMO
We determined expression and localization of the anti-apoptotic cellular FLICE inhibitory protein (cFLIP) in the porcine corpora lutea (CL) and corpus albicans (CA) during estrous and pregnancy. The CL and CA were collected at different stages of estrous to determine cFLIP immunolocalization, and mRNA and protein expression. The mRNA expression of the short cFLIP isoform (cFLIPS) was higher at the early and mid CL stages, and lower by the late CL stage (P < 0.01); mRNA expression of the long cFLIP isoform (cFLIPL) was higher at the mid CL stage, and lower at the early and late CL stages (P < 0.01). Levels of cFLIPS and cFLIPL were steady and high during the early and mid CL stages, and had significantly decreased (P < 0.01) by the late stage. The cFLIP protein was highly expressed in the early and mid CL stages of estrous, but weakly ex-pressed in the late stage. Expression of cFLIPS showed no significant difference between preovulatory corpus albicans (CA1) and corpus albicans (CA2) coexistent with the CL from the previous estrus, but cFLIPL mRNA expression was higher during CA1 than CA2. The expression of cFLIPS showed no significant difference between CA1 and CA2, but cFLIPL was not detected. The cFLIP protein was weak-ly expressed in the CA. Expression of cFLIPS and cFLIPL mRNA and proteins was observed in the CL, and the cFLIP protein was highly expressed during pregnancy. We propose that cFLIPS/L acts as a survival factor, and performs an anti-apoptotic function in the porcine CL.
Assuntos
Apoptose/genética , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/biossíntese , Corpo Lúteo/metabolismo , Ciclo Estral/genética , Animais , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Ciclo Estral/metabolismo , Feminino , Regulação da Expressão Gênica , Gravidez , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/genética , RNA Mensageiro/genética , SuínosRESUMO
BACKGROUND: Serotonin type 3 receptor (5-HT3 R) antagonists are potentially useful therapeutic agents for diarrhea-predominant irritable bowel syndrome (IBS-D). To identify biomarkers predicting effectiveness of the 5-HT3 R antagonist (ramosetron) in IBS-D. METHODS: Irritable bowel syndrome-D Japanese subjects received 2.5 or 5 µg of ramosetron once daily for 4 weeks. Colonic mucosal S100A and tryptophan hydroxylase (TPH) mRNA expression levels were measured before treatment. Genomic DNA was extracted from blood and polymorphisms of TPH1 and TPH2 were analyzed. KEY RESULTS: Forty-two patients (27 men and 15 women, mean age 42 years) with IBS-D were included for analysis. Improvement of IBS symptoms was seen in 26 (61.9%). Baseline S100A10 (p = 0.02) and TPH1 (p = 0.02) expression were significantly higher in the ramosetron responders than in the non-responders. The frequencies of the TPH1 rs4537731G allele in linkage disequilibrium with the TPH1 rs7130929 T allele (11.5% vs 50%, p = 0.003; OR: 12; 95% CI: 2.1-69) along with TPH1 rs211105 C allele (3.8% vs 43.8%, p = 0.0003; OR: 19; 95% CI: 2.1-181) were significantly lower in the responders than in the non-responders. The mean scores of diarrhea at baseline were significantly higher (5.2 vs 3.7, p = 0.005) in patients with TPH1 rs211105 T/T than those with the G allele. CONCLUSIONS & INFERENCES: TPH1 gene polymorphisms and S100A10 expression, which correlate with 5-HT signaling were associated with ramosetron effectiveness in IBS-D, and may possibly lead to prospective identification of the resistance to treatment.
Assuntos
Anexina A2/metabolismo , Benzimidazóis/administração & dosagem , Biomarcadores Farmacológicos , Diarreia/tratamento farmacológico , Síndrome do Intestino Irritável/tratamento farmacológico , Proteínas S100/metabolismo , Antagonistas do Receptor 5-HT3 de Serotonina/administração & dosagem , Triptofano Hidroxilase/genética , Adulto , Diarreia/complicações , Feminino , Humanos , Mucosa Intestinal/metabolismo , Síndrome do Intestino Irritável/complicações , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Polimorfismo de Nucleotídeo Único , Resultado do TratamentoRESUMO
BACKGROUND: Improvement in subjective symptoms has been reported in functional dyspepsia (FD) patients administered with acotiamide. Improvement was confirmed in meal-related symptoms, such as postprandial fullness, upper abdominal bloating, and early satiety. We examined the mechanism underlying the effects of acotiamide on gastric accommodation reflex (GAR) and gastroduodenal motility in FD patients. METHODS: Thirty-four FD patients (mean age, 40.4 years) were examined ultrasonographically before and after 14-18 days of acotiamide (100 mg t.i.d.) or placebo administration. To assess GAR, expansion rate in cross-sectional area of the proximal stomach was measured after every 100-mL ingestion, using a straw, of up to 400 mL of a liquid meal (consommé soup, 13.1 kcal; 400 mL) in a supine position. Next, we measured gastric emptying rate (GER), motility index (MI, antral contractions), and reflux index (RI, duodenogastric reflux) to assess gastroduodenal motility. Patients also completed a survey based on the seven-point Likert scale both before and after drug administration. KEY RESULTS: Of the 37 cases, 19 and 18 were administered with acotiamide and placebo A respectively, significant difference was observed in GAR between the acotiamide and placebo groups (21.7%vs 4.4%) after 400 mL ingestion. GER significantly accelerated after treatment in the acotiamide group (P = 0.012), no significant differences were observed in MI and RI between the two groups. Improvement rates were 35.3 and 11.8% for the acotiamide and placebo groups. CONCLUSIONS & INFERENCES: Acotiamide significantly enhances GAR and GER in FD patients. Acotiamide may have therapeutic potential for FD patients.
Assuntos
Benzamidas/uso terapêutico , Dispepsia/tratamento farmacológico , Esvaziamento Gástrico/efeitos dos fármacos , Período Pós-Prandial/efeitos dos fármacos , Estômago/efeitos dos fármacos , Tiazóis/uso terapêutico , Adulto , Idoso , Benzamidas/farmacologia , Método Duplo-Cego , Dispepsia/diagnóstico por imagem , Feminino , Motilidade Gastrointestinal/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Estômago/diagnóstico por imagem , Decúbito Dorsal , Tiazóis/farmacologia , Resultado do Tratamento , UltrassonografiaRESUMO
Despite normal eucrasia, mating desire and semen quality, sire bulls sometimes have spermatozoa with poor freezing tolerance. This study assessed effects of the addition of linoleic acid albumin (LAA) and long-term (LT) equilibrium to frozen semen on their sperm freezing tolerance. Immediately after collection using an artificial vagina and a breeding mount, semen was diluted with yolk citrate buffer; then, it was cooled slowly to 4°C during more than 5 h. Equilibrium treatment at 4°C was applied using the same extender supplemented with glycerol. Semen of bull A, with low sperm freezing tolerance, was treated with 1 mg/ml of LAA added to the first extender. The equilibrium treatment at 4°C was prolonged to 30 h. Significantly higher motility rates were obtained for the LT + LAA-treated sperm before and after freezing-thawing. However, for semen of bulls B and C with normal sperm freezing tolerance, the LT + LAA treatment barely exhibited a small effect on the motility rate. Almost no difference was found among bulls A, B and C in the motility rates of LT + LAA-treated sperm after freezing-thawing. No difference of fertility was apparent on LT + LAA-treated frozen sperm in comparison with normal sperm in embryonic collection and in vitro fertilization. It was not an aberration of fertility in vivo or in vitro. In addition, the conception rate of artificial insemination did not have a difference, and a normal calf was obtained. Results show that addition of LAA to an extender for frozen bovine spermatozoa and 30 h of low-temperature equilibrium might improve the motility of freezing-thawing spermatozoa with poor freezability. Sperm exhibited normal fertilization capability and ontogenic capability.
Assuntos
Albuminas/administração & dosagem , Bovinos , Criopreservação/veterinária , Ácido Linoleico/administração & dosagem , Preservação do Sêmen/veterinária , Animais , Criopreservação/métodos , Crioprotetores , Fertilização in vitro/veterinária , Masculino , Preservação do Sêmen/métodos , Soluções , Motilidade dos Espermatozoides , Espermatozoides/fisiologiaRESUMO
Nonerosive reflux disease (NERD) is the most common form of gastroesophageal reflux disease. Patients with NERD have a lower response rate to proton pump inhibitors (PPIs) than patients with erosive esophagitis when gauged from relief of heartburn. Sodium alginate decreases the acidity of refluxate and protects the esophageal mucosa. However, whether the addition of sodium alginate to PPI therapy can improve NERD symptoms remains unknown. Accordingly, the aim of this study was to evaluate the efficacy of adding sodium alginate to basal PPI therapy for NERD. Patients who had experienced heartburn on at least 2 days per week during the 1-month period before entering the study and had no endoscopic mucosal breaks (grade M or N according to Hoshihara's modification of the Los Angeles classification) were randomized to one of two treatments for 4 weeks: omeprazole (20 mg once daily) plus sodium alginate (30 mL four times a day) (group A) or omeprazole (20 mg once daily) alone (group B). Eighty-seven patients were enrolled, and 76 patients were randomly assigned to group A (n = 36) or group B (n = 40). Complete resolution of heartburn for at least 7 consecutive days by the end of treatment was significantly more common in group A (56.7%) than in group B (25.7%). One patient from group A had mild drug-related diarrhea that was not clinically serious. In conclusion, omeprazole combined with sodium alginate was better than omeprazole alone in Japanese patients with NERD.
Assuntos
Alginatos/uso terapêutico , Refluxo Gastroesofágico/tratamento farmacológico , Fármacos Gastrointestinais/uso terapêutico , Omeprazol/uso terapêutico , Inibidores da Bomba de Prótons/uso terapêutico , Adulto , Idoso , Transtornos de Deglutição/tratamento farmacológico , Transtornos de Deglutição/etiologia , Quimioterapia Combinada , Feminino , Refluxo Gastroesofágico/complicações , Ácido Glucurônico/uso terapêutico , Azia/tratamento farmacológico , Azia/etiologia , Ácidos Hexurônicos/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do TratamentoRESUMO
BACKGROUND: Impaired vagal function has been reported to be important in some patients with functional dyspepsia (FD). However, the pathophysiologic mechanisms influencing the cephalic phase of vagal activity in FD are incompletely understood. The aim of this study was to investigate the gastric response to modified sham feeding (MSF) on ultrasound and cardiovascular autonomic function in FD patients. METHODS: Nineteen patients with postprandial distress syndrome (PDS, 11 men and eight women; mean age: 48.2 years) and 26 healthy subjects (HS, 13 men and 13 women; mean age: 45.0 years) were studied prospectively. Firstly, cardiovascular autonomic function was assessed by spectral analysis of RR interval variability. Antral contraction was then evaluated by ultrasonography after MSF was performed to stimulate the cephalic phase of vagal activity. KEY RESULTS: Spectral analysis of RR interval variability showed that the high-frequency component was significantly smaller in the patients than in the HS (P<0.01). The frequency of antral contraction in response to MSF over 15 min was also significantly lower in the PDS patients than in the HS. The 15-min integrated antral contractile response (area under the contraction vs time curve) was significantly smaller in the PDS patients than in the HS (P<0.01). Univariate analysis revealed a modest correlation between the high-frequency component of RR interval variability and the area under the contraction vs time curve (n=46, r=0.49, P<0.01). CONCLUSIONS & INFERENCES: Autonomic abnormalities affecting the cephalic phase of vagal activity may be important in the pathogenesis of FD.
Assuntos
Dispepsia/fisiopatologia , Ingestão de Alimentos/fisiologia , Esvaziamento Gástrico/fisiologia , Período Pós-Prandial/fisiologia , Estômago/fisiologia , Adolescente , Adulto , Idoso , Animais , Feminino , Frequência Cardíaca/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Contração Muscular/fisiologia , Estômago/diagnóstico por imagem , Estômago/inervação , Ultrassonografia , Nervo Vago/fisiologia , Nervo Vago/fisiopatologia , Adulto JovemRESUMO
BACKGROUND: Proton pump inhibitors (PPIs) are less effective in non-erosive reflux disease (NERD) patients than in reflux oesophagitis patients. Whether the addition of prokinetics to PPIs improves NERD patients' symptoms remains unknown. AIM: To evaluate the efficacy of mosapride in NERD patients when used with PPI. METHODS: A total of 200 NERD patients were randomised to one of two arms: omeprazole (10 mg once daily) plus mosapride citrate (5 mg three times a day) (treatment arm) and omeprazole plus placebo (placebo arm). The primary endpoint was the rate of responders [visual analogue scale (VAS) was zero or <1 cm] after 4 weeks of treatment. The secondary endpoints were changes in the VAS score and the safety profile. RESULTS: There was no significant difference between the rates of responders in both arms in intent-to-treat (ITT) and per-protocol (PP) analysis. The change in the VAS score in treatment arm was significantly better than placebo arm in PP analysis (-4.0 ± 0.2 and -3.3 ± 0.2, mean ± S.E.M.) (N.S. in ITT analysis). The rate of adverse events was similar in both groups. CONCLUSION: The addition of mosapride to omeprazole was not more effective than omeprazole alone.
Assuntos
Antiulcerosos/administração & dosagem , Benzamidas/administração & dosagem , Refluxo Gastroesofágico/tratamento farmacológico , Fármacos Gastrointestinais/administração & dosagem , Morfolinas/administração & dosagem , Inibidores da Bomba de Prótons/administração & dosagem , Adulto , Idoso , Método Duplo-Cego , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Omeprazol/administração & dosagem , Qualidade de Vida , Índice de Gravidade de Doença , Estatística como Assunto , Inquéritos e Questionários , Resultado do TratamentoRESUMO
BACKGROUND: The short-term effects of methylnaltrexone (MNTX), a peripherally acting mu-opioid receptor antagonist, on gastrointestinal and colonic transit remain unclear. AIM: To compare the effects of placebo, codeine, subcutaneous (s.c.) MNTX and codeine with s.c. MNTX on gastrointestinal and colonic transit of solids in healthy humans. METHODS: In a randomized, parallel-group, double-blind, placebo-controlled trial of 48 healthy volunteers, effects of 6 consecutive days of placebo [s.c. and p.o. (orally), n = 8], codeine (p.o. 30 mg q.d.s., n = 8), MNTX (s.c. 0.30 mg/kg, n = 16) and combined MNTX and codeine (same doses and routes, n = 16) on gastrointestinal and colonic transit were assessed. A validated scintigraphic method was used to measure transit during the last 48 h of treatment. Bowel function was estimated during treatment as well as 1 week preceding treatment using standard diaries. Analysis of covariance was used to assess treatment effects. RESULTS: Codeine delayed colonic transit [geometric centre at 24 h (P = 0.04) and ascending colon t(1/2) (P = 0.02)] and reduced stool frequency (P = 0.002), but had no effect on stool form. MNTX did not affect transit, stool frequency or stool form, either alone or with codeine (P > 0.3). No drug interaction effects were detected (P > 0.15). CONCLUSION: Methylnaltrexone does not alter gastrointestinal or colonic transit and does not reverse acute codeine-associated delayed gut transit in health.
Assuntos
Analgésicos Opioides/uso terapêutico , Codeína/uso terapêutico , Trânsito Gastrointestinal/efeitos dos fármacos , Naltrexona/análogos & derivados , Antagonistas de Entorpecentes/uso terapêutico , Receptores Opioides mu/antagonistas & inibidores , Adolescente , Adulto , Método Duplo-Cego , Combinação de Medicamentos , Feminino , Trato Gastrointestinal/efeitos dos fármacos , Humanos , Masculino , Pessoa de Meia-Idade , Naltrexona/uso terapêutico , Compostos de Amônio Quaternário/uso terapêutico , Receptores Opioides mu/uso terapêutico , Adulto JovemRESUMO
BACKGROUND Abnormalities of colonic motility were reported in relatively small studies of patients with lower functional gastrointestinal disorders (FGID) including irritable bowel syndrome (IBS). The influence of gender and body mass on the observed motor pathophysiology is unclear. We sought to compare colonic transit in patients within different lower FGID subgroups and healthy controls, controlling for gender and BMI, and to determine whether BMI independently influences colonic motility. METHODS We evaluated a scintigraphic gastrointestinal and colonic transit database of 287 lower FGID patients associated with constipation (IBS-C, or functional constipation, n = 118), diarrhoea (IBS-D or functional diarrhoea, n = 139) or mixed bowel function (IBS-M, n = 30) and 170 healthy controls. We measured colon filling at 6 h (CF 6 h), and overall colonic transit at 8, 24 and 48 h. KEY RESULTS Colon filling at 6 h did not differentiate health from FGID. Colonic transit was abnormal at 24 h (GC24 of <1.50 or >3.86) in 29.7% of all lower FGID patients. There was a significant overall association between colonic transit and subject group (healthy controls and FGID subgroups) at 8 (P = 0.01), 24 (P < 0.001) and 48 h (P < 0.001) in particular for those with diarrhoea or constipation at 24 and 48 h (P < 0.05), even after adjusting for age, gender and BMI. In addition, BMI was associated with colonic transit after adjusting for age, gender and subject group. CONCLUSIONS & INFERENCES Abnormal transit is documented non-invasively with scintigraphy in 30% of lower FGID patients; transit measurement may help document pathophysiology and inform selection of therapy in lower FGID.
Assuntos
Colo/fisiopatologia , Doenças Funcionais do Colo/fisiopatologia , Trânsito Gastrointestinal/fisiologia , Adulto , Distribuição de Qui-Quadrado , Estudos de Coortes , Constipação Intestinal/fisiopatologia , Feminino , Esvaziamento Gástrico/fisiologia , Humanos , Masculino , Seleção de Pacientes , Fatores SexuaisRESUMO
To clarify the toxic effects and fate of zearalenone (ZEA) in ruminants, we studied histopathological changes and toxicokinetic profiles in goats administered with a single intravenous (iv) injection of ZEA at doses of 2.4 mg/kg bw and 1.2 mg/kg bw, respectively. The expression of the mRNA of estrogen receptor (ER) alpha and beta in tissues was also investigated. The histopathological study revealed that ZEA caused hepatocellular swelling and lymphocytic infiltration in the liver, kidney, and uterus. The expression of ERalpha mRNA was enhanced by ZEA in association with the histopathological changes, indicating the possible involvement of ERalpha in the toxic effects of ZEA. For toxicokinetic profiles, blood plasma, urine, and feces were collected consecutively after iv injection of ZEA and analyzed for ZEA and its metabolites with high performance liquid chromatography (HPLC). alpha-Zearalenol (ZOL) and beta-ZOL were detected with ZEA, but alpha-zearalanol (ZAL), beta-ZAL, and zearalanone were below the detection limits. The distribution half-life (t(1/2alpha)) and elimination half-life (t(1/2beta)) of ZEA were 3.15 and 28.58h, respectively. ZEA, alpha-ZOL, and beta-ZOL were excreted in urine and feces, with beta-ZOL being the predominant metabolite. The ZEA and ZOL in urine were largely in their glucuronide and/or sulphate conjugated forms, while those in feces were largely in their free forms. This study showed the toxic effect of zearalenone and its metabolites, and their pharmacokinetic characteristics in goats.
Assuntos
Estrogênios não Esteroides/toxicidade , Zearalenona/toxicidade , Animais , Doença Hepática Induzida por Substâncias e Drogas/patologia , Cromatografia Líquida de Alta Pressão , Receptor alfa de Estrogênio/biossíntese , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/biossíntese , Receptor beta de Estrogênio/genética , Estrogênios não Esteroides/administração & dosagem , Estrogênios não Esteroides/farmacocinética , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/genética , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Cabras , Meia-Vida , Hidrólise , Injeções Intravenosas , Fígado/patologia , Masculino , RNA/biossíntese , RNA/isolamento & purificação , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Distribuição Tecidual , Inclusão do Tecido , Zearalenona/administração & dosagem , Zearalenona/farmacocinéticaRESUMO
BACKGROUND: The mechanism of action of bisacodyl in the unprepared human colon remains unclear. AIM: To evaluate the effect of oral bisacodyl on the overall and regional colonic transit in humans. METHODS: In a double-blind, randomized, placebo-controlled study of 25 healthy participants, effects of oral bisacodyl (5 mg p.o. per day) and placebo on colonic transit were compared. A validated scintigraphy method was used to measure colonic transit. The primary transit endpoints, ascending colon emptying t(1/2) and geometric centre of colon isotope at 24 h (overall transit), were compared (Wilcoxon rank sum test). RESULTS: There were significant treatment effects on ascending colon t(1/2), with the bisacodyl group demonstrating accelerated emptying [median 6.5 h, interquartile range 5.0-8.0 h] relative to the placebo group [11.0 h (7.0-17.1); P = 0.03]. Numerical differences in colonic geometric centre 24 h [bisacodyl median 3.0 (2.2-3.8), placebo 4.0 (3.1-4.6)] were not significant (P = 0.19). There were no significant differences observed in geometric centre 4 h. CONCLUSION: Oral 5 mg bisacodyl accelerates ascending colon in the unprepared colon in healthy adults; this action may contribute to the drug's efficacy in constipation.
Assuntos
Bisacodil/uso terapêutico , Catárticos/uso terapêutico , Colo Ascendente/efeitos dos fármacos , Trânsito Gastrointestinal/efeitos dos fármacos , Adolescente , Adulto , Colo Ascendente/diagnóstico por imagem , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cintilografia , Resultado do Tratamento , Adulto JovemRESUMO
Several hundred thousand primordial follicles are present in the mammalian ovary, however, only a limited number develop to the pre-ovulatory stage, and then finally ovulate. The others, more than 99%, will be eliminated through a degenerative process called 'atresia'. The endocrinological regulatory mechanisms involved in follicular development and atresia have been characterized to a large extent, but the precise temporal and molecular mechanisms involved in the regulation of these events have remained unknown. From many recent studies, it is suggested that the apoptosis in ovarian granulosa cells plays a crucial role in follicular atresia. Notably, death ligand-receptor interaction and subsequent intracellular signalling have been demonstrated to be the key mechanisms regulating granulosa cell apoptosis. In this review, we provide an overview of granulosa cell apoptosis regulated by death ligand-receptor signalling. The roles of death ligands and receptors [Fas ligand (FasL)-Fas, tumour necrosis factor (TNF)alpha-TNF receptor (TNFR), and TNFalpha-related apoptosis-inducing ligand (TRAIL)-TRAIL receptor (TRAILR)] and intracellular death-signal mediators [Fas-associated death domain protein (FADD), TNF receptor 1-associated death domain protein (TRADD), caspases, apoptotic protease-activating factor 1 (Apaf1), TNFR-associated factor 2 (TRAF2), and cellular FLICE-like inhibitory protein (cFLIP), etc.] in granulosa cells will be discussed.
Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/fisiologia , Atresia Folicular/fisiologia , Células da Granulosa/fisiologia , Suínos , Animais , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/metabolismo , Proteína Ligante Fas/metabolismo , Proteína de Domínio de Morte Associada a Fas/metabolismo , Feminino , Células da Granulosa/citologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Folículo Ovariano , Receptores de Morte Celular/fisiologia , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/fisiologiaRESUMO
Although the use of organs from alpha1,3-galactosyltransferase gene knockout pigs may prolong xenograft survival, resulting in overcoming antibody-mediated hyperacute rejection, pig xenografts will be destroyed directly by cell-mediated immunity, such as NK cells, macrophages, and CD8+ cytotoxic T lymphocytes (CTLs). Therefore, conquering cell-mediated immunity, especially of human CD8+ CTLs, is of particular importance to the success of long-term xenograft survival. We have previously reported that the cytotoxicity of human CD8+ CTLs is strong against pig endothelial cells (PEC) and mediated in major part by the Fas/FasL apoptotic pathway. Cellular FLICE inhibitory protein (c-FLIP) was originally identified as a potent inhibitor of death-receptor signaling through binding competition with caspase-8 for recruitment to Fas-associated via death domain (FADD). Two major c-FLIP variants result from alternative mRNA splicing: a short, 26-kDa protein (c-FLIP S) and a long, 55-kDa form (c-FLIP L). The present study demonstrated that overexpression of c-FLIP S/L genes in PEC markedly suppressed human CD8+ CTL-mediated xenocytotoxicity; moreover, the cytoprotective effects of c-FLIP L appeared to be significantly stronger than those of c-FLIP S. Furthermore, to prove the in vivo prolongation effects of xenograft survival, we transplanted PEC transfectants with c-FLIP(S/L) genes under the rat kidney capsule. Prolonged survival was displayed by xenografts of FLIP S/L PEC transfectants, whereas xenografts of parental PEC were completely rejected by day 5 posttransplantation. Thus, intracellular blocking of death receptor-mediated apoptotic signals by overexpression of c-FLIP S/L in xenograft cells may prevent innate cellular attacks against xenografts opening the window of opportunity for long-term xenograft survival.
Assuntos
Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/imunologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T Citotóxicos/imunologia , Transplante Heterólogo/imunologia , Animais , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Linhagem Celular , Sobrevivência Celular/imunologia , Fragmentação do DNA , Endotélio Vascular , Expressão Gênica , Humanos , Plasmídeos , SuínosRESUMO
Fingertip blood flow (FTBF) as measured by laser Doppler flowmetry (LDF) measurement is considered an indicator of sympathetic nerve function. We evaluated autonomic function in patients with irritable bowel syndrome (IBS) by assessing FTBF with both LDF and continuous-wave (cw) Doppler sonography. Firstly, the two methods were compared in 40 healthy volunteers. Next, 59 patients with IBS as well as 118 healthy volunteer controls were studied. In the supine position, FTBF in the right index finger was measured with cw Doppler sonography, whereas FTBF in the left index finger was assessed with LDF. After baseline measurement for at least 5 min, the volunteers received sympathetic stimulation from cold stress applied without notification in the form of an icebag (0 degrees C) upon the left forearm for 1 min. The new cw Doppler sonography method can be used in place of the old LDF method for clinical purposes. FTBF velocity before stimulation (V(pre)) was significantly lower in the IBS group than that in the healthy volunteers (P < 0.01). In addition, the time required for FTBF to return to V(pre) after stimulation was significantly longer in the IBS group than that in the control group. (P = 0.02). Thus, measurement of FTBF with cw Doppler sonography can be useful in the assessment of sympathetic nerve function. The IBS patients showed an abnormal FTBF response suggesting the presence of excess sympathetic activity.
Assuntos
Doenças do Sistema Nervoso Autônomo/fisiopatologia , Dedos/irrigação sanguínea , Síndrome do Intestino Irritável/fisiopatologia , Adolescente , Adulto , Sistema Nervoso Autônomo/irrigação sanguínea , Sistema Nervoso Autônomo/fisiologia , Doenças do Sistema Nervoso Autônomo/diagnóstico , Velocidade do Fluxo Sanguíneo/fisiologia , Feminino , Humanos , Síndrome do Intestino Irritável/diagnóstico , Fluxometria por Laser-Doppler/métodos , Masculino , Pessoa de Meia-IdadeRESUMO
To clarify whether enzymes involved in aflatoxin B1 (AFB1) metabolism in pigs respond to antioxidant agents, the effect of feeding piglets with diets containing green tea extracts (Sunphenon) and coumarin on in vitro AFB1 metabolism by their liver and intestinal tissues was studied. The results showed that coumarin reduced AFB1-DNA adduct formation by both liver and intestinal microsomes, while Sunphenon did not have any effects. Both coumarin and Sunphenon enhanced the glutathione S-transferase (GST) activity to conjugate AFB1 to glutathione GSH in the intestine, although no effects were noted in the liver. Changes of the expression of mRNA of GSTA2 and GSTO1 were not in parallel with the observed changes of GST activity, suggesting that other GST subtypes are involved in the GST activity toward AFB1. As for lipophilic-free AFB1 metabolites, coumarin reduced the liver microsomal conversion of AFB1 to aflatoxin M1 (AFM1) and aflatoxin Q1 (AFQ1), but Sunphenon exerted no effects. Both coumarin and Sunphenon enhanced the conversion of AFB1 to aflatoxicol in the liver. All the results suggest that feeding with a diet containing coumarin affects AFB1 metabolism to enhance AFB1 detoxification through the suppression of P450 enzyme activity in the liver and the enhancement of GST activity in the intestine. Feeding with a diet containing Sunphenon enhances AFB1 detoxification, but the effects are noted mainly in the intestine.
Assuntos
Aflatoxina B1/metabolismo , Camellia sinensis/química , Cumarínicos/farmacologia , Extratos Vegetais/farmacologia , Ração Animal , Animais , Antioxidantes/farmacologia , Adutos de DNA , Dieta/veterinária , Feminino , Regulação da Expressão Gênica , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Mucosa Intestinal/metabolismo , Fígado/metabolismo , Fitoterapia , Extratos Vegetais/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade por Substrato , SuínosRESUMO
The proto-oncoproteins ETS1 and growth factor independent-1 (GFI1) are implicated in cell growth and differentiation in various types of cells, and their deregulated expression is involved in malignant transformation. Here, we report that ETS1 and GFI1 interact and affect gene expression through their cross-talk. Co-immunoprecipitation analyses and glutathione-S-transferase pull-down assays revealed that ETS1 bound directly to GFI1 via its Ets domain, and GFI1 bound to ETS1 via its zinc-finger domain. Luciferase (Luc) assays using artificial reporters showed that GFI1 repressed ETS1-mediated transcriptional activation and ETS1 repressed GFI1-mediated transcriptional activation, in a dose-dependent manner. However, in the Bax promoter where the Ets- and Gfi-binding sites (EBS and GBS) are adjacent, ETS1 and GFI1 cooperatively reduced activation. Site-directed mutagenesis on the EBS and GBS of the Bax promoter showed that both binding sites were necessary for full repression. Chromatin immunoprecipitation analyses confirmed that an ETS1-GFI1 complex formed on the Bax promoter even when either EBS or GBS was mutated. Introduction of small interfering RNA against ETS1 and/or GFI1 enhanced endogenous Bax gene expression. Our results suggest that the interaction between ETS1 and GFI1 facilitates their binding to specific sites on the Bax promoter and represses Bax expression in vivo.
Assuntos
Proteínas de Ligação a DNA/metabolismo , Proteína Proto-Oncogênica c-ets-1/metabolismo , Fatores de Transcrição/metabolismo , Proteína X Associada a bcl-2/genética , Sítios de Ligação , Células Cultivadas , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica , Humanos , Luciferases/genética , Luciferases/metabolismo , Complexos Multiproteicos , Mutação , Regiões Promotoras Genéticas , Proteína Proto-Oncogênica c-ets-1/genética , Interferência de RNA , Linfócitos T/metabolismo , Fatores de Transcrição/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
Human CD8+ cytotoxic T lymphocyte (CTL)-mediated cytotoxicity in xenograft recipients is an important obstacle for successful xenotransplantation of pig organs into humans. In our previous study, we demonstrated that xenocytotoxicity of human CD8+ CTL detrimental to pig endothelial cells (PEC) is mediated mainly by the Fas/FasL apoptotic pathway. Furthermore, we developed new methods to prevent this CTL killing by extracellular remodeling using overexpression of human decoy Fas antigen and membrane-bound human FasL on pig xenograft cells. The cellular FLICE-inhibitory protein (c-FLIP), a caspase-8 inhibitor that lacks the cysteine domain, is a negative regulator of death receptor-mediated apoptosis. c-FLIP proteins exist as long (c-FLIP(L)) and short (c-FLIPs) splice variants, both capable of protecting cells from death receptor-mediated apoptosis. In this report, we have demonstrated that both pig c-FLIPs and pig c-FLIP(L) significantly inhibit human CD8+ CTL-mediated xenocytotoxicity toward stably transfected PEC, although the expression level of pig Fas antigen on cell surface was not changed. These data suggested that intracellular remodeling with overexpression of pig c-FLIP in xenograft cells may decrease the innate cellular responses against xenografts, facilitating long-term xenograft survival.
Assuntos
Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD/genética , Citotoxicidade Imunológica , Endotélio Vascular/imunologia , Linfócitos T Citotóxicos/imunologia , Animais , Linhagem Celular , Primers do DNA , DNA Complementar , Endotélio Vascular/citologia , Endotélio Vascular/fisiologia , Regulação da Expressão Gênica/imunologia , Sobrevivência de Enxerto , Humanos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos , Transfecção , Transplante Heterólogo/imunologiaAssuntos
Endoscopia do Sistema Digestório/instrumentação , Endoscopia do Sistema Digestório/métodos , Endossonografia/métodos , Pólipos Intestinais/cirurgia , Síndrome de Peutz-Jeghers/cirurgia , Adolescente , Humanos , Pólipos Intestinais/patologia , Intestino Delgado/diagnóstico por imagem , Intestino Delgado/cirurgia , Masculino , Síndrome de Peutz-Jeghers/patologiaRESUMO
BACKGROUND: Lower gastrointestinal bleeding is a frequent cause of hospitalization, but diagnostic methods for this condition are not fully established. Transabdominal ultrasound is a widely accepted diagnostic tool in bowel diseases. AIM: To evaluate the usefulness of transabdominal ultrasound for lower gastrointestinal bleeding. METHODS: We reviewed the medical records of consecutive patients who underwent transabdominal ultrasound as the first diagnostic procedure for acute haematochezia during the period June 1999 to June 2004. The study group comprised 111 patients and all underwent colonoscopy thereafter. Detection and diagnosis of lower gastrointestinal bleeding by ultrasonographic examination were evaluated by comparing the ultrasound diagnosis with the colonoscopic findings and final diagnosis. RESULTS: The bleeding site was localized by colonoscopy in 90 of the 111 patients (81%). The bleeding site was localized by ultrasound in 59 of the 90 patients (66%). When the bleeding site was in the rectum, ultrasonographic detectability was 30% (10/33); ultrasonographic detectability was 82-100% when the bleeding site was elsewhere. Rectal bleeding and diverticular bleeding were difficult to diagnose by ultrasound, but for the other diseases, diagnosis by ultrasonographic examination was possible in 91-100% of cases. CONCLUSIONS: Ultrasonographic examination may be an effective screening method for lower gastrointestinal bleeding.