RESUMO
During orthodontic tooth movement, osteoclasts resorb the alveolar bone at the compress side of periodontium. Reactive oxygen species (ROS) works as intracellular signaling molecules of RANKL during osteoclastogenesis, although ROS has cytotoxicity against cells such as lipid oxidation. To deal with oxidative stress, cells have a defense system that is scavenging ROS by augmented antioxidative stress enzymes via transcriptional regulation with nuclear factor E2-related factor 2 (Nrf2). Previously, we reported that augmented antioxidative stress enzymes by Nrf2-gene transfer inhibited bone destruction. In the present study, we examined the effects of Nrf2 activation on osteoclastogenesis and, thereby, orthodontic tooth movement and orthodontic relapse. Mouse macrophage cell line RAW264.7 cells were used as osteoclast progenitor cells and stimulated with recombinant RANKL (100 ng/mL) with or without Nrf2 activator sulforaphane (SFN) and epigallocatechin gallate (EGCG) or ROS scavenger catechin. Osteoclastogenesis, resorption activity, and osteoclast marker gene expression were examined. Intracellular ROS was analyzed by flow cytometry. Maxillary first molars of C57BL6 male mice were moved palatally with 0.012-inch NiTi wire (100-mN force); SFN or EGCG was injected into the palatal gingiva once a week; and phosphate buffered saline was injected on the contralateral side. Tooth movement was monitored using a stone model with precise impression, and the amount of the tooth movement was compared among groups. SFN and EGCG significantly, but catechin weakly, inhibited RANKL-mediated osteoclastogenesis in vitro. Western blot analysis revealed that SFN and EGCG augmented the nuclear translocation of Nrf2 and the expression of anti-oxidative stress enzymes such as HO-1, although catechin did not. SFN and EGCG significantly, but catechin weakly, attenuated the intracellular ROS. Finally, animal experiment revealed that both SFN and EGCG successfully inhibited the orthodontic tooth movement. Additionally, SFN inhibited the relapse. These results suggest that Nrf2 activation could be therapeutic target for the anchorage enforcement in orthodontic treatment and pharmacologic retention against relapse.
Assuntos
Fator 2 Relacionado a NF-E2/fisiologia , Osteoclastos/fisiologia , Técnicas de Movimentação Dentária/métodos , Animais , Antioxidantes/farmacologia , Catequina/análogos & derivados , Catequina/farmacologia , Linhagem Celular , Sequestradores de Radicais Livres/farmacologia , Heme Oxigenase-1/análise , Isotiocianatos/farmacologia , Macrófagos/efeitos dos fármacos , Masculino , Proteínas de Membrana/análise , Camundongos , Camundongos Endogâmicos C57BL , Fator 2 Relacionado a NF-E2/efeitos dos fármacos , Osteoclastos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ligante RANK , Espécies Reativas de Oxigênio/análise , Espécies Reativas de Oxigênio/antagonistas & inibidores , Recidiva , Transdução de Sinais/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , SulfóxidosRESUMO
Thirty-eight women with Takayasu arteritis were studied using thallium-201 stress myocardial scintigraphy to assess the prevalence and pathophysiology of the perfusion abnormality. Twenty (53%) had abnormal scintigraphic findings (group A). Abnormal scans were divided into 3 groups: permanent defects in 6, reversible defects in 7 and slow washout in 7. The remaining 18 patients had normal scintigrams (group N). Group A had a tendency to be older and to have a high prevalence of complicated significant aortic regurgitation. Interventricular thickness plus left ventricular posterior wall thickness (26 +/- 7 vs 17 +/- 2 mm, p less than 0.01) and left ventricular mass (267 +/- 121 vs 133 +/- 39 g, p less than 0.01) were all greater in group A on echocardiography. The mean value of the central aortic pressure in systole was 170 +/- 15 mm Hg in the 7 catheterized patients in group A. Coronary ostial stenoses were present in 2 group A patients who showed reversible defects on scintigrams. These data indicate that the abnormal perfusion detected by imaging in patients with Takayasu arteritis was responsible for a decrease in coronary reserve or myocardial damage, or both, due to long-standing systemic hypertension or aortic regurgitation. Coronary artery disease should be considered if a reversible defect is present.
Assuntos
Arterite de Takayasu/diagnóstico por imagem , Radioisótopos de Tálio , Tomografia Computadorizada de Emissão , Adulto , Fatores Etários , Idoso , Pressão Sanguínea/fisiologia , Doença das Coronárias/diagnóstico por imagem , Doença das Coronárias/fisiopatologia , Dipiridamol/farmacologia , Ecocardiografia , Ecocardiografia Doppler , Eletrocardiografia , Teste de Esforço , Feminino , Ventrículos do Coração/patologia , Humanos , Pessoa de Meia-Idade , Arterite de Takayasu/fisiopatologia , Função Ventricular Esquerda/fisiologiaRESUMO
A poly(3-hydroxybutyric acid) (PHB) depolymerase gene of Comamonas testosteroni YM1004 was cloned on Sau3AI fragment from genomic DNA into Escherichia coli DH5. Nucleotide sequence analysis dedicated a 1539 bp open reading frame encoding a protein 513 amino acid with a putative 25 residue signal peptide for secretion. The deduced amino acid sequence was very similar to that of PHB depolymerase of Comamonas sp. In order to understand the characteristics of substrate-binding domain of the depolymerase, we constructed its glutathione S-transferase (GST) fusion protein and investigated the ability of adsorption on PHB single crystals by using gold-conjugated antibody and transmission electron microscopy. The fusion protein adsorbed on PHB single crystals tightly and homogeneously, suggesting that binding domain contributes to the adsorption of enzyme on solid PHB without site specificity.
Assuntos
Hidrolases de Éster Carboxílico/química , Hidrolases de Éster Carboxílico/genética , Bacilos e Cocos Aeróbios Gram-Negativos/enzimologia , Bacilos e Cocos Aeróbios Gram-Negativos/genética , Sequência de Aminoácidos , Hidrolases de Éster Carboxílico/metabolismo , Clonagem Molecular , DNA Bacteriano/análise , Glutationa Transferase/genética , Imuno-Histoquímica , Microscopia Eletrônica , Dados de Sequência Molecular , Ligação Proteica/fisiologia , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/metabolismo , Análise de Sequência de DNARESUMO
Serum amyloid P component (SAP) is a normal plasma constituent that is observed both in senile plaque and in neurofibrillary tangle in brains of patients with Alzheimer's disease (AD). In this study, we evaluated the SAP levels in cerebrospinal fluid (CSF) of 72 patients with AD, 11 frontotemporal dementia and nine normal control subjects. There was no significant difference in the SAP levels between the AD group and other groups. However, among AD patients, cognitive function was rated using the Mini-Mental State Examination and was correlated with the SAP level (R = 0.38, P < 0.05). Our results suggest that measurement of the SAP levels in CSF can be useful for assessing the degree of cognitive impairment in AD patients.
Assuntos
Doença de Alzheimer/líquido cefalorraquidiano , Componente Amiloide P Sérico/líquido cefalorraquidiano , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/psicologia , Cognição , Demência/líquido cefalorraquidiano , Humanos , Entrevista Psiquiátrica Padronizada , Pessoa de Meia-Idade , Valores de Referência , Proteínas tau/líquido cefalorraquidianoRESUMO
Cyclin D1 overexpression is remarkably frequent in several human carcinomas and is believed to be a critical event in oncogenesis. We examined cyclin D1 expression, p53 expression, and the Ki-67 labeling index by immunostaining in human gallbladder mucosa in conditions varying from normal to malignant tissue. We also examined K-ras codon 12 mutations in these tissues with a two-step polymerase chain reaction. Nuclear cyclin D1 overexpression was observed in 48% of carcinomas occurring independently of adenoma, but not in adenomas, carcinomas arising in adenomas, or nonneoplastic lesions. Cytoplasmic cyclin D1 overexpression was observed in about 15% of abnormal specimens, irrespective of the type of epithelial abnormality. Carcinomas showing nuclear cyclin D1 overexpression had significantly higher Ki-67 labeling indexes than those with no overexpression. Moderately to poorly differentiated adenocarcinomas showed a higher incidence of nuclear cyclin D1 overexpression than papillary to well differentiated carcinomas. Specimens with cyclin D1 overexpression showed a high incidence of lymph permeation, venous permeation, and lymph node metastasis. We conclude that nuclear cyclin D1 overexpression is a critical event importantly associated with cell proliferation and invasive growth in gallbladder carcinogenesis, and that cyclin D1 immunostaining may become a useful marker for evaluating gallbladder carcinomas.
Assuntos
Adenocarcinoma/metabolismo , Adenoma/metabolismo , Núcleo Celular/metabolismo , Ciclina D1/biossíntese , Neoplasias da Vesícula Biliar/metabolismo , Adenocarcinoma/genética , Adenocarcinoma/patologia , Adenoma/genética , Adenoma/patologia , Biomarcadores Tumorais , Divisão Celular , Códon , DNA de Neoplasias/análise , Neoplasias da Vesícula Biliar/genética , Neoplasias da Vesícula Biliar/patologia , Genes ras/genética , Humanos , Técnicas Imunoenzimáticas , Antígeno Ki-67/metabolismo , Mutação Puntual , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Proteína Supressora de Tumor p53/biossínteseRESUMO
Alkanediazohydroxides are common key intermediates in carcinogenesis and mutagenesis of N-nitroso compounds, which are widely found in human environment. Mutagenicity of (E)- and (Z)-potassium alkanediazotates, as precursors of corresponding alkanediazohydroxides were evaluated to investigate the effect of geometric isomerism and also the effect of alkyl groups on their biological activity. Mutagenicity of N-nitroso-N-alkylureas which spontaneously produce alkanediazohydroxides after non-enzymatic hydrolysis were also tested in comparison to that of the corresponding diazotates and other activated chemical species of N-nitrosamines. When the mutagenicity was assayed in three microbial strains, Salmonella typhimurium TA1535, and Escherichia coli WP2 and WP2 uvrA, the order of mutagenic potency of the compounds with the same alkyl group was as follows; (E)-diazotates > (Z)-diazotates > nitrosoureas. The effect of alkyl groups on the mutagenic potency was different in Salmonella strain and in E. coli strains, and this result could be explained by the efficiency of O6-alkylguanine-DNA alkyltransferase. In each bacterial strain, this effect of alkyl groups was similar in mutagenicity induced by (E)- and (Z)-diazotates, N-nitroso-N-alkylureas and other activated N-nitrosodialkylamines such as alpha-hydroxy nitrosamines. The geometrical isomerism affected the mutagenicity of (E)- and (Z)-potassium alkanediazotates, and the result suggested that alkanediazohydroxides react through diazonium ions in a cage rather than through free alkyldiazonium ions which have no geometrical isomerism. Our results confirmed that (E)-potassium alkanediazotates, (Z)-potassium alkanediazotates and N-nitroso-N-alkylureas all decomposed through diazohydroxides, and that alkanediazohydroxides are the active alkylating species of N-nitroso compounds, and also that the geometrical isomerism is important for carcinogenic N-nitroso compounds to show their biological activity.
Assuntos
Alquilantes/farmacologia , Compostos Azo/farmacologia , Carcinógenos/farmacologia , Testes de Mutagenicidade , Mutagênicos/farmacologia , Compostos Nitrosos/farmacologia , Poluentes Ambientais , Escherichia coli/efeitos dos fármacos , Humanos , Isomerismo , Salmonella typhimurium/efeitos dos fármacosRESUMO
BACKGROUND: Although frozen section is recommended to prevent tumor dissemination following laparoscopic cholecystectomy (LC) for unsuspected gallbladder cancer, there are no reports concretely demonstrating its effectiveness and outcome. METHODS: Frozen section during LC was performed in 990 patients with gallstones. The sensitivity, specificity of frozen section, and false-negative cases were evaluated in comparison with postoperative entire cross sections. RESULTS: In frozen section, 983 cases were diagnosed as benign and 7 cases as malignant. Of the benign cases, cancer was discovered in 4 patients postoperatively in which frozen section was diagnosed as regenerative epithelial severe atypia. Sensitivity was 64% and specificity was 100%. Concerning the results of frozen section by p-TNM classification, cancer was diagnosed in 40% of Tis lesions, whereas it was found in 83% of T2 or T3 lesions. CONCLUSION: Frozen section is effective in cases with T2 or greater lesions for which conversion to radical surgery should be required.
Assuntos
Colecistectomia Laparoscópica/métodos , Secções Congeladas/métodos , Neoplasias da Vesícula Biliar/diagnóstico , Neoplasias da Vesícula Biliar/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Secções Congeladas/normas , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
The intraperitoneal injection of glycogen in the mouse resulted, shortly thereafter, in the accumulation of 14-23 million neutrophils in the peritoneal cavity and a four-fold increase in the numbers of circulating neutrophils. Preceding the influx of leukocytes, the exudation of plasma proteins and the chemotactic activity for mouse neutrophil in vitro increased in the peritoneal fluid. Among various protease inhibitors examined, chymostatin alone suppressed the plasma protein exudation. Indomethacin and dexamethasone reduced the accumulation of white cells and protein exudation. These nonsteroidal and steroidal antiinflammatory drugs were equally effective whether given simultaneously with or 60 min before glycogen or whether administered intraperitoneally or orally. Colchicine showed a suppressive effect on the leukocyte accumulation but enhanced the protein exudation.
Assuntos
Anti-Inflamatórios/uso terapêutico , Glicogênio/farmacologia , Inflamação/induzido quimicamente , Inibidores de Proteases/uso terapêutico , Animais , Glicogênio/administração & dosagem , Glicogênio/metabolismo , Inflamação/tratamento farmacológico , Inflamação/patologia , Inflamação/fisiopatologia , Cinética , Camundongos , Fatores de TempoRESUMO
The kinetics of adsorption and hydrolysis by an extracellular PHB depolymerase from Alcaligenes faecalis were studied at 37 degrees C on the surface of five types of polyhydroxyalkanoate (PHA) films. The films of poly[(R)-3-hydroxybutyrate] (P(3HP)), poly(3-hydroxypropionate) (P(3HP)), and poly(4-hydroxybutyrate)(P(4HB)) were hydrolyzed by the enzyme, while the films of poly[(S)-2-hydroxypropionate)(P(2HP)) and poly(6-hydroxyhexanoate)(P(6HH)) were not eroded. The PHB depolymerase with binding and catalytic domains adsorbed on the surface of all PHA films used, and the adsorption kinetics were found to obey the Langmuir isotherm. The cross-area per one molecule of enzyme binding to the surface of PHA film was estimated to be 17 +/- 8 (nm2/molecule). It has been concluded that the binding domain of enzyme is non-specific for the binding to the surface of PHA film, while the active site in a catalytic domain is specific for the hydrolysis of PHA molecules.
Assuntos
Hidrolases de Éster Carboxílico/química , Poliésteres/química , Adsorção , Alcaligenes/enzimologia , Sítios de Ligação , Hidrólise , Hidroxibutiratos/química , Cinética , Pseudomonas/enzimologia , Fatores de TempoRESUMO
The substrate specificities of three extracellular polyhydroxybutyrate (PHB) depolymerases from Alcaligenes faecalis (PhaZ Afa), Pseudomonas stutzeri (PhaZ Pst), and Comamonas acidovorans (PhaZ Cac), which are grouped into types A and B based on the position of a lipase box sequence in the catalytic domain, were examined for films of 12 different aliphatic polyesters. Each of these PHB depolymerases used was capable of hydrolyzing poly(3-hydroxybutyrate) (P(3HB)), poly(3-hydroxypropionate) (P(3HP)), poly(4-hydroxybutyrate) (P(4HB)), poly(ethylene succinate) (PESU), and poly(ethylene adipate) (PEA) but could not hydrolyze another seven polyesters. In addition, the binding characteristics of substrate binding domains from PhaZ Afa, PhaZ Cac, and PHB depolymerase from Comamonas testosteroni (PhaZ Cte) were studied by using fusions with glutathione S-transferase (GST). All of fusion proteins adsorbed strongly on the surfaces of polyester granules of P(3HB), P(3HP), and poly(2-hydroxypropionate) (P(2HP)) which was not hydrolyzed by the PHB depolymerases used in this study, while they did not bind on Avicel and chitin granules. The adsorption kinetics of the fusion proteins to the surface of P(3HB) and P(2HP) granules were found to obey the Langmuir isotherm. The cross-area per molecule of fusion protein bound to P(3HB) granules was estimated to be 12+/-4 nm2/molecule. It has been suggested that the active sites in catalytic domains of PHB depolymerases have a similar conformational structure, and that several amino acids in substrate-binding domains of PHB depolymerases interact specifically with the surface of polyesters.
Assuntos
Alcaligenes/enzimologia , Hidrolases de Éster Carboxílico/metabolismo , Bacilos e Cocos Aeróbios Gram-Negativos/enzimologia , Pseudomonas/enzimologia , Adsorção , Sequência de Aminoácidos , Sítios de Ligação , Hidrolases de Éster Carboxílico/química , Hidrolases de Éster Carboxílico/genética , Hidrólise , Hidroxibutiratos/metabolismo , Dados de Sequência Molecular , Poliésteres/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Homologia de Sequência de Aminoácidos , Especificidade por SubstratoRESUMO
A microdialysis sampling technique for the intracerebral measurement of somatostatin (SS) in extracellular fluid was examined in the goat. The microdialysis probe (70-mm shaft, 0.5 mm outer diameter) contained at its tip a 4-mm length of copolymer dialysis membrane (20 kDa cut-off). Artificial cerebrospinal fluid (artificial CSF) was pumped through the probe tip at a rate of 4 microliters/min with a batter-driven syringe pump, and effluent fractions of dialysate (120 microliters) were collected every 30 min. An in vitro recovery test showed that changes in the SS concentration in dialysate were highly correlated (r = 0.95, P < 0.01) with those in the external medium, and the relative recovery averaged 2.0%. As a validation for in vivo microdialysis, trails were conducted with conscious behaving goats wherein the inflow dialysate was changed transiently from artificial CSF with low potassium (2.5 mM) to a solution of 300 mM KCl. Potassium-induced depolarization around the probe tip located in the preoptic area and in the hypothalamus induced an increase in SS concentrations in dialysate at each location. In the most remarkable response, the concentrations of SS were increased 6-fold and 11-fold in the first and second 30-min fractions, respectively, compared with prepotassium concentrations. These results suggest that intracerebral SS levels in extracellular fluid could be estimated from conscious behaving goats by the use of our intracerebral microdialysis system.
Assuntos
Cabras/metabolismo , Hipotálamo/química , Microdiálise/veterinária , Área Pré-Óptica/química , Somatostatina/análise , Animais , Feminino , Hipotálamo/metabolismo , Hipotálamo/fisiologia , Microdiálise/métodos , Microdiálise/normas , Potássio/farmacologia , Área Pré-Óptica/metabolismo , Área Pré-Óptica/fisiologia , Radioimunoensaio/veterinária , Somatostatina/metabolismoRESUMO
We performed the aortic stop flow and hypoxic perfusion chemotherapy for unresectable gallbladder cancer under general anesthesia and made the perfusion only in the abdominal cavity by clamping the abdominal aorta and inferior vena cava near the site of diaphragma. Although temporary hypertension occurred just after clamping the aorta, no severe problem could be seen during operation. After surgery the patient had no abnormalities such as liver or kidney dysfunction, and tumor markers gradually came to decrease. Furthermore, the effectiveness of this therapy is greatly anticipated for the unresectable abdominal cancer, evaluating the selection of anti-cancer drugs and their administration dosage etc.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Quimioterapia do Câncer por Perfusão Regional/métodos , Neoplasias da Vesícula Biliar/tratamento farmacológico , Anestesia Geral , Aorta Abdominal , Fluoruracila/administração & dosagem , Neoplasias da Vesícula Biliar/patologia , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Mitomicina/administração & dosagem , Oxigênio/fisiologiaRESUMO
Ten patients with liver metastases from advanced gastric cancer received percutaneous ethanol injection therapy (PEI) and chemotherapy by hepatic arterial infusion (HAI) via implantable reservoir. A 90% ethanol solution including 10%. Lipiodol was injected in the liver as PEI.5-FU, EPIR and MMC were used as the regimen for HAI chemotherapy. We have performed this therapy (PEI + HAI) for ten patients with liver metastases since February, 1997. These patients have received this therapy for 4-36 months and three patients died within 16 months. However, three patients did not develop any liver failure after this therapy. The median survival rate was 25.2 months. There are statistically significant differences between upto ss and over se of invasion, and between INF alpha and gamma (p = 0.005).
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Etanol/administração & dosagem , Bombas de Infusão Implantáveis , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/secundário , Neoplasias Gástricas/patologia , Idoso , Epirubicina/administração & dosagem , Fluoruracila/administração & dosagem , Artéria Hepática , Humanos , Infusões Intra-Arteriais , Injeções Intralesionais , Óleo Iodado/administração & dosagem , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Mitomicina/administração & dosagem , Taxa de SobrevidaRESUMO
There are few reports about the methods, amounts, and kinds of dosage about intermittent intra-arterial chemotherapy of liver metastases from primal pathological type's squamous cell carcinoma. Because they are less than liver metastases from adenocarcinoma of colon or stomach. Although it is important of other factors about the operative method of primary focus and metastases of the other parts, it is possible that those cases obtained the good prognosis and protected liver failure, if those liver metastases could be controlled well. In our department from January 1987 to December 1989, 9 cases of inoperative liver metastases of squamous cell carcinoma (esophagus: 4 cases, larynx: 3 cases and cervix of uterus 2 cases) were treated of intra-arterial infusion chemotherapy of FAM (5Fu 500 mg/week, ADM 30 mg/4 weeks and MMC 4 mg/2 weeks) and CDDP methods (only CDDP 10 mg/week). Cases of esophagus carcinoma were treated with FAM method. On the CT-scan one of the cases showed the reduction rate of more than 50% and was a Progressive Response (PC), and the SCC tumor marker decreased in 2 cases. However, 2 other cases died of liver failure. Cases of larynx were treated with FAM and CDDP methods. However, on the CT-scan all of the cases showed No Change (NC) nor decrease in SCC. But thinking of prognosis FAM was better than CDDP. Cases of cervix of uterus were treated with the FAM and CDDP methods. FAM was not different than the CDDP in the prognosis and effect.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Carcinoma de Células Escamosas/tratamento farmacológico , Neoplasias Esofágicas/patologia , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Faríngeas/patologia , Neoplasias Uterinas/patologia , Idoso , Carcinoma de Células Escamosas/secundário , Cisplatino/administração & dosagem , Doxorrubicina/administração & dosagem , Esquema de Medicação , Feminino , Fluoruracila/administração & dosagem , Humanos , Infusões Intra-Arteriais , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Mitomicina , Mitomicinas/administração & dosagemRESUMO
Multifocal, raised, ulcerated firm nodules accompanied by an intussuscepted area were detected in the jejunum of an 8-year-old Holstein cow. The cut surfaces of the nodules were yellow-white. Microscopically, the lamina propria was expanded by an intense infiltration of epithelioid cells, multinucleate giant cells, macrophages, lymphocytes and neutrophils. Numerous bacteria were found within the granulomatous lesions. These were argyrophilic, gram-positive, periodic acid-Schiff-positive, segmented, rarely branched, elongate filamentous bacteria (2-28 µm in length, 0.2-0.35 µm in diameter). Ultrastructurally, a cell wall with an electron-transparent zone was detected. The present pathogen was clearly different from the argyrophilic, gram-negative, non-segmented, filamentous bacterium previously reported in a Holstein cow with jejunal granuloma. Comparative 16S rDNA gene sequencing analysis revealed that the organism was an unpublished species (GenBank accession number AB539875). This is the first report of bovine jejunal granuloma associated with an argyrophilic gram-positive segmented filamentous bacterium.
Assuntos
Enterite/patologia , Infecções por Bactérias Gram-Positivas/microbiologia , Infecções por Bactérias Gram-Positivas/patologia , Infecções por Bactérias Gram-Positivas/veterinária , Doenças do Jejuno/patologia , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/patologia , Enterite/microbiologia , Bactérias Gram-Positivas/genética , Bactérias Gram-Positivas/isolamento & purificação , Doenças do Jejuno/microbiologiaRESUMO
A 26-month-old thoroughbred colt with a 4-month history of continuous diarrhoea and weight loss was subject to necropsy examination. The small intestinal mucosa was thickened and this change particularly affected the terminal ileum. Microscopical examination revealed multifocal epithelial hyperplasia, with multifocal granulomas and marked lymphocytic infiltration of the lamina propria. Numerous gram-negative argyrophilic curved bacilli were observed within the cytoplasm of affected enterocytes. Macrophages and epithelioid cells forming the granulomas had abundant, lightly eosinophilic, foamy cytoplasm, with occasional large, clear vacuoles containing gram-positive coccobacilli. Immunohistochemical studies suggested that the argyrophilic bacilli were Lawsonia intracellularis and the gram-positive coccobacilli were Rhodococcus equi. L. intracellularis-specific DNA fragments were amplified from the affected ileocaecal mucosa by polymerase chain reaction. Virulent R. equi (VapA positive) was isolated in pure culture from the liver and mesenteric lymph nodes. These results suggested that the two intracytoplasmic organisms had induced multifocal proliferative and granulomatous enteritis accompanied by severe and extensive lymphocytic infiltration.
Assuntos
Infecções por Actinomycetales/veterinária , Infecções por Desulfovibrionaceae/veterinária , Doenças dos Cavalos/microbiologia , Lawsonia (Bactéria) , Rhodococcus equi , Infecções por Actinomycetales/microbiologia , Infecções por Actinomycetales/patologia , Animais , Infecções por Desulfovibrionaceae/microbiologia , Infecções por Desulfovibrionaceae/patologia , Doenças dos Cavalos/patologia , Cavalos , Íleo/microbiologia , Íleo/patologia , Imuno-Histoquímica , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Fígado/microbiologia , Fígado/patologia , Reação em Cadeia da PolimeraseAssuntos
Transplante de Células/métodos , Hepatócitos/transplante , Falência Hepática/terapia , Fígado Artificial , Animais , Aspartato Aminotransferases/sangue , Células Cultivadas , Modelos Animais de Doenças , Encefalopatia Hepática/sangue , Encefalopatia Hepática/terapia , Hepatócitos/citologia , Humanos , Falência Hepática/sangue , Antígeno Nuclear de Célula em Proliferação/análise , Ratos , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Phagocytic cells disposing of dying granulosa cells in the atretic ovarian follicles were morphologically studied in guinea pig ovaries at various stages of estrous cycle. Epon embedded semithin sections stained with toluidine blue were observed with a light microscope, and ultrathin sections were examined under a transmission electron microscope. Frozen sections were processed for acid phosphatase histochemistry and MR-1 (a monoclonal antibody against guinea pig macrophages) immunohistochemistry. In animals during the estrus period (days 1 and 2) as well as the second half of the estrous cycle (days 11 and 16), there were numerous mature follicles in which massive groups of granulosa cells were undergoing apoptosis. Two kinds of phagocytic cells were identified in these follicles of the initial stage of atresia: one was intact granulosa cells ingesting neighboring dead granulosa cells, and the other was large round cells identified as macrophages due to their strong acid phosphatase activity and MR-1 immunoreactivity. Mature follicles of the advanced stage of atresia were frequently recognized during the metestrus period (days 4 and 5). Small stellate cells were regarded as surviving granulosa cells, while large round cells showing intense reactions for acid phosphatase and MR-1 were identified as macrophages. This study demonstrates that both intact granulosa cells and macrophages participate in the elimination of apoptotic granulosa cells in atretic mature follicles of the guinea pig ovary, and remain even in the advanced stages of atresia.