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1.
J Anim Sci ; 95(9): 3949-3960, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28992019

RESUMO

The objective of this study was to characterize the composition of the forestomach and fecal microbiota in Japanese Black calves with white scours. Forestomach fluid, feces, and peripheral blood were collected from healthy calves ( = 5; age 10 ± 2 d) and scouring calves ( = 5; age 10 ± 1 d) on the day on which white scours occurred. The pH and concentrations of VFA, lactic acid, and ammonia nitrogen (NH-N) of the forestomach fluids were determined. Microbiota composition and gene copy numbers in the forestomach fluid and feces were analyzed by 454 pyrosequencing and quantitative real-time PCR (qPCR), respectively. The cytokine mRNA level in peripheral leukocytes was evaluated by qPCR. The pH of the forestomach fluid of the scouring calves tended to be higher than that of the healthy calves ( = 0.056). No significant difference was detected in the total VFA, lactic acid, or NH-N concentrations in the forestomach fluids of the 2 groups. Firmicutes, Bacteroidetes, and Proteobacteria were the predominant phyla in the forestomach fluid and feces. At the genus level, the relative abundance of in the forestomach fluid was significantly higher in the scouring calves ( < 0.05) and the relative abundance of in the feces was significantly higher than that in the forestomach in the healthy calves ( < 0.05). Furthermore, the bacterial diversity indices of feces were lower in the scouring calves. Quantitative PCR amplification using some of the primer pairs failed in the forestomach fluid and feces in both groups. These results suggested that fermentation in the forestomach may affect the occurrence of white scours, resulting in changes in the composition and diversity of the forestomach fluid and fecal microbiota in Japanese Black calves.


Assuntos
Bactérias/classificação , Doenças dos Bovinos/microbiologia , Diarreia/veterinária , Fezes/microbiologia , Microbiota , Rúmen/microbiologia , Animais , Bactérias/genética , Bactérias/isolamento & purificação , Bacteroidetes/classificação , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Bovinos , Doenças dos Bovinos/metabolismo , Diarreia/metabolismo , Diarreia/microbiologia , Fermentação , Firmicutes/classificação , Firmicutes/genética , Firmicutes/isolamento & purificação , Suco Gástrico/microbiologia , Proteobactérias/classificação , Proteobactérias/genética , Proteobactérias/isolamento & purificação , Rúmen/metabolismo , Análise de Sequência de DNA/veterinária
2.
Gene Ther ; 16(2): 297-302, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18800152

RESUMO

Adenovirus (Ad) serotype 35 (Ad35) vectors have attracted remarkable attention as alternatives to conventional Ad serotype 5 (Ad5) vectors. In a previous study, we showed that intravenously administered Ad35 vectors exhibited a safer profile than Ad5 vectors in cynomolgus monkeys, which ubiquitously express CD46, an Ad35 receptor, in a pattern similar to that in humans. However, the Ad35 vectors poorly transduced the organs. In this study, we examined the transduction properties of Ad35 vectors after local administration into organs of cynomolgus monkeys. The vectors transduced different types of cells depending on the organ. Hepatocytes and microglia were mainly transduced after the vectors were injected into the liver and cerebrum, respectively. Injection of the vectors into the femoral muscle resulted in the transduction of cells that appeared to be fibroblasts and/or macrophages. Conjunctival epithelial cells showed transgene expression following infusion into the vitreous body of the eyeball. Transgene expression was limited to areas around the injection points in most of the organs. In contrast, Ad35 vector-mediated transgene expression was not detected in any of the organs not injected with Ad35 vectors. These results suggest that Ad35 vectors are suitable for gene delivery by direct administration to organs.


Assuntos
Adenoviridae/genética , Vetores Genéticos/genética , Transdução Genética , Adenoviridae/classificação , Administração Tópica , Animais , Regulação da Expressão Gênica/genética , Vetores Genéticos/administração & dosagem , Injeções , Macaca fascicularis , Proteína Cofatora de Membrana/metabolismo , Distribuição Tecidual/genética , Transgenes/genética , beta-Galactosidase/biossíntese , beta-Galactosidase/genética
3.
Brain Res ; 922(1): 30-41, 2001 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-11730699

RESUMO

Age-related changes in PS-1 localization were examined in the brains of 22 cynomolgus monkeys ranging in age from embryonic day 87 to 35 years. In embryonic monkey brains, anti-PS-1 antibody N12, which recognizes the PS-1 N-terminal fragment (Ntf) and holo protein, stained immature neuronal cells. In juvenile monkeys, N12 stained large pyramidal neurons, cerebral neocortical neurons, and cerebellar Purkinje's cells. Cytoplasmic staining of these cells was granular in appearance. In aged monkeys, N12 stained neurons in all layers of the neocortex. In contrast, regardless of the age of the animals examined, M5, an anti-PS-1 antibody that specifically recognizes only the PS-1 C-terminal fragment (Ctf), stained neurons in all layers of the neocortex and neurons in the cerebellum. M5 also stained neuropil and white matter, and in aged monkeys, M5 stained swollen neurites of mature senile plaques. Age-related changes in PS-1 expression were further examined using Western blot analysis of mitochondrial, myelin, microsomal, nuclear, synaptosomal, and cytosol fractions isolated from 10 monkey brains ranging in age from embryonic day 87 to 32 years. In all brains, Ntf and Ctf were expressed most abundantly in the microsome fraction. The amount of PS-1 in the nuclear fraction dramatically increased with age. We conclude that the transport of PS-1 diminished with age and that PS-1 fragments accumulated in endoplasmic reticulum (ER) associated with the nuclear membrane.


Assuntos
Envelhecimento/metabolismo , Química Encefálica/fisiologia , Encéfalo/crescimento & desenvolvimento , Proteínas de Membrana/metabolismo , Animais , Western Blotting , Encéfalo/embriologia , Centrifugação com Gradiente de Concentração , Feminino , Imuno-Histoquímica , Macaca fascicularis , Emaranhados Neurofibrilares/metabolismo , Neurônios/metabolismo , Neurônios/ultraestrutura , Inclusão em Parafina , Gravidez , Presenilina-1 , Frações Subcelulares/metabolismo
4.
Clin Cancer Res ; 7(12): 4027-32, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11751497

RESUMO

BACKGROUND: The matrix-degrading proteinases are believed to play an important role in the invasion and metastasis of hepatocellular carcinoma (HCC), but no one has ever seen the in situ matrix-degrading activity in HCCs. PURPOSE: To demonstrate the cellular localization of actual gelatinolytic activity and to investigate the invasive potential of human HCC. EXPERIMENTAL DESIGN: HCC cases (30) were subjected to in situ gelatin zymography and SDS-gelatin gel zymogram. RESULTS: In situ gelatin zymography revealed a heterogeneous gelatinolytic activity in HCC cells, as well as stromal cells of noncancerous livers. The gelatinolytic intensity was stronger in 15 HCC nodules than in the corresponding noncancerous livers and was significantly associated with the cancer invasion to the capsule of the HCCs and to the portal veins. An intense gelatinolytic activity was detected in HCC cells in the front of tumor invasion. SDS-gelatin gel zymogram revealed gelatinases A and B that were mostly in latent forms. CONCLUSIONS: The present study demonstrates high gelatinolytic activity at the invasive front of HCCs at a cellular level and that HCC has an invasive potential with the gelatin (matrix)-degrading metalloproteinases. Furthermore, it suggests the importance of the activation mechanism of gelatinolytic enzymes in the invasion and metastasis of HCCs.


Assuntos
Carcinoma Hepatocelular/patologia , Gelatinases/análise , Neoplasias Hepáticas/patologia , Invasividade Neoplásica/patologia , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/cirurgia , Diferenciação Celular , Eletroforese em Gel de Poliacrilamida , Gelatina , Hepacivirus/isolamento & purificação , Humanos , Isoenzimas/análise , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/cirurgia , Neoplasias Hepáticas/virologia , Estadiamento de Neoplasias , Veia Porta/patologia , Células Estromais/patologia
5.
Oncogene ; 20(42): 6095-101, 2001 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-11593417

RESUMO

DNA multiploidy may involve specific DNA ploidy states with respect to genetic alterations such as oncogenes, tumor suppressor gene mutation and microsatellite instability. To clarify the role of DNA multiploidy in colorectal cancer, we analysed allelic imbalance involving the ATM gene, localized to chromosome 11q22-23 and thought to be involved in genetic stability, in a series of multiploid colorectal carcinomas. In addition, p53 gene mutation (exons 5-8) and allelic imbalance at 11q24 loci distal to the ATM locus were also examined. The crypt isolation technique coupled with DNA cytometric sorting and polymerase chain reaction assay using 10 microsatellite markers tightly linked to the ATM gene were used to study ATM allelic imbalance in 55 colorectal carcinomas (15 diploid, 13 aneuploid, 27 multiploid). While allelic imbalance at the ATM locus was rarely observed in diploid and aneuploid carcinomas, multiploid carcinomas exhibited a high frequency of ATM allelic imbalance. In multiploid carcinoma samples, diploid subpopulations showed a smaller range of allelic imbalance at the loci tested compared to aneuploid subpopulations that demonstrated allelic imbalance over a relatively large region. Also, the frequency of AI at 11q24 showed a similar tendency to that at the ATM locus for each DNA ploidy state. An association between p53 gene mutation and ATM allelic imbalance in multiploid carcinoma was also observed. Our results suggest that ATM allelic imbalance and p53 gene mutations occur during the progression from diploid to aneuploid cell populations in multiploid colorectal carcinomas.


Assuntos
Desequilíbrio Alélico , Carcinoma/genética , Neoplasias Colorretais/genética , Poliploidia , Proteínas Serina-Treonina Quinases/genética , Proteínas Mutadas de Ataxia Telangiectasia , Proteínas de Ciclo Celular , DNA de Neoplasias/análise , Proteínas de Ligação a DNA , Frequência do Gene , Genes p53 , Humanos , Mutação , Reação em Cadeia da Polimerase , Proteínas Supressoras de Tumor
6.
Acta Med Okayama ; 55(6): 349-55, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11779097

RESUMO

Hepatic encephalopathy is one of the major complications in decompensated liver cirrhosis. The current study was conducted to clarify the mechanisms of zinc deficiency in liver cirrhosis and its involvement in hepatic encephalopathy via ammonia metabolism. Ten patients each with compensated or decompensated liver cirrhosis and 11 healthy volunteers were enrolled in the study. Serum zinc levels and its daily urinary excretion were measured, an oral zinc-tolerance test was performed to examine zinc malabsorption, and the effects of diuretics on zinc excretion and of zinc supplementation on ammonia metabolism in the skeletal muscle were studied. The mean serum zinc levels in patients with decompensated liver cirrhosis were found to be significantly lower than the levels in controls and patients with compensated liver cirrhosis. The serum zinc levels were inversely correlated with blood ammonia in the fasting state. In the oral zinc-tolerance test, the percent increase in serum zinc levels 120 and 180 min after ingestion was less in cirrhotic patients than in controls. A diuretic administration resulted in a significant reduction in serum zinc levels. An increased uptake of ammonia by and an increased release of glutamine from leg skeletal muscle after oral supplementation of zinc sulfate were evident. Taken together, zinc deficiency in decompensated cirrhotic patients appears to be due to low absorption and to high urinary excretion, for which excessive diuretic administration is, in part, responsible, and zinc supplementation might play an important role in the prevention of hepatic encephalopathy by activating glutamine synthetase.


Assuntos
Amônia/metabolismo , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/metabolismo , Zinco/deficiência , Zinco/uso terapêutico , Amônia/sangue , Glutamina/metabolismo , Humanos , Músculo Esquelético/metabolismo , Zinco/sangue , Zinco/urina
7.
J Agric Food Chem ; 48(11): 5440-3, 2000 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11087498

RESUMO

Suppression of the furylfuramide-induced SOS response by 25 kinds monoterpenoids (hydrocarbons, alcohols, ketones, and aldehydes) with a p-menthane skeleton was studied. Suppression of the SOS-inducing activity by monoterpenoids was determined in the umu test using Salmonella typhimurium TA1535/pSK1002. The terpene alcohols, ketones, and aldehydes had potent suppressive effects, but the hydrocarbons did not. Especially, (+)-menthol, (+)-pulegone, piperitenone, and cuminaldehyde were shown to have the most potent suppressive effects, and the ID(50) (dose for 50% inhibition) was 0.52 micromol/mL.


Assuntos
Furilfuramida/farmacologia , Mentol/análogos & derivados , Mentol/farmacologia , Resposta SOS em Genética/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , Terpenos/farmacologia , Álcoois/farmacologia , Mentol/química , Estrutura Molecular , Testes de Mutagenicidade , Salmonella typhimurium/genética , Relação Estrutura-Atividade , Terpenos/química
8.
J Agric Food Chem ; 48(9): 4377-80, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10995366

RESUMO

The recently isolated paeonol (2-hydroxy-4-methoxyacetophenone), as one of the antimutagenic compounds from Discorea japonica, was used as a lead compound for detailed structure-activity relationship studies. Nine acetophenones (2-hydroxy-4-methoxy, 2-hydroxy-5-methoxy, 2-hydroxy-6-methoxy, 4-hydroxy-3-methoxy, o-methoxy, m-methoxy, p-methoxy, and 2,5-dimethoxyacetophenone and acetophenone) were investigated for their ability of suppression of furylfuramide-induced SOS response using Salmonella typhimurium TA1535/pSK1002 in the umu test, against the mutagen, 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The results showed that 2-hydroxy-6-methoxyacetophenone displayed the strongest activity (EC(50) = 0.6 micromol/mL), and a hydroxyl group at C-2 is necessary feature for acetophenone derivatives to show the suppressive effects of furylfuramide-induced SOS response.


Assuntos
Acetofenonas/farmacologia , Furilfuramida/farmacologia , Resposta SOS em Genética , Salmonella typhimurium/genética
9.
Pathol Int ; 50(8): 610-5, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10972858

RESUMO

Formalin-fixed paraffin-embedded sections of three cases of ameloblastic fibrodentinoma (AFD) were studied by the avidin-biotin-peroxidase complex method using antibodies against neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and S100 protein and the results were compared with those in ameloblastic fibroma (AF). A striking histopathological characteristic of AFD was the formation of abortive dentin with various degrees of maturation at the epithelial-mesenchymal tissue interface. Central cells of enamel organ-like epithelia with various stages of abortive dentin induction in AFD were generally positive for NSE. Dental lamina-like epithelial cells also showed positive staining in some areas. No cells were positive for NSE in AF. Positive staining for GFAP was observed in the juxta-epithelial mesenchymal tissue of the formation stage of immature dentin with various numbers of entrapped cells in AFD, but GFAP staining was negative in other mesenchymal and epithelial tissues at other stages. In AF, no GFAP-positive cells were found. There were a few S100 protein-positive cells found in the foci of epithelial components in both AFD and AF. Mesenchymal cells showing a dendritic or spindle shape were positive for S100 protein in some areas of AFD and AF. Although such cells in the mesenchymal component of pigmented AFD were more numerous than in non-pigmented AFD and AF, their distribution pattern in the former condition was basically similar to that in the latter. Although the present results, obtained from conventional immunohistochemical procedures, do not directly reflect the expression of neural crest-derived cells in the dentinogenesis of AFD, such results do not disprove the possibility of the expression of neural proteins probably related to neural crest-derived cells in dentinogenesis under certain pathologic conditions in odontogenic mixed tumors. Such a phenomenon may also occur during dentinogenesis in other odontogenic mixed tumors and in normal tooth differentiation, but at an undetectable level.


Assuntos
Ameloblastoma/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Neoplasias Mandibulares/metabolismo , Neoplasias Maxilares/metabolismo , Odontoma/metabolismo , Fosfopiruvato Hidratase/metabolismo , Proteínas S100/metabolismo , Adulto , Ameloblastoma/patologia , Ameloblastoma/cirurgia , Biomarcadores Tumorais/metabolismo , Criança , Pré-Escolar , Dentina/patologia , Feminino , Humanos , Técnicas Imunoenzimáticas , Masculino , Neoplasias Mandibulares/patologia , Neoplasias Mandibulares/cirurgia , Neoplasias Maxilares/patologia , Neoplasias Maxilares/cirurgia , Proteínas de Neoplasias/metabolismo , Odontoma/patologia , Odontoma/cirurgia
10.
Circulation ; 102(6): 670-6, 2000 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-10931808

RESUMO

BACKGROUND: Triglyceride-rich lipoproteins (TGLs) are atherogenic. However, their cellular mechanisms remain largely unexplained. This study examined the effects of isolated remnant-like lipoprotein particles (RLPs) on the expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and tissue factor (TF), proatherothrombogenic molecules, in cultured human endothelial cells. METHODS AND RESULTS: RLPs were isolated from plasma of hypertriglyceridemic patients by use of the immunoaffinity gel mixture of anti-apoA-1 and anti-apoB-100 monoclonal antibodies. The incubation of cells with RLPs significantly upregulated mRNA and protein expression of these molecules. Total TGLs (d<1.006) and LDL had fewer or minimal effects on expression of these molecules compared with RLPs. RLPs increased intracellular oxidant levels, as assessed with an oxidant-sensitive probe. Combined incubation with alpha-tocopherol or N-acetylcysteine, both antioxidants, suppressed RLP-induced increase in expression of these molecules. In patients with higher plasma levels of RLPs, plasma levels of soluble forms of ICAM-1 and VCAM-1 were significantly higher than in patients with lower RLP levels. Treatment with alpha-tocopherol for 1 month decreased levels of the soluble adhesion molecules concomitantly with an increase in resistance of RLPs to oxidative modification in patients with high RLP levels. CONCLUSIONS: RLPs upregulated endothelial expression of ICAM-1, VCAM-1, and TF, proatherothrombogenic molecules, partly through a redox-sensitive mechanism. RLPs may have an important role in atherothrombotic complications in hypertriglyceridemic patients.


Assuntos
Arteriosclerose/etiologia , Endotélio Vascular/metabolismo , Lipoproteínas/fisiologia , Arteriosclerose/tratamento farmacológico , Células Cultivadas , Meios de Cultura/metabolismo , DNA/metabolismo , Endotélio Vascular/citologia , Humanos , Hipertrigliceridemia/sangue , Molécula 1 de Adesão Intercelular/sangue , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Peróxidos Lipídicos/metabolismo , Lipoproteínas/sangue , NF-kappa B/metabolismo , Oxirredução , Fragmentos de Peptídeos/sangue , Fragmentos de Peptídeos/fisiologia , RNA Mensageiro/metabolismo , Solubilidade , Tromboplastina/genética , Tromboplastina/metabolismo , Molécula 1 de Adesão de Célula Vascular/sangue , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/metabolismo , Vitamina E/uso terapêutico
11.
Pathol Int ; 50(6): 514-9, 2000 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10886731

RESUMO

A rare case of carcinoma characterized by extensive chondroid elements at a site of primary esophageal and metastatic lesion is reported. The patient was a 67-year-old man complaining of dysphagia due to an ulcerative lesion at the lower middle esophagus. He underwent irradiation treatment prior to surgery. Histologically, the tumor consisted of both carcinomatous and chondroid elements and had invaded deeply into the esophageal wall. The carcinomatous cells had gradually become chondroid cells embedded within an extensive extracellular matrix. In addition, the metastatic lesion showed findings similar to those of the primary lesion. Immunohistochemistry revealed that both carcinomatous and chondroid elements were immunostained with cytokeratin and epithelial membrane antigen, suggesting an epithelial nature to the chondroid cells. Conversely, only chondroid cells were positively stained for S-100 protein. Furthermore, bone morphogenetic proteins (BMP) were positive for chondroid cells and their surrounding carcinomatous cells. Given the apparent transition between carcinomatous and chondroid cells based on microscopy and immunohistochemical findings in the present case, we concluded that the chondroid cells were derived from carcinomatous cells. In addition, our findings suggest that BMP produced by carcinomatous cells lead to chondroid differentiation of the carcinoma cells.


Assuntos
Carcinoma de Células Escamosas/patologia , Condrócitos/patologia , Neoplasias Esofágicas/patologia , Idoso , Proteínas Morfogenéticas Ósseas/análise , Carcinoma de Células Escamosas/metabolismo , Diferenciação Celular , Condrócitos/metabolismo , Neoplasias Esofágicas/metabolismo , Humanos , Imuno-Histoquímica , Queratinas/análise , Masculino , Mucina-1/análise , Proteínas S100/análise
12.
Spine (Phila Pa 1976) ; 25(5): 537-42, 2000 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-10749628

RESUMO

STUDY DESIGN: A study of the relation between the development of mechanical allodynia and the reorganization of primary afferent terminals in the sensory lamina of the rat spinal cord dorsal horn after partial dorsal root ganglion injury in rats. OBJECTIVES: To investigate the pathologic mechanisms of mechanical allodynia after partial dorsal root ganglion injury. SUMMARY OF BACKGROUND DATA: After experimental peripheral nerve injury causing neuropathic pain, myelinated afferent fibers sprout into lamina II of the dorsal horn. This lamina is associated with nociceptive-specific neurons that generally are not stimulated by myelinated fiber input from mechanical receptors. These morphologic changes are suggested to have significance in the pathogenesis of chronic mechanical allodynia, although it is not known whether this kind of morphologic change occurs after dorsal root ganglion injury. METHODS: After partial dorsal root ganglion crush injury, the mechanical force causing footpad withdrawal was measured with von Frey hairs, and myelinated primary afferents were labeled with cholera toxin B subunit horseradish peroxidase, a selective myelinated fiber tracer that identifies transganglionic synapses. RESULTS: After partial dorsal root ganglion injury, mechanical allodynia developed in the corresponding footpad within 3 days and persisted throughout the experimental period. At 2 and 4 weeks after the injury, B subunit horseradish peroxidase-positive fibers, presumably myelinated afferents, were observed to be sprouting into lamina II of the dorsal horn on the injured side, but not on the contralateral control side. CONCLUSIONS: Morphologic change in spinal cord dorsal horn lamina II occurs after partial dorsal root ganglion injury. This change may have significance in the pathogenesis of chronic mechanical allodynia after partial dorsal root ganglion injury.


Assuntos
Gânglios Espinais/lesões , Gânglios Espinais/patologia , Nociceptores/patologia , Células do Corno Posterior/patologia , Vias Aferentes/patologia , Animais , Comportamento Animal/fisiologia , Contagem de Células , Toxina da Cólera , Peroxidase do Rábano Silvestre , Compressão Nervosa , Fibras Nervosas/patologia , Fibras Nervosas/fisiologia , Plasticidade Neuronal/fisiologia , Dor/patologia , Dor/fisiopatologia , Células do Corno Posterior/ultraestrutura , Ratos , Ratos Sprague-Dawley , Tato/fisiologia
13.
Gastroenterology ; 118(5): 835-41, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10784582

RESUMO

BACKGROUND & AIMS: Microsatellite instability (MSI) in mitochondrial DNA (mtDNA) is observed in some colorectal carcinomas. We attempted to determine if mitochondrial MSI (mtMSI) and mutations occur in gastric carcinomas and if the mtMSI phenotype underlies specific clinicopathologic profiles. METHODS: Sixty-two gastric carcinomas (34 intestinal and 28 diffuse types) were investigated. Coding mutations in 8 different mitochondrial genes, mtMSI in a noncoding (C)n tract, and p53 gene mutations were examined by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis. MSI in nuclear DNA (nMSI) and loss of the p53 gene were examined using microsatellite markers. RESULTS: Ten of 62 (16%) carcinomas showed the mtMSI phenotype. Mitochondrial gene mutation was detected in 5 carcinomas, 4 of which also showed the mtMSI phenotype. There was a positive correlation between mtMSI and nMSI status. In intestinal carcinomas, mtMSI, nMSI, and p53 gene alterations were frequently detected from early to advanced stages. In diffuse carcinomas, both kinds of MSI were found in only advanced (subserosal or serosal invasion) carcinomas. Six of 7 carcinomas with the nMSI phenotype and all 5 carcinomas with mitochondrial coding mutations had a considerable intestinal-type tumor cell component. CONCLUSIONS: Mitochondrial gene mutations, which are associated with the mtMSI phenotype, may play a specific role in the tumorigenesis of intestinal-type gastric carcinomas.


Assuntos
DNA Mitocondrial/genética , DNA de Neoplasias/genética , Repetições de Microssatélites , Neoplasias Gástricas/genética , Núcleo Celular/genética , Mutação da Fase de Leitura , Humanos , Perda de Heterozigosidade , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , Neoplasias Gástricas/patologia , Proteína Supressora de Tumor p53/genética
14.
J Surg Oncol ; 73(3): 158-63, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10738270

RESUMO

BACKGROUND AND OBJECTIVES: The genetic alterations involved in extrahepatic bile duct (EHBD) cancer are poorly understood. Our aim was to identify aberrations of the K-ras, p53, and APC genes in EHBD cancer. METHODS: We investigated aberrations of these genes in 52 EHBD cancers using polymerase chain reaction (PCR) single-strand conformation polymorphism analysis, followed by direct sequence determination and a PCR restriction fragment length polymorphism assay. RESULTS: The K-ras, p53, and APC genes were mutated in 9.6%, 32.7%, and 0% of EHBD cancers, respectively. Loss of heterozygosity at the p53 and APC gene loci was identified in 15.6% and 38.5% of EHBD cancers, respectively. CONCLUSIONS: Our results suggest that an unknown suppressor gene on 5q other than the APC gene may be responsible for EHBD cancer.


Assuntos
Neoplasias dos Ductos Biliares/genética , Ductos Biliares Extra-Hepáticos , Genes APC/genética , Genes p53/genética , Genes ras/genética , Mutação Puntual , Idoso , Neoplasias dos Ductos Biliares/mortalidade , Neoplasias dos Ductos Biliares/patologia , Feminino , Humanos , Perda de Heterozigosidade , Metástase Linfática , Masculino , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Polimorfismo Conformacional de Fita Simples , Taxa de Sobrevida
15.
Lung Cancer ; 27(3): 189-97, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10699692

RESUMO

Small cell lung cancer (SCLC) frequently metastasizes to bone and bone marrow. Skeletal scintigraphy and bone marrow cytology or biopsy, are incorporated into the staging procedures to examine these organs. However, skeletal scintigraphy is not highly specific to metastases, and only one or two bone marrow sites can be examined by cytology or biopsy. We have already reported that magnetic resonance imaging (MRI) could improve the sensitivity in detecting bone marrow metastases. The result of the bone marrow MRI was an independent prognostic factor of SCLC patients [9]. In the present study, we analyzed the results of skeletal scintigraphy and bone marrow aspiration with special reference to the results of MRI examination. We also analyzed the relationship between bone marrow lesions and bone lesions. For this purpose, we visualized bone marrow metastases with MRI and determined their anatomical locations and sizes. Approximately half of bone marrow lesions stayed in bone marrow during follow-up period ranging from 57 to 154 days, whereas about half of them were accompanied by hot spots in follow-up skeletal scintigraphy, which indicates the destruction of osseous structure. Additionally, 87.5% of osteolytic changes that newly appeared in skeletal scintigraphy were preceded by adjacent bone marrow lesions. All new lesions that appeared in follow-up skeletal scintigraphy within 3 months after the initial presentation had the preceding bone marrow lesions. These results mean that almost all lesions in skeletal scintigraphy derived from bone marrow metastases. Furthermore, appreciable volume of cancer cells is present in bone marrow before osteolytic changes appear in skeletal scintigraphy.


Assuntos
Neoplasias da Medula Óssea/patologia , Carcinoma de Células Pequenas/patologia , Neoplasias Pulmonares/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia por Agulha , Neoplasias da Medula Óssea/diagnóstico por imagem , Neoplasias da Medula Óssea/secundário , Carcinoma de Células Pequenas/diagnóstico por imagem , Carcinoma de Células Pequenas/secundário , Progressão da Doença , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Cintilografia
16.
Arch Pathol Lab Med ; 124(3): 382-6, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10705390

RESUMO

BACKGROUND: Contamination of nontumor tissue makes genetic analysis difficult. For this reason, it is important to obtain pure tumor tissue to ensure accurate genetic analysis. OBJECTIVE: To accurately assess the incidence of mutation of tumor suppressor genes (p53: exon 5-8; APC: mutated cluster region; NF-2 gene: all exons) in 45 colorectal carcinomas. METHODS: We developed an application of the polymerase chain reaction-single-strand conformation polymorphism and DNA sequence by coupling them with crypt isolation. RESULTS: Mutations of p53 and APC genes were found in 24 and 22 of 45 colorectal carcinomas, respectively. No mutation of the NF-2 gene was observed in this cancer. Single-strand conformation polymorphism using a crypt isolation technique showed a clear migrating band and no false-positive data. CONCLUSIONS: The crypt isolation technique is a useful method for accurately analyzing genetic alterations. Furthermore, our proposed method confirmed the morphological findings obtained before the genetic analysis.


Assuntos
Adenocarcinoma/genética , Neoplasias Colorretais/genética , Análise Mutacional de DNA/métodos , Genes Supressores de Tumor/genética , Adenocarcinoma/patologia , Adulto , Idoso , Colo/patologia , Neoplasias Colorretais/patologia , DNA de Neoplasias/análise , Feminino , Genes APC/genética , Genes da Neurofibromatose 2/genética , Genes p53/genética , Humanos , Mucosa Intestinal/patologia , Perda de Heterozigosidade , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples
17.
J Agric Food Chem ; 47(4): 1346-9, 1999 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10563978

RESUMO

Two isoflavones, daidzein (1) and genistein (2), were isolated from soybean hypocotyls. Daidzein and genistein showed a suppressive effect on umu gene expression of the SOS response in Salmonellatyphimurium TA1535/pSK1002 against the mutagen 3-amino-1, 4-dimethyl-5H-pyrido[4,3b]indole (Trp-P-1), which requires liver metabolizing enzymes. Compound 1 suppressed 73% of the SOS-inducing activity at concentrations <0.74 micromol/mL, and the ID(50) value was 0.37 micromol/mL. Compound 2 suppressed 95% of the SOS-inducing activity at concentrations <0.74 micromol/mL, and the ID(50) value was 0.17 micromol/mL. Compounds 1 and 2 were also assayed with the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide) and activated Trp-P-1. In addition to the antimutagenic activities of daidzein and genistein against Trp-P-1, frylfuramide and activated Trp-P-1 were assayed by an Ames test using S. typhimurium TA100.


Assuntos
Antimutagênicos/farmacologia , Genisteína/farmacologia , Glycine max/química , Isoflavonas/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Sementes/química , Antimutagênicos/química , Antimutagênicos/isolamento & purificação , Carbolinas/farmacologia , Genisteína/química , Genisteína/isolamento & purificação , Hipocótilo , Isoflavonas/química , Isoflavonas/isolamento & purificação , Testes de Mutagenicidade , Mutagênicos/farmacologia , Resposta SOS em Genética/efeitos dos fármacos , Salmonella typhimurium/genética
18.
Pathol Int ; 49(7): 617-25, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10504522

RESUMO

Although numerous studies have assessed the biologic parameters of tumors, measurement of these parameters has had, to date, little impact on histologic diagnosis. Furthermore, analysis of a single parameter is insufficient to evaluate tumor malignant potential. In the present study, cell proliferation, DNA ploidy and p53 product were analyzed to objectify the tumor malignant potential in colorectal adenomas and intramucosal carcinomas. Sixty-one adenomas and 49 intramucosal carcinomas were studied using immunohistochemical analysis of Ki-67 and p53, silver-staining nucleolar organizer region (AgNOR) stain and DNA ploidy in fresh samples. Intramucosal carcinoma exhibited a greater Ki-67-positive rate and AgNOR count than the adenomas, although these parameters varied widely among samples. The incidence of aneuploidy and p53 over-expression in colorectal intramucosal carcinomas was significantly higher than in colorectal adenomas. These results indicate that DNA aneuploidy and p53 accumulation are the most reliable parameters for distinguishing benign and malignant lesions.


Assuntos
Adenocarcinoma/diagnóstico , Adenoma/diagnóstico , Neoplasias Colorretais/diagnóstico , DNA de Neoplasias/análise , Ploidias , Proteína Supressora de Tumor p53/análise , Adenocarcinoma/química , Adenoma/química , Adenoma/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Divisão Celular , Neoplasias Colorretais/química , Neoplasias Colorretais/genética , Feminino , Citometria de Fluxo , Humanos , Técnicas Imunoenzimáticas , Mucosa Intestinal/química , Mucosa Intestinal/patologia , Antígeno Ki-67/análise , Masculino , Pessoa de Meia-Idade , Região Organizadora do Nucléolo/química , Coloração pela Prata
19.
Spine (Phila Pa 1976) ; 23(17): 1853-8; discussion 1859, 1998 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-9762742

RESUMO

STUDY DESIGN: Lumbar peripheral nerves were examined to determine whether they were responsive to electrical stimulation of the ventral portion of the lumbar disc in anesthetized rats. OBJECTIVES: To confirm by electrophysiologic means the neural correspondence between the ventral portion of the lumbar disc and the groin. SUMMARY OF BACKGROUND DATA: Patients with a degenerated lumbar disc occasionally report groin pain. However, its pathogenesis has not been investigated. The authors of the current study found that chemical stimulation of the ventral portion of rat lumbar disc caused cutaneous plasma extravasation in the groin, and thereby hypothesize the neural relation between the lumbar disc and the groin. METHODS: The ventral portion of rat L5-L6 disc was electrically stimulated, and the elicited action potentials were recorded from the iliohypogastric, genitofemoral, lateral femoral cutaneous, sural, and sciatic nerves. The roles of the lumbar sympathetic trunks and spinal cord in the generation of the action potentials were examined. RESULTS: Action potentials were elicited principally in the genitofemoral nerve; the action potentials of the genitofemoral nerve were not influenced by transection of the cervical spinal cord, whereas they disappeared immediately after death, which indicates that they are induced by a spinal reflex. The action potentials were reduced considerably after destruction of the lumbar sympathetic trunks, suggesting that they comprise an afferent path of the reflex. CONCLUSIONS: The ventral portion of the lumbar disc had spatial relation to the groin area via a spinal reflex. Such a relation suggests that a disorder in the ventral portion of the lumbar disc may be a possible source of groin referred pain.


Assuntos
Virilha/inervação , Deslocamento do Disco Intervertebral/fisiopatologia , Disco Intervertebral/fisiologia , Dor/fisiopatologia , Reflexo/fisiologia , Potenciais de Ação/fisiologia , Animais , Estimulação Elétrica , Eletrofisiologia , Disco Intervertebral/inervação , Deslocamento do Disco Intervertebral/complicações , Vértebras Lombares , Masculino , Dor/diagnóstico , Dor/etiologia , Ratos , Ratos Wistar , Nervos Espinhais/fisiologia
20.
J Gastroenterol ; 33(1): 80-5, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9497226

RESUMO

We report a rare case of multiple gastric plasmacytomatous lesions at an early stage found incidentally in a stomach resected because of gastric cancer. The three lesions had different macroscopic features, showing depressed, submucosal, and nodular tumor-forming types. The smallest, a depressed lesion, produced IgG of the lambda and kappa types, and had a plasma-cell granuloma-like appearence, whereas the largest, a submucosal tumor, was formed by the monoclonal proliferation of atypical plasma cells containing IgA-kappa type immunoglobulin, and the nodular lesion exhibited histological features intermediate between those of the other two lesions. From their microscopic features and the profile of immunoglobulin production, we believe that these lesions may represent different stages in the possible course of development of plasmacytoma from the early stage of plasma cell granuloma. All three lesions were located far from the gastric carcinoma and it was unclear whether they had any causal relationship with it.


Assuntos
Adenocarcinoma/complicações , Adenocarcinoma/patologia , Granuloma de Células Plasmáticas/complicações , Granuloma de Células Plasmáticas/patologia , Neoplasias Primárias Múltiplas/patologia , Plasmocitoma/complicações , Plasmocitoma/patologia , Neoplasias Gástricas/complicações , Neoplasias Gástricas/patologia , Adenocarcinoma/imunologia , Idoso , Granuloma de Células Plasmáticas/imunologia , Humanos , Masculino , Neoplasias Primárias Múltiplas/imunologia , Plasmocitoma/imunologia , Neoplasias Gástricas/imunologia
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