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1.
Clin Radiol ; 64(3): 291-7, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19185659

RESUMO

AIM: To evaluate the diagnostic role of additional oblique coronal and oblique sagittal magnetic resonance imaging (MRI) for an anterior cruciate ligament (ACL) tear. MATERIALS AND METHODS: A total of 101 patients who had undergone preoperative knee MRI examinations with orthogonal and two sets of oblique images were enrolled in the study. Two radiologists evaluated the MRI images by the use of four methods: orthogonal images only (method A); orthogonal and additional oblique coronal images (method B); orthogonal and oblique sagittal images (method C); and orthogonal images with oblique coronal and sagittal images (method D). The status of the ACL (normal or tear) was determined by consensus. The sensitivity, specificity, and accuracy for an ACL tear with the use of each method were calculated in comparison with arthroscopy as the reference standard, and values were statistically analysed using the McNemar test. The diagnostic accuracies were compared using receiver operating characteristic (ROC) analysis. RESULTS: Arthroscopy identified 10 partial ACL tears and 30 complete ACL tears. The specificities and accuracies for methods B, C, and D were significantly higher than the specificities and accuracies for method A (p<0.05). There was no significant difference in the sensitivity, specificity, and accuracy for methods B, C, and D. Diagnostic ability was not significantly different for each method, as determined by ROC analysis (p>0.05). CONCLUSIONS: Additional oblique imaging for an ACL tear improved the specificity. Either of the oblique imaging methods is sufficient, and no further improvement in the diagnostic efficacy was achieved by simultaneous use.


Assuntos
Lesões do Ligamento Cruzado Anterior , Imageamento por Ressonância Magnética/métodos , Adolescente , Adulto , Ligamento Cruzado Anterior/patologia , Artroscopia , Criança , Feminino , Humanos , Traumatismos do Joelho/diagnóstico , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto Jovem
2.
Acta Radiol ; 48(8): 869-74, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17924218

RESUMO

BACKGROUND: Magnetic resonance (MR) imaging has proved to be an excellent tool in diagnosing injuries of the cruciate ligaments and menisci. However, multiple planes and sometimes optimal oblique or double-oblique scan planes are needed due to the variability in the positioning of important structures, which means there is a lower throughput and longer scanning time. PURPOSE: To compare the performance of a 1-mm-thickness fast spin-echo (FSE) proton-density-weighted (PDW) MR imaging technique with multiplanar reconstruction (MPR) in diagnosing tears of the menisci and cruciate ligaments with that of conventional MR imaging. MATERIAL AND METHODS: Twenty-five consecutive patients underwent preoperative conventional and 1-mm-thickness FSE PDW MR imaging with subsequent knee arthroscopic surgery. Two musculoskeletal radiologists evaluated the status of the cruciate ligaments and menisci using two sets of MR images (method A: conventional images including seven sequences, taking 26 min; method B: 1-mm-thickness FSE PDW images with MPR, taking 7 min 20 s). The diagnostic efficacies of both methods for tears of the cruciate ligament and menisci were calculated and compared. RESULTS: Arthroscopic surgery revealed 10 anterior cruciate ligament (ACL) tears, one posterior cruciate ligament (PCL) tear, and 26 meniscal tears. The diagnostic values of both methods were 100% for a cruciate ligament tear. The diagnostic values (sensitivity, specificity, accuracy, positive predictive value, and negative predictive value) for meniscal tears were 90%, 100%, 96%, 100%, and 94% for method A, and 95%, 100%, 98%, 100%, and 97% for method B, respectively. There were no significant differences in the diagnostic values between methods A and B. CONCLUSION: 1-mm-slice-thickness FSE PDW imaging with MPR showed comparable performance in diagnosing tears of the cruciate ligaments and menisci to conventional sequences but the scan time was much shorter. Therefore, this technique (method B) might improve the throughput of a 3T MR imaging system.


Assuntos
Ligamento Cruzado Anterior/patologia , Processamento de Imagem Assistida por Computador/métodos , Traumatismos do Joelho/diagnóstico , Imageamento por Ressonância Magnética/métodos , Meniscos Tibiais/patologia , Ligamento Cruzado Posterior/patologia , Adolescente , Adulto , Lesões do Ligamento Cruzado Anterior , Artroscopia , Criança , Feminino , Humanos , Traumatismos do Joelho/cirurgia , Articulação do Joelho/patologia , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Ligamento Cruzado Posterior/lesões , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Lesões do Menisco Tibial
3.
Mol Immunol ; 19(9): 1199-202, 1982 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-6183581

RESUMO

Hybrid cell lines producing monoclonal antibodies against human prostatic acid phosphatase (E. C. 3.1.3.2) were prepared by the fusion of mouse myeloma cells with the spleen cells of BALB/c mice and Lewis rats immunized with prostatic acid phosphatase (PAP). Approximately 14% of the hybrid cell microcultures which produced specific antibodies were cloned, and 6 eventually yielded stable cell lines. The monoclonal antibodies produced by these 6 hybridomas were characterized for their isotypes, isoelectric points, concentrations and affinities. The specificity of these monoclonal antibodies was further investigated by radioimmunoassay and immunohistochemical methods. All of the 6 monoclonal antibodies exhibited strict specificity for prostatic acid phosphatase.


Assuntos
Fosfatase Ácida/imunologia , Anticorpos Monoclonais/imunologia , Animais , Afinidade de Anticorpos , Especificidade de Anticorpos , Reações Cruzadas , Epitopos/imunologia , Humanos , Células Híbridas , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Próstata/enzimologia , Ratos , Ratos Endogâmicos Lew
4.
Semin Oncol ; 3(2): 107-13, 1976 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-935881

RESUMO

The possibility of altering the course of prostatic cancer by immunologic means requires a clear understanding of the host-tumor relationship in this disease. Available data suggest that prostatic tumors contain both prostate-specific and tumor-specific antigens, although evidence on the latter is still debatable. Patients with prostatic cancer often show nonspecific depression of their cell-mediated immunocompetence, as do patients with many other forms of cancer. The question of whether prostatic cancers are immunogenic; that is, whether they elicit a specific immunologic response, remains unanswered.


Assuntos
Neoplasias da Próstata/imunologia , Fosfatase Ácida/análise , Anticorpos Antineoplásicos/análise , Formação de Anticorpos , Antígenos de Neoplasias/análise , Sobrevivência Celular , Humanos , Imunidade Celular , Imunoterapia , Masculino , Modelos Biológicos , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/terapia , Testes Cutâneos
5.
J Immunol Methods ; 84(1-2): 105-16, 1985 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-2415632

RESUMO

Monoclonal antibodies against human prostatic acid phosphatase (PAPase) were produced by immunization of human primary spleen cell cultures. Dissociated spleen cells were cultured for 5-8 days in the presence of 100 ng/ml of PAPase and pokeweed mitogen (1:5000). Following immunization, B cells were isolated and infected with Epstein-Barr virus (EBV). Two weeks after EBV-transformation, cells were fused with either mouse myeloma cells (SP2/OAg14) or human/mouse heteromyeloma cells (SHM-D33). Hybrid clones were screened for anti-PAPase production. In 7 independent immunizations, the average fusion frequency was 3.6 per 10(6) lymphocytes. 18-32% of the hybridomas produced anti-PAPase; approximately 75% of these secreted IgM and 25% secreted IgG. Antibody specificity was determined by immunoassay and immunohistological studies. The procedures described here may be suitable for the production of human monoclonal antibody of a useful specificity.


Assuntos
Fosfatase Ácida/imunologia , Anticorpos Monoclonais/isolamento & purificação , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Linfócitos B/imunologia , Fusão Celular , Herpesvirus Humano 4 , Humanos , Hibridomas/imunologia , Técnicas Imunoenzimáticas , Masculino , Camundongos , Mieloma Múltiplo , Mitógenos de Phytolacca americana/farmacologia , Hiperplasia Prostática/enzimologia , Baço/citologia
6.
Immunol Lett ; 78(1): 13-20, 2001 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-11470146

RESUMO

There are many mutations in the gene encoding Hepatitis B virus (HBV) core antigen of chronic active hepatitis patients, and such mutations are most likely to be related to the severity of disease. Here, we constructed plasmids containing wild-type and deletion type of HBV core gene (HBc) to develop an experimental DNA vaccine and to compare immunogenicity of two types of HBc vaccine. Twenty-nine wild-types and seven deletion types of HBc were detected in sera of 32 Korean patients with chronic active hepatitis. Four wild-types (W1, W2, W4, W6) and two deletion types (D3, D4) of HBc were cloned into the pcDNA3 vector. Intramuscular immunization with wild-type HBc efficiently increased serum anti-HBc antibody response in a dose-dependent manner. Anti-HBc antibody response in mice injected with W6 increased 14 days after immunization, and peaked after 30 days and was maintained at least up to 50 days. W6 immunization induced a specific cytotoxic T lymphocyte response to W6-transfected 3LL (3LL-W6), and reduced the sizes of tumor mass of mice challenged with 3LL-W6 or 3LL transfected with D4. However, intramuscular immunization with D3 and D4 did not show antibody response at all. D3 and D4 have 157 bp (from 331 to 491 bp) and 122 bp (from 327 to 448 bp) gene deletion, respectively, and these encode class II MHC-restricted T-cell epitope. Altogether, these results suggest that mutant virus that has deleted HBc gene may evade immune systems due to loss of T-cell epitope.


Assuntos
DNA Viral/administração & dosagem , Antígenos do Núcleo do Vírus da Hepatite B/administração & dosagem , Vírus da Hepatite B/imunologia , Hepatite B Crônica/imunologia , Vacinas de DNA/administração & dosagem , Animais , Modelos Animais de Doenças , Deleção de Genes , Anticorpos Anti-Hepatite B/análise , Anticorpos Anti-Hepatite B/biossíntese , Antígenos do Núcleo do Vírus da Hepatite B/genética , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Vírus da Hepatite B/genética , Hepatite B Crônica/prevenção & controle , Hepatite B Crônica/virologia , Humanos , Injeções Intramusculares , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Plasmídeos , Linfócitos T Citotóxicos/imunologia , Fatores de Tempo , Células Tumorais Cultivadas , Vacinação
7.
Schizophr Res ; 67(1): 71-4, 2004 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-14741326

RESUMO

The reduced incidence of cancer observed in schizophrenia patients may be related to differences in genetic background. It has been suggested that genetic predisposition towards schizophrenia is associated with reduced vulnerability to lung cancer, and p53 gene is one of the candidate genes. We tested the genetic association between schizophrenia and lung cancer by analyzing polymorphic sites in the p53 gene. Genotype and allele frequencies at two polymorphic sites in the p53 gene (BstUI and MspI restriction sites in exon 4 and intron 6, respectively) were studied in Korean schizophrenia (n=179) and lung cancer patients (n=104). Comparisons of the genotype and allele frequencies of the MspI polymorphism revealed significant differences between schizophrenia and lung cancer patients. The results suggest that the p53 polymorphism specifically found in schizophrenia patients may be associated with reduced vulnerability to lung cancer.


Assuntos
Genes p53/genética , Neoplasias Pulmonares/etnologia , Neoplasias Pulmonares/genética , Polimorfismo Genético/genética , Esquizofrenia/etnologia , Esquizofrenia/genética , Adulto , Feminino , GTP Fosfo-Hidrolases/genética , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Coreia (Geográfico) , Masculino , Pessoa de Meia-Idade
8.
Neurochem Int ; 36(1): 19-26, 2000 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10566955

RESUMO

We have investigated the expression of neuropeptide Y (NPY) in C6 glioma cells after the glutamatergic stimulation by the in situ RT-PCR and immunocytochemical techniques. The expression of NPY mRNA correlated well with immunocytological findings in each series of experiments. NPY protein expression was enhanced by glutamate (1, 10, 50, 100 microM, and 1 mM) dose-dependently, and its expression was slightly increased by N-methyl-D-aspartate (NMDA; 1, 10, 100, 500 microM, and 1 mM) and kainic acid (1, 10, 100, 300 microM, and 1 mM). We pretreated the cells with dopamine, haloperidol, pentylenetetrazol, and muscimol before each stimulation. The pentylenetetrazol and muscimol did not significantly alter the patterns of NPY expression induced by the glutamatergic stimulation. On the other hand, the dopamine and haloperidol pretreatment significantly elevated the levels of NPY expression that were induced by NMDA and kainic acid. Our results indicate that NPY release is closely related to glutamatergic stimulation, and it could be dynamically mediated by GABAergic and dopaminergic costimulation.


Assuntos
Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Ácido Glutâmico/farmacologia , Ácido Caínico/farmacologia , Neuropeptídeo Y/genética , Transcrição Gênica/efeitos dos fármacos , Animais , Dopamina/farmacologia , Glioma , Haloperidol/farmacologia , Muscimol/farmacologia , Pentilenotetrazol/farmacologia , RNA Mensageiro/genética , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas
9.
Ann N Y Acad Sci ; 390: 16-26, 1982.
Artigo em Inglês | MEDLINE | ID: mdl-6178343

RESUMO

Human prostatic acid phosphatase [PAP] is antigenically uniquely different from acid phosphatases of other tissue origins. Nevertheless, a small degree of antigenic cross-reactivity between PAP and other lysosomal acid phosphatase(s) [LAP] has been suspected. In order to resolve this question, we have adopted two approaches: one involving structural studies by peptide mapping, and the other involving topological mapping through the use of uniquely defined antibodies. Purified PAP was dissociated into subunits and was further cleaved by chemical and enzymological methods. The limited digestion of PAP by submaxillary protease yielded three fragments [Sp-1, 2, and 3]. One of the fragments, Sp-3 [Mr = 11,000-12,000], was shown to regain catalytic activity after interaction with anti-PAP antibodies. This along with other data suggested that the active site is localized in the Sp-3 fragment. These submaxillary protease fragments were also used in the antigenic studies. For the detailed antigenic mapping studies, we prepared 12 monoclonal anti-PAP antibodies. These monoclonal anti-PAP antibodies exhibited a remarkably specific binding to PAP, particularly to the Sp-1 fragment, without binding to other acid phosphatase preparations. We also prepared lysosomal acid phosphatase [LAP] and raised anti-LAP antibodies in rabbits. The anti-LAP antibodies were fractionated into subpopulations by the preparative isoelectric focusing method. Three anti-LAP antibody subpopulations [pI 5.2, 6.9, and 7.5] exhibited specific binding to LAP. However, two anti-LAP subpopulations [pI 5.3 and and 6.8] showed binding to the Sp-3 fragment, an active site fragment of PAP. Thus, the PAP molecule seems to consist of three domains, namely, Sp-1, Sp-3, and Sp-2. Sp-3, which is the active site domain, is an antigenically cross-reactive region. The Sp-1 domain represents an antigenically unique region of PAP, whereas none of the antibodies studied thus far bind to the Sp-2 fragment.


Assuntos
Fosfatase Ácida/imunologia , Anticorpos Monoclonais/biossíntese , Epitopos/análise , Lisossomos/imunologia , Próstata/enzimologia , Fosfatase Ácida/análise , Fosfatase Ácida/genética , Anticorpos Monoclonais/análise , Formação de Anticorpos , Especificidade de Anticorpos , Ligação Competitiva , Cromatografia em Gel , Reações Cruzadas , Epitopos/genética , Humanos , Focalização Isoelétrica , Lisossomos/enzimologia , Masculino , Fragmentos de Peptídeos/análise , Peptídeo Hidrolases
10.
Neurosci Lett ; 292(1): 60-2, 2000 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-10996450

RESUMO

Several studies have suggested that the activity of nitric oxide synthase (NOS) may be involved in the regulation of food intake in the genetically obese Zucker rats. In the present study, we investigated the expression of NOS in various hypothalamic regions of obese and lean Zucker rats using nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. Obese Zucker rats showed significantly lower staining intensities of NADPH-diaphorase-positive neurons in the paraventricular nucleus (PVN), lateral hypothalamic area (LHA) and ventromedial hypothalamic nucleus (VMH) than lean Zucker rats did. The differences in staining intensities between obese and lean Zucker rats were large in both the PVN and LHA, but such differences were relatively small in the VMH.


Assuntos
Di-Hidrolipoamida Desidrogenase/biossíntese , Hipotálamo/enzimologia , Obesidade/enzimologia , Animais , Feminino , Histocitoquímica , Região Hipotalâmica Lateral/enzimologia , Região Hipotalâmica Lateral/patologia , Hipotálamo/patologia , Neurônios/enzimologia , Neurônios/patologia , Óxido Nítrico Sintase/metabolismo , Obesidade/patologia , Núcleo Hipotalâmico Paraventricular/enzimologia , Núcleo Hipotalâmico Paraventricular/patologia , Ratos , Ratos Mutantes , Ratos Zucker , Coloração e Rotulagem , Núcleo Hipotalâmico Ventromedial/enzimologia , Núcleo Hipotalâmico Ventromedial/patologia
11.
Neurosci Lett ; 302(2-3): 109-12, 2001 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-11290399

RESUMO

The homozygous anorexia mutant (anx/anx) mice present with premature death during the third or fourth postnatal week: this phenotype is caused by a lethal mutation, anx, on chromosome 2, which has an autosomal recessive mode of inheritance. These animals also present phenotypically with decreased food intake, weight loss, and neurological deficits such as hyperactivity, body tremors, uncoordinated gait, and head weaving. In order to investigate changes in the occurrence of cell proliferation and apoptosis in the dentate gyrus of the hippocampus of anx/anx mice, 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay were performed in this study. In addition, the volume of the dentate gyrus was estimated via stereological analysis. anx/anx mice showed significantly higher numbers of both BrdU- and TUNEL-positive cells in the dentate gyrus than those of the control mice. Furthermore, the volume of the dentate gyrus of anx/anx mice was significantly reduced compared to that of the control mice.


Assuntos
Anorexia/fisiopatologia , Apoptose/genética , Divisão Celular/genética , Giro Denteado/patologia , Animais , Anorexia/genética , Bromodesoxiuridina/farmacocinética , Contagem de Células , Giro Denteado/metabolismo , Giro Denteado/fisiopatologia , Ingestão de Alimentos/genética , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Mutantes Neurológicos , Fenótipo , Inanição/genética , Inanição/fisiopatologia
12.
Phys Ther ; 79(6): 558-66, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-10372867

RESUMO

BACKGROUND AND PURPOSE: Computerized 3-dimensional (3-D) motion measurement systems are used by those interested in human motion. The purposes of this study were (1) to determine the limits of accuracy in determining intersegmental angles during pendular motion at varying speeds and (2) to determine changes in accuracy introduced by autodigitization and digitization by experienced manual raters. METHODS: Angular speed of a T-shaped pendulum was systematically increased by releasing the pendulum from 4 angles (0 degrees [no movement], 45 degrees, 90 degrees, and 120 degrees). Twelve reference angles calculated from markers placed on the pendulum were estimated over 20 frames for 10 trials at each release position. RESULTS: Mean errors across trials and frames for intersegmental angles reconstructed by a 3-D motion measurement system were within +/- 1 degree across all release positions. An analysis of variance and a post hoc Tukey test revealed that the mean error for the autodigitized trials was larger than that for the manually digitized trials. For the autodigitized trials, the static trials (release position=0 degrees) produced less mean error than the trials with movement produced. The ICCs showed a high degree of consistency among all raters, ranging from .707 to .999. CONCLUSION AND DISCUSSION: Our findings support the conclusion that under carefully controlled conditions, a 3-D motion measurement system can produce clinically acceptable measurements of accuracy across a range of angular speeds. Furthermore, acceptable accuracy is possible regardless of the digitization method.


Assuntos
Marcha , Processamento de Imagem Assistida por Computador , Modalidades de Fisioterapia/instrumentação , Humanos , Padrões de Referência , Gravação em Vídeo
15.
Exp Clin Endocrinol Diabetes ; 118(5): 333-7, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20213597

RESUMO

Metabolic syndrome (MetS) is rapidly growing into one of the major public health issues worldwide. Interleukin 1 receptor antagonist (IL1Ra) functions as a competitor of proinflammatory cytokines and has an important role in metabolic functions, including insulin secretion. To identify the relationship between the interleukin 1 receptor antagonist gene (IL1RN) and MetS, we genotyped nine single nucleotide polymorphisms (SNPs) in the gene using direct sequencing in 66 MetS patients and 346 normal subjects in the Korean population. Among the nine polymorphisms, after adjusting for age and sex, rs928940 (G>T) showed a significant association with MetS in the codominant ( P= 0.023) and recessive models ( P= 0.011). Also, rs315952 (C>T) exhibited a significant association with MetS in the codominant model ( P= 0.046). The results suggest that the IL1RN polymorphisms may be associated with MetS in the Korean population.


Assuntos
Proteína Antagonista do Receptor de Interleucina 1/genética , Síndrome Metabólica/genética , Adulto , Glicemia/metabolismo , Estatura , Índice de Massa Corporal , Peso Corporal , DNA/genética , DNA/isolamento & purificação , Éxons , Genes Dominantes , Genes Recessivos , Hemoglobinas Glicadas/metabolismo , Humanos , Coreia (Geográfico) , Desequilíbrio de Ligação , Pessoa de Meia-Idade , Razão de Chances , Polimorfismo Genético , Polimorfismo de Nucleotídeo Único , Valores de Referência
16.
Br J Radiol ; 81(970): e238-41, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18796550

RESUMO

Pulmonary Langerhans cell histiocytosis is a rare pulmonary disease that typically affects cigarette smokers from 30-40 years of age onwards. It is very rare in children, especially for those under 15 years of age. We report an atypical radiological manifestation of isolated pulmonary Langerhans cell histiocytosis in a 12-year-old girl that showed multifocal consolidation and multiple small nodules on an initial chest radiograph, and gradual fibrotic change with multiple cysts on follow-up chest radiographs and CT scans.


Assuntos
Tosse/etiologia , Histiocitose de Células de Langerhans/diagnóstico por imagem , Pulmão/diagnóstico por imagem , Tuberculose Pulmonar/diagnóstico por imagem , Criança , Terapia Combinada , Diagnóstico Diferencial , Feminino , Histiocitose de Células de Langerhans/complicações , Histiocitose de Células de Langerhans/tratamento farmacológico , Humanos , Poluição por Fumaça de Tabaco/efeitos adversos , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Tuberculose Pulmonar/tratamento farmacológico
17.
Pharmacogenomics J ; 6(5): 327-32, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16415917

RESUMO

The use of topiramate (TPM) in the treatment of binge-eating disorder, bulimia nervosa, and antipsychotic-induced weight gain has recently increased, however, the exact molecular basis for its effects on body weight reduction and improved glucose homeostasis, is yet to be elucidated. Here we investigated the effect and signaling pathway of TPM on glucose uptake in L6 rat skeletal muscle cells, which account for >70% of glucose disposal in the body. Intriguingly, we found that TPM (10 microM) stimulated the rate of glucose uptake up to twofold increase. And TPM-stimulated glucose transport was inhibited with the overexpression of dominant-negative form of AMP-activated protein kinase (AMPK), an important mediator in glucose transport, implicating that AMPK-mediated pathway is involved. The TPM-stimulated glucose transport was blocked by SB203580, a specific inhibitor of AMPK downstream mediator, p38 mitogen-activated protein kinase (MAPK) protein. LY294002, an inhibitor of phosphatidylinositol (PI) 3-kinase, which is another crucial mediator in independent glucose transport pathway, did not inhibit TPM-stimulated glucose transport. We also found that TPM increased the phosphorylation level of AMPK and p38 MAPK, whereas no effect on the activity of PI 3-kinase of TPM, when assessed by PI 3-kinase assay, was observed. These results together suggest that TPM stimulates glucose transport, not via PI 3-kinase mediated, but via AMPK-mediated pathway in skeletal muscle cells, thereby contributing to the body weight regulation and glucose homeostasis.


Assuntos
Fármacos Antiobesidade/farmacologia , Frutose/análogos & derivados , Glucose/metabolismo , Complexos Multienzimáticos/metabolismo , Músculo Esquelético/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Quinases Ativadas por AMP , Animais , Transporte Biológico , Linhagem Celular , Relação Dose-Resposta a Droga , Frutose/farmacologia , Homeostase , Imidazóis/farmacologia , Fibras Musculares Esqueléticas/efeitos dos fármacos , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Fosforilação , Inibidores de Proteínas Quinases/farmacologia , Piridinas/farmacologia , Ratos , Topiramato , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
18.
Gerontology ; 22(1-2): 89-108, 1976.
Artigo em Inglês | MEDLINE | ID: mdl-1245299

RESUMO

This review discusses the molecular and cytokinetic aspects of cellular aging of human diploid fibroblasts. Despite a large amount of data, the basis of their limited life span has not been defined. A replicative defect in aging cells lies in the mechanism(s) that initiates DNA synthesis, and the control of this mechanism(s) is mediated through an alteration in the sythesis of specific RNA and protein molecules. Two major groups of hypotheses have been offered to explain these findings. The differentiation hypotheses are based on the concept of a 'biological clock', while error hypotheses presuppose defects in genetic transcription or translation.


Assuntos
Envelhecimento , Células Cultivadas/metabolismo , Animais , Membrana Celular/metabolismo , Cricetinae , DNA/biossíntese , Diploide , Fibroblastos/metabolismo , Genética Médica , Humanos , Células Híbridas , Técnicas In Vitro , Camundongos , Proteínas/metabolismo
19.
Cancer Chemother Rep ; 59(1): 147-9, 1975.
Artigo em Inglês | MEDLINE | ID: mdl-805656

RESUMO

We have (a) examined culture conditions for prostatic cells, (b) devised a procedure for the selective enrichment of epithelial cells, and (c) developed a way to identify prostatic epithelial cells by immunologic methods.


Assuntos
Células Epiteliais , Epitélio , Próstata/citologia , Fosfatase Ácida/análise , Animais , Células Cultivadas , Testes Imunológicos de Citotoxicidade , Epitélio/imunologia , Fibroblastos , Haplorrinos/imunologia , Humanos , Masculino , Métodos , Próstata/enzimologia , Tripsina
20.
Proc Natl Acad Sci U S A ; 79(16): 5061-5, 1982 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-6956915

RESUMO

Hybrid cell lines producing monoclonal antibodies against human prostatic acid phosphatase [PAPase; orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) were prepared by the fusion of mouse myeloma cells with the spleen cells of PAPase-immunized BALB/c mice. Approximately 23% of the hybrid cells initially plated after cell fusion produced specific antibodies: 34 microcultures were cloned, and 8 eventually yielded stable cell lines. The monoclonal antibodies produced by these eight hybridomas were characterized for their isotypes, isoelectric points, concentrations, and affinities. All of the eight monoclonal antibodies exhibited strict specificity for PAPase as determined by radioimmunoassay and immunohistochemical methods. These antibodies were used as probes for the antigenic mapping of this enzyme, and three nonoverlapping determinants were recognized. Further binding studies with PAPase fragments, generated by cleavage with a submaxillaris protease, showed that those three determinants are clustered on one fragment of PAPase. These monoclonal antibodies may be useful in refinement of clinical immunoassays of PAPase or immunohistological study of PAPase-synthesizing cells.


Assuntos
Fosfatase Ácida/imunologia , Próstata/enzimologia , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Humanos , Isoenzimas/imunologia , Fígado/enzimologia , Lisossomos/enzimologia , Masculino , Pâncreas/enzimologia
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