RESUMO
The mutations (Trp8 --> Arg and Ile15 --> Thr) in the human LHbeta -subunit caused by nucleotide point mutations in the LHbeta gene were reported in women with immunologically anomalous LH and menstrual disorders. To estimate the effects of the mutations on LH bioactivity in vitro and in vivo, we constructed a LHbeta gene containing this nucleotide mutation in each site or in both sites by site- directed mutagenesis and analyzed the bioactivities of the mutant LH expressed in Chinese hamster ovary cells. Although no alterations were noted in the receptor-binding activity of LH due to the mutations, the LHs containing the mutations at Trp8 --> Arg and at both sites in the beta-subunit showed a higher biopotency of progesterone production in vitro. The clearance rate from the circulation was faster in vivo in all mutant LHs, which were induced primarily by the mutation at Trp8 --> Arg. However, the mutations did not affect the ability of LH to induce ovulation in vivo. These results indicate that LH consisting of the mutant beta-subunit exhibits hyperbioactivity on steroidogenesis and has a short turnover rate. This may affect the endocrine pathway among women with the mutant LH and lead to menstrual disorders.
Assuntos
Hormônio Luteinizante/genética , Receptores do LH/metabolismo , Animais , Sequência de Bases , Células CHO , Cricetinae , Feminino , Técnicas de Transferência de Genes , Humanos , Hormônio Luteinizante/metabolismo , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Mutação Puntual , Ratos , Ratos WistarRESUMO
To analyze the structure of LH in three patients with immunologically anomalous LH, the whole coding region of the LH beta-subunit gene was examined. These patients were infertile, and their serum LH levels could not be measured with an immunoassay kit. Immunoblotting of the LH beta-subunit showed no marked changes in the molecular size of LH beta. Genomic DNA was extracted from peripheral lymphocytes of the patients and normal controls, and LH beta genes were amplified by the polymerase chain reaction technique, using primer pairs that are capable of specifically amplifying only the LH beta gene without interference by the CG beta genes. No deletions were observed in the coding regions of the LH beta gene of the patients. Nucleotide sequencing revealed two nucleotide substitutions in the LH beta gene of the patients, which cause amino acid replacements from Trp8 (TGG) to Arg8 (CGG) and Ile15 (ATC) to Thr15 (ACC). Restriction fragment length polymorphism analysis in three families indicated that the affected probands were homozygous, and their family members were heterozygous, except for their husbands. The heterozygotes showed reduced detectability with the LH immunoassay kit. These results suggest that these amino acid replacements are responsible for this immunologically anomalous variant.
Assuntos
Genes , Doenças do Sistema Imunitário/genética , Hormônio Luteinizante/genética , Hormônio Luteinizante/imunologia , Mutação Puntual , Adulto , Sequência de Bases , Bioensaio , Western Blotting , Feminino , Humanos , Imunoensaio , Hormônio Luteinizante/análise , Dados de Sequência Molecular , Peso Molecular , Linhagem , Polimorfismo de Fragmento de RestriçãoRESUMO
A 13-year-old boy was the victim of attempted strangulation. His condition had returned to normal by the sixth day after the assault; however, from the seventh day, choreoathetosis, dystonia, and marked pseudobulbar paralysis developed in the boy. The computed tomographic scans and T2-weighted magnetic resonance images that were obtained at this time revealed low-density and high-signal intensities in the region of the bilateral putamen and caudate nucleus. These symptoms and the changes in his computed tomographic scans and magnetic resonance images subsided gradually during a 2-month period. Sequential analysis of the cerebrospinal fluid for gamma-aminobutyric acid and dopamine concentrations during his illness revealed reciprocal changes with normal recovery. Because of the delayed onset of neurological changes and the cerebrospinal fluid showing reversible symptoms, the delayed encephalopathy after strangulation had been related to the biochemical alterations that followed anoxia in the vulnerable basal ganglia.
Assuntos
Asfixia/complicações , Encefalopatias/etiologia , Hipóxia Encefálica/etiologia , Adolescente , Asfixia/fisiopatologia , Encéfalo/patologia , Encéfalo/fisiopatologia , Encefalopatias/líquido cefalorraquidiano , Encefalopatias/diagnóstico , Núcleo Caudado/patologia , Crime , Dopamina/líquido cefalorraquidiano , Eletroencefalografia , Humanos , Hipóxia Encefálica/líquido cefalorraquidiano , Hipóxia Encefálica/diagnóstico , Hipóxia Encefálica/fisiopatologia , Imageamento por Ressonância Magnética , Masculino , Putamen/patologia , Tomografia Computadorizada por Raios X , Ácido gama-Aminobutírico/líquido cefalorraquidianoRESUMO
Nonsensitized (DA x PVG/c) F1 hybrid female rats regularly accepted male skin isografts. However, almost all F1 females sensitized with male F1 bone marrow cells rejected male skin isografts. Spleen cells taken from sensitized F1 females, either before or after the rejection of male skin isografts, conferred resistance against graft-versus-host (GVH) reactions produced in F1 females by parental PVG/c lymph node cells, but only when the cell donors were male. Repeated injection with bone marrow cells from F1 males failed to cause rejection of established male skin grafts by F1 females. Additional male skin grafts transferred after bone marrow cell injection were also retained. However, if established male skin isografts had been deliberately removed from F1 females before injection of male bone marrow cells, subsequently placed male skin grafts were rejected. Neither spleen cells collected from F1 females bearing established male skin isografts after repeated injection with male bone marrow cells, nor spleen cells obtained from F1 females that had received four consecutive male skin isografts were observed to confer resistance against GVH reactions produced by male PVG/c lymph node cells in F1 female recipients. Splenectomy of prospective F1 female recipients did not confer on them the capacity to reject male skin isografts. These results imply that the placement of a male skin isograft interfered with subsequent attempts to stimulate F1 hybrid females to reject male grafts and to resist GVH reactions produced by lymphocytes from PVG/c males.
Assuntos
Sobrevivência de Enxerto/fisiologia , Transplante de Pele/imunologia , Quimeras de Transplante , Transplante Isogênico/imunologia , Animais , Transplante de Medula Óssea/imunologia , Transplante de Medula Óssea/patologia , Cruzamentos Genéticos , Feminino , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/imunologia , Masculino , Ratos , Ratos Endogâmicos , Fatores de TempoRESUMO
OBJECTIVE: To evaluate the clinical significance of the LH consisting of a mutant beta-subunit (Trp8 to Arg8 and Ile15 to Thr15). DESIGN: Clinical and biochemical studies. SETTING: Fertility center at the University Hospital and its research laboratory. PATIENTS: Fifty-one patients with menstrual disorders and three homozygote cases and two heterozygote cases of the mutant LH who were reported previously. INTERVENTIONS: Nucleotide mutations of the LH beta gene in patients with menstrual disorders were screened using techniques of the polymerase chain reaction and restriction fragment length polymorphism. Immunologic and biologic activities of the mutant LH and endocrinologic profiles in the affected women were evaluated. MAIN OUTCOME MEASURES: Serum LH levels measured with different immunoassay kits; serum FSH and LH on the GnRH test; serum thyroid-stimulating hormone, PRL, T, and androstenedione; ultrasound examination of the ovaries; clinical hyperandrogenic symptoms; and biologic activity of LH. RESULTS: Two cases of homozygotes and four of heterozygotes affected by the LH beta gene mutations were discovered in the current study through screening of patients with menstrual disorders. Serum LH levels in the homozygote cases were undetectable using a LH immunoassay kit, whereas levels in the heterozygote cases showed reduced detectability with the kit. However, the ratio of the mutant LH values in the bioassay to those in the immunoassay was higher in the homozygote group than that in the control subjects. Response patterns of serum gonadotropins to GnRH in the homozygote were similar to those in patients with polycystic ovary syndrome. CONCLUSION: The mutations of LH beta-subunit might be related to menstrual disorder in some patients.
Assuntos
Hormônio Luteinizante/genética , Distúrbios Menstruais/genética , Mutação , Adolescente , Adulto , Sequência de Bases , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Liberador de Gonadotropina , Heterozigoto , Homozigoto , Humanos , Imunoensaio , Hormônio Luteinizante/sangue , Dados de Sequência Molecular , Síndrome do Ovário Policístico/sangue , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Prolactina/sangueRESUMO
The injectable cephalosporin cefpirome (CPR) was launched in Japan in 1993. It has widely been used in the various infectious diseases. We therefore studied the clinical and bacteriological efficacy of CPR against infections caused by Bacteroides species, Prevotella species and Porphyromonas species frequently isolated from the obstetric and gynecologic infections. Thirteen institutions were involved in this study which ran from March 1994 to January 1999. The administration dosage of CPR was 2 to 4 gram per day administered by drip infusion or intravenous infusion. The duration of treatment was from 3 to 15 days. The evaluations were performed before and after the treatment. CPR was administered to 194 patients with obstetric and gynecologic infections, and 146 of 194 cases were acceptable for the evaluation of drug efficacy. Bacteroides species were identified in 102 patients. Clinical efficacy in 146 cases was excellent in 12 patients, good in 110, fair in 9 and poor in 15 patients. The eradication rate for Bacteroides species could be in 37 cases out of 54 evaluable cases; Prevotella species in 38 out of 49; and Porphyromonas species in 5 out of 5. The overall assessment of bacteriological efficacy was "eradicated" in 91 cases out of 133 (68.4%). Adverse reactions including abnormal findings in laboratory tests were seen in 8 patients (4.76%). Based on these results, CPR promises efficacy and safety in the treatment of obstetric and gynecologic infections due to Bacteroides species.
Assuntos
Infecções por Bacteroidaceae/tratamento farmacológico , Bacteroides/efeitos dos fármacos , Cefalosporinas/uso terapêutico , Doenças dos Genitais Femininos/tratamento farmacológico , Porphyromonas/efeitos dos fármacos , Prevotella/efeitos dos fármacos , Adulto , Idoso , Infecções por Bacteroidaceae/microbiologia , Bacteroides/isolamento & purificação , Cefalosporinas/farmacologia , Resistência Microbiana a Medicamentos , Feminino , Doenças dos Genitais Femininos/microbiologia , Humanos , Pessoa de Meia-Idade , Porphyromonas/isolamento & purificação , Prevotella/isolamento & purificação , Resultado do Tratamento , CefpiromaAssuntos
Analgésicos , Anestesia Caudal , Anestesia Epidural , Adolescente , Adulto , Fatores Etários , Idoso , Analgésicos/administração & dosagem , Criança , Espaço Epidural/fisiologia , Feminino , Humanos , Injeções , Masculino , Pessoa de Meia-Idade , PressãoRESUMO
We report a case of left-sided horizontal gaze palsy, ipsilateral adduction weakness, and left peripheral facial weakness, all of which indicate the lesion in the left median pontine tegmentum. The enhanced MRIs revealed a discrete left median pontine tegmental lesion, involving the abducens nucleus, MLF, and facial nerve knee. This lesion spared the area of the left PPRF. Among these structures, the area of the abducens nucleus seems to be responsible for the unilateral horizontal gaze palsy. We are not aware of any previous precise neuroradiological documentation of unilateral paralysis of conjugate gaze due to a lesion of the abducens nucleus by sagittal and horizontal MRIs.
Assuntos
Tronco Encefálico , Oftalmoplegia/etiologia , Idoso , Idoso de 80 Anos ou mais , Encefalopatias/complicações , Encefalopatias/diagnóstico , Meios de Contraste , Eletroculografia , Feminino , Gadolínio DTPA , Humanos , Imageamento por Ressonância Magnética , Nistagmo Optocinético , Compostos Organometálicos , Ácido PentéticoRESUMO
This study reports a case of MELAS with epileptic seizure, and reviews the characteristics of seizures in patients with this syndrome. They are characterized by: (1) generalized and/or partial seizures, (2) frequent association with visual symptoms and hemiparesis, and (3) posteriorly predominant EEG abnormalities.
Assuntos
Acidose Láctica/fisiopatologia , Encefalopatias/fisiopatologia , Transtornos Cerebrovasculares/fisiopatologia , Eletroencefalografia , Epilepsias Parciais/fisiopatologia , Mitocôndrias Musculares , Doenças Neuromusculares/fisiopatologia , Adulto , Córtex Cerebral/fisiopatologia , Potenciais Evocados/fisiologia , Humanos , Masculino , Mitocôndrias Musculares/fisiologia , Estado Epiléptico/fisiopatologia , SíndromeRESUMO
BACKGROUND/AIMS: Matrix metalloproteinase-13, one of the principal neutral proteinases capable of cleaving native fibrillar collagens, is important in the degradation and remodeling of extracellular matrix. However, its precise expression in liver injury has not been characterized. We examined the kinetics of the expression of matrix metalloproteinase-13 and one of its specific inhibitors, tissue inhibitor of metalloproteinase-1, in acute liver injury in rats. METHODS: Acute liver injury was induced by administration of carbon tetrachloride or two different doses of D-galactosamine hydrochloride in Wistar rats. Hepatic matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 mRNA levels were then examined by Northern blotting. RESULTS: All rats survived after liver injury induced by carbon tetrachloride or low doses of D-galactosamine hydrochloride. However, rats died 5 days after induction of liver injury by high doses of D-galactosamine hydrochloride. In carbon tetrachloride-induced liver injury, matrix metalloproteinase-13 mRNA was transiently increased between 6 h and 1 day after injury. Tissue inhibitor of metalloproteinase-1 mRNA expression was increased between 6 h and 3 days after the peak of matrix metalloproteinase-13 expression. Similar patterns of matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 expression were observed in low-dose D-galactosamine hydrochloride-induced liver injury. In contrast, in high-dose D-galactosamine hydrochloride-induced liver injury, tissue inhibitor of metalloproteinase-1 expression peaked before matrix metalloproteinase-13 expression, which was increased 2 days after injury. Both mRNA levels continued to increase until death. CONCLUSIONS: Transient expression of matrix metalloproteinase-13, followed by that of tissue inhibitor of metalloproteinase-1, was observed during recovery from acute liver injury induced by carbon tetrachloride and low-dose D-galactosamine hydrochloride. In contrast, disordered expression of matrix metalloproteinase-13 was observed in fatal liver injury caused by high-dose D-galactosamine hydrochloride. These results indicate that matrix metalloproteinase13 plays an important role in the early phase of recovery from liver injury.
Assuntos
Colagenases/biossíntese , Fígado/enzimologia , Inibidores Teciduais de Metaloproteinases/biossíntese , Animais , Northern Blotting , Tetracloreto de Carbono/toxicidade , Fígado/patologia , Masculino , Metaloproteinase 13 da Matriz , RNA Mensageiro/análise , Ratos , Ratos WistarRESUMO
BACKGROUND/AIMS: Tissue inhibitor of metalloproteinase-1, a specific inhibitor of matrix metalloproteinases, plays an important role in the pathogenesis of fibrosis and tumor progression. However, the precise expression of tissue inhibitor of metalloproteinase-1 messenger RNA in human hepatic fibrosis has not yet been defined. We investigated the spatial distribution of tissue inhibitor of metalloproteinase-1 messenger RNA in chronic human liver disease. METHODS: Northern and in situ hybridization of probes to tissue inhibitor of metalloproteinase-1 messenger RNA were performed in specimens from 16 surgically resected human livers. Immunohistochemical staining of sections for tissue inhibitor of metalloproteinase-1 and immunoelectron microscopy were also performed. RESULTS: Northern hybridization demonstrated that expression of tissue inhibitor of metalloproteinase-1 messenger RNA was increased 3.9-fold in mild chronic hepatitis, 6.8-fold in moderate chronic hepatitis, and 6.4-fold in cirrhosis, compared with control liver. In situ hybridization showed the expression of tissue inhibitor of metalloproteinase-1 messenger RNA in spindle-shaped cells in the fibrous septa and lobules in chronic hepatitis and cirrhosis; these cells were immunohistochemically positive for a-smooth muscle actin. Immunoelectron microscopy revealed localization of tissue inhibitor of metalloproteinase-1 in between fibers, to the rough endoplasmic reticula of stellate cells located in the lobules and periportal areas, and to fibroblasts in the fibrous septa. These results indicate that tissue inhibitor of metalloproteinase-1 was produced mainly by stellate cells in the specimens of chronic liver diseases. CONCLUSIONS: Expression of tissue inhibitor of metalloproteinase-1 messenger RNA is increased in hepatic fibrosis and stellate cells are involved primarily in its expression.
Assuntos
Hepatopatias/metabolismo , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Adulto , Idoso , Biomarcadores , Northern Blotting , Doença Crônica , Feminino , Humanos , Imuno-Histoquímica , Hepatopatias/fisiopatologia , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/análise , RNA Mensageiro/biossínteseRESUMO
To clarify whether apoptosis is involved in endometriosis, we obtained eutopic endometrial tissues along with endometriotic tissues from the uterus (adenomyosis) (n = 12) and from the ovary (n = 12) from patients undergoing gynaecological surgery. Apoptosis-induced DNA fragmentation was detected by the TdT-mediated dUTP-biotin nick-end labelling method, and immunostaining with a monoclonal antibody against the Fas, Le(y) or B-cell leukaemia/lymphoma-2 (bcl-2) was also performed using the same tissue section. Analysis showed that apoptosis was occurring in all the samples of ovarian endometriotic tissue but in only two of the 12 adenomyotic and in five of the 24 eutopic endometrial tissue samples. In none of these cases was apoptosis correlated with phases of the menstrual cycle. The expression of bcl-2 in the eutopic endometrial and adenomyotic tissues was limited to the proliferative phase, and was observed in only one of the 12 cases of ovarian endometriosis. Fas and Ley were expressed randomly across a wide range in both the eutopic and ectopic endometrial tissues. These results suggest that the features of ovarian endometriosis are different from those of adenomyosis and eutopic endometrium in terms of the involvement of apoptosis. In addition, the regulatory mechanism involved in ovarian endometriosis may differ from that in other endometrial cells.
Assuntos
Apoptose , Endometriose/patologia , Endométrio/patologia , Adulto , Dano ao DNA , Endométrio/metabolismo , Endométrio/ultraestrutura , Feminino , Humanos , Imuno-Histoquímica , Microscopia Eletrônica , Proteínas Proto-Oncogênicas c-bcl-2/metabolismoRESUMO
BACKGROUND/AIMS: Plasminogen activators and plasminogen activator inhibitors are important regulators of the balance between the proteolytic and antiproteolytic activities that determine extracellular matrix turnover. We examined the expression of plasminogen activator-plasmin system components in experimental liver fibrosis of rats. METHODS: Liver fibrosis was produced in rats by injecting carbon tetrachloride for 6 to 12 weeks. Gene expression for plasminogen activator inhibitor-1 (PAI-1), urokinase and tissue plasminogen activators (uPA and tPA), urokinase plasminogen activator receptor (uPAR), and transforming growth factor-beta1 (TGF-beta1) was examined by Northern analysis. Western analysis was performed to detect protein expression of PAI-1, uPA and uPAR. An immunohistochemical study was performed to detect the localization of PAI-1. Additionally, primary cultured liver cells were examined by Northern and Western analyses for this protein with or without prior incubation with TGF-beta1. RESULTS: At 6 weeks, when fibrosis had occurred, uPA and uPAR mRNAs had increased 2.8-fold and 1.8-fold, respectively; PAI-1 and tPA mRNA levels were unchanged. At the cirrhotic stage (9 to 12 weeks), mRNA levels for PAI-1, uPA, uPAR and tPA were all increased. Western analysis also showed increased uPA and uPAR expressions in fibrotic liver, and increased PAI-1, uPA and uPAR expressions in cirrhotic liver. PAI-1 protein was also demonstrated immunohistochemically along sinusoids, vessels, and bile duct cells of normal and fibrotic liver. In liver cell cultures, Kupffer cells, hepatocytes, and especially stellate cells, expressed PAI-1. Expression was enhanced in stellate cells cultured from fibrotic or cirrhotic liver or stimulated in vitro with TGF-beta1. CONCLUSION: Though increased uPA and uPAR may act on matrix degradation in fibrotic liver, increased PAI-1 together with uPA, uPAR and tPA are associated with overall inhibition of matrix degradation in cirrhotic liver. Hepatic stellate cells are an important source of PAI-1 during liver fibrosis.
Assuntos
Cirrose Hepática Experimental/etiologia , Cirrose Hepática Experimental/metabolismo , Fígado/metabolismo , Ativadores de Plasminogênio/metabolismo , Inativadores de Plasminogênio/metabolismo , Animais , Northern Blotting , Western Blotting , Células Cultivadas , Imuno-Histoquímica , Fígado/patologia , Cirrose Hepática Experimental/patologia , Masculino , Inibidor 1 de Ativador de Plasminogênio/biossíntese , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores de Ativador de Plasminogênio Tipo Uroquinase , Fator de Crescimento Transformador beta/genética , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/metabolismoRESUMO
We have previously reported increased expression of matrix metalloproteinase-2 (MMP-2) using a rat model of liver fibrosis. However we did not clarify how the precursor of MMP-2 (proMMP-2) was activated. Therefore, we used human liver specimens with chronic hepatitis (CH) and liver cirrhosis (LC) to examine expression of membrane-type-1-MMP (MT1-MMP), which has recently been determined to activate proMMP-2. Northern hybridization studies showed a 5.4- and 1.4-fold increase in MMP-2 expression in CH and LC, respectively, as compared with normal liver. MT1-MMP gene expression simultaneously increased 4.0- and 1.4-fold in CH and LC, respectively. In situ hybridization using 35S-cRNA probes of MMP-2 and MT1-MMP showed prominent silver granules in elongated cells found in the lobules, periportal areas, and fibrous septa of CH and LC samples. These elongated cells expressed alpha-smooth muscle actin by immunohistochemistry. Immunoelectron microscopic examination localized MMP-2 and MT1-MMP to the rough endoplasmic reticulum of stellate cells located in the lobules and periportal areas, or to fibroblasts in the fibrous septa, suggesting that MMP-2 and MT1-MMP were produced by these cells. In addition, cytoplasmic and membranous immunodeposits of both MMPs were found in endothelial cells, Kupffer cells, capillary endothelial cells, and lymphocytes, indicating that activation of proMMP-2 occurs locally. Increased expression of MMP-2 and MT1-MMP was detected in CH and LC, while dual over-expression was found in stellate cells and fibroblasts, possibly resulting in the increase of active MMP-2 in and around these cells. These findings suggest that activated MMP-2 may remodel liver parenchyma during the process of liver fibrosis.
Assuntos
Gelatinases/metabolismo , Cirrose Hepática/enzimologia , Metaloendopeptidases/metabolismo , Adulto , Idoso , Northern Blotting , Retículo Endoplasmático Rugoso/enzimologia , Retículo Endoplasmático Rugoso/ultraestrutura , Feminino , Hepatite Crônica/enzimologia , Hepatite Crônica/patologia , Humanos , Imuno-Histoquímica , Hibridização In Situ , Fígado/enzimologia , Fígado/ultraestrutura , Cirrose Hepática/patologia , Masculino , Metaloproteinase 2 da Matriz , Metaloproteinases da Matriz Associadas à Membrana , Microscopia Imunoeletrônica , Pessoa de Meia-Idade , RNA/análiseRESUMO
Matrix metalloproteinase-II (MMP-II, 72-kd type IV collagenase, or gelatinase) is one of the gene families of zinc enzymes capable of degrading extracellular matrix molecules, and specifically of degrading type IV and V collagens, gelatin, fibronectin, and elastin. In this study, we used both the liver fibrosis model and the reversibility model of experimental cirrhosis to clarify how MMP-II participates in liver fibrosis of rats. To produce fibrosis model, rats received subcutaneous injections of CCl4 twice weekly for 7, 9, or 14 weeks. For the reversibility model, rats were treated with CCl4 three times a week for 8 weeks and killed at 3, 7, 14, 28, or 42 days after discontinuation of treatment. MMP-II gene expression was studied by Northern hybridization technique, and gelatinase activity of MMP-II was examined by zymography using gelatin substrate. At the same time, an immunohistochemical study using anti-type IV collagen antibody was carried out. In liver fibrosis model, nodule formation was established at 14 weeks. Immunodeposit of type IV collagen was increased in wide fibrous septa and was clearly observed along sinusoidal wall. Gene expression of MMP-II increased up to 7 to 12 times compared with that of controls, with the expression rate being maximum at an intermediate stage of fibrosis. Zymography showed the expressions of both 65-kd latent MMP-II, which is confirmed to be activated by adding p-aminophenylmercuric acetate, and 62-kd active MMP-II during fibrosis. The expression of both forms increased 13 to 28 times as the fibrosis progressed.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Gelatinases/metabolismo , Cirrose Hepática Experimental/metabolismo , Metaloendopeptidases/metabolismo , Animais , Northern Blotting , Colágeno/metabolismo , Gelatinases/genética , Imuno-Histoquímica , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática Experimental/patologia , Masculino , Metaloproteinase 2 da Matriz , Metaloendopeptidases/genética , RNA Mensageiro/metabolismo , Ratos , Ratos WistarRESUMO
BACKGROUND: The prognoses of patients with squamous cell carcinoma of the ovary are quite poor. However, preoperative diagnosis is difficult due to the rarity of this tumor and its similarity to mature cystic teratoma (MCT). The objective of this study was to assess the value of tumor markers and clinical characteristics in making a differential diagnosis between MCT and squamous cell carcinoma arising from MCT. METHODS: Between September 1979 and June 1996, 37 patients with ovarian squamous cell carcinoma arising from MCT were treated by the Tokai Ovarian Tumor Study Group. The authors evaluated tumor markers, tumor size, and age as parameters for differentiation between MCT and squamous cell carcinoma arising from MCT. Diagnostic efficiency was calculated as the sensitivity multiplied by the specificity. RESULTS: There were significant differences (P < or = 0.0002) in age, tumor size, and levels of squamous cell carcinoma antigen (SCC), CA125, and CEA, as well as a significant difference (P < or = 0.0396) in the CA19-9 level between MCT and squamous cell carcinoma arising from MCT. Diagnostic efficiency was highest for SCC (63.0%), followed by CA125 (50.7%). Receiver operating characteristic (ROC) curves demonstrated that CEA was the best screening marker for squamous cell carcinoma arising from MCT, whereas age and tumor size were better markers than CA125 or CA19-9. The optimal cutoff values for age and tumor size were 45 years and 99 mm, respectively, according to ROC analysis. CONCLUSIONS: These findings demonstrate that age and tumor size are important factors in making a differential diagnosis. In addition, SCC and CEA levels should be measured in patients age 45 years or older who have an MCT-like ovarian tumor larger than 99 mm in greatest dimension.
Assuntos
Carcinoma de Células Escamosas/diagnóstico , Neoplasias Ovarianas/diagnóstico , Serpinas , Teratoma/diagnóstico , Adulto , Fatores Etários , Idoso , Antígenos de Neoplasias/análise , Antígeno Ca-125/análise , Antígeno CA-19-9/análise , Antígeno Carcinoembrionário/análise , Carcinoma de Células Escamosas/patologia , Diagnóstico Diferencial , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias Ovarianas/patologia , Teratoma/patologiaRESUMO
We investigated the molecular species of sulfated sialyl Lewis X determinants, the putative L-selectin ligand, expressed on high endothelial venules (HEV) in human lymph nodes. Comparison of the reactivity pattern of HEV with the reactivity of the pure 6-sulfo, 6'-sulfo, or 6,6'-bissulfo sialyl Lewis X determinant with hitherto known anti-sialyl Lewis X antibodies strongly suggested 6-sulfo sialyl Lewis X to be the best candidate for the major sulfated sialyl Lewis X determinant on HEV, followed by 6,6'-bissulfo sialyl Lewis X, whereas 6'-sulfo sialyl Lewis X was unlikely. We newly generated monoclonal antibodies (mAbs) G152 and G72 directed against 6-sulfo sialyl Lewis X, which intensely labeled HEV in immunohistochemical examination and inhibited binding of recombinant L-selectin-IgG to HEV, suggesting that the determinant serves as a ligand for L-selectin. To test the concomitant expression of 6, 6'-bissulfo sialyl Lewis X, specific mAbs (G2706, G27011, G27037, and G27039) were generated, but all antibodies failed to react to HEV. Next, we established mAbs (AG97 and AG273) directed against 6-sulfo Lewis X, the asialo form of 6-sulfo sialyl Lewis X. The antibodies were not reactive to untreated HEV, but strongly reacted to sialidase-treated HEV. This indicated the predominance of the sialylated form of 6-sulfo sialyl Lewis X and minimal expression of its asialo form, corroborating that it was synthesized by fucosyltransferase VII, the isoenzyme that preferentially produces the sialylated form of the determinant.