RESUMO
BACKGROUND: CD5-positive (CD5+) diffuse large B-cell lymphoma (DLBCL) shows poor prognosis and frequent central nervous system (CNS) relapses under anthracycline-containing chemotherapy. The aim of this study was to determine the prognosis and CNS relapse incidence of CD5+ DLBCL in the rituximab era. PATIENTS AND METHODS: We analyzed 337 patients with CD5+ DLBCL who received chemotherapy with (R-chemotherapy group; n = 184) or without (chemotherapy group; n = 153) rituximab. RESULTS: No significant difference was found in clinical background comparisons between the two groups. In the R-chemotherapy group, 60% of the patients were older than 65 years at diagnosis. Both the complete response rate and overall survival (OS) were significantly better in the R-chemotherapy group (P = 0.0003 and P = 0.002, respectively). Multivariate analysis confirmed that chemotherapy without rituximab was associated with unfavorable OS. However, the probability of CNS relapse did not differ between the two groups (P = 0.89). The CNS relapse was strongly associated with short OS (P < 0.0001). In the R-chemotherapy group, 83% of patients who experienced CNS relapse had parenchymal disease. CONCLUSIONS: Our results indicate that rituximab improves the OS of patients with CD5+ DLBCL but does not decrease the CNS relapse rate. More effective treatments with CNS prophylaxis are needed for CD5+ DLBCL patients.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Encefálicas/tratamento farmacológico , Antígenos CD5/metabolismo , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Recidiva Local de Neoplasia/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais Murinos/administração & dosagem , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/secundário , Ciclofosfamida/administração & dosagem , Doxorrubicina/administração & dosagem , Feminino , Seguimentos , Humanos , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Prednisona/administração & dosagem , Indução de Remissão , Estudos Retrospectivos , Rituximab , Taxa de Sobrevida , Resultado do Tratamento , Vincristina/administração & dosagem , Adulto JovemRESUMO
We present here seven cases of idiopathic multicentric Castleman's disease (MCD) showing effusion at the initial clinical presentation. This series includes a high proportion of middle-aged and elderly females (5/7). Various autoantibodies were detected in six cases. Anemia (Hb < 10 g/dl) was detected in four cases, leukocytosis (WBC > 10 × 10(9)/l) in three and thrombycytopenia (<100 × 10(9)/l) in five. Positivity for C-reactive protein or elevated erythrocyte sedimentation rate was recorded in all seven cases. Elevated serum IgG level (>2000 mg/dl) was recorded in only three cases. Elevated serum interleukin-6 level was recorded in all four cases examined. At the onset of disease, four cases were associated with idiopathic thrombocytic purpura. During the course of disease, one case each was diagnosed as systemic sclerosis + Sjögren's syndrome (SJS) and SJS. Histologically, five lesions exhibited a mixed type of Castleman's disease, and one case each exhibited a hyaline-vascular type and plasma cell type. The non-neoplastic nature of the B-lymphocytes was demonstrated by immunohistochemistry and polymerase chain reaction. There were no human herpes type-8 virus-positive cells in any of the seven lesions. Good responsiveness to glucocorticoid therapy has been seen in all six cases treated. From a therapeutic perspective, it is important to discriminate this subtype of MCD.
Assuntos
Hiperplasia do Linfonodo Gigante , Exsudatos e Transudatos , Adulto , Idoso , Hiperplasia do Linfonodo Gigante/diagnóstico , Hiperplasia do Linfonodo Gigante/imunologia , Hiperplasia do Linfonodo Gigante/patologia , Hiperplasia do Linfonodo Gigante/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Indução de RemissãoRESUMO
This study was undertaken to establish reliable factors in order to identify chlamydial cervicitis among suspicious patients. Between January and December 2007, 406 patients who were suspected to have cervicitis due to clinical symptoms, were tested with polymerase chain reaction (PCR) for Chlamydia trachomatis (CT), vaginal pH and Nugent score (NS) in our University hospital and related clinics. During the same period, 67 patients who were diagnosed as having other sexually transmitted diseases (Neisseria gonorrhoeae (NG), Trichomonas vaginalis, Condyloma acuminatum and genital herpes) were also made to participate in this study. Eighty-nine women (22%) were positive for CT PCR. Bacterial vaginosis (BV)-positive women were tested positive for CT PCR (75/288), significantly higher than those without BV (6/66, P = 0.01). In addition, under 20-years old women were positive for CT PCR (24/57), significantly higher than those who were over 30 years old (16/113, P = 0.001). The proportion of patients with high NS (>7) in CT, NG and T. vaginalis cases were 75/89 (84.3%), 22/27 (81.5%) and 11/14 (78.6%), respectively. Whereas the high NS of the C. acuminatum and genital herpes groups were recorded at 7/14 (50%) and 4/12 (33.3%), respectively. Younger women with BV could be at a higher risk for STDs, especially for CT cervicitis.
Assuntos
Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/isolamento & purificação , Infecções Sexualmente Transmissíveis/diagnóstico , Cervicite Uterina/diagnóstico , Vaginose Bacteriana/diagnóstico , Adulto , Infecções por Chlamydia/epidemiologia , Infecções por Chlamydia/microbiologia , Infecções por Chlamydia/fisiopatologia , Chlamydia trachomatis/classificação , Chlamydia trachomatis/genética , Feminino , Humanos , Concentração de Íons de Hidrogênio , Reação em Cadeia da Polimerase , Infecções Sexualmente Transmissíveis/epidemiologia , Infecções Sexualmente Transmissíveis/etiologia , Infecções Sexualmente Transmissíveis/microbiologia , Cervicite Uterina/epidemiologia , Cervicite Uterina/microbiologia , Cervicite Uterina/fisiopatologia , Vagina/microbiologia , Vagina/fisiologia , Vaginose Bacteriana/epidemiologia , Vaginose Bacteriana/microbiologia , Adulto JovemRESUMO
Cadmium induces the expression of the 70 kDa heat shock protein (HSP70) and metallothionein (MT), both of which are considered to be associated with intracellular glutathione (GSH) metabolism in the cellular protection mechanism against cadmium-induced cellular injury. We determined the effects of N-acetyl-L-cysteine (NAC), which increases the intracellular GSH levels, on the induction of HSP70 and MT gene expression in a cultured cell line of human amniotic cells (WISH) exposed to CdCl2. The mRNA level of MT-II, a major isoform of MT genes, was more prominently increased than that of HSP70 when WISH cells were exposed to CdCl2 (5-15 microM, for 6 h). The treatment of WISH cells with 1.5 and 30 mM NAC for 2 h increased the intracellular GSH levels by 1.4- and 3.1-fold, respectively. Pretreatment of cells with 30 mM NAC significantly reduced both HSP70 and MT-II mRNA levels in the cells exposed to 50 microM CdCl2. This concentration of NAC also efficiently suppressed the cadmium-induced lethality. On the contrary, pretreatment with 1.5 mM NAC suppressed only the induction of HSP70 gene expression in the 50 microM CdCl2-treated cells, and did not inhibit the metal toxicity. However, this low concentration of NAC efficiently suppressed lipid peroxidation which was increased by 50 microM CdCl2. Furthermore, this low concentration of NAC also decreased the CdCl2-induced gene expression of HSP32 which represents a general response to oxidative stress. Taken together, NAC seems to have at least two concentration-dependent functions in WISH cells exposed to CdCl2; the low concentration of NAC can suppress the induction of HSP70 gene expression as well as the increase of lipid peroxidation via an antioxidant pathway, while the high concentration of NAC can suppress the induction of MT-II mRNA as well as cadmium-induced cell death. Our present data suggest that changes in intracellular redox status, as reflected by GSH concentration, have more important effects on the induction of HSP70 mRNA rather than that of MT-II mRNA in human amniotic cells exposed to cadmium.
Assuntos
Acetilcisteína/farmacologia , Cádmio/toxicidade , Proteínas de Choque Térmico HSP70/genética , Metalotioneína/genética , Acetilcisteína/administração & dosagem , Âmnio , Linhagem Celular , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Humanos , Peroxidação de Lipídeos/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The induction mechanism of heat shock 70 (Hsp70) gene by cadmium was investigated. In human amniotic WISH cells, Hsp 70 was induced by cadmium in a dose- and time-dependent manner. Cadmium-induced Hsp70 mRNA levels were enhanced 3- to 4-fold after depletion of intracellular glutathione (GSH) by either diethylmaleate or buthionine sulfoximine. Under these conditions, hydrogen peroxide might increase in the absence of substrate for glutathione peroxidase. We found that exogenous hydrogen peroxide alone induced Hsp70 which was further enhanced significantly after GSH-depletion by diethylmaleate. On the other hand, treatment of cells by diethyldithiocarbamate, an inhibitor of superoxide dismutase, induced Hsp70 2-fold over the level of control. This induction was further stimulated by cadmium even in the presence of GSH. Furthermore, a 4-fold increase of intracellular GSH by the treatment of cells with glutathione isopropyl ester did not diminish the cadmium-induced Hsp70. Gel mobility shift assays of nuclear extracts, from these differently treated cells, with oligonucleotide containing a promoter region of Hsp70 gene revealed that the levels of Hsp70 mRNA observed in the present study corresponded to the changes of transcription. These results imply that the induction of Hsp70 mRNA by cadmium is mediated at least partly via reactive oxygen species and attenuated by cellular GSH and that some part of cadmium-induced Hsp70 can not be eliminated by GSH, suggesting that multiple signals are functioning for this induction.
Assuntos
Cádmio/farmacologia , Cloretos/farmacologia , Glutationa/farmacologia , Proteínas de Choque Térmico HSP70/genética , RNA Mensageiro/biossíntese , Sequência de Bases , Cloreto de Cádmio , Linhagem Celular , Radicais Livres , Humanos , Peróxido de Hidrogênio/farmacologia , Dados de Sequência Molecular , Superóxidos/metabolismoRESUMO
PURPOSE: RPR 109881A is a new semisynthetic taxoid compound that has a similar mechanism of action to docetaxel. The purpose of this phase I study was to characterize the maximum-tolerated dose (MTD), toxicity profile, pharmacokinetic profile, and antitumor effects of this agent. PATIENTS AND METHODS: Nineteen eligible patients with advanced solid tumors were enrolled. RPR 109881A was administered as a 1-hour intravenous infusion every 3 weeks at doses ranging from 15 to 75 mg/m(2). Pharmacokinetic evaluation was performed at the first cycle. RESULTS: Neutropenia (febrile neutropenia) and fatigue were dose-limiting toxicities at doses of 60 and 75 mg/m(2) and seemed to be dose-related. Both thrombocytopenia and anemia were infrequent. Nonhematologic toxicities were generally mild. Pharmacokinetic studies indicated that RPR 109881A plasma disposition was bi- or triphasic, with a high total plasma clearance, a large volume of distribution, and a long terminal half-life. The area under the concentration-time curve (AUC) and the peak concentration of RPR 109881A seemed to increase with increasing dose proportionally, suggesting linear pharmacokinetics. Urinary excretion over 48 hours was low, with a mean of 0.8 +/- 0.36% of the administered dose. A significant relationship existed between the percentage decrease of neutrophil counts and the AUC of RPR 109881A. Among 18 assessable patients, two partial and two minor responses were documented. CONCLUSION: RPR 109881A was found to be a well-tolerated and promising taxoid agent. The MTD was 75 mg/m(2), and the recommended dose for phase II study was 60 mg/m(2) as a 1-hour infusion every 3 weeks.
Assuntos
Antineoplásicos Fitogênicos/efeitos adversos , Antineoplásicos Fitogênicos/farmacocinética , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Paclitaxel/efeitos adversos , Paclitaxel/farmacocinética , Taxoides , Adulto , Idoso , Antineoplásicos Fitogênicos/administração & dosagem , Relação Dose-Resposta a Droga , Esquema de Medicação , Feminino , Humanos , Infusões Intravenosas , Masculino , Pessoa de Meia-Idade , Paclitaxel/administração & dosagem , Paclitaxel/análogos & derivadosRESUMO
The objectives of the present study were to evaluate whether a schedule-dependent pharmacokinetic and/or pharmacodynamic interaction exists between two sequences of docetaxel and doxorubicin administration and to determine the maximal tolerated dose (MTD) of this combination. Patients with chemotherapy-naïve metastatic or recurrent advanced breast cancer were enrolled. In the crossover design, tandem dose escalation of docetaxel and doxorubicin was performed. Docetaxel, in doses ranging from 50-70 mg/m2, was administered for 1 h by drip infusion either just before or after a 5-min bolus i.v. injection of doxorubicin at dosages from 40-50 mg/ m2. The sequence of drug administration was switched after the first course in each patient, and the sequence of drug administration thereafter depended on the patient's choice. Twenty-five patients were initially assessable for toxicity. The MTD in the sequence of doxorubicin after docetaxel was 40 and 50 mg/m2, respectively, with the dose-limiting toxicity of neutropenia. On the other hand, the MTD of the sequence of docetaxel after doxorubicin was 70 and 50 mg/m2, respectively. The dose-limiting toxicities in this sequence were neutropenia and diarrhea. Duration of grade 4 neutropenia in the sequence of docetaxel followed by doxorubicin was significantly longer than that in the alternate sequence (P = 0.0062). However, there was no difference in pharmacokinetic parameters of docetaxel, doxorubicin, and doxorubicinol between the two sequences. The sequence of 50 mg/m2 doxorubicin followed by 60 mg/m2 docetaxel is recommended for subsequent clinical trials for practical reasons.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/administração & dosagem , Doxorrubicina/uso terapêutico , Paclitaxel/análogos & derivados , Paclitaxel/administração & dosagem , Paclitaxel/uso terapêutico , Taxoides , Adulto , Idoso , Análise de Variância , Protocolos de Quimioterapia Combinada Antineoplásica/farmacocinética , Protocolos de Quimioterapia Combinada Antineoplásica/toxicidade , Área Sob a Curva , Neoplasias da Mama/sangue , Cromatografia Líquida de Alta Pressão , Estudos Cross-Over , Docetaxel , Relação Dose-Resposta a Droga , Doxorrubicina/farmacocinética , Doxorrubicina/toxicidade , Feminino , Humanos , Dose Máxima Tolerável , Pessoa de Meia-Idade , Neutropenia , Paclitaxel/farmacocinética , Paclitaxel/toxicidade , Fatores de TempoRESUMO
Introduction of an acyl group to the 3-O-position of erythromycin A derivatives instead of L-cladinose led to a novel class of macrolide antibiotics that we named "acylides". The 3-O-nitrophenylacetyl derivative TEA0777 showed significantly potent activity against not only erythromycin-susceptible Gram-positive pathogens but also inducibly macrolides-lincosamides-streptogramin B (MLS(B))-resistant Staphylococcus aureus and efflux-resistant Streptococcus pneumoniae. These results indicated that acylides have potential as next-generation macrolide antibiotics.
Assuntos
Antibacterianos/síntese química , Eritromicina/análogos & derivados , Eritromicina/síntese química , Animais , Antibacterianos/química , Antibacterianos/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecium/efeitos dos fármacos , Eritromicina/química , Eritromicina/farmacologia , Camundongos , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/mortalidade , Staphylococcus aureus/efeitos dos fármacos , Streptococcus pneumoniae/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
We have previously shown that thioredoxin and thioredoxin reductase were immunohistochemically localized in cytotrophoblasts, decidua and stromal cells in the stem villi of human placenta and that the addition of exogenous thioredoxin and thioredoxin reductase to mitochondrial fractions from human placenta displayed a protective effect on fumarase activity against oxidative stress. In this study, to investigate further the roles of thioredoxin and thioredoxin reductase in protecting pregnancy against oxidative stress, we examined the effect of lipopolysaccharide (LPS), which induces a variety of cytokines and produces radical oxygen species, on the expression of thioredoxin and thioredoxin reductase in mouse placenta. We focused on the placental protective effect in the second trimester, when the onset of placental dysfunction might occasionally lead to a critical state for the fetus. Thus we analysed placentae from mice on day 13 of pregnancy at various time points after they were injected with LPS (50 microg/kg i.p.) or saline as a control. The expressions of thioredoxin and thioredoxin reductase were evaluated by Western blotting and immunohistochemistry. Western blot analysis revealed that LPS approximately quadrupled the expression of both thioredoxin and thioredoxin reductase in the placentae of pregnant mice. When both proteins were localized immunohistochemically, it was found that the decidua and the diploid trophoblasts in the basal zone were intensively stained. Furthermore, the expression of 4-hydroxy-2-nonenal (HNE)-modified proteins, which are markers of oxidative stress, was enhanced in placenta by LPS. Our study suggests that the induced thioredoxin and thioredoxin reductase might protect the placenta from the stress induced by LPS.
Assuntos
Lipopolissacarídeos/farmacologia , Placenta/química , Tiorredoxina Dissulfeto Redutase/análise , Tiorredoxinas/análise , Aldeídos/farmacologia , Animais , Western Blotting , Escherichia coli , Feminino , Idade Gestacional , Immunoblotting , Imuno-Histoquímica , Peroxidação de Lipídeos , Camundongos , Camundongos Endogâmicos ICR , Gravidez , Proteínas/análiseRESUMO
Recent studies have indicated that oxidative stress is involved in the pathogenesis of pre-eclampsia. Oxidative stress damages systemic tissues, and placental damage may result in intrauterine growth retardation and fetal distress. Thus, this study attempted to elucidate the placental localization of thioredoxin and thioredoxin reductase, substances that may reduce oxidative stress. Furthermore, it studied the defence mechanism of the thioredoxin-thioredoxin reductase system against oxidative stress in mitochondria of normal human placenta where reactive oxygen species are primarily produced. The examination of thioredoxin reductase activity in subcellular fractions of human placenta indicated that thioredoxin reductase was located not only in cytoplasm, but also in mitochondria. The existence of thioredoxin and thioredoxin reductase in human placenta was confirmed immunologically using antibodies raised against thioredoxin and thioredoxin reductase. Thioredoxin and thioredoxin reductase were localized histochemically in cytotrophoblasts, decidua, and stromal cells in the stem villi. The addition of exogenous thioredoxin and thioredoxin reductase to fumarase in mitochondria of human placenta displayed a protective effect against oxidative stress. In conclusion, this study confirmed the intracellular localization and the tissue distribution of thioredoxin and thioredoxin reductase in human placenta. Moreover, the complete thioredoxin-thioredoxin reductase system in human placenta may protect the placenta from damage caused by oxidative stress.
Assuntos
Estresse Oxidativo , Placenta/química , Tiorredoxina Dissulfeto Redutase/análise , Tiorredoxinas/análise , Western Blotting , Citosol/enzimologia , Feminino , Fumarato Hidratase/metabolismo , Idade Gestacional , Humanos , Peróxido de Hidrogênio/farmacologia , Imuno-Histoquímica , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/enzimologia , Placenta/ultraestrutura , Gravidez , Tiorredoxina Dissulfeto Redutase/fisiologiaRESUMO
Complete hydatidiform mole and coexistent fetus (CMCF) is a rare occurrence and is associated with an increased risk of persistent gestational trophoblastic diseases. The aim of this study was to reveal a potential risk factor and to determine optimum management of CMCF cases. Molar tissues are cytogenetically divided into two types, homozygous and heterozygous. The molar tissue of our case showed a 46, XY heterozygous complete mole. Genomic DNA was analyzed by the polymerase chain reaction using sets of unlabelled forward and Cy-5-labelled reverse primers for DNA marker loci. The patient developed persistent trophoblastic disease (PTD) with lung metastasis. Since 1980 there have been 13 reports (including our case) that cytogenetically revealed CMCF and clarified the clinical outcome. Nine of the 16 CMCF cases before 21 weeks of gestation and seven of the 12 CMCF cases after 22 weeks of gestation developed PTD. The incidence of PTD from CMCF was not related to the gestational age at termination or delivery. There were 10 case reports that analyzed the zygosity of a mole, heterozygous or homozygous. Two of six homozygous and three of four heterozygous moles in CMCF cases developed PTD. A heterozygous mole is thought to be a high risk factor for the incidence of PTD. Cytogenetic study is clinically useful for the optimum management of CMCF cases.
Assuntos
Mola Hidatiforme/diagnóstico , Adulto , Antineoplásicos/uso terapêutico , Gonadotropina Coriônica Humana Subunidade beta/sangue , DNA/análise , Feminino , Genótipo , Idade Gestacional , Heterozigoto , Humanos , Mola Hidatiforme/complicações , Mola Hidatiforme/genética , Cariotipagem , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/secundário , Masculino , Metotrexato/uso terapêutico , Reação em Cadeia da Polimerase , Gravidez , Gravidez Múltipla , Tomografia Computadorizada por Raios X , Neoplasias Trofoblásticas/etiologia , Gêmeos , Ultrassonografia Pré-Natal , Neoplasias Uterinas/etiologiaRESUMO
Recent studies have indicated that pre-eclampsia is closely associated with oxidative stress both in maternal circulation and in the placenta. Protein thiol/disulphide oxidoreductases, such as thioredoxin, glutaredoxin, and protein disulphide isomerase have recently been found to eliminate reactive oxygen species (ROS) and regenerate oxidatively damaged proteins. Protein thiol/disulphide oxidoreductases may also play a role in combating pre-eclampsia. In this study, we examined the accumulation of 4-hydroxy-2-nonenal (HNE)-modified proteins, which are markers of lipid peroxidation, in human placentae of normal and pre-eclamptic subjects. We also examined the protein levels of thioredoxin, glutaredoxin, and protein disulphide isomerase in placentae. Immunoblotting and immunohistochemistry showed that HNE-modified proteins accumulated to a greater extent in pre-eclamptic placentae than in normal placentae. In both normal and pre-eclamptic placentae, thioredoxin, glutaredoxin, and protein disulphide isomerase were detected in the trophoblasts of the floating villi. The levels of these proteins were increased approximately 2- to 3-fold in the pre-eclamptic placentae compared to the normal placentae. These results indicated that the pre-eclamptic placentae were exposed to oxidative stress and that the protein thiol/disulphide oxidoreductases were adaptively induced in pre-eclamptic placentae, suggesting possible roles for thioredoxin, glutaredoxin, and protein disulphide isomerase in protecting placental functions against oxidative stress caused by pre-eclampsia.
Assuntos
Oxirredutases , Placenta/enzimologia , Pré-Eclâmpsia/enzimologia , Proteína Dissulfeto Redutase (Glutationa)/metabolismo , Adulto , Aldeídos/metabolismo , Feminino , Idade Gestacional , Glutarredoxinas , Humanos , Immunoblotting , Imuno-Histoquímica , Peroxidação de Lipídeos , Estresse Oxidativo , Placenta/química , Gravidez , Isomerases de Dissulfetos de Proteínas/análise , Isomerases de Dissulfetos de Proteínas/metabolismo , Proteínas/análise , Proteínas/metabolismo , Tiorredoxinas/análise , Tiorredoxinas/metabolismoRESUMO
The aim of this study was to elucidate whether a novel mitochondrial antioxidant protein, SP-22, is localized in placenta and whether its expression is induced in placenta of lipopolysaccharide (LPS)-exposed mouse. Western blot analysis of normal human placenta indicated that the SP-22 protein was located in the mitochondrial fraction. Immunohistochemical analysis of SP-22 in normal placenta showed that immunoreactive SP-22 was distributed mostly in cytotrophoblastic cells but with almost no signal in syncytiotrophoblasts. The positive signals were also detected in the decidual cells and stromal cells in stem villi of normal placenta. We also examined LPS-mediated inflammatory placenta on day 13 of pregnancy at various time points after LPS injection (50 microg/kg, intraperitoneally). Western blot analysis indicated that LPS approximately quadrupled the expression of SP-22 in placenta of LPS-exposed mouse. When the SP-22 protein was localized immunohistochemically, the decidua and the diploid trophoblasts in the basal zone were intensively stained in placenta of LPS-exposed mouse compared to the control. The localization and inducible expression of SP-22 protein in placenta suggest a possible role in antioxidant system in mitochondria of normal and inflammatory placentae.
Assuntos
Inflamação/metabolismo , Mitocôndrias/química , Peroxidases/análise , Doenças Placentárias/metabolismo , Placenta/ultraestrutura , Animais , Western Blotting , Decídua/química , Feminino , Idade Gestacional , Humanos , Imuno-Histoquímica , Inflamação/induzido quimicamente , Lipopolissacarídeos , Camundongos , Peroxirredoxina III , Peroxirredoxinas , Placenta/química , Doenças Placentárias/induzido quimicamente , Gravidez , Células Estromais/química , Trofoblastos/químicaRESUMO
A growing body of evidence indicates that the pathogenesis of pre-eclampsia is closely associated with oxidative stress occurring in mitochondria. In the present study, we evaluated the degree of mitochondrial lipid peroxidation by assessing the accumulation of 4-hydroxy-2-nonenal (HNE)-modified proteins and examined the expression of mitochondrial antioxidant protein peroxiredoxin III/SP-22 in normal and pre-eclamptic human placentae. The accumulation of HNE-modified proteins increased to a greater extent in both the mitochondria and cytosol of pre-eclamptic placentae than in those of normal placentae. Moreover, the accumulation of HNE-modified proteins was much more evident in the mitochondria than in the cytosol, indicating that lipid peroxidation occurred mainly in the mitochondria of pre-eclamptic placentae. The mRNA expression of peroxiredoxin III/SP-22 was increased about 2-fold in pre-eclamptic placentae compared to normal placentae. The protein levels of peroxiredoxin III/SP-22 were approximately 4-fold higher in pre-eclamptic placentae than in normal placentae. Immunohistochemistry of placental tissues showed that the levels of peroxiredoxin III/SP-22 protein were increased in the trophoblasts of floating villi, stromal cells of stem villi, and decidual cells in pre-eclamptic placentae. These results indicate that peroxiredoxin III/SP-22 plays a crucial role in the protection of placental function from oxidative stress occurring in mitochondria of pre-eclamptic placentae.
Assuntos
Mitocôndrias/metabolismo , Estresse Oxidativo , Peroxidases/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Adulto , Aldeídos/metabolismo , Feminino , Idade Gestacional , Humanos , Técnicas Imunoenzimáticas , Peroxidação de Lipídeos , Peroxidases/genética , Peroxirredoxina III , Peroxirredoxinas , Placenta/patologia , Pré-Eclâmpsia/patologia , Gravidez , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaRESUMO
OBJECTIVE: To examine whether or not estrogens induced the expression of protein thiol/disulfide oxidoreductases such as protein disulfide isomerase (PDI), thioredoxin (Trx), Trx reductase, and glutaredoxin (Grx) in vascular endothelial cells. METHODS: The regenerative effects of the protein thiol/disulfide oxidoreductases, PDI, Trx and Grx, on oxidatively damaged proteins were assayed using H2O2-inactivated glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a reporter enzyme. The induction of protein thiol/disulfide oxidoreductases and the accumulation of protein adducts generated by lipid peroxidation were examined by Western blotting in estrogen-treated bovine aortic endothelial cells (BAECs). RESULTS: Reduced PDI, Trx and Grx regenerated the H2O2-inactivated GAPDH in vitro. The levels of these protein disulfide oxidoreductases in BAECs were increased by pretreatment with 0.01-10 micromol/l 17beta-estradiol, the largest increase (about fourfold of the control) being found for PDI. Other sex hormones such as progesterone and testosterone did not affect the contents of these oxidoreductases in BAECs. 4-Hydroxy-2-nonenal (HNE)-protein adducts, which are generated by lipid peroxidation, were accumulated in BAECs exposed to paraquat, whereas the pretreatment of BAECs with 17beta-estradiol suppressed their accumulation. CONCLUSIONS: The estrogen-mediated induction of the protein thiol/disulfide oxidoreductases such as PDI, Trx, Trx reductase and Grx suggested a possible involvement of these oxidoreductases in the antioxidant protection of estrogen observed in the vascular system.
Assuntos
Antioxidantes/farmacologia , Endotélio Vascular/efeitos dos fármacos , Estradiol/farmacologia , Estresse Oxidativo , Oxirredutases , Proteína Dissulfeto Redutase (Glutationa)/biossíntese , Animais , Aorta , Bovinos , Células Cultivadas , Endotélio Vascular/metabolismo , Glutarredoxinas , Isomerases de Dissulfetos de Proteínas/metabolismo , Proteínas/metabolismo , Tiorredoxina Dissulfeto Redutase/metabolismo , Tiorredoxinas/metabolismoRESUMO
Normal human diploid fibroblasts (HDF) have a finite life span in vitro and have been used as a model system for the study of in vivo aging. Little is known about how changes in gene expression may affect the immortalization of human fibroblasts. We looked for cDNA clones whose mRNAs were differentially expressed between mortal senescent SV40-transformed human fibroblasts (B-32) and the immortal counterparts (B-32F) derived from B-32 cells. We identified three cDNA isolates by subtractive differential hybridization with 32P-labeled cDNA probes from B-32 cells and B-32F cells. Nucleotide sequence analysis of these cDNA clones revealed that they were homologous to the human vimentin, a human mitochondrial gene and a human gene of unknown nature. Slot blot and Northern blot analyses demonstrated that the former two were preferentially expressed in senescent B-32 cells and the last one was less expressed in B-32F immortal cells.
Assuntos
Senescência Celular/genética , DNA Complementar/genética , Vimentina/genética , Química Encefálica/genética , Linhagem Celular Transformada , Transformação Celular Viral , Células Cultivadas , Clonagem Molecular , Sondas de DNA , DNA Mitocondrial/genética , Fibroblastos/fisiologia , Expressão Gênica , Biblioteca Gênica , Humanos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico/métodos , RNA Mensageiro/genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Vírus 40 dos Símios , Vimentina/fisiologiaRESUMO
The purpose of the present study was to investigate the correlation between tumor size and prognosis in stage IB and II cervical cancer and to elucidate the adequacy of new FIGO staging system for cervical cancer. The subjects included 128 patients with cervical cancer (stage IB = 86, IIA = 18, and IIB = 24) who had undergone radical hysterectomy. The largest tumor size of the pathology specimen was measured in two dimensions, and the correlation between tumor size and prognosis was investigated. In addition, tumor size of the pathology specimen was compared with the largest tumor diameter measured by MRI in stage IB cancers. Patients with a tumor size greater than 3 cm2 had a significantly worse 5-year survival rate (63%) when compared to those with tumor size no greater than 3 cm2 (96%) (P < 0.0001). Multivariate analysis revealed that independent prognostic factors were tumor size (P = 0.003) and lymph node metastasis (P = 0.015). By regression analysis, the largest tumor size of the pathology specimen was relatively well correlated with the largest tumor diameter by MRI in stage IB cancers; 3 cm2 of tumor size in the pathology specimen corresponded to 3.4 cm of tumor diameter by MRI. The adequacy of new FIGO staging system was considered relatively acceptable.
RESUMO
The novel 6-O-methyl tricyclic ketolides TE-802 and its analogs were synthesized by two successive cyclization reactions, 11,12-cyclic carbamate formation by intramolecular Michael addition and 9,11-diazaheptene ring construction by intramolecular dehydration reaction. These new tricyclic ketolides exhibited good in vitro antibacterial activity against not only erythromycin-susceptible strains but also erythromycin-resistant Staphylococcus aureus and Streptococcus pneumoniae, which are problematic pathogens of nosocomial and community-acquired respiratory tract infections, respectively.
Assuntos
Antibacterianos/síntese química , Cetolídeos , Macrolídeos , Antibacterianos/química , Antibacterianos/farmacologia , Ciclização , Resistência Microbiana a Medicamentos , Eritromicina/farmacologia , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Staphylococcus aureus/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
Although erythromycin A contains five hydroxyl groups, regioselective methylation at the C-6 hydroxyl group was achieved to the extent of 90% when a 9-O-substituted erythromycin A 9-oxime was employed as substrate. The methylation and its selectivity are dependent on an O-protecting group at the 9-oxime, solvent, base, and methylating reagent. In particular, the use of a polar aprotic solvent is indispensable for the methylation. Among the 9-oxime derivatives, 2'-O,3'-N-bis(benzyloxycarbonyl)-N-demethylerythromycin A 9-[O-(2-chlorobenzyl)oxime] was the most important intermediate for the synthesis of clarithromycin (6-O-methylerythromycin A).
Assuntos
Claritromicina/química , Claritromicina/síntese química , Eritromicina/análogos & derivados , Eritromicina/química , Eritromicina/síntese química , MetilaçãoRESUMO
We describe siblings with the non-salt-losing form of 21-hydroxylase deficiency who had hypersecretion of aldosterone and plasma renin activity (PRA). Blood pressure and serum electrolytes in both cases were normal despite the aldosterone hypersecretion. Aldosterone secretion was elevated markedly with ACTH administration and with sodium deprivation and/or volume depletion during ACTH suppression by dexamethasone. With suppression by dexamethasone, aldosterone hypersecretion was decreased with lowering of the steroids proximal to the block in the biosynthetic pathway. However, urinary sodium excretion was decreased. These results suggest that the biosynthetic pathway for aldosterone production was preserved. Furthermore, aldosterone hypersecretion and high PRA may serve to compensate for the sodium loss which results in turn from the overproduction of the sodium-losing steroids, such as progesterone and 17 alpha-hydroxyprogesterone which are aldosterone antagonists.