RESUMO
The assignment of 1H-NMR signals of the heme methyl and propionate groups of cytochrome c3 of D. vulgaris Miyazaki F was performed. The heme assignment was revised for hemes 2 and 3 (sequential heme numbering). Namely, heme 4 is mainly reduced at first with hemes 1, 2 and 3 following it in this order. The p2H titration of heme methyl signals in four macroscopic oxidation states was performed in the p2H range of 5.2 to 9.0. While the heme methyl resonances in the fully oxidized state showed just small changes with p2H, most resonances in the intermediate oxidation states revealed clear p2H dependence. In particular, the methyl resonances of heme 1 shifted significantly in the acidic region. Then, the chemical shifts of beta-CH2 (next to the carboxyl group) of all propionate groups in the fully oxidized state were observed at various p2H in the range of 4.5 to 9.0. Only the propionate group at C-13 (IUPAC-IUB nomenclature) of heme 1 showed a clear change in this p2H range, its titration curve being similar to those of the methyl resonances of heme 1 in the intermediate oxidation states. pKa of the propionate group was 5.95 +/- 0.05. Analysis of the microscopic formal redox potentials was carried out for the observations at p2H 5.2, 7.1 and 9.0. The redox potentials of heme 1 showed the most remarkable p2H dependence, resulting in the change of the order of the redox potentials of four hemes. A significant change was also found in the interacting potential between hemes 1 and 2. In the light of the p2H-titration experiments, the propionate at C-13 of heme 1 was identified as the most plausible ionizable group responsible for the p2H dependence of microscopic redox potentials of heme 1 in the acidic region.
Assuntos
Grupo dos Citocromos c/química , Desulfovibrio vulgaris/química , Heme/química , Grupo dos Citocromos c/metabolismo , Desulfovibrio vulgaris/metabolismo , Transporte de Elétrons , Heme/metabolismo , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Oxirredução , Propionatos/química , PrótonsRESUMO
A gene encoding rubredoxin from Desulfovibrio vulgaris (Miyazaki F) was cloned and overexpressed in Escherichia coli. A 1.1-kilobase pair DNA fragment, isolated from D. vulgaris (Miyazaki F) by double digestion with SmaI and SalI, contained two genes, the rubredoxin gene (rub) and the desulfoferrodoxin gene (rbo) which was situated upstream of rub. The deduced amino acid sequence of desulfoferrodoxin was homologous to those from other strains and Cys residues that are responsible to bind irons were also conserved. The expression system for rub was constructed under the control of the T7 promoter in E. coli. The purified protein was soluble and had a characteristic visible absorption spectrum. Inductively coupled plasma-atomic emission analysis and electron paramagnetic resonance analysis of the recombinant rubredoxin revealed the presence of an iron ion in a distorted tetrahedral geometry that was the same as native D. vulgaris rubredoxin. In vitro NADH reduction analysis indicated that recombinant rubredoxin was active, and its redox potential was determined as -5 mV.
Assuntos
Desulfovibrio vulgaris/genética , Genes Bacterianos , Rubredoxinas/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Espectroscopia de Ressonância de Spin Eletrônica , Escherichia coli/genética , Ferredoxinas/genética , Vetores Genéticos , Ferro/análise , Dados de Sequência Molecular , NADH NADPH Oxirredutases/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Mapeamento por Restrição , Rubredoxinas/biossíntese , Rubredoxinas/isolamento & purificação , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Espectrofotometria , Zinco/análiseRESUMO
PURPOSE: To determine the distribution of matrix metalloproteinase-1 (MMP-1) in the uveoscleral outflow pathway and other anterior segment tissues of normal human eyes. METHODS: Normal human eyes were fixed in methacarn and sectioned and immunostained using a specific polyclonal antibody to MMP-1. Immunoreactivity was visualized using diaminobenzidine. To compare the staining intensity in various tissues, the mean optical density within the ciliary body, mid-iris stroma, iris root, uveal trabecular meshwork, cornea, and sclera was determined using imaging densitometry. To determine the cellular distribution of MMP-1 in ciliary muscle, additional sections were double-immunostained using antibodies to MMP-1 and calponin. These sections were examined by confocal laser scanning microscopy. Specificity of the antibody to MMP-1 in ocular tissues was confirmed by western blot analysis with uveal tract homogenates. RESULTS: Moderate-to-strong MMP-1 immunoreactivity was observed in ciliary muscle, iris, sclera, corneal endothelium, and ciliary nonpigmented epithelium. Lighter immunoreactivity was observed in corneal epithelium, blood vessels, trabecular meshwork, Schlemm's canal, and associated collector channels. Confocal microscopy showed that ciliary muscle MMP-1 was primarily inside ciliary muscle cells. Densitometry showed that net optical density was approximately fivefold greater in ciliary muscle, iris root, and sclera than in trabecular meshwork. CONCLUSIONS: MMP-1 was prominently identified in regions of the anterior segment of normal human eyes associated with the uveoscleral outflow pathway and in the iris, corneal endothelium, and ciliary nonpigmented epithelium. These data support the hypothesis that MMP-1 activity is involved in regulating uveoscleral outflow facility.
Assuntos
Segmento Anterior do Olho/enzimologia , Humor Aquoso/metabolismo , Colagenases/análise , Esclera/enzimologia , Úvea/enzimologia , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Densitometria , Humanos , Técnicas Imunoenzimáticas , Metaloproteinase 1 da Matriz , Microscopia Confocal , Pessoa de Meia-IdadeRESUMO
PURPOSE: To investigate the expression of functional gap junctions and the effect of protein kinase C (PKC) on such junctions in confluent cultures of bovine trabecular meshwork (TM) cells. METHODS: Expression of the gap junction protein connexin43 in TM cells was examined by immunofluorescence microscopy. Intercellular communication by gap junctions was assessed by observing the diffusion of fluorescent dye from an individual cell injected with lucifer yellow. The phosphorylation of connexin43 was evaluated by immunoblot analysis with a monoclonal antibody to this protein. RESULTS: Immunofluorescence staining revealed that connexin43 was localized to sites of contact between adjacent TM cells. Exposure of cells to the PKC activator phorbol 12-myristate 13-acetate (PMA; 10 nM, 1 hour) had no marked effect on the pattern of connexin43 immunofluorescence. Injection of a TM cell with lucifer yellow resulted in the spread of the dye into neighboring cells. Dye coupling was inhibited by PMA in a dose- and time-dependent manner, and this inhibition was prevented by pretreatment of cells with the PKC inhibitor bisindolylmaleimide I. Immunoblot analysis of control TM cell lysates yielded connexin43 bands corresponding to the nonphosphorylated protein (43 kDa) and three phosphorylated forms (47, 48, and 49 kDa). Cells exposed to PMA (10 nM, 1 hour) yielded an additional band corresponding to a 44-kDa form of phosphorylated connexin43 and showed a decrease in the intensity of the band corresponding to the nonphosphorylated protein and an increase in the intensity of the 47-kDa band. CONCLUSIONS: TM cells communicate with each other through gap junctions, and the communication is inhibited by PKC, probably, at least in part, through phosphorylation of connexin43.
Assuntos
Comunicação Celular/fisiologia , Conexina 43/metabolismo , Malha Trabecular/metabolismo , Actinas/metabolismo , Animais , Câmara Anterior/metabolismo , Bovinos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Técnica Indireta de Fluorescência para Anticorpo , Corantes Fluorescentes/metabolismo , Junções Comunicantes/fisiologia , Isoquinolinas/metabolismo , Microscopia de Fluorescência , Fosforilação , Proteína Quinase C/antagonistas & inibidores , Proteína Quinase C/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Fatores de Tempo , Malha Trabecular/citologia , Malha Trabecular/efeitos dos fármacosRESUMO
PURPOSE: Mutations in the trabecular meshwork inducible glucocorticoid response (TIGR) gene, also known as myocilin, have recently been linked to some forms of glaucoma. Recent studies have shown that TIGR protein also is expressed in the ciliary muscle. Because uveoscleral outflow, which traverses the ciliary muscle, is increased by prostaglandins (PGs), the present study assessed whether topical PGs alter the amount of TIGR protein within the ciliary muscle. METHODS: Vehicle was topically applied to one eye, and 2 microg PGF(2alpha)-isopropyl ester (PGF(2alpha)-IE) was applied to the other eye of cynomolgus monkeys twice daily for 5 days. Pressure reductions of 5 mm Hg in the PGF(2alpha)-IE-treated eyes were confirmed. The eyes were then fixed and paraffin sections were cut from each eye. The distribution of TIGR protein in the ciliary muscle was determined by confocal scanning laser microscopy. Additional sections were immunostained with a polyclonal antibody to recombinant TIGR protein or with a polyclonal antibody to a synthetic peptide corresponding to the leucine zipper region within the TIGR protein. Staining intensity in the ciliary muscle was assessed by measuring optical density (OD) along two line segments overlying the ciliary muscle, by using a high-resolution imaging densitometer. RESULTS: TIGR protein immunoreactivity was observed in ciliary muscle fibers throughout the ciliary muscle. Extracellular TIGR immunoreactivity colocalized with collagen type IV immunoreactivity. Intracellular staining also was present. Immunoreactivity was less intense in the sections from the PGF(2alpha)-IE-treated eyes compared with the vehicle-treated eyes. This was reflected in the reduction of mean OD scores in each monkey. Overall, the reduction of mean OD scores in the treated eyes was 42.1% +/- 9.9% (P < 0.005) with the anti-recombinant TIGR antibody and 27.3% +/- 10.4% with the anti-TIGR peptide antibody (P < 0.005). CONCLUSIONS: TIGR protein immunoreactivity was present both intracellularly and extracellularly in the ciliary muscle of the cynomolgus monkey. This suggests that extracellular TIGR protein is in contact with aqueous humor in the uveoscleral outflow pathway. Moreover, IOP-lowering topical PGF(2alpha)-IE treatment decreases the amount of TIGR protein in the ciliary muscle.
Assuntos
Corpo Ciliar/efeitos dos fármacos , Proteínas do Citoesqueleto/metabolismo , Dinoprosta/farmacologia , Proteínas do Olho/metabolismo , Glicoproteínas/metabolismo , Músculo Liso/efeitos dos fármacos , Administração Tópica , Animais , Corpo Ciliar/metabolismo , Dinoprosta/análogos & derivados , Técnica Indireta de Fluorescência para Anticorpo , Pressão Intraocular/efeitos dos fármacos , Macaca fascicularis , Microscopia Confocal , Músculo Liso/metabolismoRESUMO
PURPOSE: To investigate in monkey ciliary muscle the relationship between the extent of anterior segment inflammation and alterations of collagen type I as determined by quantitative imaging densitometry. METHODS: Anterior segment inflammation was induced in one eye of five cynomolgus monkey by cannulation of the anterior chamber, by anterior chamber injection of bovine serum albumin, or by disruption of the iris and anterior lens capsule with a needle. Increases in inflammatory cells were scored in hematoxylin and eosin-stained sections. Parallel eye sections were immunostained for collagen type I and developed using diaminobenzidine. Optical density (OD) was measured along two line segments overlying the immunostained ciliary muscle using two-dimensional imaging densitometry. To assess antibody labeling of ciliary muscle structures, additional sections were double-immunostained using antibodies to collagen type I and calponin and examined by confocal microscopy. RESULTS: In each of the inflamed eyes, hematoxylin and eosin-stained sections showed signs of chronic inflammation including lymphocytes and macrophages dispersed among ciliary muscle fibers and in the iris. Double label confocal microscopy showed collagen type I immunoreactivity in the interstitial extracellular matrix between bundles of ciliary smooth muscle fibers. Collagen type I OD scores in each of the inflamed eyes were less by 16% to 55%, compared with the contralateral control eyes. The mean of the OD scores for all inflamed eyes was 39%+/-7% less than the mean of the control eye scores (mean +/- SEM, P < 0.001). Regression analysis showed a close correlation between inflammatory cell scores in the treated eyes and the reduction of OD scores (r = 0.94, P = 0.02). CONCLUSIONS: These results indicate that the density of collagen type I in the extracellular matrix (ECM) of monkey ciliary muscle is reduced during anterior segment inflammation and support the view that reduction of ciliary muscle ECM may contribute to increased uveoscleral outflow facility during anterior segment inflammation.
Assuntos
Corpo Ciliar/metabolismo , Colágeno/metabolismo , Músculo Liso/metabolismo , Uveíte Anterior/metabolismo , Animais , Proteínas de Ligação ao Cálcio/metabolismo , Doença Crônica , Corpo Ciliar/patologia , Densitometria , Matriz Extracelular/metabolismo , Proteínas do Olho/metabolismo , Feminino , Técnicas Imunoenzimáticas , Macaca fascicularis , Masculino , Proteínas dos Microfilamentos , Microscopia Confocal , Proteínas Musculares/metabolismo , Músculo Liso/patologia , Soroalbumina Bovina , Uveíte Anterior/induzido quimicamente , Uveíte Anterior/patologia , CalponinasRESUMO
A case of primary malignant fibrous histiocytoma of the diaphragm which, to our knowledge, is the first record in the world literature, is presented. It occupied almost the entire left hemidiaphragm and was surgically removed with the left lower pulmonary lobe, stomach, colon, and spleen. The entire left hemidiaphragm then was replaced with Marlex mesh. Prosthetic replacement of the entire hemidiaphragm, which has not been previously reported, was successfully performed with preservation of pulmonary function on the affected side. The patient died of brain metastases of malignant fibrous histiocytoma more than 4 months postoperatively. Diagnosis of diaphragmatic tumors and a technique which involves the use of a prosthesis are discussed.
Assuntos
Diafragma/cirurgia , Histiocitoma Fibroso Benigno/cirurgia , Próteses e Implantes/normas , Neoplasias Encefálicas/secundário , Colectomia , Gastrectomia , Histiocitoma Fibroso Benigno/patologia , Histiocitoma Fibroso Benigno/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Pneumonectomia , Esplenectomia , Telas CirúrgicasRESUMO
OBJECTIVE: To investigate the effects of topical prostaglandin F(2 alpha)--isopropyl ester (PGF(2 alpha)-IE) administration on immunoreactivity of matrix metalloproteinases (MMPs) 1, 2, and 3 within the anterior segment tissues of monkey eyes. METHODS: Eight eyes from 4 cynomolgus monkeys were evaluated. One eye from each monkey was treated with 2 mg of PGF(2 alpha)-IE twice daily for 5 days, and intraocular pressure reduction was measured. After fixation and processing, deparaffinized sections of anterior segments were immunostained using antibodies to MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), or MMP-3 (stromelysin-1). Optical density along 2 line segments overlying the iris root, ciliary muscle, and adjacent sclera and perpendicular to their long axes was measured using imaging densitometry. RESULTS: Compared with the contralateral vehicle-treated eyes, statistically significant increases in optical density scores were observed in the iris root, ciliary muscle, and adjacent sclera for all 3 MMPs (P<.01). In these tissues, MMP-1 immunoreactivity was increased by a mean +/- SD of 89% +/- 16%, 61% +/- 8%, and 66% +/- 57%, respectively; MMP-2 immunoreactivity by 129% +/- 53%, 82% +/- 27%, and 267% +/- 210%, respectively; and MMP-3 immunoreactivity by 207% +/- 84%, 83% +/- 49%, and 726% +/- 500%, respectively. CONCLUSIONS: Treatment of monkey eyes with PGF(2 alpha)-IE induces elevation of MMP-1, MMP-2, and MMP-3 in tissues of the uveoscleral outflow pathway. These increases suggest that MMPs might play an important role in the increased uveoscleral outflow observed with topical prostaglandin treatment. CLINICAL RELEVANCE: Immunoreactivity of MMPs in tissues of the monkey uveoscleral outflow pathway is increased after topical treatment with PGF(2 alpha)-IE. This response also might be involved in the intraocular pressure--lowering effect of other prostanoids used to treat glaucoma.
Assuntos
Segmento Anterior do Olho/efeitos dos fármacos , Dinoprosta/administração & dosagem , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Esclera/efeitos dos fármacos , Úvea/efeitos dos fármacos , Administração Tópica , Animais , Segmento Anterior do Olho/enzimologia , Corpo Ciliar/efeitos dos fármacos , Corpo Ciliar/enzimologia , Dinoprosta/análogos & derivados , Técnicas Imunoenzimáticas , Pressão Intraocular/efeitos dos fármacos , Iris/efeitos dos fármacos , Iris/enzimologia , Macaca fascicularis , Músculo Liso/efeitos dos fármacos , Músculo Liso/enzimologia , Soluções Oftálmicas/administração & dosagem , Esclera/enzimologia , Úvea/enzimologiaRESUMO
BACKGROUND: Topical prostaglandin F2alpha isopropyl ester increases uveoscleral outflow in monkeys and humans. OBJECTIVE: To investigate the effects of prostaglandin F2alpha isopropyl ester with topical administration on collagen types I, III, and IV within the anterior segment tissue of monkey eyes. METHODS: Eight eyes of 4 cynomolgus monkeys were evaluated. One eye of each monkey was treated with 2 microg of prostaglandin F2alpha isopropyl ester twice daily for 5 days, and intraocular pressure reduction was confirmed. These eyes were fixed in methacarn, and paraffin sections were immunostained using antibodies to collagen types I, II, or IV. To measure staining intensity, optical density (OD) was determined using 2-dimensional imaging densitometry. Mean OD scores along line segments placed over the ciliary muscle were determined. RESULTS: Mean+/-SD OD scores for collagen types I, III, and IV were less in the ciliary muscle of prostaglandin-treated eyes than in vehicle-treated eyes by 52%+/-7%, 45%+/-6%, and 45%+/-5%, respectively. In the sclera adjacent to the ciliary body, mean OD scores for collagen types I and III were less in prostaglandin-treated eyes, by 43%+/-32% and 45%+/-13%, respectively. The scleral stroma was minimally immunoreactive for collagen type IV. All differences were significant by the paired Student t test (P<.05). CONCLUSIONS: This study shows reduced collagen types I, III, and IV immunoreactivity in the ciliary muscle and adjacent sclera following topical prostaglandin F2alpha isopropyl ester treatment. These reductions may contribute to the increased uveoscleral outflow observed with topical prostaglandin treatment. CLINICAL RELEVANCE: The cellular mechanism by which certain prostaglandins lower intraocular pressure is not known. The present study provides immunohistochemical data demonstrating that intraocular pressure reduction that occurs with topical prostaglandin F2alpha is associated with a reduction of collagens within the uveoscleral outflow pathway.
Assuntos
Humor Aquoso/metabolismo , Corpo Ciliar/efeitos dos fármacos , Colágeno/metabolismo , Dinoprosta/análogos & derivados , Músculo Liso/efeitos dos fármacos , Esclera/efeitos dos fármacos , Administração Tópica , Animais , Corpo Ciliar/metabolismo , Corpo Ciliar/patologia , Dinoprosta/administração & dosagem , Dinoprosta/farmacologia , Técnicas Imunoenzimáticas , Pressão Intraocular/efeitos dos fármacos , Macaca fascicularis , Músculo Liso/metabolismo , Músculo Liso/patologia , Soluções Oftálmicas/administração & dosagem , Soluções Oftálmicas/farmacologia , Esclera/metabolismo , Esclera/patologiaRESUMO
The gene encoding a flavodoxin of Desulfovibrio vulgaris (Miyazaki F) was cloned, and overexpressed in Escherichia coli. A 1.6-kbp DNA fragment, isolated from D. vulgaris (Miyazaki F) by double digestion with SalI and EcoRI, contained the flavodoxin gene and its regulatory region. An expression system for the flavodoxin gene under control of the T7 promoter was constructed in E. coli. The purified protein was soluble and exhibited a characteristic visible absorption spectrum. HPLC analysis of the recombinant flavodoxin revealed the presence of an identical FMN to that found in the native D. vulgaris flavodoxin, and its dissociation constant with FMN was determined to be 0.38 nM. In vitro H2 reduction analysis indicated that the recombinant flavodoxin is active, and its redox potential was determined to be E1 = -434 and E2 = -151 mV using methyl viologen and 2-hydroxy-1,4-naphthoquinone, respectively. Its redox behavior was also examined with the recombinant flavodoxin adsorbed onto a graphite electrode. The mutant, A16E, was also produced, which revealed the feature of a conserved Glu residue at the surface of the molecule.
Assuntos
Clonagem Molecular , Desulfovibrio vulgaris/genética , Flavodoxina/biossíntese , Expressão Gênica , Genes Bacterianos , Sequência de Bases , Eletroquímica , Eletrodos , Eletroforese em Gel de Poliacrilamida , Escherichia coli/metabolismo , Mononucleotídeo de Flavina/metabolismo , Flavodoxina/química , Flavodoxina/genética , Flavodoxina/isolamento & purificação , Grafite , Immunoblotting , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Oxirredução , Ligação Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Espectrofotometria UltravioletaRESUMO
To determine the factors influencing the prognosis of variant angina, the clinical characteristics and long-term prognosis of 158 consecutive Japanese patients were examined and compared with those in previous major western studies (Pisa, Montreal, and Duke studies). The Japanese patients were characterized by relatively low prevalences of coronary risk factors, significant coronary stenoses and previous myocardial infarction. Survival and survival without myocardial infarction for the entire group or for the subpopulation with significant coronary artery disease were significantly better in the Japanese population than in the western populations; however, in the subpopulation without significant coronary artery disease, the prognosis was excellent in all four studies. If the prevalence of coronary artery disease was corrected for the Japanese population, there would be no difference in the prognosis between the Japanese and the western populations. It is concluded: (1) the overall prognosis of variant angina may be better in Japanese patients, and (2) coronary artery disease appears to be the strongest prognostic factor for assessing the differences in the prognosis between the Japanese and the western populations.
Assuntos
Angina Pectoris Variante/mortalidade , Comparação Transcultural , Adulto , Idoso , Angina Pectoris Variante/terapia , Bloqueadores dos Canais de Cálcio/uso terapêutico , Ponte de Artéria Coronária , Eletrocardiografia , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/mortalidade , Nitroglicerina/uso terapêutico , PrognósticoRESUMO
The differentiation of the rat stria vascularis (SV) was investigated by light- and electron microscopy and by immunocytochemistry. Loss of the basal lamina at the epithelial-mesenchymal interface of SVs as indicated by immunoreactions of laminin and fibronectin induces the formation of vascular feet by basal infoldings of the marginal cells (MCs), and the development of the strial capillaries (SCs) by mesenchymal cells in a manner of vasculogenesis is progressing at the same time. The production of fibronectin in the rough endoplasmic reticulum of mesenchymal cells and the involvement of this glycoprotein in a mechanical linkage between the vasoformative mesenchymal cells and endothelial ones of the SCs are indicated by immunocytochemistry. The plasma membrane of the marginal cells (MCs) begins to show immunoreactions of Na+.K+ ATPase at postnatal day 5 and is conjugated to each other by tight junctions at postnatal day 14. The apical tubules of the differentiating MCs do not seem to be involved in the endocytotic activity but are involved in the plasma membrane supply for the rapid differentiation.
Assuntos
Estria Vascular/citologia , Animais , Diferenciação Celular , Membrana Celular/enzimologia , Ducto Coclear/citologia , Ducto Coclear/embriologia , Ducto Coclear/ultraestrutura , Retículo Endoplasmático/ultraestrutura , Fibronectinas/metabolismo , Imuno-Histoquímica , Laminina/metabolismo , Mesoderma/citologia , Microscopia Eletrônica , Microscopia Eletrônica de Varredura , Microscopia Imunoeletrônica , Ratos , Ratos Wistar , ATPase Trocadora de Sódio-Potássio/metabolismo , Estria Vascular/ultraestruturaRESUMO
Mutation of the adenosine 3',5'-cyclic monophosphate receptor protein gene (crp) of Escherichia coli induced by protons, ionizing radiation of charged particles, was analyzed to determine the specificity of the mutational spectrum. The majority, 44 of 49 mutations detected, were base substitutions, and three frameshifts and two gross structural changes were also found. Base substitutions included 35 transversions and nine transitions. G:C to T:A transversions were the dominant type of base substitution, followed by G:C to C:G and A:T to T:A transversions. Almost all transitions were eight G:C to A:T changes. The spectrum of proton mutagenesis was quite different from that of X-ray mutagenesis of the crp gene, in which G:C to A:T transitions dominated.
Assuntos
Proteína Receptora de AMP Cíclico/genética , Escherichia coli/efeitos da radiação , Genes Bacterianos/efeitos da radiação , Mutagênese , Prótons/efeitos adversos , Sequência de Bases , Análise Mutacional de DNA , DNA Bacteriano/efeitos da radiação , Escherichia coli/genética , Mutação da Fase de Leitura , Dados de Sequência Molecular , Plasmídeos/efeitos da radiação , Mutação PuntualRESUMO
OBJECTIVES: This study investigated the possibility of a relationship between blood pressure level and rotating 3-shift work in a prospective follow-up of workers in a zipper and aluminum sash factory in Japan. METHODS: Altogether 1551 men aged 18-49 years were followed prospectively for 5 years, and the cumulative incidence of hypertension among 3-shift workers was compared with that of day workers. A multiple logistic analysis was used for adjusting for base-line characteristics such as age, body mass index, blood pressure, and drinking habit. RESULTS: In the younger age group, the relative risk of the rotating 3-shift workers during the observational period was increased compared with that of day workers after adjustment for the confounding factors. In the older group, the cumulative incidence of hypertension was not higher for workers who had continued shift work. However, a relatively high risk of hypertension was found for workers who converted from 3-shift work to day work when compared with those who remained on shift work and day work. CONCLUSIONS: It is suggested that there is an association between 3-shift work and blood pressure.
Assuntos
Hipertensão/etiologia , Indústrias , Doenças Profissionais/etiologia , Tolerância ao Trabalho Programado , Adolescente , Adulto , Pressão Sanguínea , Fatores de Confusão Epidemiológicos , Seguimentos , Humanos , Incidência , Descrição de Cargo , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Risco , Fatores de Risco , Viés de Seleção , Tolerância ao Trabalho Programado/fisiologia , Tolerância ao Trabalho Programado/psicologiaRESUMO
The role of the submandibular glands in the excretion of parenterally administered bis (tributyltin) oxide (TBTO) was studied. Fine structural alterations of the submandibular glands were observed with an electron microscope. Accumulation sites of TBTO were determined with an X-ray microanalyzer and tin concentrations in saliva and blood were measured by atomic absorption spectrophotometry.
Assuntos
Glândula Submandibular/metabolismo , Compostos de Trialquitina/metabolismo , Animais , Microanálise por Sonda Eletrônica , Masculino , Microscopia Eletrônica , Ratos , Ratos Wistar , Saliva/química , Espectrofotometria Atômica , Glândula Submandibular/efeitos dos fármacos , Glândula Submandibular/ultraestrutura , Estanho/análise , Estanho/sangue , Compostos de Trialquitina/farmacologiaRESUMO
To efficiently store and retrieve cardiac catheterization records, we have developed a computer-assisted database, which comprises a 16-bit microcomputer with dual floppy disk drives, a 20 MB random-access memory, hard disk drive, and a line printer. All programmings were accomplished using a relational database management system (R:base 5000, Microrim, Inc.). Data inquiry procedures could be performed with direct operational commands of the system as well as with preprogrammed command files, and final results of searches were printed out with a line printer. The major advantages of the present system described in this report include: (1) the relatively easy and rapid creation of the database, (2) ease of modification of the database structures even after the system design is finished, (3) operational commands in combination with conditional operator(s) are flexible and powerful enough to allow the end user to retrieve data based on various kinds of criteria, (4) a high-level programming language provided by the R:base automates a series of database procedures with relative ease, (5) relational capabilities of the database management system can enhance the possibility of reconstruction of a new data file from a single or several preexisting data files, and (6) the system can be realized at reasonable cost.
Assuntos
Cateterismo Cardíaco/instrumentação , Computadores , Sistemas de Gerenciamento de Base de Dados , Registros Médicos Orientados a Problemas , Prontuários Médicos , Microcomputadores , Software , HumanosRESUMO
BACKGROUND: In Japan ABO-incompatible liver transplantation has been done on >100 occasions up to 2003. However, <30% are cases involving adults. The difficultly of ABO-incompatible liver transplantation is associated with the high frequency of humoral rejection and local disseminated intravascular coagulation (DIC), leading to many postoperative complications. We report a successful case of adult ABO-incompatible liver transplantation with the use of an intrahepatic artery infusion. METHODS: A 36-year-old man with Wilson disease, underwent living donor liver transplantation from an ABO-incompatible donor. The immunosuppressive therapy included multiple perioperative plasmaphereses, splenectomy, and treatment with tacrolimus, methylprednisolone, and cyclophosphamide. The dose and blood level of tacrolimus were the same as in ABO-compatible cases. In addition to these therapies, we administered an intrahepatic arterial infusion with prostaglandin (PG) E1 alone. RESULTS: After perioperative plasmapheresis and cyclophosphamide, antidonor blood group antibody titers remained undiluted and without vascular complications throughout the postoperative course, but there was a tendency for bleeding that continued for 10 days after transplantation. On postoperative day 10, a reexploration was performed for intraabdominal bleeding. During another operation on postoperative day 59 a biloma was found and drained. The patient has now survived for 120 days after transplantation with normal liver function. CONCLUSIONS: Beneficial effect of intrahepatic artery infusion with PGE1 seems to be useful in adult ABO-incompatible liver transplantation.
Assuntos
Sistema ABO de Grupos Sanguíneos , Degeneração Hepatolenticular/cirurgia , Infusões Intra-Arteriais , Transplante de Fígado/métodos , Adulto , Incompatibilidade de Grupos Sanguíneos , Quimioterapia Combinada , Artéria Hepática , Degeneração Hepatolenticular/sangue , Humanos , Imunossupressores/uso terapêutico , Cuidados Intraoperatórios , Testes de Função Hepática , Transplante de Fígado/imunologia , Doadores Vivos , Masculino , Plasmaferese , Esplenectomia , Resultado do TratamentoRESUMO
This article presents an evaluation of the factor structures of the Japanese version of the Maslach Burnout Inventory (MBI). The MBI is a widely used psychometric instrument for measuring 'burnout' developed by Maslach and her co-workers. The MBI consists of four subscales: Emotional Exhaustion, Personal Accomplishment, Depersonalization, and Involvement. The MBI was translated into Japanese along with a back-translation and was administered to a sample of 267 nurses. Various psychometric analyses showed that the Japanese version of the MBI has high reliability for the 22 items scored for the frequency dimension. The factor analysis using principal factoring with an oblique rotation resulted in three factor structures that had different implications from the MBI: Emotional Exhaustion/Depersonalization, Personal Accomplishment, and Physical Exhaustion. The correlationship between the MBI and the General Health Questionnaire (GHQ), measures of depression, showed that burnout was a unique phenomenon.
Assuntos
Esgotamento Profissional/diagnóstico , Enfermeiras e Enfermeiros/psicologia , Psicometria/métodos , Adulto , Depressão/diagnóstico , Análise Fatorial , Feminino , Humanos , Japão , Idioma , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
A 67-year-old man was hospitalized with a diagnosis of nephrotic syndrome. Physical findings at admission were generalized edema and macroglossia. Urinalysis showed massive proteinuria, + +occult blood, and granular and broad casts. Ig A lambda monoclonal gammopathy was noted in the serum. There was no evidence of myeloma in the bone marrow aspirate, scintigram or X-ray of the bone. A biopsy specimen of the kidney showed massive deposits of structureless material in the glomeruli. Marked cell infiltration was also observed in the interstitium. Multinucleated giant cells were occasionally seen in the Bowman's capsules and the interstitium. There were reactive changes in the Bowman's capsule adjacent to the giant cell. The deposits were proved to be amyloid by positive staining with Congo red and apple-green birefringence by polarized light. In addition, microfibrills seen on electron microscopy displayed deposits. Amyloid depositions were observed in other tissues such as gingiva, skin and tongue. Staining of amyloid with Congo red was resistant to potassium permanganate, and amyloid was positively stained with lambda-light chain of immunoglobulin. These findings indicated that the patient had primary amyloidosis. Infiltration of the multinucleated giant cell has been reported only in patients with familial amyloidosis and secondary amyloidosis associated with rheumatoid arthritis. To our knowledge the present case is a first report of the giant cell infiltration in a Bowman's capsule in primary amyloidosis.
Assuntos
Amiloidose/patologia , Células Gigantes/patologia , Nefropatias/patologia , Idoso , Amiloide/metabolismo , Humanos , Nefropatias/metabolismo , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , MasculinoRESUMO
The discrimination of mono-syllable words (67S word-list) pronounced by a male and a female speaker was investigated in noise in 39 normal hearing subjects. The subjects listened to the test words at a constant level of 62 dB together with white or weighted noise in four S/N conditions. By processing the data with logit transformation, S/N-discrimination curves were presumed for each combination of a speech material and a noise. Regardless of the type of noise, the discrimination scores for the female voice started to decrease gradually at a S/N ratio of +10 dB, and reached 10 to 20% at-10 dB. For the male voice in white noise, the discrimination curve was similar to those for the female voice. On the contrary, the discrimination score for the male voice in weighted noise declined rapidly from a S/N ratio of +5 dB, and went below 10% at -5 dB. The discrimination curves seem to be shaped by the interrelations between the spectrum of the speech material and that of the noise.