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1.
Int J Cosmet Sci ; 42(4): 346-358, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32251525

RESUMO

OBJECTIVES: To evaluate improved protein extraction and two-dimensional electrophoresis (2DE) separation methods with Japanese reference human hair (JRH); to determine whether fibre curvature is related to protein composition in curly and straight Japanese women's human hair (JHH) samples; and to identify proteins from JRH 2DE maps and expression differences between curly and straight JHH. METHODS: Hair keratin and keratin-associated proteins (KAPs) were extracted intact with dithiothreitol or tris(2-carboxyethyl) phosphine from JRH or from curved or straight JHH. Extracted proteins were isoelectric-focused on first-dimensional pH gradient gel strips, then separated by molecular weight on laboratory-made, second-dimension, large format gels. The software compared protein abundance between duplicate 2DE gels of curved and straight JHH. Thirty-eight proteins from a JRH 2DE gel were enzyme-cleaved for MALDI-TOF-MS analysis to determine peptide composition, and where possible, de novo sequencing gave peptide sequence data. An in-house human hair protein database incorporating ninety-eight annotated protein sequences assisted MS analysis. RESULTS: 2DE gels of tris(2-carboxyethyl) phosphine-extracted JRH improved keratin and KAP resolution and number compared to those of dithiothreitol-extracted JRH and published commercially made second-dimensional gels. Silver-stained 2DE gels of the straight or curved JHH sets were remarkably similar. Over-staining to reveal basic proteins caused poor resolution of the major acidic protein classes. Software comparisons of fifty-nine resolved proteins revealed two were significantly different in abundance between curved and straight hairs but in insufficient amounts for MS analysis. MS identified twelve proteins from a JRH CBBG-stained 2DE gel: six type II keratins, three type I keratins and three high sulphur proteins. A further eight were potential conformational isoforms and isoelectric variants of the identified proteins bringing the total to twenty identified or partially identified proteins. CONCLUSION: Root-end human hair extraction with tris(2-carboxyethyl) phosphine improves protein resolution and visualizes more proteins on large format 2DE gels. The two minor protein differences between duplicate straight or curved JHH 2DE gels were unlikely to change fibre structure from straight to curved hair. MS results confirmed that multiple isoforms exist of various hair proteins. Low sequence coverage prevented distinction between members in rows of homologous protein spots of similar molecular weight.


OBJECTIFS: évaluer l'amélioration de l'extraction de protéines et les méthodes de séparation bidimensionnelle par électrophorèse (2DE) avec des cheveux humains de référence Japonais (JRH), déterminer si la courbure de la fibre est liée à la composition protéique dans les échantillons de cheveux humains des Japonaises (JHH) bouclés et raides et identifier les protéines issues des cartes JRH 2DE et les différences d'expression entre les JHH bouclés et raides. MÉTHODES: la kératine des cheveux et les protéines associées à la kératine (KAP) ont été extraites intactes avec du dithiothréitol ou du tris (2-carboxyéthyl) phosphine des JRH ou des JHH bouclés ou raides. Les protéines extraites ont subi une focalisation isoélectrique sur des bandes de gel à gradient de pH unidimensionnelles, puis ont été séparées par poids moléculaire sur des gels bidimensionnels de grand format, fabriqués en laboratoire. Le logiciel a comparé l'abondance des protéines entre les deux duplicatas de gels 2DE des JHH bouclés et raides. Trente-huit protéines provenant d'un gel 2DE JRH ont été clivés par enzyme pour l'analyse MALDI-TOF-MS afin de déterminer la composition des peptides, et dans la mesure du possible, un séquençage de novo a donné des données de séquence des peptides. Une base de données interne des protéines capillaires humaines incorporant 98 séquences de protéines annotées a aidé l'analyse MS. RÉSULTATS: les gels 2DE de JRH extraits par le tris (2-carboxyéthyl) ont amélioré la résolution et le nombre de la kératine et du KAP par rapport à ceux du JRH extrait par le dithiothréitol et des gels bidimensionnels fabriqués commercialement. Les gels 2DE à coloration argentée des ensembles de JHH raides ou bouclés étaient remarquablement similaires. La sur-coloration pour révéler les protéines de base a provoqué une mauvaise résolution des principales classes de protéines acides. Les comparaisons logicielles des 59 protéines résolues ont révélé que deux présentaient une différence significative d'abondance entre les cheveux bouclés et raides, mais en quantités insuffisantes pour une analyse MS. La MS a identifié douze protéines provenant d'un gel 2DE coloré CBBG JRH : six kératines de type II, trois kératines de type I et trois protéines à forte teneur en soufre. Huit autres étaient des isoformes conformationnels potentiels et des variantes isoélectriques des protéines identifiées, ramenant le total à 20 protéines identifiées ou partiellement identifiées. CONCLUSION: l'extraction des cheveux humains à la racine avec du tris (2-carboxyéthyl) phosphine améliore la résolution des protéines et permet de visualiser plus de protéines sur les gels 2DE grand format. Les deux différences de protéines mineures entre les duplicatas des gels 2DE JHH raides ou bouclés étaient peu susceptibles de changer la structure des fibres de cheveux raides à bouclés. Les résultats de la MS ont confirmé qu'il existe plusieurs isoformes de diverses protéines capillaires. Une faible couverture de séquence a empêché la distinction entre les protéines homologues de poids moléculaire similaire.


Assuntos
Povo Asiático , Eletroforese em Gel Bidimensional/métodos , Cabelo/química , Proteínas/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Espectrometria de Massas em Tandem/métodos , Sequência de Aminoácidos , Humanos , Japão , Peso Molecular , Proteínas/isolamento & purificação
2.
J Appl Microbiol ; 126(1): 31-39, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30221812

RESUMO

AIMS: This study was conducted to assess the presence and extent of differences in the gut microbiota of common bottlenose dolphins depending on rearing facilities. METHODS AND RESULTS: Faecal samples were collected from 16 common bottlenose dolphins at three aquaria in Japan. After extracting DNA from the faeces, the V3-V4 region of bacterial 16S rRNA was amplified and sequenced using Illumina MiSeq platform. The constituent phyla of the gut microbiota were similar among aquaria; however, the most dominant phylum differed depending on the facility, and the compositions of microbiota were remarkably varied at the family level among aquaria. The alpha diversity indices tended to differ among aquaria. Some bacterial families observed in terrestrial mammalian carnivores or carnivorous fish were detected, as well as several bacterial species suspected of being pathogenic in dolphins. CONCLUSION: Our findings indicate that captive environmental conditions including prey and housing types may contribute to differences in the gut microbiota of the dolphins. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study revealing the differences in gut microbiota of captive dolphins among facilities. Our findings will provide valuable information for improving the health management of dolphins.


Assuntos
Bactérias/isolamento & purificação , Golfinho Nariz-de-Garrafa/microbiologia , Microbioma Gastrointestinal , Animais , Bactérias/classificação , Bactérias/genética , Golfinho Nariz-de-Garrafa/crescimento & desenvolvimento , DNA Bacteriano/genética , Fezes/microbiologia , Hidrobiologia , Japão , RNA Ribossômico 16S/genética
3.
Int J Cosmet Sci ; 41(2): 132-146, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30719733

RESUMO

OBJECTIVE: A new hair-care process has been specifically developed for the straightening of curved Japanese woman's hair . The process included sodium 2-naphthalene sulfonate (SNS) in the reduction and oxidation steps of a conventional perming process. Our objective was to develop an understanding of how this process caused hair straightening by measuring the changes to morphology and ultrastructure between untreated, conventionally permed and SNS permed hair. Untreated and SNS permed Merino wool fibres were used to confirm structural changes. METHODS: Japanese hair samples were measured for single-fibre curvature before and after perming treatments. A silver staining method was developed to stain hair fibres without changing fibre curvature so that transmission electron microscopy could be used to measure changes in the lateral dimensions of all structural components from the cellular to protein filament level. Electron tomography determined intermediate filament slopes and slope changes after SNS perming relative to the central longitudinal axis of the fibre. RESULTS: SNS perming was found to cause greater lateral swelling than conventional perming of: the paracortical cells of wool; the cuticle, the cuticular cell membrane complex and the macrofibrillar centre-to-centre distance of hair; and of the intermediate filaments in wool and hair. In curved hair, SNS perming caused the intermediate filaments of the helical macrofibrils to simultaneously swell and to tilt further, resulting in the slight longitudinal contraction of the macrofibrils. The overall swelling and tilting was greatest in the helical macrofibrils of Type B cortical cells predominately located in the convex fibre half. The presence of a higher percentage of helical macrofibrils in the convex fibre half than in the concave fibre half caused a contraction differential between the two halves leading to straighten of the curved fibre. A mechanical model was proposed to explain how SNS perming straightened curly hair. CONCLUSION: The effects of conventional and SNS perming on the morphological and ultrastructural components of curved Japanese hair and high-curl Merino wool fibres have given clear insights into understanding the mechanism of fibre curvature change.


OBJECTIF: Un nouveau procédé de soin des cheveux a été spécialement conçu pour lisser les cheveux ondulés des Japonaises[1]. Le procédé utilise le sulfonate de naphthalène-2 sodium (SNS) dans les étapes de réduction et d'oxydation du procédé conventionnel de permanente. Notre objectif était de comprendre la façon dont ce procédé induisait le lissage des cheveux en mesurant les différences de changement morphologique et ultrastructural entre les cheveux non traités et ceux soumis à une permanente conventionnelle et une permanente à base de SNS. Des fibres de laine de mérinos non traitées et soumises à une permanente à base de SNS ont été utilisées pour confirmer les changements structurels. MÉTHODES: Des échantillons de cheveux japonais ont été utilisés pour mesurer la courbure d'une fibre isolée avant et après le traitement de permanente. Une méthode de coloration argent a été mise au point pour colorer les fibres de cheveux sans changer la courbure des fibres afin de pouvoir utiliser la microscopie électronique en transmission pour mesurer les modifications des dimensions en largeur de tous les composants structurels du filament, de la cellule aux protéines. Une tomographie électronique a déterminé les pentes intermédiaires et les changements de pente des filaments après permanente à base de SNS par rapport à l'axe longitudinal central de la fibre. RÉSULTATS: On a constaté que la permanente à base de SNS induisait un gonflement en largeur plus important que la permanente classique des cellules paracorticales de la laine; de la cuticule, du complexe de la membrane cellulaire cuticulaire et de la distance centre à centre des macrofibrilles du cheveu; et des filaments intermédiaires dans la laine et les cheveux. Dans les cheveux ondulés, la permanente à base de SNS a provoqué à la fois un gonflement et une inclinaison des filaments intermédiaires des macrofibrilles hélicoïdales, entraînant une légère contraction longitudinale des macrofibrilles. Au total, le gonflement et l'inclinaison étaient plus importants dans les macrofibrilles hélicoïdales des cellules corticales de type B situées principalement dans la moitié convexe de la fibre. La présence d'un pourcentage plus élevé de macrofibrilles hélicoïdales dans la moitié convexe par rapport à la moitié concave de la fibre a entraîné une contraction différentielle entre les deux moitiés qui a entraîné le redressement de la fibre courbée. Un modèle mécanique a été proposé pour expliquer comment la permanente à base de SNS lissait les cheveux bouclés. CONCLUSION: Les effets de la permanente conventionnelle et à base de SNS sur les composants morphologiques et ultrastructuraux des cheveux japonais ondulés et des fibres de laine très frisés de mérinos ont permis de mieux comprendre le mécanisme du changement de courbure des fibres.


Assuntos
Preparações para Cabelo , Cabelo/química , Microscopia Eletrônica de Transmissão/métodos , Naftalenossulfonatos/química , Tomografia/métodos , Animais , Povo Asiático , Feminino , Cabelo/ultraestrutura , Humanos , Japão , Ovinos
4.
Phys Rev Lett ; 115(7): 077001, 2015 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-26317741

RESUMO

Quantum spin liquids, which are spin versions of quantum matter, have been sought after in systems with geometrical frustration. We show that disorder drives a classical magnet into a quantum spin liquid through conducting NMR experiments on an organic Mott insulator, κ-(ET)_{2}Cu[N(CN)_{2}]Cl. Antiferromagnetic ordering in the pristine crystal, when irradiated by x rays, disappears. Spin freezing, spin gap, and critical slowing down are not observed, but gapless spin excitations emerge, suggesting a novel role of disorder that brings forth a quantum spin liquid from a classical ordered state.

5.
Kyobu Geka ; 64(7): 574-8, 2011 Jul.
Artigo em Japonês | MEDLINE | ID: mdl-21766711

RESUMO

A 79-year-old female who had been observed for an intrapericardial cyst for 3 years was admitted due to severe dyspnea and back pain. During 3 years observation, there were no symptoms and no growth of the cyst. Computed tomography scans showed rapid expansion of the cyst compared with an magnetic resonance imaging (MRI) taken 9-days before when she visited our hospital for the 1st time. Urgent complete extirpation of the tumor was successfully done and a histological diagnosis found it was an intrapericardial bronchogenic cyst. This is a 1st report of intrapericardial bronchogenic cyst of which rapid expansion is clearly demonstrated by radiology. It is suggested that complete surgical excision of intrapericardial bronchogenic cysts at the time of diagnosis even if asymptomatic.


Assuntos
Cisto Broncogênico/complicações , Cisto Broncogênico/patologia , Insuficiência Respiratória/etiologia , Idoso , Feminino , Humanos , Pericárdio
6.
Acta Virol ; 54(2): 91-8, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20545436

RESUMO

UNLABELLED: Phylogenetic analysis of nucleotide sequences or deduced amino acid sequences of phosphoprotein (P protein), matrix (M) protein, and glycoprotein (G protein) genes of 18 Chinese isolates of Rabies virus (RABV) from 2003-2007 showed that these isolates formed a separate monophyletic lineage consisting of sub-lineages A and B. Compared with laboratory-fixed strains, recent Chinese isolates of sub-lineage B contained Val or Ala instead of Met69 in P protein, which is involved in generating truncated P proteins. In addition, one of these isolates CHpg3 had Pro instead of Ser63 and Leu instead of Ser64. Importantly, all functional domains of P and M proteins of all recent Chinese isolates were similar to those of laboratory-fixed strains. This study showed that although the recent Chinese RABV isolates belonged to a distinct lineage, their functional domains of P and M proteins were highly conserved. KEYWORDS: rabies virus; glycoprotein; phosphoprotein; matrix protein; China.


Assuntos
Genes Virais , Fosfoproteínas/genética , Vírus da Raiva/genética , Vírus da Raiva/isolamento & purificação , Proteínas da Matriz Viral/genética , Proteínas Estruturais Virais/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , China , Primers do DNA/genética , DNA Viral/genética , Cães , Evolução Molecular , Chaperonas Moleculares , Dados de Sequência Molecular , Filogenia , Raiva/veterinária , Raiva/virologia , Vírus da Raiva/classificação , Homologia de Sequência de Aminoácidos , Suínos , Fatores de Tempo
7.
Kyobu Geka ; 62(5): 385-7, 2009 May.
Artigo em Japonês | MEDLINE | ID: mdl-19425379

RESUMO

Published data has suggested that low body weight is risk factor for surgical intervention in congenital heart disease. We report a case of a low birth weight infant with ventricular septal defect (VSD) and pulmonary hypertension (PH). A female infant was born at 31 weeks' gestation. The birth weight was 904 g. At birth she was suffering from congestive heart failure. We diagnosed VSD and PH. We performed pulmonary artery banding at 23 days of age. She accepted the band at a circumference of 15.5 mm. And we performed VSD patch closure and de-banding at 175 days of age. After operation she was on good course and discharged at 19 postoperative day.


Assuntos
Comunicação Interventricular/cirurgia , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Feminino , Humanos , Recém-Nascido , Masculino
8.
Kyobu Geka ; 62(9): 822-5, 2009 Aug.
Artigo em Japonês | MEDLINE | ID: mdl-19670787

RESUMO

We report a case of ventricular septal defect (VSD) associated with infective endocarditis complication of aortic and pulmonary valvular regurgitation. He received treatment for pyelonephritis and after 2 months he was suffering from congestive heart failure. Echocardiography showed vegetation on the aortic valve and the anterior semilunar cusp of the pulmonary valve. He was referred to our department for surgical repair. Aortic valvular replacement, pulmonary valvular repair and direct closure of VSD were performed. The postoperative course was uneventful and the patient discharged at 28 postoperative day.


Assuntos
Insuficiência da Valva Aórtica/cirurgia , Endocardite/complicações , Comunicação Interventricular/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência da Valva Pulmonar/cirurgia
9.
Kyobu Geka ; 62(7): 535-7, 2009 Jul.
Artigo em Japonês | MEDLINE | ID: mdl-19588822

RESUMO

Noonan syndrome is one of the most common nonchromosomal syndromes seen in children with congenital heart disease. The major cause of death is congestive heart failure. We report a case of Noonan syndrome with hypertrophic cardiomyopathy and arterial septal defect (ASD). Since 10 years old he had been suffering from congestive heart failure. ASD patch closure with a fenestrated flap valve patch was performed at 12 years of age. After operation, left-to-right shunt flow through the patch gradually decreased and he was discharged at 19 postoperative day in good condition.


Assuntos
Cardiomiopatia Hipertrófica/complicações , Comunicação Interatrial/complicações , Síndrome de Noonan/complicações , Criança , Humanos , Masculino
10.
Dis Esophagus ; 21(4): 355-63, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18477259

RESUMO

The Los Angeles classification system is the most widely employed criteria associated with the greatest interobserver agreement among endoscopists. In Japan, the Los Angeles classification system has been modified (modified LA system) to include minimal changes as a distinct grade of reflux esophagitis, rather than as auxiliary findings. This adds a further grading M defined as minimal changes to the mucosa, such as erythema and/or whitish turbidity. The modified LA system has come to be used widely in Japan. However, there have been few reports to date that have evaluated the interobserver agreement in diagnosis when using the modified LA classification system incorporating these minimal changes as an additional grade. A total of 100 endoscopists from university hospitals and community hospitals, as well as private practices in the Osaka-Kobe area participated in the study. A total of 30 video clips of 30-40 seconds duration, mostly showing the esophagocardiac junction, were created and shown to 100 endoscopists using a video projector. The participating endoscopists completed a questionnaire regarding their clinical experience and rated the reflux esophagitis as shown in the video clips using the modified LA classification system. Agreement was assessed employing kappa (kappa) statistics for multiple raters. The kappa-value for all 91 endoscopists was 0.094, with a standard error of 0.002, indicating poor interobserver agreement. The endoscopists showed the best agreement on diagnosing grade A esophagitis (0.167), and the poorest agreement when diagnosing grade M esophagitis (0.033). The kappa-values for the diagnoses of grades N, M, and A esophagitis on identical video pairs were 0.275-0.315, with a standard error of 0.083-0.091, indicating fair intraobserver reproducibility among the endoscopists. The study results consistently indicate poor agreement regarding diagnoses as well as fair reproducibility of these diagnoses by endoscopists using the modified LA classification system, regardless of age, type of practice, past endoscopic experience, or current workload. However, grade M reflux esophagitis may not necessarily be irrelevant, as it may suggest an early form of reflux disease or an entirely new form of reflux esophagitis. Further research is required to elucidate the pathophysiological basis of minimal change esophagitis.


Assuntos
Esofagite Péptica/classificação , Esofagite Péptica/diagnóstico , Esofagoscopia , Variações Dependentes do Observador , Adulto , Idoso , Esofagite Péptica/patologia , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade
11.
Acta Virol ; 50(1): 33-7, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16599183

RESUMO

Thirty-four rabies virus (RV) isolates from foxes (8), insectivore bats (9), cattle (14), sheep (1), a goat (1) and a donkey (1) from Paraiba state, northeastern Brazil, were genetically characterized. Sequences of 890 nts of nucleoprotein (N) genes of these isolates were analyzed and compared with those of other Brazilian isolates characterized earlier. Phylogenetic analysis revealed three genetical lineages of RV co-existing in this region. Each lineage was found to be associated with particular host species and to circulate independently of each other. The first lineage was found in foxes (Dusicyon sp.) and could be discriminated from domestic carnivore isolates from Sao Paulo, Goias and Minas Gerais in the southern and central Brazil. The second lineage was associated with insectivorous bats (Molossus spp.) and differed from vampire bat-associated RV isolates. The third lineage was found in livestock and clustered with vampire bat-associated RV isolates from Sao Paulo, Tocantins, Goias and Matto Grosso. These results indicate that RV of these genetic lineages are cocirculating in the Paraiba state and that livestock in this region are infected with vampire bat-associated RV, suggesting that the vampire bat is the main reservoir of livestock rabies in this region.


Assuntos
Vírus da Raiva/classificação , Vírus da Raiva/genética , Animais , Sequência de Bases , Bovinos/virologia , Quirópteros/virologia , Equidae/virologia , Raposas/virologia , Cabras/virologia , Filogenia , RNA Viral/análise , RNA Viral/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Ovinos/virologia
12.
Dev Comp Immunol ; 23(3): 213-9, 1999 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-10402208

RESUMO

Using an antibody against the synthetic peptide corresponding to the COOH-terminal region of human cytochrome b558 large subunit, a broad band was specifically detected in neutrophil lysates from 6 marine fish and 2 freshwater fish by western blotting. Immunofluorescence assay showed that the antibody recognized the epitopes in eel and tilapia neutrophils permeabilized with detergent. These results suggest that the cytochrome b large subunit universally exists in fish neutrophils and that the epitopes are exposed to the cytoplasmic side of fish neutrophils as well as human neutrophils. Furthermore, a synthetic peptide corresponding to the COOH-terminus of the large subunit apparently blocked superoxide production in a specific and dose-dependent fashion in eel and tilapia neutrophils, indicating that the region equivalent to the COOH-terminus of cytochrome b large subunit is responsible for superoxide generation in fish neutrophils.


Assuntos
Grupo dos Citocromos b/fisiologia , Peixes , NADPH Oxidases , Neutrófilos/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Grupo dos Citocromos b/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Immunoblotting , Dados de Sequência Molecular , Peptídeos/síntese química , Peptídeos/metabolismo , Superóxidos/metabolismo
13.
Dev Comp Immunol ; 22(4): 433-7, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9699488

RESUMO

When western blotting analysis of Japanese eel neutrophil lysate was performed using antibody against the synthetic peptide corresponding to the carboxyl terminal region of human cytochrome b558 large subunit, a broad band was specifically detected at approximately 90 kDa. The antibody recognized the epitope present in eel neutrophils only after the cells were permeabilized with detergent. These results indicate that the large subunit of cytochrome b exists in fish neutrophils and the epitope is exposed to the cytoplasmic side of membrane, and suggest that cytochrome b is conserved across species.


Assuntos
Grupo dos Citocromos b/metabolismo , Enguias , Neutrófilos/enzimologia , Animais , Western Blotting , Epitopos , Técnica Indireta de Fluorescência para Anticorpo , NADPH Oxidases/metabolismo , Fragmentos de Peptídeos/imunologia
14.
Dev Comp Immunol ; 20(5): 323-30, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-9016386

RESUMO

Two oxygen radicals, superoxide and hydrogen peroxide, were measured by ferricytochrome C reduction and scopoletin fluorescent assays to investigate oxygen metabolism during the respiratory burst in Japanese eel neutrophils. Maximal superoxide production was obtained using 0.1 microgram/mL of PMA. Oxygen consumption was almost equivalent to superoxide production, indicating that the consumed oxygen was almost fully converted to superoxide in eel neutrophils. Hydrogen peroxide production was approximately half that of oxygen consumption or superoxide production in neutrophils elicited by killed bacteria or casein. The amounts of superoxide and hydrogen peroxide production, by neutrophils elicited by irritants, were about 11.5 and 5.0 nM/10(7) cells/ min, respectively, which were significantly higher than those of unelicited neutrophils (2.4 and 0.3 nM/10(7) cells/min, respectively).


Assuntos
Anguilla/metabolismo , Neutrófilos/metabolismo , Oxigênio/análise , Oxigênio/metabolismo , Explosão Respiratória/fisiologia , Anguilla/classificação , Animais , Cinética
15.
Dev Comp Immunol ; 25(1): 47-53, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-10980319

RESUMO

Oxygen radical generation by stimulation with phorbol myristate acetate (PMA) was evaluated in bottlenose dolphin neutrophils. A Cypridina luciferin analog-dependent chemiluminescent assay demonstrated that dolphin neutrophils generate superoxide by the addition of PMA, and that its superoxide-forming activity is completely suppressed by diphenylene iodonium, a specific inhibitor of NADPH oxidase. These results indicate that dolphin neutrophils possess NADPH oxidase activity. Furthermore, the NADPH oxidase activity (hydrogen peroxide production) in dolphin neutrophils, as well as in human neutrophils, was greater at 37 degrees C than at a lower temperature. RT-PCR with specific primers revealed that dolphin neutrophils expressed the mRNAs of the major NADPH oxidase components, which included membrane-associated flavocytochrome b (gp91(phox) and p22(phox)) and cytosolic factors (p40(phox), p47(phox), and p67(phox)), implying the existence of these protein homologues in dolphin neutrophils.


Assuntos
Golfinhos/genética , NADPH Oxidases/genética , Neutrófilos/enzimologia , Superóxidos/sangue , Animais , Bovinos , Células Cultivadas , Golfinhos/sangue , Inibidores Enzimáticos/farmacologia , Radicais Livres/sangue , Humanos , Peróxido de Hidrogênio/metabolismo , Cinética , Camundongos , Neutrófilos/efeitos dos fármacos , Oniocompostos/farmacologia , RNA Mensageiro/metabolismo , Temperatura , Acetato de Tetradecanoilforbol/farmacologia
16.
Int J Hematol ; 67(1): 27-35, 1998 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9594442

RESUMO

Adeno-associated virus (AAV)-based vector is a promising gene transfer vehicle by virtue of the characteristics of wild-type AAV:tropism to a wide range of human tissues and locus-specific integration at chromosome 19q13.3. To elucidate the nature of the recombinant AAV (rAAV), transduction of neomycin phosphotransferase enzyme gene (NeoR gene) into seven human leukemia cell lines was performed. Transduction efficiencies were assessed by colony formation assay and limiting dilution assay. The results suggested that both assays are comparable. Transduction efficiencies of the NeoR gene into K-562, MEG-O1, Raji, MOLT-3, HL-60, U937 and NKM-1 at a multiplicity of infection (MOI) of 0.1 were 0.27, 0.25, 0.015, 0.009, < 0.0025 and < 0.0025%, respectively. After purification and concentration of rAAV, 27% efficiency was observed in K562 at an MOI of 7 and a linear relationship between MOI and efficiency was confirmed, suggesting that this system may be useful for gene transduction into leukemia cells. Integration of the NeoR gene into the host genome was detected by Southern blotting analysis, which showed various sizes of digested fragments. A fluorescent in situ hybridization (FISH) study was carried out on 11 clones, in all of which the NeoR gene was integrated out of chromosome 19q13.3. In five of the clones, whole chromosome painting probes revealed that the integration sites were chromosomes 1q, 2q, 2q, 11p, 12p and 13q.


Assuntos
Dependovirus/genética , Técnicas de Transferência de Genes , Leucemia/genética , Recombinação Genética , Humanos , Células Tumorais Cultivadas
17.
Diabetes Res Clin Pract ; 25(1): 13-7, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7835207

RESUMO

To determine the effect of glycemic control on vitamin B12 (B12) metabolism in diabetes mellitus, we studied B12 metabolism in 19 diabetic patients with poor glycemic control and 15 normal individuals. The diabetic patients had significantly higher total B12 binding capacity (3303 +/- 963 pg/ml), higher serum B12 levels (1173 +/- 503 pg/ml) and unsaturated B12 binding capacity (2131 +/- 902 pg/ml) when compared with the normal controls, but there was no difference in R-binder levels and the B12 binding ratio between the two groups. During a 2-week admission to establish glycemic control, the fructosamine levels in the diabetic patients decreased from 556 to 428 mumol/l and the total B12 binding capacity as well as unsaturated B12 binding capacity were significantly improved to the normal range (P < 0.01), but serum B12 levels, R-binder levels and the B12 binding ratio were not changed. There was a significant association between serum fructosamine levels and the total B12 binding capacity in poorly controlled diabetic patients and the decrease of fructosamine was correlated significantly with the change of total B12 binding capacity and serum B12 levels in diabetic patients. These results indicate the effects of glycemic control on B12 metabolism in diabetes mellitus.


Assuntos
Glicemia/metabolismo , Diabetes Mellitus/sangue , Hexosaminas/sangue , Transcobalaminas/metabolismo , Vitamina B 12/sangue , Adulto , Análise de Variância , Biomarcadores/sangue , Diabetes Mellitus/urina , Feminino , Frutosamina , Glicosúria , Humanos , Masculino , Valores de Referência , Transcobalaminas/análise , Vitamina B 12/metabolismo
18.
J Neurosurg ; 95(1 Suppl): 122-7, 2001 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-11453413

RESUMO

The authors report on a 19-year-old man with an acquired tonsillar herniation caused by a craniocervical junction injury in which serial magnetic resonance (MR) images demonstrated patent and isolated segments of the central canal participating in the dilation and then formation of a cervical syrinx. The patient was involved in a motor vehicle accident; he developed tonsillar herniation as a complication of subarachnoid and epidural hemorrhage, predominantly observed around the cisterna magna and upper cervical canal. Repeated MR images obtained over an 11-month period indicated the for mation and acute enlargement of the syrinx. Ten months after the accident, the patient presented with sensory disturbance in both upper extremities and spasticity due to syringomyelia. He underwent craniocervical decompressive surgery and doraplasty, which reduced the size of syringomyelia. The authors postulate that the patent central canal may play a role in determining the location of a syrinx remote from a focus of cerebrospinal fluid obstruction.


Assuntos
Doenças Cerebelares/etiologia , Encefalocele/etiologia , Traumatismos Cranianos Fechados/complicações , Lesões do Pescoço/complicações , Canal Medular/patologia , Siringomielia/etiologia , Adulto , Malformação de Arnold-Chiari/diagnóstico , Malformação de Arnold-Chiari/etiologia , Malformação de Arnold-Chiari/cirurgia , Doenças Cerebelares/diagnóstico , Doenças Cerebelares/cirurgia , Cerebelo/patologia , Cerebelo/cirurgia , Descompressão Cirúrgica , Dilatação Patológica/diagnóstico , Dilatação Patológica/cirurgia , Progressão da Doença , Encefalocele/diagnóstico , Encefalocele/cirurgia , Seguimentos , Traumatismos Cranianos Fechados/diagnóstico , Traumatismos Cranianos Fechados/cirurgia , Humanos , Imageamento por Ressonância Magnética , Masculino , Lesões do Pescoço/diagnóstico , Lesões do Pescoço/cirurgia , Canal Medular/cirurgia , Siringomielia/diagnóstico , Siringomielia/cirurgia
19.
Vet Microbiol ; 66(4): 301-11, 1999 May.
Artigo em Inglês | MEDLINE | ID: mdl-10384891

RESUMO

One hundred and seventeen rectal fecal specimens were collected in 1995 and 1996 from calves with diarrhea in Kagoshima Prefecture in Japan. The bovine group A rotavirus was detected by enzyme immunoassay in 43 of 117 specimens and isolated from 33 of the 43 specimens that were positive. G serotype, P serotype, and P genotype of 33 isolates were identified by reverse transcription-polymerase chain reaction, and 20 of 33 isolates (60.6%) were identified as G serotype 8. Thus, we discovered that calf diarrhea caused by bovine group A rotavirus carrying G serotype 8 specificity was prevalent in this research area during this research period. To our knowledge, this is the first report on the prevalence of calf diarrhea caused by the bovine group A rotavirus carrying G serotype 8 specificity.


Assuntos
Doenças dos Bovinos/virologia , Diarreia/veterinária , Infecções por Rotavirus/veterinária , Rotavirus/patogenicidade , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Efeito Citopatogênico Viral , Primers do DNA/química , Diarreia/epidemiologia , Diarreia/virologia , Eletroforese em Gel de Ágar/veterinária , Eletroforese em Gel de Poliacrilamida , Fezes/microbiologia , Japão/epidemiologia , Prevalência , RNA Viral/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Rotavirus/química , Rotavirus/classificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Nitrato de Prata/química , Coloração pela Prata/veterinária
20.
Vet Microbiol ; 86(4): 343-9, 2002 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-11955784

RESUMO

G8 bovine group A rotavirus was the most predominant serotype in calf diarrheal fecal specimens examined between 1995 and 1996 in Japan [Vet. Microbiol. 66 (1999) 301]. To date, no evidence that G8 was the most predominant in the typeable specimens has been observed outside Japan. To investigate whether G8 continues to be as common as G6 and G10, the incidence of the main serotypes was determined in the same area (Kagoshima prefecture) between 1997 and 1998 by reverse transcription-polymerase chain reaction. From a total of 104 rotavirus-positive specimens, we successfully identified G type in 79 (76.0%) and P type in 71 (68.3%). The combination of G and P types varied periodically; in 1995, G10P[11] was most common. The most predominant types changed drastically in 1996, and G8 and mixed P types were the most predominant. A dramatic shift of the most predominant type occurred again in 1997 when G6P[5] was most common. This frequency of G6P[5] also continued in 1998. These results suggest that the serotypes prevailing in certain areas change periodically.


Assuntos
Doenças dos Bovinos/epidemiologia , Diarreia/veterinária , Infecções por Rotavirus/veterinária , Rotavirus/genética , Animais , Bovinos , Doenças dos Bovinos/virologia , Diarreia/epidemiologia , Diarreia/virologia , Fezes/virologia , Japão/epidemiologia , Prevalência , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Rotavirus/isolamento & purificação , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/virologia , Sorotipagem
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