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1.
J Dairy Sci ; 103(4): 3656-3667, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32089297

RESUMO

The objective of this study was to evaluate effects of butyrate supplementation on plasma concentration of glucagon-like peptide-2 (GLP-2), apparent total-tract digestibility, and responses to a grain challenge of lactating dairy cows fed diets differing in starch content. Eight Holstein cows averaging 58.6 ± 9.96 d in milk (4 primiparous cows fitted with rumen cannula and 4 multiparous intact cows) were blocked by parity and assigned to one of two 4 × 4 Latin squares balanced for carryover effects with a 2 × 2 factorial arrangement of treatments. Treatments were dietary starch content [20.6 vs. 27.5%, respectively, for low starch (LS) and high starch (HS)] and butyrate supplementation (butyrate vs. control) with 21-d periods. Butyrate was provided as Gustor BP70 WS (Norel, S.A., Madrid, Spain), containing 70% sodium butyrate and 30% fatty acid mixture, at 2% of dietary dry matter (providing butyrate at 1.1% of dietary dry matter), and control premix contained 70% wheat bran and 30% fatty acid mixture. Feeds, orts, and fecal samples were collected from d 17 to 19 to determine apparent total-tract nutrient digestibility. Blood and rumen fluid samples were collected on d 19. The baseline of dry matter intake (DMI) was determined as average DMI from d 17 to 19 for each cow, and cows were feed-restricted at 60% of the baseline DMI on d 20, and a grain challenge was conducted by providing steam-flaked corn grain at 0.6% of body weight, on an as-fed basis, in addition to each treatment diet on d 21, and blood and ruminal fluid samples were collected. The interaction of dietary starch content by butyrate supplementation was significant for plasma GLP-2 concentration, being greater for cows fed butyrate with the HS diet than those fed the other 3 diets. Cows fed butyrate increased n-butyrate concentration in the ruminal fluid and tended to increase dry matter and organic matter digestibility compared with the control. During the grain challenge, rumen endotoxin concentration increased over time and was higher for cows fed the HS diets compared with those fed LS diets. However, response variables related to inflammation were not affected by the grain challenge. However, serum haptoglobin, lipopolysaccharide-binding protein, and serum amyloid-A concentrations were greater for cows fed butyrate with the LS diet, but not for those fed the HS diet. These results indicate that butyrate supplementation may increase plasma GLP-2 concentration for cows fed HS diets, and total-tract digestibility regardless of dietary starch content. However, butyrate supplementation did not mitigate inflammation in this study.


Assuntos
Ração Animal , Butiratos/farmacologia , Dieta/veterinária , Trato Gastrointestinal/efeitos dos fármacos , Peptídeo 2 Semelhante ao Glucagon/metabolismo , Amido/metabolismo , Animais , Bovinos , Digestão/efeitos dos fármacos , Ácidos Graxos/metabolismo , Feminino , Fermentação , Lactação , Rúmen/metabolismo
2.
J Intellect Disabil Res ; 62(3): 217-224, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29193472

RESUMO

BACKGROUND: Previous studies have suggested that ID influences the depth of general anaesthesia (GA) and delays emergence from GA. In this retrospective cohort study, we investigated whether ID affects the time taken to emerge from GA. METHODS: We selected dental patients who underwent GA at the Department of Dental Anaesthesiology, Okayama University Hospital, using predefined inclusion and exclusion criteria, before dividing the selected participants into ID and non-ID (control) groups. Relevant data were collected from electronic anaesthesia records. Emergence time, the time from the discontinuation of propofol and remifentanil to tracheal extubation, was recorded for each patient. We compared the data of the ID group and control group. The association between ID and the emergence time was tested for statistical significance. Multivariate linear regression analysis was used to control for confounders. RESULTS: A total of 97 cases (control = 50, ID = 47) were included in the study. The emergence time was significantly longer in the ID group (ID group: 15.8 ± 6.6 min, control group: 10.8 ± 3.6 min). The ID group included more men and lower propofol and remifentanil infusion rates. The treatment time was longer, and the mean bispectral index was lower in the ID group. Sevoflurane inhalation was used only for anaesthesia induction in the ID group. In the multivariate linear regression analysis, ID was found to be significantly associated with a longer emergence time. CONCLUSION: Our results suggest that ID is associated with a longer emergence time from GA.


Assuntos
Anestesia Geral/estatística & dados numéricos , Anestesia Intravenosa/estatística & dados numéricos , Anestésicos Gerais/administração & dosagem , Estado de Consciência/efeitos dos fármacos , Deficiência Intelectual , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Bucais/estatística & dados numéricos , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo , Adulto Jovem
3.
Soft Matter ; 11(23): 4572-5, 2015 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-25947305

RESUMO

Ultrasmall-angle synchrotron X-ray scattering measurements showed that a three-dimensional polymer lattice of a few 100 nm order with body-centered cubic O(8-) symmetry was formed in a polymer-stabilized blue phase liquid crystal. We obtained clear experimental evidence that the polymer chains condensed selectively in the disclinations within the blue phase during photo-polymerization of monomers in the blue phase.

4.
Insect Mol Biol ; 23(4): 466-74, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24698498

RESUMO

Salp16, a 16-kDa tick salivary gland protein, is known to be the molecule involved in the transmission of Anaplasma phagocytophilum, an obligate intracellular pathogen causing zoonotic anaplasmosis, from its mammalian hosts to Ixodes scapularis. Recently, the presence of A. phagocytophilum was documented in Japan and Ixodes persulcatus was identified as one of its vectors. The purpose of this study was to identify Salp16 genes in I. persulcatus and characterize their function. Two cDNA clones encoding the Salp16-like sequences were obtained from the salivary glands of fed female I. persulcatus ticks and designated Salp16 Iper1 and Iper2. Gene expression analyses showed that the Salp16 Iper genes were expressed specifically in the salivary glands and were up-regulated by blood feeding. These proteins attenuated the oxidative burst of activated bovine neutrophils and inhibited their migration induced by the chemoattractant interleukin-8 (IL-8). These results demonstrate that Salp16 Iper proteins contribute to the establishment of blood feeding as an immunosuppressant of neutrophil, an essential factor in innate host immunity. Further examination of the role of Salp16 Iper in the transmission of pathogens, including A. phagocytophilum, will increase our understanding of the tick-host-pathogen interface.


Assuntos
Anaplasma phagocytophilum/crescimento & desenvolvimento , Anaplasmose/transmissão , Ixodes/imunologia , Ixodes/microbiologia , Neutrófilos/imunologia , Neutrófilos/microbiologia , Glândulas Salivares/metabolismo , Proteínas e Peptídeos Salivares/genética , Sequência de Aminoácidos , Anaplasmose/imunologia , Animais , Vetores Artrópodes , Sequência de Bases , Bovinos/imunologia , DNA Complementar , Feminino , Dados de Sequência Molecular , Glândulas Salivares/microbiologia , Proteínas e Peptídeos Salivares/metabolismo
5.
Nat Cell Biol ; 3(4): 425-8, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11283618

RESUMO

Kinesin is a molecular motor that moves processively by regular 8-nm steps along microtubules. The processivity of this movement is explained by a hand-over-hand model in which the two heads of kinesin work in a coordinated manner. One head remains bound to the microtubule while the other steps from the alphabeta-tubulin dimer behind the attached head to the dimer in front. The overall movement is 8 nm per ATPase cycle. To investigate elementary processes within the 8-nm step, we have developed a new assay that resolves nanometre displacements of single kinesin molecules with microsecond accuracy. Our data show that the 8-nm step can be resolved into fast and slow substeps, each corresponding to a displacement of approximately 4 nm. The substeps are most probably generated by structural changes in one head of kinesin, leading to rectified forward thermal motions of the partner head. It is also possible that the kinesin steps along the 4-nm repeat of tubulin monomers.


Assuntos
Cinesinas/metabolismo , Animais , Bovinos , Microtúbulos/metabolismo
6.
J Cell Biol ; 110(1): 53-62, 1990 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-2153147

RESUMO

Muscle needs an elastic framework to maintain its mechanical stability. Removal of thin filaments in rabbit skeletal muscle with plasma gelsolin has revealed the essential features of elastic filaments. The selective removal of thin filaments was confirmed by staining with phalloidin-rhodamine for fluorescence microscopy, examination of arrowhead formation with myosin subfragment 1 by electron microscopy, and analysis by SDS-PAGE. Thin section electron microscopy revealed the elastic fine filaments (approximately 4 nm in diameter) connecting thick filaments and the Z line. After removal of thin filaments, both rigor stiffness and active tension generation were lost, but the resting tension remained. These observations indicate that the thin filament-free fibers maintain a framework composed of the serial connections of thick filaments, the elastic filaments, and the Z line, which gives passive elasticity to the contractile system of skeletal muscle. The resting tension that remained in the thin filament-free fibers was decreased by mild trypsin treatment. The only protein component that was digested in parallel with the decrease in the resting tension and the disappearance of the elastic filaments was alpha-connectin (also called titin 1), which was transformed from the alpha to the beta form (from titin 1 to 2, respectively). Thus, we conclude that the main protein component of the elastic filaments is alpha-connectin (titin 1).


Assuntos
Citoesqueleto de Actina/ultraestrutura , Proteínas de Ligação ao Cálcio , Proteínas de Ligação ao Cálcio/sangue , Citoesqueleto/ultraestrutura , Proteínas dos Microfilamentos , Proteínas dos Microfilamentos/sangue , Músculos/ultraestrutura , Miofibrilas/ultraestrutura , Actinas/análise , Actinas/metabolismo , Animais , Plaquetas , Proteínas de Ligação ao Cálcio/isolamento & purificação , Bovinos , Elasticidade , Gelsolina , Proteínas dos Microfilamentos/isolamento & purificação , Microscopia Eletrônica , Contração Muscular , Subfragmentos de Miosina/metabolismo , Coelhos , Sarcômeros/ultraestrutura , Tripsina
7.
J Cell Biol ; 101(6): 2167-72, 1985 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-3905821

RESUMO

In an earlier study connectin, an elastic protein of striated muscle, was found to be associated with "gap filaments" originating from the thick filaments in the myofibril, but it was not clear whether it extends to Z lines or not (Maruyama, K., H. Sawada, S. Kimura, K. Ohashi, H. Higuchi, and Y. Umazume, 1984, J. Cell Biol., 99:1391-1397). In the present immunoelectron microscopic study using polyclonal antibodies against native connectin, we have concluded that the connectin structures are directly linked to Z lines from the thick (myosin) filaments in myofibrils of skinned fibers of frog skeletal muscle. There were five distinct antibody-binding stripes in each half of the A band and two stripes in the A-I junction region. Deposits of antibodies were recognized in I bands and Z lines. We suggest that connectin filaments run alongside the thick filaments, starting from a region approximately 0.15 micron from the center of the A band.


Assuntos
Proteínas Musculares/metabolismo , Músculos/ultraestrutura , Miofibrilas/ultraestrutura , Proteínas Quinases , Sarcômeros/ultraestrutura , Animais , Conectina , Técnicas Imunológicas , Microscopia Eletrônica , Relaxamento Muscular , Concentração Osmolar , Rana catesbeiana
8.
J Cell Biol ; 99(4 Pt 1): 1391-7, 1984 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-6384237

RESUMO

Indirect immunofluorescence microscopy of highly stretched skinned frog semi-tendinous muscle fibers revealed that connectin, an elastic protein of muscle, is located in the gap between actin and myosin filaments and also in the region of myosin filaments except in their centers. Electron microscopic observations showed that there were easily recognizable filaments extending from the myosin filaments to the I band region and to Z lines in the myofibrils treated with antiserum against connectin. In thin sections prepared with tannic acid, very thin filaments connected myosin filaments to actin filaments. These filaments were also observed in myofibrils extracted with a modified Hasselbach-Schneider solution (0.6 M KCl, 0.1 M phosphate buffer, pH 6.5, 2 mM ATP, 2 mM MgCl2, and 1 mM EGTA) and with 0.6 M Kl. SDS PAGE revealed that connectin (also called titin) remained in extracted myofibrils. We suggest that connectin filaments play an important role in the generation of tension upon passive stretch. A scheme of the cytoskeletal structure of myofibrils of vertebrate skeletal muscle is presented on the basis of our present information of connectin and intermediate filaments.


Assuntos
Citoesqueleto/ultraestrutura , Proteínas Musculares/análise , Músculos/ultraestrutura , Proteínas Quinases , Animais , Conectina , Eletroforese em Gel de Poliacrilamida , Imunofluorescência , Soros Imunes , Miosinas/análise , Rana catesbeiana
9.
J Cell Biol ; 120(3): 711-24, 1993 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8425898

RESUMO

To clarify the full picture of the connectin (titin) filament network in situ, we selectively removed actin and myosin filaments from cardiac muscle fibers by gelsolin and potassium acetate treatment, respectively, and observed the residual elastic filament network by deep-etch replica electron microscopy. In the A bands, elastic filaments of uniform diameter (6-7 nm) projecting from the M line ran parallel, and extended into the I bands. At the junction line in the I bands, which may correspond to the N2 line in skeletal muscle, individual elastic filaments branched into two or more thinner strands, which repeatedly joined and branched to reach the Z line. Considering that cardiac muscle lacks nebulin, it is very likely that these elastic filaments were composed predominantly of connectin molecules; indeed, anti-connectin monoclonal antibody specifically stained these elastic filaments. Further, striations of approximately 4 nm, characteristic of isolated connectin molecules, were also observed in the elastic filaments. Taking recent analyses of the structure of isolated connectin molecules into consideration, we concluded that individual connectin molecules stretched between the M and Z lines and that each elastic filament consisted of laterally-associated connectin molecules. Close comparison of these images with the replica images of intact and S1-decorated sarcomeres led us to conclude that, in intact sarcomeres, the elastic filaments were laterally associated with myosin and actin filaments in the A and I bands, respectively. Interestingly, it was shown that the elastic property of connectin filaments was not restricted by their lateral association with actin filaments in intact sarcomeres. Finally, we have proposed a new structural model of the cardiac muscle sarcomere that includes connectin filaments.


Assuntos
Citoesqueleto de Actina/ultraestrutura , Actinas/fisiologia , Miosinas/fisiologia , Músculos Papilares/ultraestrutura , Sarcômeros/ultraestrutura , Actinas/isolamento & purificação , Animais , Anticorpos Monoclonais , Elasticidade , Eletroforese em Gel de Poliacrilamida , Técnica de Congelamento e Réplica , Immunoblotting , Microscopia Eletrônica , Microscopia Imunoeletrônica , Modelos Estruturais , Miosinas/isolamento & purificação , Coelhos
10.
Transplant Proc ; 51(5): 1382-1386, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31027828

RESUMO

BACKGROUND: Rituximab (RIT) is effective as a part of the desensitization therapy before ABO-incompatible kidney transplantation (ABOi-KTx), and a single dose of RIT at 375 mg/m2 or less is recommended. However, adequate RIT dose recommendations have not yet been established for individual recipients. Therefore, we evaluated the relationship between the proportion of B cells in peripheral blood and acute antibody-mediated rejection (AAMR). METHODS: Forty-four consecutive ABOi-KTx recipients were enrolled in this retrospective study. Before transplantation, subjects were treated with RIT at various doses, ranging from 65 to 400 mg/body (46-263 mg/m2), followed by plasmapheresis and intravenous immunoglobulin as a desensitization therapy. The percentage of CD19+ cells in the total peripheral blood lymphocytes population (%CD19) was determined the day before transplantation. Transplant recipients were divided into 2 groups according to pretransplant %CD19, as follows: low %CD19 group, ≤ 1.2% (n = 35) and high %CD19 group, > 1.2% (n = 9). The relationship between %CD19 and incidence of AAMR was evaluated, and the predicting factors for AAMR incidence were determined by univariate and multivariate analyses. RESULTS: The incidence of AAMR was significantly higher in the high %CD19 group than in the low %CD19 group (44.4% vs 5.7%, P = .006). Furthermore, multivariate analysis showed that %CD19 > 1.2% was the only independent factor to predict AAMR, with an odds ratio of 14.31 (P = .038). CONCLUSION: High %CD19 values after rituximab administration in ABOi-KTx recipients implies insufficient depletion of B cells, which can lead to AAMR.


Assuntos
Antígenos CD19/sangue , Incompatibilidade de Grupos Sanguíneos/tratamento farmacológico , Rejeição de Enxerto/prevenção & controle , Transplante de Rim/métodos , Rituximab/administração & dosagem , Adulto , Feminino , Rejeição de Enxerto/sangue , Rejeição de Enxerto/imunologia , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Incidência , Linfócitos/imunologia , Masculino , Pessoa de Meia-Idade , Plasmaferese/métodos , Estudos Retrospectivos , Transplantados
11.
J Bone Joint Surg Br ; 90(3): 324-9, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18310754

RESUMO

We prospectively examined the physical and imaging findings, including MRI, of 23 patients with spontaneous osteonecrosis of the knee after obtaining informed consent to acquire tissue specimens at surgery. There were four men and 19 women, with a mean age of 67.5 years (58 to 77). Plain radiographs were designated as stages 1, 2, 3 or 4 according to the classification of Koshino. Five knees were classified as stage 1, five as stage 2, seven as stage 3 and six as stage 4. The histological specimens were stained with haematoxylin and eosin and tetrachrome. In the early stages of the condition, a subchondral fracture was noted in the absence of any features of osteonecrosis, whereas in advanced stages, osteonecrotic lesions were confined to the area distal to the site of the fracture which showed impaired healing. In such cases, formation of cartilage and fibrous tissue, occurred indicating delayed or nonunion. These findings strongly suggest that the histopathology at each stage of spontaneous osteonecrosis is characterised by different types of repair reaction for subchondral fractures.


Assuntos
Necrose da Cabeça do Fêmur/patologia , Articulação do Joelho/patologia , Idoso , Medula Óssea/diagnóstico por imagem , Medula Óssea/patologia , Cartilagem Articular/diagnóstico por imagem , Cartilagem Articular/patologia , Progressão da Doença , Feminino , Necrose da Cabeça do Fêmur/diagnóstico por imagem , Humanos , Articulação do Joelho/diagnóstico por imagem , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Radiografia , Coloração e Rotulagem
12.
Anesth Prog ; 55(3): 73-7, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18788841

RESUMO

Benzodiazepines in intravenous sedation are useful, owing to their outstanding amnesic effect when used for oral surgery as well as dental treatments on patients with intellectual disability or dental phobia. However, compared with propofol, the effect of benzodiazepine lasts longer and may impede discharge, especially when it is administered orally because of fear of injections. Although flumazenil antagonizes the effects of benzodiazepine quickly, its effect on the equilibrium function (EF) has never been tested. Since EF is more objective than other tests, the purpose of this study is to assess the sedation level and EF using a computerized static posturographic platform. The collection of control values was followed by the injection of 0.075 mg/kg of midazolam. Thirty minutes later, 0.5 mg or 1.0 mg of flumazenil was administered, and the sedation level and EF were measured until 150 minutes after flumazenil administration. Flumazenil antagonized sedation, and there was no apparent resedation; however, it failed to antagonize the disturbance in EF. This finding may be due to differences in the difficulty of assessing the sedation level and performing the EF test, and a greater amount of flumazenil may effectively antagonize the disturbance in EF.


Assuntos
Anestesia Dentária/métodos , Sedação Consciente/métodos , Flumazenil/administração & dosagem , Moduladores GABAérgicos/administração & dosagem , Hipnóticos e Sedativos/farmacologia , Midazolam/farmacologia , Equilíbrio Postural/efeitos dos fármacos , Adulto , Recuperação Demorada da Anestesia , Diagnóstico por Computador , Relação Dose-Resposta a Droga , Método Duplo-Cego , Humanos , Hipnóticos e Sedativos/antagonistas & inibidores , Injeções Intravenosas , Midazolam/antagonistas & inibidores , Estudos Prospectivos , Adulto Jovem
13.
J Clin Invest ; 99(5): 867-78, 1997 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-9062344

RESUMO

Previous studies have indicated that nitric oxide (NO) released from Kupffer cells modulates biological viability of cocultured hepatoma cells. This study was designed to evaluate the mechanisms by which Kupffer cells synthesize and release NO in reponse to cocultured hepatoma cells. Kupffer cells isolated from male Wistar rats were cocultured with rat hepatoma cell line, AH70 cells. The sum of nitrite and nitrate levels increased in the culture medium of Kupffer cells with AH70 cells as compared with those of Kupffer cells or AH70 cells alone. Increased expressions of iNOS and iNOS mRNA in Kupffer cells cocultured with AH70 cells were detected by an immunofluorescence staining and a fluorescence in situ hybridization study, respectively. A fluorescence in situ DNA-protein binding assay revealed that NF-kappaB activation occurs in Kupffer cells and activated NF-kappaB moved into the nuclei preceding to an increased production of NO. Oxidative stress indicated by dichlorofluorescein fluorescence was observed in Kupffer cells cocultured with AH70 cells. An increased calcium mobilization indicated as increased fluo-3-associated fluorescence was also induced in Kupffer cells after coculture with AH70 cells. Monoclonal antibodies directed against rat CD18 and ICAM-1, as well as TMB-8, a calcium inhibitor, prevented the calcium mobilization, active oxygen production, and NF-kappaB activation in addition to the increased production of NO. Pyrrolidine dithiocarbamate, an inhibitor of oxidative NF-kappaB activation, diphenylene iodonium, an NADPH oxidase inhibitor, and quinacrine, a phospholipase A2 inhibitor, significantly attenuated the increase in dichlorofluorescein fluorescence, NF-kappaB activation, and NO production. Therefore, this study suggests that CD18/ICAM-1-dependent cell-to-cell interaction with hepatoma cells causes calcium mobilization and oxidative activation of NF-kappaB, which may lead to the increased production of NO in Kupffer cells.


Assuntos
Antígenos CD18/fisiologia , Técnicas de Cocultura , Molécula 1 de Adesão Intercelular/fisiologia , Células de Kupffer/metabolismo , NF-kappa B/fisiologia , Óxido Nítrico/metabolismo , Animais , Anticorpos Monoclonais/imunologia , Transporte Biológico , Compostos de Bifenilo/farmacologia , Antígenos CD18/imunologia , Cálcio/metabolismo , Comunicação Celular , Núcleo Celular/metabolismo , Células Cultivadas , Meios de Cultura/análise , Técnica Indireta de Fluorescência para Anticorpo , Ácido Gálico/análogos & derivados , Ácido Gálico/imunologia , Ácido Gálico/farmacologia , Hibridização In Situ , Molécula 1 de Adesão Intercelular/imunologia , Masculino , Microscopia Confocal , NF-kappa B/metabolismo , Nitratos/análise , Nitratos/metabolismo , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase/imunologia , Óxido Nítrico Sintase/metabolismo , Nitritos/análise , Nitritos/metabolismo , Oniocompostos/farmacologia , Estresse Oxidativo , Pirrolidinas/farmacologia , Quinacrina/farmacologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Tiocarbamatos/farmacologia , Fatores de Tempo , Células Tumorais Cultivadas
14.
Mol Cell Biol ; 10(7): 3717-26, 1990 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-2355920

RESUMO

During spermatogenesis, several genes are expressed in a germ cell-specific manner. Previous studies have demonstrated that rat and mouse spermatogenic cells produce a 1,700-nucleotide proenkephalin RNA, while somatic cells that express the proenkephalin gene contain a 1,450-nucleotide transcript. Using cDNA cloning, RNA protection, and primer extension analyses, we showed that transcription of the rat and mouse spermatogenic-cell RNAs is initiated downstream from the proenkephalin somatic promoter in the first somatic intron (intron As). In both species, the germ cell cap site region consists of multiple start sites distributed over a length of approximately 30 base pairs. Within rat and mouse intron As, the region upstream of the germ cell cap sites is GC rich and lacks TATA sequences. A consensus binding site for the transcription factor SP1 was identified in intron As downstream of the proenkephalin germ cell cap site region. These features are characteristic of several previously described promoters that lack TATA sequences. Homologies were also identified between the proenkephalin and rat cytochrome c spermatogenic-cell promoters, including the absence of a TATA box, a multiple start site region, and several common sequences. This promoter motif thus may be shared with other genes expressed in male germ cells.


Assuntos
Encefalinas/genética , Genes , Fígado/metabolismo , Precursores de Proteínas/genética , Testículo/metabolismo , Transcrição Gênica , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA/genética , DNA/isolamento & purificação , Sondas de DNA , Biblioteca Genômica , Íntrons , Masculino , Camundongos , Dados de Sequência Molecular , Especificidade de Órgãos , Sondas RNA , Ratos , Mapeamento por Restrição , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Espermatogênese
15.
Vet Res Commun ; 31(8): 985-91, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17285250

RESUMO

Complement receptor type 3 (CR3)- and Fc receptor (FcR)-mediated metalloproteinase-9 (MMP-9) secretion and their intracellular signalling of bovine neutrophils were evaluated. Relative density of MMP-9 secreted by neutrophils stimulated with opsonized zymosan (OPZ, stimulant for CR3) was significantly (p < 0.05) increased when the OPZ concentration was increased from 0 to 0.4 mg/ml. Similar results were obtained for neutrophils stimulated with heat-aggregated IgG (Agg-IgG, stimulant for Fc receptor) at concentrations from 0 to 0.40 mg/ml. Preincubation of neutrophils with 1-30 nmol/L wortmannin (phosphoinositide 3-kinase inhibitor) resulted in inhibition of MMP-9 secretion induced by stimulation with OPZ and Agg-IgG in a concentration-dependent manner, 30 nmol/L wortmannin causing complete inhibition. Similarly, preincubation of neutrophils with 0-100 mumol/L genistein (tyrosine kinase inhibitor) also resulted in inhibition of OPZ- and Agg-IgG-induced MMP-9 secretion in a concentration-dependent manner, with 100 micromol/L genistein causing complete inhibition. Significant (p < 0.05) positive correlations were found between MMP-9 and luminal-dependent chemiluminescent response (LDCL) in the case of stimulation with OPZ (r = 0.754) and in the case of stimulation with Agg-IgG (r = 0.728). Our findings suggested that CR3 and FcR play a critical role in production of MMP-9 and may be regulated by intracellular signal transduction, including that by phosphoinositide 3-kinase (PI3K) and tyrosine kinase (TK).


Assuntos
Bovinos/imunologia , Metaloproteinase 9 da Matriz/metabolismo , Neutrófilos/imunologia , Receptores de Complemento/imunologia , Receptores Fc/imunologia , Androstadienos/farmacologia , Animais , Eletroforese em Gel de Poliacrilamida/veterinária , Genisteína/farmacologia , Imunoglobulina G/imunologia , Medições Luminescentes/veterinária , Metaloproteinase 9 da Matriz/imunologia , Inibidores de Metaloproteinases de Matriz , Ativação de Neutrófilo/efeitos dos fármacos , Ativação de Neutrófilo/imunologia , Neutrófilos/enzimologia , Inibidores de Proteínas Quinases/farmacologia , Receptores de Complemento/metabolismo , Receptores Fc/metabolismo , Transdução de Sinais , Estatísticas não Paramétricas , Wortmanina , Zimosan/imunologia
16.
JDR Clin Trans Res ; 2(2): 158-167, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30931781

RESUMO

Intravenous sedation is useful for dental treatment in patients with intellectual disabilities. However, it is often necessary to manage such patients with deep sedation because their cooperation cannot be obtained. During deep sedation, undetected hypoventilation can lead to severe complications, such as hypoxia. Recently, capnographic monitoring has been advocated as a useful technique for preventing hypoxia during sedation. This randomized control trial evaluated whether the use of capnography reduces the incidence of hypoxia during the deep sedation of patients for dental treatment. This study involved patients with intellectual disabilities who underwent dental treatment under sedation. The subjects were randomized to the intervention group (I-group) or control group (C-group). All of the patients underwent routine monitoring, as well as bispectral index (BIS) and capnographic monitoring; however, only an independent observer had access to the patients' capnographic data during the dental procedures. Sedation was maintained at a BIS of 50 to 70 by administration of propofol. In the I-group, the independent observer signaled to the dental anesthesiologist if the capnogram indicated that the patient had been suffering from alveolar hypoventilation or apnea for >15 s. In the C-group, the observer signaled to the dental anesthesiologist if the capnogram indicated that the patient had been suffering from alveolar hypoventilation or apnea for >60 s. In both groups, the dental anesthesiologists responded to the signals using appropriate airway management strategies. The primary endpoint of this study was the incidence of hypoxia during dental treatment, which was defined as oxygen saturation of <95%. Hypoxemic episodes occurred in 13.4% and 34.8% of cases in the I-group and C-group, respectively. The incidence of hypoxia was significantly lower in the I-group. These results suggest that capnographic monitoring during deep sedation for dental treatment prevents hypoxemic episodes by allowing the early detection of hypoventilation. Knowledge Transfer Statement: This is the first randomized controlled trial to examine whether the use of capnography reduces the incidence of hypoxia during deep sedation for dental treatment. The findings of this study can be used by clinicians to aid decision-making regarding dental sedation standards at individual clinics. Moreover, they can be used as high-level evidence during the production or updating of clinical guidelines for dental sedation by leading associations.

17.
Transplant Proc ; 49(1): 84-87, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28104165

RESUMO

BACKGROUND: De novo donor-specific antibody (dnDSA), especially against class II HLA, correlates with chronic active antibody-mediated rejection (CAAMR), which eventually leads to graft loss. It would be helpful if we could identify the patients at high risk of dnDSA development in terms of histocompatibility. Structure-based matching strategy assessing mismatched epitopes/eplets by comparing polymorphic amino acid sequences can predict the risk of development of dnDSA and CAAMR. However, it has not been evaluated in Japanese patients whose diversity in HLA is limited. PATIENTS AND METHODS: We retrospectively studied 55 living related kidney transplant patients and ascertained donor and recipient HLA-A, -B, -DRB1, and -DQB1. The number of mismatched eplets was determined using an algorithm, HLAMatchmaker version 3. The relationship between characteristics of mismatched eplets and development of CAAMR was evaluated. RESULTS: There were 8 patients in the CAAMR group and 47 in the control group. The numbers of mismatched HLAs (3.6 ± 1.2 in CAAMR and 3.7 ± 2.0 in control groups), mismatched eplets (32.2 ± 10.4 in CAAMR and 34.4 ± 19.8 in control groups), mismatched DRB1 eplets (11.2 ± 4.3 in CAAMR and 11.5 ± 7.9 in control groups), and mismatched DQB1 eplets (9.2 ± 4.3 in CAAMR and 10.5 ± 7.3 in control groups) were not significantly different. Significantly more patients had at least one highly immunogenic mismatched eplet (62.5% in CAAMR and 25.5% in control groups; P = .024 by χ2 test). CONCLUSIONS: The presence of highly immunogenic mismatched eplets is associated with development of CAAMR.


Assuntos
Anticorpos/imunologia , Rejeição de Enxerto/imunologia , Cadeias beta de HLA-DQ/imunologia , Transplante de Rim/efeitos adversos , Imunologia de Transplantes/imunologia , Sequência de Aminoácidos , Formação de Anticorpos , Epitopos/imunologia , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Doadores de Tecidos
18.
Cancer Res ; 47(1): 56-61, 1987 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-2431767

RESUMO

Six mouse monoclonal antibodies reactive with carcinoembryonic antigen (CEA) were prepared and used for the analysis of the antigenic heterogeneity of CEA in patient sera. Their reaction specificity and the chemical nature of antigenic epitopes recognized by them were analyzed by radioimmunoassay on the basis of reactivities with different preparations of CEA, normal fecal antigen 2, and nonspecific cross-reacting antigen 2 before and after chemical and/or enzymatic treatment. Two antibodies, F3-30 and F4-82, raised with CEA were reactive with different peptide epitopes on the antigen molecules and revealed a quite universal reactivity with all CEA, normal fecal antigen 2, or nonspecific cross-reacting antigen 2 preparations tested. The serum CEA values obtained with these antibodies were highly correlated with those obtained with conventional radioimmunoassays for CEA. The other four antibodies (F4-11 and F33-37 raised with CEA, F8-52 with normal fecal antigen 2, and F48-60 with nonspecific cross-reacting antigen 2) were found to recognize carbohydrate epitopes with different specificities and revealed very heterogeneous reactivities. The serum CEA values estimated with these four antibodies were highly variable depending on the antibody used, suggesting that the expression of carbohydrate epitopes on the CEA molecules in patient sera was quite heterogeneous. The antigenic heterogeneity of the carbohydrate epitopes was detected even in a single patient serum by affinity chromatography. The causes that give rise to the difference in CEA values between the Roche and the Daiichi kits were analyzed on the basis of reactivities of three groups of patient sera, which showed extremely different ratios for the Roche and Daiichi kits, with monoclonal anti-carbohydrate antibodies. The results obtained suggest that, at least in part, the diversity of antigenic expression on carbohydrate chains on the CEA molecules in patient sera and the variation in specificity or quantity of anti-carbohydrate antibodies in the polyclonal antibody preparations used for the respective assay systems may result in the differences in the estimated CEA values.


Assuntos
Anticorpos Monoclonais/imunologia , Carboidratos/imunologia , Antígeno Carcinoembrionário/imunologia , Antígeno Carcinoembrionário/análise , Epitopos/análise , Humanos , Radioimunoensaio , Kit de Reagentes para Diagnóstico
19.
Cancer Res ; 60(22): 6396-402, 2000 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-11103804

RESUMO

Viral fusogenic membrane glycoproteins (FMGs) are candidates for gene therapy of solid tumors because they cause cell fusion, leading to formation of lethal multinucleated syncytia. However, the cellular mechanisms mediating cell death after FMG-induced cell fusion remain unclear. The present study was designed to examine the mechanisms by which FMG expression in hepatocellular carcinoma cells lead to cell death. Transfection of Hep3B cells with the Gibbon Ape leukemia virus hyperfusogenic envelope protein (GALV-FMG) resulted in the formation of multinucleated syncytia that reached a maximum 5 days after transfection (100 nuclei/syncytia). The syncytia were viable for a period of 2 days and then rapidly lost viability by day 5. Mitochondrial dysfunction occurred in GALV-FMG-induced syncytia prior to loss of viability with loss of the mitochondrial membrane potential, cellular ATP depletion, and release of mitochondrial cytochrome c-GFP into the cytosol. The pan-caspase inhibitor, Z-VAD-fmk, did not prevent cell death. However, glycolytic generation of ATP with fructose effectively increased cellular ATP and preserved syncytial viability. These data suggest that expression of FMG in hepatoma cells results in the formation of multinucleated syncytia, causing mitochondrial failure with ATP depletion, a bioenergetic form of cell death with necrosis. This form of cell death should be effective in vivo and enhance the bystander effect, suggesting that FMG-based gene therapy deserves further study for the treatment of hepatocellular and other cancers.


Assuntos
Carcinoma Hepatocelular/terapia , Terapia Genética/métodos , Células Gigantes/patologia , Vírus da Leucemia do Macaco Gibão/genética , Neoplasias Hepáticas/terapia , Proteínas Virais de Fusão/genética , Trifosfato de Adenosina/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Inibidores de Caspase , Caspases/metabolismo , Morte Celular/fisiologia , Fusão Celular/métodos , Metabolismo Energético/fisiologia , Frutose/farmacologia , Células Gigantes/metabolismo , Células Gigantes/virologia , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Hepáticas/fisiologia , Necrose , Fosforilação Oxidativa , Células Tumorais Cultivadas , Proteínas Virais de Fusão/biossíntese
20.
Biomed Pharmacother ; 59 Suppl 1: S54-7, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16275509

RESUMO

The effect of aging on blood pressure (BP) and heart rate (HR) was investigated in a cross-sectional study in the high-altitude community of Leh, Ladakh (altitude: 3524 m) and a Japanese community in U town, Hokkaido (altitude: 25 m). BP and HR were obtained in a sitting position from 332 subjects 13-81 years of age in Ladakh, and from 216 Japanese citizens, 24-79 years of age. Measurements were taken after a 2-min rest, using a semi-automated BP device (UA-767 PC, A and D Co. LTD, Tokyo). High-altitude people showed higher diastolic BP and HR values than lowland people (83.2 vs. 76.9 mmHg and 78.6 vs. 69.2 bpm, P < 0.001), but no difference in systolic BP. Highland people also showed a steeper BP increase with age than the lowland people (systolic BP: 0.7476 vs. 0.3179 mmHg/year, P < 0.0005; diastolic BP: 0.3196 vs. 0.0750 mmHg/year, P < 0.001). This chronoecologic investigation in Ladakh examined the circulation as a physiological system at high-altitude. Our data indicate the need for a more comprehensive cardiovascular assessment for a better diagnosis and a more fruitful treatment. Longitudinal observations of effects of socio-ecologic factors on the cardiovascular system should help prevent strokes and other cardiovascular events, especially at high altitude.


Assuntos
Envelhecimento/fisiologia , Altitude , Pressão Sanguínea/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Estudos Transversais , Feminino , Humanos , Índia/epidemiologia , Japão/epidemiologia , Modelos Lineares , Masculino , Pessoa de Meia-Idade
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