RESUMO
This study aimed to investigate effects of a 100-km ultramarathon on cardiac markers of exercise-induced-hypertensive marathoners. 10 marathoners with exercise-induced hypertension and 10 normal marathoners participated in the study. Their blood samples were collected before starting, at 50 km, and after finishing the course (100 km). Creatinine kinase was more significantly increased in the exercise-induced-hypertensive group than in the normal group at 100 km (P<0.05). N-terminal pro-brain nutriuretic peptide was significantly increased in the exercise-induced-hypertensive group at 50 km and 10 km (P<0.05) which was significant being doubled compared to the normal group (P<0.05). Exercise-induced-hypertensive marathoners showed a significant triple-increase in C-Reactive protein at 100 km (P<0.05). In conclusion, although the exercise-induced-hypertensive runners did not have myocardial damage during the 100 km ultramarathon, they had higher myocardial stress and more damage in active muscles due to a bloodstream disability.
Assuntos
Exercício Físico/fisiologia , Hipertensão/etiologia , Resistência Física/fisiologia , Corrida/fisiologia , Adulto , Biomarcadores/metabolismo , Proteína C-Reativa/metabolismo , Creatina Quinase/metabolismo , Teste de Esforço , Humanos , Masculino , Pessoa de Meia-Idade , Peptídeo Natriurético Encefálico/metabolismo , Fragmentos de Peptídeos/metabolismoRESUMO
Edwardsiella tarda is a freshwater pathogen that may cause mild to invasive infections with high mortality in humans. We describe two patients with serious E. tarda infections. The first patient was a woman with a tuboovarian abscess (TOA) and bilateral salpingitis requiring surgical resection and drainage. Her hospital course was complicated by a postoperative wound infection. TOA fluid as well as surgical wound culture revealed pure growth of E. tarda resistant to several antibiotics. The second patient was a man with a bloodstream E. tarda infection and cholangitis who recently traveled to Ecuador. He presented with hypoxia and further workup revealed choledocholithiasis and common bile duct benign polyps. Both patients made a full recovery.
Assuntos
Abscesso Abdominal , Edwardsiella tarda/fisiologia , Infecções por Enterobacteriaceae/complicações , Sepse , Infecção da Ferida Cirúrgica , Abscesso Abdominal/tratamento farmacológico , Abscesso Abdominal/etiologia , Abscesso Abdominal/cirurgia , Adulto , Idoso , Antibacterianos/uso terapêutico , Infecções por Enterobacteriaceae/tratamento farmacológico , Feminino , Humanos , Masculino , Sepse/tratamento farmacológico , Sepse/etiologia , Infecção da Ferida Cirúrgica/tratamento farmacológico , Infecção da Ferida Cirúrgica/etiologiaRESUMO
PURPOSE: To compare the influence of two different regions of interest (ROIs) on diffusion tensor metrics in dystrophic thigh muscles using a custom-made (whole muscle) ROI including and a selective ROI excluding areas of fatty replacement. METHODS: Diffusion tensor imaging (DTI) and chemical-shift-encoded water-fat magnetic resonance imaging (MRI) of the thigh was conducted on a 3-Tesla system in 15 cases with muscular dystrophy and controls. The ROIs were chosen according to patterns of fatty replacement on co-registered axial DTI and gradient echo sequence (GRE) images. Fractional anisotropy (FA), apparent diffusion coefficient (ADC), fiber track length (FTL), and muscle fat fractions (MFF) were compared between both ROI segmentations. These comparisons, muscle-specific correlation coefficients, and the influence of ROI localization on tensor metrics were derived based on linear mixed effects regression models. RESULTS: In the cases a high correlation was observed for ADC and FA with MFF using a custom ROI. The correlation was weaker but still significant with a selective ROI method. Using the custom ROI, FTL correlated significantly with MFF in 3 out of 4 muscles (râ¯≤ -0.51). A correlation was not found for the selective ROI method. Interaction analysis revealed that the association of ADC and FA with MFF was not significantly influenced by the ROI localization. For FTL the ROI localization significantly reduced the negative association with MFF. CONCLUSION: The DTI metrics and FTL of custom ROI segmentation are significantly influenced by MFF. Contrary to ADC and FA, the effect of MFF on FTL is significantly reduced when applying selective ROI segmentation, which could therefore be a better option for MR tractography.
Assuntos
Músculo Esquelético/patologia , Distrofias Musculares/patologia , Tecido Adiposo/patologia , Adulto , Idoso , Água Corporal , Imagem de Tensor de Difusão/métodos , Feminino , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Coxa da Perna , Adulto JovemRESUMO
Human erythroid dematin is a cytoskeletal protein capable of bundling actin filaments in vitro. The carboxyl terminal domain of dematin is homologous to the headpiece domain of villin, an actin-binding protein of the brush border cytoskeleton. Here we report the complete structure of the dematin gene located on human chromosome 8p21.1, a region frequently deleted in prostate cancer. The dematin gene is composed of 15 exons spanning approximately 15 kb. We also report two novel isoforms of dematin derived from alternative splicing of the dematin gene in the brain.
Assuntos
Processamento Alternativo , Proteínas Sanguíneas/genética , Cromossomos Humanos Par 8 , Fosfoproteínas , Sequência de Aminoácidos , Sequência de Bases , DNA Complementar , Células Precursoras Eritroides/metabolismo , Humanos , Proteínas dos Microfilamentos , Dados de Sequência Molecular , Homologia de Sequência de AminoácidosRESUMO
Human p55, the major palmitoylated protein associated with the cytoplasmic face of the erythrocyte membrane, is believed to modulate interactions between protein 4.1 and glycophorin C. It is the prototype of a newly described family of signaling molecules that includes hD1g, the human homologue of the Drosophila discs-large tumor suppressor protein. Chronic myeloid leukemia is characterized by transformation to a fulminating acute leukemia, heralded by evolution of the Philadelphia chromosome positive genotype (Ph +) to further abnormalities. RT-PCR of p55 mRNA from a patient with acute megakaryoblastic CML revealed a 69 base pair deletion in the PDZ domain, corresponding to exon 5 of the p55 gene. The deletion of constitutive exon 5 not only marks the first abnormality of the p55 cDNA in human disease but also the first abnormality of a PDZ domain in human disease and may represent another genetic abnormality associated with CML in blast crisis.
Assuntos
Crise Blástica/genética , Éxons , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Núcleosídeo-Fosfato Quinase/genética , Domínios de Homologia de src , Sequência de Aminoácidos , Guanilato Quinases , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Dados de Sequência Molecular , Núcleosídeo-Fosfato Quinase/químicaRESUMO
Dematin is a cytoskeletal protein that bundles actin filaments in a phosphorylation-dependent manner. The primary structure of dematin is organized into an N-terminal core domain of unknown function and a C-terminal domain that is homologous to the "headpiece" domain of villin. We have previously localized the dematin gene on human chromosome 8p21.1, a region distal to the ankyrin locus for hereditary spherocytosis. Radiation hybrid mapping now places dematin between D8S258 and D8S137, two microsatellite markers frequently deleted in prostate cancer. The 8p21.1 region is also deleted in prostate, breast, colon, and bladder cancers, suggesting the presence of a tumor suppressor gene(s). Using laser-capture microdissection technique and fluorescence in situ hybridization (FISH), we demonstrate loss of heterozygosity (LOH) of the dematin gene in a majority of chromosomal region 8p21-linked prostate tumors. One allele of dematin was also deleted in the established prostate adenocarcinoma cell line PC-3, which displays a classic oncogenic phenotype. Overexpression of wild-type dematin in PC-3 cells resulted in the restoration of a more polarized, epithelial-like phenotype. Conversely, the heterologous expression of dominant negative mutants of dematin perturbed normal cell morphology of NIH 3T3 fibroblasts. These results suggest a biological function of dematin in the regulation of cell shape, with implications in the pathobiology of prostate tumorigenesis.
Assuntos
Proteínas Sanguíneas/fisiologia , Cromossomos Humanos Par 8 , Perda de Heterozigosidade , Fosfoproteínas , Neoplasias da Próstata/genética , Células 3T3 , Animais , Proteínas Sanguíneas/genética , Citoplasma/metabolismo , Proteínas do Citoesqueleto , DNA Antissenso/metabolismo , Epitélio/metabolismo , Humanos , Masculino , Camundongos , Proteínas dos Microfilamentos , Modelos Genéticos , Fenótipo , Mapeamento Físico do Cromossomo , Próstata/metabolismo , Células Tumorais CultivadasRESUMO
Relatively little is known about the contractile behavior of the human articular chondrocyte. Other connective tissue cells are known to express a contractile actin isoform, alpha-smooth muscle actin, in response to injury, at selected stages of wound healing, and in certain pathological conditions. This and recent work demonstrating contractile behavior in adult canine articular chondrocytes in vitro prompted the present study of the distribution of alpha-smooth muscle actin-containing chondrocytes in human articular cartilage. Approximately 75% of the chondrocytes in the superficial region of cartilage expressed alpha-smooth muscle actin as demonstrated by immunohistochemistry. In contrast, only approximately 10% of the cells in the deep region stained for this contractile actin isoform. There was no correlation of the percentage of alpha-smooth muscle actin-positive cells in either region with Mankin grade or with age. This is the first report of a contractile potential for human articular chondrocytes. The roles of alpha-smooth muscle actin in these cells warrant further investigation. The question of whether it is necessary to refer to these cells as myochondrocytes is considered.
Assuntos
Actinas/metabolismo , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Músculo Liso/metabolismo , Idoso , Idoso de 80 Anos ou mais , Cartilagem Articular/citologia , Contagem de Células , Condrócitos/citologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Articulação do Quadril/metabolismo , Articulação do Quadril/patologia , Articulação do Quadril/cirurgia , Humanos , Técnicas Imunoenzimáticas , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Osteoartrite/patologia , Osteoartrite/cirurgiaRESUMO
For each patient who requires removal of anterior teeth, there are a multitude of treatment considerations. Cosmetic demands, functional needs, treatment sequencing, timeliness, and affordability are some primary concerns that must be addressed on an individual basis. A patient will generally want a cosmetic and functional prosthesis at the earliest possible opportunity. Providing the most appropriate interim prosthesis for a given patient is both challenging and rewarding. The numerous clinical techniques for immediate interim tooth replacement are reviewed, and previously unreported methods are presented to assist the clinician in the selection of interim prosthesis design.
Assuntos
Planejamento de Dentadura , Prótese Parcial Imediata , Resinas Acrílicas , Dente Canino , Dente Suporte , Prótese Parcial Fixa , Prótese Parcial Removível , Humanos , Incisivo , Dente ArtificialAssuntos
Citoplasma/química , Proteínas do Citoesqueleto , Proteínas de Membrana/classificação , Proteínas de Membrana/metabolismo , Proteínas dos Microfilamentos , Neuropeptídeos , Animais , Sítios de Ligação , Proteínas Sanguíneas/química , Proteínas Sanguíneas/classificação , Proteínas Sanguíneas/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Citoplasma/metabolismo , Humanos , Proteínas de Membrana/química , Nomes , Fosfoproteínas/química , Fosfoproteínas/classificação , Fosfoproteínas/metabolismo , Proteínas/química , Proteínas/classificação , Proteínas/metabolismoRESUMO
IFN-gamma activates macrophages to kill diverse intracellular pathogens, but does not activate human macrophages to kill virulent Mycobacterium tuberculosis. We tested the hypothesis that this is due to inhibition of IFN-gamma signaling by M. tuberculosis and found that M. tuberculosis infection of human macrophages blocks several responses to IFN-gamma, including killing of Toxoplasma gondii and induction of FcgammaRI. The inhibitory effect of M. tuberculosis is directed at transcription of IFN-gamma-responsive genes, but does not affect proximal steps in the Janus kinase-STAT pathway, as STAT1alpha tyrosine and serine phosphorylation, dimerization, nuclear translocation, and DNA binding are intact in M. tuberculosis-infected cells. In contrast, there is a marked decrease in IFN-gamma-induced association of STAT1 with the transcriptional coactivators CREB binding protein and p300 in M. tuberculosis-infected macrophages, indicating that M. tuberculosis directly or indirectly disrupts this protein-protein interaction that is essential for transcriptional responses to IFN-gamma. Gamma-irradiated M. tuberculosis and isolated cell walls reproduce the effects of live bacteria, indicating that the bacterial component(s) that initiates inhibition of IFN-gamma responses is constitutively expressed. Although lipoarabinomannan has been found to exert effects on macrophages, it does not account for the inhibitory effects of cell walls. These results indicate that one mechanism for M. tuberculosis to evade the human immune response is to inhibit the IFN-gamma signaling pathway, and that the mechanism of inhibition is distinct from that reported for Leishmania donovani or CMV, in that it targets the interaction of STAT1 with the basal transcriptional apparatus.
Assuntos
Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/metabolismo , Interferon gama/antagonistas & inibidores , Interferon gama/fisiologia , Mycobacterium tuberculosis/imunologia , Transativadores/antagonistas & inibidores , Transativadores/metabolismo , Transcrição Gênica/imunologia , Animais , Anticorpos Monoclonais/farmacologia , Transporte Biológico , Proteína de Ligação a CREB , Núcleo Celular/metabolismo , Células Cultivadas , Humanos , Fator Gênico 3 Estimulado por Interferon , Macrófagos/imunologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Macrófagos/parasitologia , Mycobacterium tuberculosis/crescimento & desenvolvimento , Proteínas Nucleares/metabolismo , Fosforilação , RNA Mensageiro/biossíntese , Receptores de IgG/antagonistas & inibidores , Receptores de IgG/biossíntese , Receptores de IgG/genética , Fator de Transcrição STAT1 , Serina/metabolismo , Transdução de Sinais/imunologia , Frações Subcelulares/imunologia , Frações Subcelulares/microbiologia , Toxoplasma/crescimento & desenvolvimento , Toxoplasma/imunologia , Fatores de Transcrição/antagonistas & inibidores , Fatores de Transcrição/metabolismo , Ativação Transcricional/imunologia , Fator de Crescimento Transformador beta/antagonistas & inibidores , Fator de Crescimento Transformador beta/imunologia , Tirosina/antagonistas & inibidores , Tirosina/metabolismoRESUMO
Human p55 is an abundantly palmitoylated phosphoprotein of the erythroid membrane. It is the prototype of a newly discovered family of membrane-associated proteins termed MAGUKs (membrane-associated guanylate kinase homologues). The MAGUKs interact with the cytoskeleton and regulate cell proliferation, signaling pathways, and intercellular junctions. Here, we report the complete intron-exon map of the human erythroid p55 gene (HGMW-approved symbol MPP1). The structure of the p55 gene was determined from cosmid clones isolated from a cosmid library specific for the human X chromosome. There is a single copy of the p55 gene, composed of 12 exons and spanning approximately 28 kb in the q28 region of the human X chromosome. The exon sizes range from 69 (exon 5) to 203 (exon 10) bp, whereas the intron sizes vary from 280 bp (intron 2) to approximately 14 kb (intron 1). The intron-exon boundaries conform to the donor/acceptor consensus sequence, GT-AG, for splice junctions. Several of the exon boundaries correspond to the boundaries of functional domains in the p55 protein. These domains include a SH3 motif and a region that binds to cytoskeletal protein 4.1. In addition, a comparison of the genomic and the primary structures of p55 reveals a highly conserved phosphotyrosine domain located between the protein 4.1 binding domain and the guanylate kinase domain. Finally, promoter activity measurements of the region immediately upstream of the p55 gene, which contains several cis-elements commonly found in housekeeping genes, suggest that a CpG island may be associated with the p55 gene expression in vivo.
Assuntos
Proteínas Sanguíneas/genética , Proteínas de Membrana/genética , Núcleosídeo-Fosfato Quinase/genética , Cromossomo X/genética , Sequência de Aminoácidos , Sequência de Bases , Mapeamento Cromossômico , Sequência Conservada , DNA/genética , Éxons , Genoma Humano , Guanilato Quinases , Células HeLa , Humanos , Íntrons , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Transcrição GênicaRESUMO
The PDZ domain, also known as the GLGF repeat/DHR domain, is an approximately 90-amino acid motif discovered in a recently identified family of proteins termed MAGUKs (membrane-associated guanylate kinase homologues). Sequence comparison analysis has since identified PDZ domains in over 50 proteins. Like SH2 and SH3 domains, the PDZ domains mediate specific protein-protein interactions, whose specificities appear to be dictated by the primary structure of the PDZ domain as well as its binding target. Using recombinant fusion proteins and a blot overlay assay, we show that a single copy of the PDZ domain in human erythrocyte p55 binds to the carboxyl terminus of the cytoplasmic domain of human erythroid glycophorin C. Deletion mutagenesis of 21 amino acids at the amino terminus of the p55 PDZ domain completely abrogates its binding activity for glycophorin C. Using an alanine scan and surface plasmon resonance technique, we identify residues in the cytoplasmic domain of glycophorin C that are critical for its interaction with the PDZ domain. The recognition specificity of the p55 PDZ domain appears to be unique, since the three PDZ domains of hDlg (human lymphocyte homologue of the Drosophila discs large tumor suppressor) do not bind the cytoplasmic domain of glycophorin C. Taken together with our previous studies, these results complete the identification of interacting domains in the ternary complex between p55, glycophorin C, and protein 4.1. Implications of these findings are discussed in terms of binding specificity and the regulation of cytoskeleton-membrane interactions.
Assuntos
Citoplasma/metabolismo , Glicoforinas/metabolismo , Núcleosídeo-Fosfato Quinase/metabolismo , Sequência de Aminoácidos , Glutationa Transferase/metabolismo , Glicoforinas/química , Guanilato Quinases , Humanos , Dados de Sequência Molecular , Mutagênese , Núcleosídeo-Fosfato Quinase/química , Núcleosídeo-Fosfato Quinase/genética , Ligação Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismoRESUMO
The vertebrate neural cell adhesion molecule NCAM mediates heterophilic adhesion to heparan sulfate proteoglycans in embryonic chick brain membranes. In this study, mouse L cells transfected with chicken NCAM were used to identify two of these ligands as agrin and the target of the 6C4 monoclonal antibody. A third heparan sulfate proteoglycan, perlecan, appeared not to support NCAM-mediated adhesion. Enzymatic degradation of chondroitin sulfates decreased adhesion in agrin-containing membrane fractions but increased adhesion if the agrin had previously been removed by immunoprecipitation, suggesting that interactions between heparan sulfate and chondroitin sulfate proteoglycans have important influences on adhesion. Our experiments support the view that NCAM can interact with multiple, but not with all, heparan sulfate and chondroitin sulfate proteoglycans in chick brain membranes in both positive and negative ways to influence cell adhesion.
Assuntos
Agrina/metabolismo , Adesão Celular/fisiologia , Proteoglicanas de Heparan Sulfato , Moléculas de Adesão de Célula Nervosa/fisiologia , Agrina/isolamento & purificação , Animais , Anticorpos Monoclonais , Química Encefálica , Adesão Celular/efeitos dos fármacos , Extratos Celulares , Membrana Celular/química , Embrião de Galinha , Condroitina Liases , Sulfatos de Condroitina/metabolismo , Heparina/farmacologia , Heparina Liase , Heparitina Sulfato/metabolismo , Heparitina Sulfato/farmacologia , Células L , Ligantes , Camundongos , Moléculas de Adesão de Célula Nervosa/genética , Polissacarídeo-Liases , Proteoglicanas/metabolismo , TransfecçãoRESUMO
LIM domains, found in over 60 proteins, play key roles in the regulation of developmental pathways. They were first identified as cysteine-rich motifs found in the three proteins Lin-11, Isl-1, and Mec-3. LIM proteins frequently contain DNA-binding homeodomains, allowing these proteins to activate transcription. LIM domains also function as protein-binding interfaces, mediating specific protein-protein interactions. Limatin is a novel LIM protein that binds to actin filaments via a domain that is homologous to erythrocyte dematin. Here we report the murine and human chromosomal localizations of limatin (LIMAB1). Limatin was mapped to mouse Chromosome 19 by restriction fragment length polymorphism analysis and to human chromosome region 10q25 by fluorescence in situ hybridization. Radiation hybrid mapping placed LIMAB1 in a 37-cR interval between markers D10S554 and D10S2390. Interestingly, 10q25 is a region of frequent loss of heterozygosity in human tumors, thus identifying limatin as a candidate tumor suppressor gene.
Assuntos
Mapeamento Cromossômico , Cromossomos Humanos Par 10 , Proteínas dos Microfilamentos/genética , Neoplasias/genética , Animais , Humanos , Hibridização in Situ Fluorescente , Proteínas com Domínio LIM , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Polimorfismo de Fragmento de RestriçãoRESUMO
OBJECTIVES: EPB4.1 has been previously mapped to human chromosome 1p33-p34.2. In contradiction to this chromosomal location, we have mapped EPB4.1-1p36 by using fluorescence in situ hybridization and radiation hybrid mapping. In neuroblastomas, deletions of the telomeric end of chromosome 1 (1p36) are the most common genetic aberration. METHODS: We investigated whether genetic aberrations of EPB4.1 can be detected in some neuroblastomas by analyzing 72 tumours for EPB4.1 mutation, expression, and alternative splicing pattern. Furthermore, EPB4.1 protein from a neuroblastoma cell line was studied for its subcellular localization. RESULTS: Sequence changes could be detected in 14 out of 72 neuroblastomas, including missense, silent, and intronic changes. Duplex RT-PCR analysis revealed a subset of 11 tumours expressing significantly low levels of EPB4.1. Significant EPB4.1 sequence changes that were detected included an exon 4 G/A missense mutation (amino acid: V/I) that was shown to be associated with absence of wild-type EPB4.1 expression (3 tumours), an exon 8 G/A missense mutation (V/M) (1 tumour), and an intronic sequence change that was shown to be associated with the presence of an aberrant transcript (1 tumour). Splicing pattern analysis revealed that all EPB4.1 transcripts from tumours exclude exon 3, a splicing pattern for generating the 135 kDa isoform. EPB4.1 cDNA cloned from a neuroblastoma cell line produced a 135-kDa protein with a cytoplasm/membrane localization. CONCLUSIONS: Out of 72 neuroblastomas we have identified 11 tumours with impaired EPB4.1 expression and 5 tumours with significant sequence changes. We also found that the 135 kDa isoform is the main EPB4.1 product in neuroblastoma. EPB4.1 cDNA from a neuroblastoma cell line produced a 135-kDa protein and displayed a cytoplasm/membrane localization in transfected cells.