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OBJECTIVE: Previous studies have used selective recall and descriptive dietary record methods, requiring considerable effort for assessing food and water intake. This study created a simplified lifestyle questionnaire to predict habitual water intake (SQW), accurately and quickly assessing the habitual water intake. We also evaluated the validity using descriptive dietary records as a cross-sectional study. SUBJECTS AND METHODS: First, we used crowdsourcing and machine learning to collect data, predict water intake records, and create questionnaires. We collected 305 lifestyle-related questions as predictor variables and selective recall methods for assessing water intake as an outcome variable. Random forests were used for the machine learning models because of their interpretability and accurate estimation. Random forest and single regression correlation analysis were augmented by the synthetic minority oversampling that trained the model. We separated the data by sex and evaluated our model using unseen hold-out testing data, predicting the individual and overall habitual water intake from various sources, including non-alcoholic beverages, alcohol, and food. RESULTS: We found a 0.60 Spearman's correlation coefficient for total water intake between the predicted and the selective recall method values, reflecting the target value to be achieved. This question set was then used for feasibility tests. The descriptive dietary record method helped to obtain a ground-truth value. We categorized the data by gender, season, and source: non-alcoholic beverages, alcohol, food, and total water intake, and the correlation was confirmed. Consequently, our results showed a Pearson's correlation coefficient of 0.50 for total water intake between the predicted and the selective recall method values. CONCLUSIONS: We hypothesize that dissemination of SQW can lead to better health management by easily determining the habitual water intake.
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Ingestão de Líquidos , Alimentos , Estudos Transversais , Bebidas , Inquéritos e Questionários , Etanol , Reprodutibilidade dos Testes , DietaRESUMO
We present a scheme for time-resolved pump-probe microscopy using intensity modulated laser diodes. The modulation frequencies of the pump and probe beams are varied up to 500 MHz with fixed frequency detuning typically set at 15 kHz. The frequency response of the pump-probe signal is detected using a lock-in amplifier referenced at the beat frequency. This frequency domain method is capable of characterizing the nanosecond to picosecond relaxation dynamics of sample species without the use of a high speed detector or a high frequency lock-in amplifier. Furthermore, as the pump-probe signal is based on the nonlinear interaction between the two laser beams and the sample, our scheme provides better spatial resolution than the conventional diffraction-limited optical microscopes. Time-resolved pump-probe imaging of fluorescence beads and aggregates of quantum dots demonstrates that this method is useful for the microscopic analysis of optoelectronic devices. The system is implemented using compact and low-cost laser diodes, and thus has a broad range of applications in the fields of photochemistry, optical physics, and biological imaging.
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Lasers Semicondutores , Microscopia/instrumentação , Razão Sinal-Ruído , Fatores de TempoRESUMO
Development of efficient ion supply of (58)Fe from (58)Fe(C5H5)2, and quick switching between therapy and material science at the Heavy-Ion Medical Accelerator in Chiba realized a new (57)Mn in-beam emission Mössbauer spectroscopy measurement system. Application to simple binary chemical compounds, MgO and NaF, proved the usefulness of the system to probe chemical and physical behaviors of trace impurities in solids. Annealing of lattice defects produced by the implantation and ß-decay of (57)Mn and/or γ-ray emission recoil was observed by a local probe.
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Íons Pesados , Canais Iônicos/química , Óxido de Magnésio/química , Manganês/química , Aceleradores de Partículas , Fluoreto de Sódio/química , Espectroscopia de MossbauerRESUMO
OBJECTIVE: A common haplotype M2 consisting of minor SNP alleles located in the ANXA5 gene promoter region has been described as a risk factor for various obstetric complications such as recurrent pregnancy loss, pre-eclampsia and pregnancy-related thrombophilic disorder. However, the question of whether it is the maternal or fetal genotype that contributes to the onset of these disorders remains to be resolved. METHODS: We analyzed ANXA5 gene variants in the blood and placental tissues from pre-eclampsia patients and normotensive controls. ANXA5 expression was examined by qRT-PCR, Western blotting and immunostaining. Results were compared between M2 and non-M2 carriers. RESULTS: The M2 haplotype was found to be significantly frequent in placentas from pre-eclamptic patients relative to the controls (25.5% versus 10%, P = 0.044), In contrast, no significant differences were observed in maternal blood (13.0% versus 11.3%, P = 0.597). The placental expression of ANXA5 mRNA was found to be lower in M2 carriers. When examined by Western blot and immunostaining, the ANXA5 protein levels were found to be affected more by the placental than the maternal genotype. Histological examination of the placentas from the pre-eclamptic patients demonstrated that a placental M2 haplotype correlated more closely than maternal M2 with the severity of perivillous fibrin deposition. CONCLUSIONS: Although preliminary, these results suggest that hypomorphic M2 alleles in the in placental ANXA5 promoter, whether transmitted maternally or paternally, might be an essential determinant of an increased risk of pre-eclampsia via local thrombophilia at the feto-maternal interface.
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Anexina A5/genética , Placenta/metabolismo , Polimorfismo Genético , Pré-Eclâmpsia/genética , Regiões Promotoras Genéticas , Adulto , Alelos , Anexina A5/metabolismo , Estudos de Casos e Controles , Cesárea , Vilosidades Coriônicas/química , Vilosidades Coriônicas/metabolismo , Vilosidades Coriônicas/patologia , Regulação para Baixo , Feminino , Feto/metabolismo , Fibrina/metabolismo , Estudos de Associação Genética , Heterozigoto , Humanos , Japão/epidemiologia , Placenta/patologia , Pré-Eclâmpsia/metabolismo , Pré-Eclâmpsia/patologia , Pré-Eclâmpsia/fisiopatologia , Gravidez , Risco , Índice de Gravidade de Doença , Propriedades de SuperfícieRESUMO
We found that Ba(2)Ti(13)O(22) with Ti(3+) (3d(1)) ions on a triangle-based lattice exhibits a phase transition at T(c)~200 K, below which the increase of electrical resistivity and decrease of magnetic susceptibility were observed. Transmission electron microscopy and optical reflectivity measurements indicate that the low-temperature phase of the present compound shares characteristics in common with a charge-density-wave state with remnant carriers, although a commensurate wave vector of the modulation and a linear temperature dependence of the magnetic susceptibility below T(c) suggest an exotic ordered state.
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Genes that enable crops to limit Na(+) accumulation in shoot tissues represent potential sources of salinity tolerance for breeding. In barley, the HvNax4 locus lowered shoot Na(+) content by between 12% and 59% (g(-1) DW), or not at all, depending on the growth conditions in hydroponics and a range of soil types, indicating a strong influence of environment on expression. HvNax4 was fine-mapped on the long arm of barley chromosome 1H. Corresponding intervals of â¼200 kb, containing a total of 34 predicted genes, were defined in the sequenced rice and Brachypodium genomes. HvCBL4, a close barley homologue of the SOS3 salinity tolerance gene of Arabidopsis, co-segregated with HvNax4. No difference in HvCBL4 mRNA expression was detected between the mapping parents. However, genomic and cDNA sequences of the HvCBL4 alleles were obtained, revealing a single Ala111Thr amino acid substitution difference in the encoded proteins. The known crystal structure of SOS3 was used as a template to obtain molecular models of the barley proteins, resulting in structures very similar to that of SOS3. The position in SOS3 corresponding to the barley substitution does not participate directly in Ca(2+) binding, post-translational modifications or interaction with the SOS2 signalling partner. However, Thr111 but not Ala111 forms a predicted hydrogen bond with a neighbouring α-helix, which has potential implications for the overall structure and function of the barley protein. HvCBL4 therefore represents a candidate for HvNax4 that warrants further investigation.
Assuntos
Hordeum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Locos de Características Quantitativas , Sódio/metabolismo , Sequência de Aminoácidos , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Transporte Biológico , Mapeamento Cromossômico , Hordeum/química , Hordeum/genética , Conformação Molecular , Dados de Sequência Molecular , Proteínas de Plantas/química , Alinhamento de SequênciaRESUMO
We found that in A2V13O22 (A=Ba, Sr), which contains a trilayer slab of VO in the sodium-chloride structure with periodically missing ions, the trimerization of V ions occurs at 290 K (A=Ba) and 380 K (A=Sr). V trimers form a three-dimensional network, but some V ions remain untrimerized in these compounds. The suppression of magnetic susceptibility with trimerization and the existence of a Curie tail at low temperatures, together with the result of NMR measurement, indicate that the V trimers are spin singlet, whereas the untrimerized V ions have a magnetic moment; i.e., there is a spontaneous separation between nonmagnetic and magnetic ions in the crystal.
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AIMS/BACKGROUND: To investigate how systemic overexpression of IL-4, IL-5 and IFN-gamma affects the severity of experimental conjunctivitis (EC) in mice. METHODS: The tibialis anterior muscle of naive BALB/c mice was electroporated with IL-4, IL-5, IFN-gamma or a control gene, and then the mice blood and conjunctivas were harvested to measure the eosinophil content in these tissues. To evaluate the effects of cytokine gene electroporation on the early-phase reaction (EPR), cytokine gene-electroporated ragweed (RW)-immunised mice were intravenously injected with Evans Blue (EB) and then challenged with RW in eye-drops. Thirty minutes later, their conjunctivas were harvested to extract EB and evaluate the EPR. Additionally, 24 h after RW challenge, conjunctivas were harvested from cytokine gene-electroporated RW-immunised mice, which had not received intravenous injection of EB, to measure conjunctival eosinophilia. RESULTS: Significantly more eosinophils were detected in the blood and conjunctivas of IL-5-electroporated mice in which EC was not induced. The intensity of the EPR was significantly greater in IFN-gamma-electroporated mice. Significantly greater eosinophil infiltration was seen in the conjunctivas of IL-5-electroporated EC-developing mice. CONCLUSIONS: It appears that systemic IL-5 and IFN-gamma play different roles in the development of EC.
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Conjuntivite Alérgica/imunologia , Eosinofilia/imunologia , Interferon gama/imunologia , Interleucina-4/imunologia , Ambrosia/imunologia , Animais , Túnica Conjuntiva/imunologia , Túnica Conjuntiva/patologia , Conjuntivite Alérgica/patologia , Modelos Animais de Doenças , Eletroporação/métodos , Eosinofilia/patologia , Feminino , Interferon gama/biossíntese , Interferon gama/genética , Interleucina-4/biossíntese , Interleucina-4/genética , Interleucina-5/biossíntese , Interleucina-5/genética , Interleucina-5/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodosRESUMO
Rye is a diploid crop species with many outstanding qualities, and is important as a source of new traits for wheat and triticale improvement. Rye is highly tolerant of aluminum (Al) toxicity, and possesses a complex structure at the Alt4 Al tolerance locus not found at the corresponding locus in wheat. Here we describe a BAC library of rye cv. Blanco, representing a valuable resource for rye molecular genetic studies, and assess the library's suitability for investigating Al tolerance genes. The library provides 6 x genome coverage of the 8.1 Gb rye genome, has an average insert size of 131 kb, and contains only ~2% of empty or organelle-derived clones. Genetic analysis attributed the Al tolerance of Blanco to the Alt4 locus on the short arm of chromosome 7R, and revealed the presence of multiple allelic variants (haplotypes) of the Alt4 locus in the BAC library. BAC clones containing ALMT1 gene clusters from several Alt4 haplotypes were identified, and will provide useful starting points for exploring the basis for the structural variability and functional specialization of ALMT1 genes at this locus.
Assuntos
Adaptação Fisiológica/genética , Alumínio/farmacologia , Cromossomos Artificiais Bacterianos/genética , Genes de Plantas , Biblioteca Genômica , Mapeamento Físico do Cromossomo/métodos , Secale/genética , Adaptação Fisiológica/efeitos dos fármacos , Southern Blotting , Cromossomos de Plantas/genética , Mapeamento de Sequências Contíguas , Sondas de DNA/metabolismo , DNA de Plantas/genética , Marcadores Genéticos , Haplótipos , Família Multigênica , Secale/efeitos dos fármacosRESUMO
To investigate whether insulin-producing cells obtained from ES cells via the nestin-positive cell-mediated method are of the pancreatic lineage, we established a pdx-1 knockout ES cell line and analyzed its differentiation into insulin-producing cells. As a result, pdx-1 knockout ES cell expressed insulin 2 gene at the final differentiated cells. Thus, our study demonstrated that pdx-1 is not essential for insulin gene expression, at least in cells differentiated from this population of nestin-expression enriched ES cells, and suggested that the insulin-producing cells derived from ES cells may be different from the pancreatic beta cells in terms of their lineage.
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Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/fisiologia , Proteínas de Homeodomínio/genética , Insulina/metabolismo , Transativadores/genética , Animais , Diferenciação Celular , Éxons , Genoma , Insulina/genética , Secreção de Insulina , Proteínas de Filamentos Intermediários/genética , Camundongos , Camundongos Knockout , Proteínas do Tecido Nervoso/genética , Nestina , Mapeamento por Restrição , Transativadores/deficiênciaRESUMO
AIMS/HYPOTHESIS: Duct cells of the pancreas are thought to include latent progenitors of islet endocrine cells that can be induced to differentiate by appropriate morphogens. Here we developed a method for isolating pancreatic ductal epithelial cells from adult mice that overcomes the shortcomings of previous methods. MATERIALS AND METHODS: Pancreatic ductal cells were grown in serum-free DMEM/F12 medium in the presence of cholera toxin or 8-bromo-cyclic adenosine monophosphate, which is known to be an intracellular cAMP generator. Single cell cloning was performed by limiting dilution in serum-free medium. RESULTS: The isolated clonal cells expressed high levels of cytokeratin and Ipf1 (formerly known as Pdx-1). Adenovirus-mediated expression of ngn3 (also known as Neurog3) and Ptf1a in these cells induced expression of insulin and somatostatin, and of carboxypeptidase A, respectively. Furthermore, albumin production was induced by dexamethasone or by long-term culture in serum-containing medium. CONCLUSIONS/INTERPRETATION: Stimulation of the cAMP-dependent signalling allowed us to isolate clonal pancreatic ductal cells from adult mice. These cells are able to partially differentiate into endocrine cells, exocrine cells and hepatocyte-like cells and are therefore considered to have the characteristics of endodermal progenitor cells.
Assuntos
AMP Cíclico/fisiologia , Ductos Pancreáticos/fisiologia , Transdução de Sinais/fisiologia , Animais , Técnicas de Cultura de Células , Linhagem Celular , Meios de Cultura Livres de Soro , Primers do DNA , Desenvolvimento Embrionário , Células Epiteliais/citologia , Células Epiteliais/fisiologia , Regulação da Expressão Gênica , Glucose-6-Fosfatase/genética , Camundongos , Camundongos Endogâmicos C57BL , Ductos Pancreáticos/citologia , Ductos Pancreáticos/embriologia , Ductos Pancreáticos/ultraestrutura , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: Idiopathic pulmonary fibrosis is a devastating disorder for which there is no effective treatment. Transforming growth factor (TGF)-beta plays a critical role in provoking fibrosis. Interleukin (IL)-10 is a potent immunosuppressive cytokine but its effect on the fibrosing process is unclear. A study was undertaken to examine whether IL-10 affects the production and activation of TGF-beta and thus can attenuate the fibrosis. METHODS: Mice were given an intratracheal injection of bleomycin. On day 1 or 14, IL-10 gene was delivered by rapid intravenous injection of Ringer's solution containing plasmid. Two weeks after the plasmid injection the mice were examined for fibrosis. The effect of IL-10 on TGF-beta production by alveolar macrophages was assessed. RESULTS: Even when delivered during the fibrosing phase, IL-10 gene significantly suppressed the pathological findings, hydroxyproline content, and production of both active and total forms of TGF-beta1 in the lung. Immunohistochemical analyses showed that alveolar macrophages were one of the major sources of TGF-beta1 and IL-10 diminished the intensity of the staining. IL-10 also suppressed the expression of alphaV beta6 integrin, a molecule that plays an important role in TGF-beta activation, on lung epithelial cells. Alveolar macrophages from bleomycin injected mice produced TGF-beta1 spontaneously ex vivo, which was significantly suppressed by treatment of the mice in vivo or by treatment of the explanted macrophages ex vivo with IL-10. CONCLUSION: IL-10 suppresses the production and activation of TGF-beta in the lung and thus attenuates pulmonary fibrosis, even when delivered in the chronic phase.
Assuntos
Bleomicina/toxicidade , Interleucina-10/administração & dosagem , Fibrose Pulmonar/terapia , Fator de Crescimento Transformador beta/antagonistas & inibidores , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Linhagem Celular , Técnicas de Transferência de Genes , Terapia Genética/métodos , Imuno-Histoquímica , Integrina alfa6/farmacologia , Interleucina-10/genética , Macrófagos Alveolares/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , PlasmídeosRESUMO
A coupling function that describes the interaction between self-sustained oscillators in a phase equation is derived and applied experimentally to Belousov-Zhabotinsky (BZ) oscillators. It is demonstrated that the synchronous behavior of coupled BZ reactors is explained extremely well in terms of the coupling function thus obtained. This method does not require comprehensive knowledge of either the oscillation mechanism or the interaction among the oscillators, both of these being often difficult to elucidate in an actual system. These facts enable us to accurately analyze the weakly coupled entrainment phenomenon through the direct measurement of the coupling function.
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BACKGROUND AND AIMS: 1.4-Dihydroxy-2-naphthoic acid (DHNA), a bifidogenic growth stimulator from Propionibacterium freudenreichii, is thought to have a beneficial effect as a prebiotic; however, its in vivo effect on intestinal inflammation remains unknown. The aim of this study was to determine whether oral administration of DHNA can ameliorate dextran sodium sulphate (DSS) induced colitis and to determine the possible underlying mechanisms. METHOD: Colitis was induced in mice by treatment with 2.0% DSS for seven days. DHNA (0.6 or 2.0 mg/kg) was given in drinking water prior to (preventive study) or after (therapeutic study) DSS administration. Colonic damage was histologically scored, and mucosal addressin cell adhesion molecule 1 (MAdCAM-1) expression and beta7 positive cell infiltration were determined by immunohistochemistry. mRNA levels of proinflammatory cytokines (interleukin (IL)-1beta, IL-6 and tumour necrosis factor alpha (TNF-alpha)) were determined by quantitative real time polymerase chain reaction. In addition, bacterial flora in the caecum, concentrations of short chain acids, and luminal pH were examined. RESULTS: DHNA improved survival rate and histological damage score in mice administered DSS in both the preventive and therapeutic studies. DHNA significantly attenuated the enhanced expression of MAdCAM-1, the increased beta7 positive cell number, and the increased mRNA levels of IL-1beta, IL-6, and TNF-alpha in DSS treated colon. In addition, the decreased number of Lactobacillus and Enterobacteriaceae induced by DSS was recovered by DHNA. Preventive effects on decrease in butyrate concentration and decrease in pH level in mice administered DSS were also observed in the DHNA preventive study. CONCLUSION: DHNA, a novel type of prebiotic, attenuates colonic inflammation not only by balancing intestinal bacterial flora but also by suppressing lymphocyte infiltration through reduction of MAdCAM-1.
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Proteínas de Bactérias/uso terapêutico , Colite/terapia , Colo , Mucosa Intestinal/metabolismo , Naftóis/uso terapêutico , Propionibacterium/fisiologia , Animais , Proteínas de Bactérias/farmacologia , Moléculas de Adesão Celular/análise , Colite/metabolismo , Colite/prevenção & controle , Colo/microbiologia , Citocinas/genética , Citocinas/metabolismo , Sulfato de Dextrana , Ácidos Graxos Voláteis/análise , Fezes/química , Expressão Gênica/efeitos dos fármacos , Imuno-Histoquímica/métodos , Cadeias beta de Integrinas/análise , Mucosa Intestinal/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Mucoproteínas , Naftóis/farmacologia , Receptores de Retorno de Linfócitos/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Organismos Livres de Patógenos Específicos , Taxa de SobrevidaRESUMO
The efficacy of sparfloxacin (SPFX) for the control of bronchial asthma was evaluated in 26 patients with suspected Chlamydia pneumoniae infection. Patients were randomly allocated to receive SPFX 200 mg/day (n = 14) or control treatment (n = 12) for 21 days. Significant improvements in serum C-reactive protein levels, and significant decreases in peripheral eosinophil counts, serum eosinophil cationic protein (ECP) and sputum ECP were observed in the SPFX-treated group at day 21. SPFX-treated patients also had a significantly reduced frequency of asthma symptoms, reduced inhalant beta2-stimulant use, and significant increases in morning peak expiratory flow. At the end of the study, C. pneumoniae was undetectable in two SPFX-treated patients who underwent polymerase chain reaction testing, but one control patient who was tested still had detectable levels of C. pneumoniae. These results suggest that SPFX could be used to control bronchial asthma in patients with suspected persistent C. pneumoniae infection.
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Asma/tratamento farmacológico , Infecções por Chlamydophila/tratamento farmacológico , Chlamydophila pneumoniae/patogenicidade , Fluoroquinolonas/uso terapêutico , Administração por Inalação , Idoso , Antiasmáticos/administração & dosagem , Antiasmáticos/uso terapêutico , Anticorpos Antibacterianos/sangue , Asma/complicações , Proteína C-Reativa/efeitos dos fármacos , Proteína C-Reativa/metabolismo , Infecções por Chlamydophila/etiologia , Infecções por Chlamydophila/microbiologia , Chlamydophila pneumoniae/imunologia , Chlamydophila pneumoniae/isolamento & purificação , Proteína Catiônica de Eosinófilo/sangue , Proteína Catiônica de Eosinófilo/efeitos dos fármacos , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Pico do Fluxo ExpiratórioRESUMO
Cytosolic phospholipase A(2)alpha (cPLA(2)alpha) plays an important role in arachidonate pathway. To investigate the contribution of cPLA(2)alpha to autoimmune diabetes, we established non-obese diabetic (NOD) mouse, an excellent model for human type 1 diabetes, deficient in cPLA(2)alpha. These mice showed severe insulitis and a higher incidence of diabetes. In their macrophages, decreased prostaglandin E(2) (PGE(2)) induced by cPLA(2)alpha deficiency, and the increase in production of tumor necrosis factor (TNF)-alpha were observed. These results suggested that cPLA(2)alpha plays a protective role in progression of insulitis and development of autoimmune diabetes by suppression of TNF-alpha production from macrophages.
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Citosol/enzimologia , Diabetes Mellitus Tipo 1/enzimologia , Fosfolipases A/fisiologia , Animais , Ácido Araquidônico/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Genótipo , Fosfolipases A2 do Grupo IV , Humanos , Insulina/metabolismo , Macrófagos/metabolismo , Macrófagos Peritoneais/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Camundongos Knockout , Repetições de Microssatélites , Fosfolipases A/química , Reação em Cadeia da Polimerase , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Free fatty acids (FFA) have generally been proposed to regulate pancreatic insulin release by an intracellular mechanism involving inhibition of CPT-1. The recently de-orphanized G-protein coupled receptor, FFA(1)R/GPR40, has been shown to be essential for fatty-acid-stimulated insulin release in MIN6 mouse insulinoma cells. The CPT-1 inhibitor, 2-bromo palmitate (2BrP), was investigated for its ability to interact with mouse FFA(1)R/GPR40. It was found to inhibit phosphatidyl inositol hydrolysis induced by linoleic acid (LA) (100 muM in all experiments) in HEK293 cells transfected with FFA(1)R/GPR40 and in the MIN6 subclone, MIN6c4. 2BrP also inhibited LA-stimulated insulin release from mouse pancreatic islets. Mouse islets were subjected to antisense intervention by treatment with a FFA(1)R/GPR40-specific morpholino oligonucleotide for 48 h. Antisense treatment of islets suppressed LA-stimulated insulin release by 50% and by almost 100% when islets were pretreated with LA for 30 min before applying the antisense. Antisense treatment had no effect on tolbutamide-stimulated insulin release. Confocal microscopy using an FFA(1)R/GPR40-specific antibody revealed receptor expression largely localized to the plasma membrane of insulin-producing cells. Pretreating the islets with LA for 30 min followed by antisense oligonucleotide treatment for 48 h reduced the FFA(1)R/GPR40 immunoreactivity to background levels. The results demonstrate that FFA(1)R/GPR40 is inhibited by the CPT-1 inhibitor, 2BrP, and confirm that FFA(1)R/GPR40 is indeed necessary, at least in part, for fatty-acid-stimulated insulin release.
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Ácidos Graxos/metabolismo , Insulina/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Animais , Linhagem Celular , Feminino , Humanos , Hipoglicemiantes/metabolismo , Secreção de Insulina , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/metabolismo , Ácido Linoleico/metabolismo , Camundongos , Oligonucleotídeos Antissenso/metabolismo , Palmitatos/metabolismo , Fosfatidilinositóis/metabolismo , Receptores Acoplados a Proteínas G/genéticaRESUMO
The aetiology of Crohn's disease (CD) remains unknown. Since SAMP1/Yit mice have been reported to develop CD-like spontaneous enteric inflammation, such mice have been studied as an animal model of CD. In this study, using this model we examined T lymphocyte migration in microvessels of intestinal mucosa in vivo and the expression of adhesion molecules by immunohistochemistry. Fluorescence-labelled T lymphocytes isolated from AKR/J (control) mice were injected into the tail veins of recipient mice, and T lymphocyte migration in the postcapillary venules of Peyer's patches, submucosal microvessels, and villus capillaries of the terminal ileum was monitored using an intravital microscope. Adhesion of T lymphocytes was significantly increased in 35 week old SAMP1/Yit mice compared with that in AKR/J or 15 week old SAMP1/Yit mice. Immunohistochemical study showed increased infiltration of CD4, CD8 and beta7-integrin-positive cells and increased expression of MAdCAM-1 and VCAM-1 in the terminal ileum of SAMP1/Yit mice. Antibodies against MAdCAM-1 and VCAM-1 significantly inhibited adhesion of T lymphocytes to microvessels of the terminal ileum, and anti-MAdCAM-1 antibody showed stronger suppressive effect than the anti-VCAM-1 antibody. Periodical administration of anti-MAdCAM-1 antibody twice a week for 7 weeks significantly ameliorated ileitis of SAMP1/Yit mice, but submucosal hypertrophy was not significantly suppressed. Anti-VCAM-1 antibody treatment failed to show significant resolution of ileitis. In addition, anti-MAdCAM-1 antibody treatment also attenuated established ileitis. The results demonstrate that, although MAdCAM-1 and VCAM-1 play an important role in T lymphocyte-endothelial cell interactions in SAMP1/Yit mice, MAdCAM-1 may be a more appropriate target for therapeutic modulation of chronic ileitis.
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Ileíte/imunologia , Imunoglobulinas/imunologia , Mucoproteínas/imunologia , Linfócitos T/imunologia , Animais , Anticorpos/administração & dosagem , Anticorpos/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Adesão Celular/imunologia , Moléculas de Adesão Celular , Movimento Celular/imunologia , Células Cultivadas , Imuno-Histoquímica/métodos , Cadeias beta de Integrinas/imunologia , Mucosa Intestinal/imunologia , Camundongos , Camundongos Endogâmicos , Molécula 1 de Adesão de Célula Vascular/imunologiaRESUMO
In Japan the incidence of atypical mycobacteriosis has steadily increased, with Mycobacterium avium-intracellulare complex (MAC) the most common infecting organism. A standard chemotherapy regimen for MAC infection has not been established because of significant resistance to anti-mycobacterial drugs. Sparfloxacin has good antimicrobial activity against several acid-fast bacteria and is expected to be an effective drug for treating mycobacteriosis. We examined the effects of adding sparfloxacin to anti-tuberculotic combination therapy in six patients with MAC pulmonary disease. Drug susceptibility was also assessed using the agar dilution method. The minimum inhibitory concentrations (MICs) for sparfloxacin, levofloxacin, isoniazid, rifampicin, streptomycin, ethambutol and clarithromycin was measured in clinical isolates from all patients; sparfloxacin showed the lowest MIC. Bacteriological and clinical improvements were observed in the four patients who completed the study. Dosing was discontinued in two patients because of pruritic skin eruptions. Sparfloxacin shows promise as an anti-mycobacterial agent for treating MAC pulmonary disease.