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1.
BMJ Open ; 10(2): e029735, 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-32102803

RESUMO

INTRODUCTION: While major depression causes substantial distress and impairment for affected individuals and society, the effectiveness of cognitive behavioural therapy (CBT) in treating the condition has been established. However, the therapeutic mechanism underlying the efficacy of CBT remains unknown. This study aimed to describe a protocol for a randomised controlled trial that will measure the CBT-induced clinical and neural changes in patients with non-psychotic major depression. METHODS AND ANALYSIS: The current study is a 16-week assessor-blinded, randomised, parallel-group trial with a 12-month follow-up as part of usual depression care at an outpatient clinic. Patients aged 20-69 years with major depressive disorder will be randomly assigned to receive either CBT in addition to their usual treatment or talking control in addition to their usual treatment for 16 weeks. The primary outcome is the functional changes in the brain areas that have been associated with future-oriented thinking at 16 weeks; secondary outcomes include changes in functional brain connectivity, severity and changes in the scores of objective and subjective clinical depression symptoms, proportion of responders and remitters and quality of life. The intention-to-treat analysis will be used. ETHICS AND DISSEMINATION: All protocols and the informed consent form are compliant with the Ethics Guideline for Clinical Research (Japanese Ministry of Health, Labour and Welfare). Ethical Review Committees at the Keio University School of Medicine have approved the study protocol (version 3, 11 September 2017). We will disseminate research findings to scientific and general audiences through national and international conference presentations as well as lay summaries to the general public, including mental health consumer and publications in international peer-reviewed psychiatry and brain imaging journals. TRIAL REGISTRATION NUMBER: UMIN Clinical Trials Registry (UMIN000018155); Pre-results.

2.
Medicine (Baltimore) ; 99(1): e18687, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31895836

RESUMO

The impact of time of day or day of week on the survival of emergency trauma patients is still controversial. The purpose of this study was to evaluate the outcomes of these patients according to time of day or day of week of emergency admission by using data from the nationwide Japan Trauma Data Bank (JTDB).This study enrolled 236,698 patients registered in the JTDB database from 2004 to 2015, and defined daytime as 09:00 AM to 16:59 PM and nighttime as 17:00 PM to 08:59 AM, weekdays as Monday to Friday, and weekends as Saturday, Sunday, and national holidays. The outcome measures were death in the emergency room (ER) and discharge to death.In total, 170,622 patients were eligible for our analysis. In a multivariable logistic regression adjusted for confounding factors, both death in the ER and death at hospital discharge were significantly lower during the daytime than at nighttime (623/76,162 [0.82%] vs 954/94,460 [1.01%]; adjusted odds ratio [AOR] 0.79; 95% confidence interval [CI] 0.71-0.88 and 5765/76,162 [7.57%] vs 7270/94,460 [7.70%]; AOR 0.88; 95% CI 0.85-0.92). In contrast, the weekdays/weekends was not significantly related to either death in the ER (1058/114,357 [0.93%] vs 519/56,265 [0.92%]; AOR 0.95; 95% CI 0.85-1.06) or death at hospital discharge (8975/114,357 [7.85%] vs 4060/56,265 [7.22%]; AOR 1.02; 95% CI 0.97-1.06).In this population of emergency trauma patients in Japan, both death in the ER and death at hospital discharge were significantly lower during the daytime than at night, but the weekdays/weekends was not associated with outcomes of these patients.


Assuntos
Serviços Médicos de Emergência/estatística & dados numéricos , Mortalidade Hospitalar/tendências , Adulto , Idoso , Serviços Médicos de Emergência/normas , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
3.
Artigo em Inglês | MEDLINE | ID: mdl-31478074

RESUMO

PURPOSE: Posttraumatic meningitis is one of the severe complications that can result in increased mortality and longer hospital stay among trauma patients. Factors such as cerebrospinal fluid (CSF) fistula and basilar skull fracture are associated with posttraumatic meningitis. However, it remains unclear whether procedures such as burr hole surgery in the emergency department and decompressive craniectomy are associated with posttraumatic meningitis. The aim of this study was to assess factors associated with posttraumatic meningitis with a nationwide hospital-based trauma registry in Japan. METHODS: This was a retrospective observational study with a 12-year study period from January 2004 to December 2015. We included trauma patients registered in the Japanese Trauma Data Bank, whose head Abbreviated Injury Scale score was ≥ 3 in this study. The main endpoint was the occurrence of meningitis during hospitalization. Multivariable logistic regression analysis was used to assess independent parameters associated with posttraumatic meningitis such as CSF fistula, burr hole surgery in the emergency department, and decompressive craniectomy. RESULTS: Among 60,390 head injury patients with head AIS score 3 or more, 284 (0.5%) patients had posttraumatic meningitis. Factors associated with posttraumatic meningitis were burr hole surgery in the emergency department (adjusted odds ratio [AOR] 2.158 [95% confidence interval (CI) 1.401-3.325]), decompressive craniectomy (AOR 2.123 [95% CI 1.506-2.993]), external ventricular drainage (AOR 1.843 [95% CI, 1.157-2.935]), CSF leakage (AOR 3.328 [95% CI 2.205-5.022]), and basilar skull fracture (AOR 1.651 [95% CI 1.178-2.314]). CONCLUSIONS: In this population of trauma patients, burr hole surgery in the emergency department and decompressive craniectomy was associated with posttraumatic meningitis.

4.
Artigo em Inglês | MEDLINE | ID: mdl-31119320

RESUMO

PURPOSE: The aim of this study was to evaluate the association between the implementation of pelvic angiography (PA) and outcome in emergency pediatric patients with pelvic fracture. METHODS: We extracted data on pelvic fracture patients aged ≤ 19 years between 2004 and 2015 from a nationwide trauma registry in Japan. The main outcome was hospital mortality. We assessed the relationship between implementation of PA and hospital mortality using one-to-one propensity-score-matching analysis to reduce potential confounding effects in comparing the PA group with the non-PA group. RESULTS: In total, 1351 patients were eligible for our analysis, with 221 patients (16.4%) included in the PA group and 1130 patients (83.6%) included in the non-PA group. For all patients, the proportion of hospital mortality was higher in the PA group than in the non-PA group [13.6% (30/221) vs 7.1% (80/1130), crude odds ratio (OR) 2.062 (95% confidence interval (CI), 1.318-3.224); p = 0.002]. In the propensity-score-matched patients, the proportion of hospital mortality was lower in the PA group than in the non-PA group [10.5% (22/200) vs 18.2% (38/200), p = 0.027]. This finding was confirmed in both the multivariable logistic regression model [adjusted OR 0.392 (95% CI, 0.171-0.896); p = 0.026] and the conditional logistic regression model [conditional OR 0.484 (95% CI, 0.261-0.896); p = 0.021]. CONCLUSION: The implementation of PA was significantly associated with lower hospital mortality among emergency pediatric patients with pelvic fractures compared with the non-implementation of PA.

5.
Endocr J ; 66(5): 459-468, 2019 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-30842364

RESUMO

Pancreatic polypeptide (PP) is a 36-amino acid peptide encoded by the Ppy gene, which is produced by a small population of cells located in the periphery of the islets of Langerhans. Owing to the high amino acid sequence similarity among neuropeptide Y family members, antibodies against PP that are currently available are not convincingly specific to PP. Here we report the development of mouse monoclonal antibodies that specifically bind to PP. We generated Ppy knockout (Ppy-KO) mice in which the Ppy-coding region was replaced by Cre recombinase. The Ppy-KO mice were immunized with mouse PP peptide, and stable hybridoma cell lines producing anti-PP antibodies were isolated. Firstly, positive clones were selected in an enzyme-linked immunosorbent assay for reactivity with PP coupled to bovine serum albumin. During the screening, hybridoma clones producing antibodies that cross-react to the peptide YY (PYY) were excluded. In the second screening, hybridoma clones in which their culture media produce no signal in Ppy-KO islets but detect specific cells in the peripheral region of wild-type islets, were selected. Further studies demonstrated that the selected monoclonal antibody (23-2D3) specifically recognizes PP-producing cells, not only in mouse, but also in human and rat islets. The monoclonal antibodies with high binding specificity for PP developed in this study will be fundamental for future studies towards elucidating the expression profiles and the physiological roles of PP.


Assuntos
Anticorpos Monoclonais , Ilhotas Pancreáticas/imunologia , Polipeptídeo Pancreático/imunologia , Animais , Camundongos , Camundongos Knockout , Neuropeptídeo Y/imunologia , Peptídeo YY/imunologia
6.
BMJ Open ; 9(1): e025350, 2019 01 29.
Artigo em Inglês | MEDLINE | ID: mdl-30700488

RESUMO

OBJECTIVE: Although it is important to assess prehospital factors associated with traffic crash fatalities to decrease them as a matter of public health, such factors have not been fully revealed. METHODS: Using data from the Japanese Trauma Data Bank, a large hospital-based trauma registry in Japan, we retrospectively analysed traffic crash patients transported to participating facilities that treated patients with severe trauma from 2004 to 2015. This study defined registered emergency patients whose systolic blood pressure was 0 mm Hg or heart rate was 0 bpm at hospital arrival as being in prehospital cardiopulmonary arrest (CPA). Prehospital factors associated with prehospital CPA due to traffic crash were assessed with multivariable logistic regression analysis. RESULTS: In total, 66 243 patients were eligible for analysis. Of them, 3390 (5.1%) patients were in CPA at hospital arrival. A multivariable logistic regression model showed the following factors to be significantly associated with prehospital CPA: ages 60-74 years (adjusted OR (AOR) 1.256, 95% CI 1.142 to 1.382) and ≥75 years (AOR 1.487, 95% CI 1.336 to 1.654), male sex (AOR 1.234, 95% CI 1.139 to 1.338), night-time (AOR 1.575, 95% CI 1.458 to 1.702), weekend including holiday (AOR 1.078, 95% CI 1.001 to 1.161), rural area (AOR 1.181, 95% CI 1.097 to 1.271), back seat passenger (AOR 1.227, 95% CI 0.985 to 1.528) and pedestrian (AOR 1.754, 95% CI 1.580 to 1.947) as types of patients. CONCLUSION: In this population, factors associated with prehospital CPA due to a traffic crash were elderly people, male sex, night-time, weekend/holiday, back seat passenger, pedestrian and rural area. These fundamental data may be of help in reducing and preventing traffic crash deaths.


Assuntos
Acidentes de Trânsito/mortalidade , Parada Cardíaca Extra-Hospitalar/mortalidade , Adolescente , Adulto , Distribuição por Idade , Idoso , Criança , Pré-Escolar , Bases de Dados Factuais , Serviço Hospitalar de Emergência/estatística & dados numéricos , Feminino , Humanos , Lactente , Recém-Nascido , Japão/epidemiologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Estudos Retrospectivos , Fatores de Risco , Distribuição por Sexo , Adulto Jovem
7.
Reprod Med Biol ; 17(4): 466-473, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30377401

RESUMO

Purpose: The purpose of this study was to examine whether preconception maternal dietary pattern is associated with in vitro fertilization (IVF) outcome among Japanese women. Methods: This prospective study included 140 Japanese women who underwent conventional-IVF/intracytoplasmic sperm injection. The patients' diets during the previous month before egg retrieval were assessed with validated brief-type self-administered diet history questionnaire. Dietary patterns from 33 predefined food groups [energy-adjusted food (g/1000 kcal)] were extracted by factor analysis. The primary outcome measure was clinical pregnancy rate after IVF. Results: Thirty-six women had confirmed clinical pregnancy. Three dietary patterns were identified: "Vegetable and seafood," "Western," and "Rice and miso soup." The "Vegetables and seafood" dietary pattern (high intakes of green and other vegetables, mushrooms, seasoning, fish, soy products, chicken, and potatoes) was not associated with clinical pregnancy ([odds ratio per one-quartile increase in dietary pattern: 0.94 (95% confidence interval: 0.67-1.32), P = 0.73]. This relationship was unaltered after controlling for potential confounders. Furthermore, no association was seen between the other two dietary patterns and clinical pregnancy. Conclusions: The three maternal preconception dietary patterns identified revealed no meaningful association with IVF outcome in Japanese women. Further studies in various populations with different dietary patterns are needed to confirm these findings.

8.
Medicine (Baltimore) ; 97(35): e12112, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30170440

RESUMO

According to guidelines from the Eastern Association for the Surgery of Trauma, computed tomography (CT) with intravenous contrast is strongly recommended to diagnose clinically significant blunt traumatic aortic injury (BTAI). However, it remains unclear whether the timing of CT scanning is associated with the prognosis of BTAI patients.We extracted data on emergency patients who suffered a BTAI in the chest and/or the abdomen from 2004 to 2015 from the Japanese Trauma Data Bank, a nationwide trauma registry. The primary outcome was death in the emergency department (ED) and secondary outcome was discharge to death. In addition, we assessed the relationship between death in the ED and the timing of CT scanning by shock status in subgroup analysis. We divided these patients into the tertile groups of early (≤26 minutes), middle (27-40 minutes), and late (≥41 minutes) phases based on the time interval from hospital arrival to start of first CT scanning, and assessed death of BTAI patients in the ED by CT scanning time with the use of a multivariable logistic regression model.In total, 421 patients who suffered BTAI in the chest and/or the abdomen were eligible for our analysis. The proportion of patients dying at hospital admission was 7.7% (11/142) in the early group, 11.1% (15/135) in the middle group, and 17.6% (25/144) in the late group. In a multivariable logistic regression adjusted for confounding factors, the adjusted odds ratio (AOR) of death in the ED was 1.833 (95% confidence interval [CI]: 0.601-5.590, P = .287) in the middle group and 2.832 (95% CI: 1.007-7.960, P = .048) in the late group compared with the early group. Compared with the early group, the late group tended to have a higher rate of discharge to death (AOR: 1.438, 95% CI: 0.735-2.813). In the patients with shock, the AOR was 3.292 (95% CI: 0.495-21.902) in the middle group and 6.039 (95% CI: 0.990-36.837) in the late group compared with the early group.This study revealed that a longer time interval from hospital arrival to CT scanning was associated with higher mortality in the ED in patients with BTAI.


Assuntos
Aorta Torácica/lesões , Tomografia Computadorizada por Raios X/métodos , Lesões do Sistema Vascular/diagnóstico por imagem , Ferimentos não Penetrantes/diagnóstico por imagem , Adulto , Idoso , Aorta Torácica/diagnóstico por imagem , Serviço Hospitalar de Emergência , Feminino , Mortalidade Hospitalar , Humanos , Escala de Gravidade do Ferimento , Japão , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Alta do Paciente/estatística & dados numéricos , Prognóstico , Sistema de Registros , Estudos Retrospectivos , Fatores de Tempo , Tomografia Computadorizada por Raios X/estatística & dados numéricos , Lesões do Sistema Vascular/mortalidade , Ferimentos não Penetrantes/mortalidade
9.
J Med Internet Res ; 20(9): e10743, 2018 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-30249583

RESUMO

BACKGROUND: Meta-analyses of several randomized controlled trials have shown that cognitive behavioral therapy (CBT) has comparable efficacy to antidepressant medication, but therapist availability and cost-effectiveness is a problem. OBJECTIVE: This study aimed to evaluate the effectiveness of Web-based CBT blended with face-to-face sessions that reduce therapist time in patients with major depression who were unresponsive to antidepressant medications. METHODS: A 12-week, assessor-masked, parallel-group, waiting- list controlled, randomized trial was conducted at 3 medical institutions in Tokyo. Outpatients aged 20-65 years with a primary diagnosis of major depression who were taking ≥1 antidepressant medications at an adequate dose for ≥6 weeks and had a 17-item GRID-Hamilton Depression Rating Scale (HAMD) score of ≥14 were randomly assigned (1:1) to blended CBT or waiting-list groups using a computer allocation system, stratified by the study site with the minimization method, to balance age and baseline GRID-HAMD score. The CBT intervention was given in a combined format, comprising a Web-based program and 12 45-minute face-to-face sessions. Thus, across 12 weeks, a participant could receive up to 540 minutes of contact with a therapist, which is approximately two-thirds of the therapist contact time provided in the conventional CBT protocol, which typically provides 16 50-minute sessions. The primary outcome was the alleviation of depressive symptoms, as measured by a change in the total GRID-HAMD score from baseline (at randomization) to posttreatment (at 12 weeks). Moreover, in an exploratory analysis, we investigated whether the expected positive effects of the intervention were sustained during follow-up, 3 months after the posttreatment assessment. Analyses were performed on an intention-to-treat basis, and the primary outcome was analyzed using a mixed-effects model for repeated measures. RESULTS: We randomized 40 participants to either blended CBT (n=20) or waiting-list (n=20) groups. All patients completed the 12-week treatment protocol and were included in the intention-to-treat analyses. Participants in the blended CBT group had significantly alleviated depressive symptoms at week 12, as shown by greater least squares mean changes in the GRID-HAMD score, than those in the waiting list group (-8.9 points vs -3.0 points; mean between-group difference=-5.95; 95% CI -9.53 to -2.37; P<.001). The follow-up effects within the blended CBT group, as measured by the GRID-HAMD score, were sustained at the 3-month follow-up (week 24) and posttreatment (week 12): posttreatment, 9.4 (SD 5.2), versus follow-up, 7.2 (SD 5.7); P=.009. CONCLUSIONS: Although our findings warrant confirmation in larger and longer term studies with active controls, these suggest that a combined form of CBT is effective in reducing depressive symptoms in patients with major depression who are unresponsive to antidepressant medications. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trials Registry: UMIN000009242; https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000010852 (Archived by WebCite at http://www.webcitation. org/729VkpyYL).


Assuntos
Terapia Cognitivo-Comportamental/métodos , Transtorno Depressivo Maior/terapia , Internet/normas , Adulto , Feminino , Humanos , Masculino , Resultado do Tratamento
10.
Diabetes Obes Metab ; 19 Suppl 1: 54-62, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28880472

RESUMO

Glucose is a primary stimulator of insulin secretion. It has been thought that glucose exerts its effect by a mechanism solely dependent on glucose metabolism. We show here that glucose induces rapid Ca2+ and cyclic AMP signals in ß-cells. These rapid signals are independent of glucose-metabolism and are reproduced by non-metabolizable glucose analogues. These results led us to postulate that glucose activates a cell-surface receptor, namely the glucose-sensing receptor. Rapid signals induced by glucose are blocked by inhibition of a sweet taste receptor subunit T1R3 and a calcium-sensing receptor subunit CaSR. In accordance with these observations, T1R3 and CaSR form a heterodimer. In addition, a heterodimer of T1R3 and CaSR is activated by glucose. These results suggest that a heterodimer of T1R3 and CaSR is a major component of the glucose-sensing receptor. When the glucose-sensing receptor is blocked, glucose-induced insulin secretion is inhibited. Also, ATP production is significantly attenuated by the inhibition of the receptor. Conversely, stimulation of the glucose-sensing receptor by either artificial sweeteners or non-metabolizable glucose analogue increases ATP. Hence, the glucose-sensing receptor signals promote glucose metabolism. Collectively, glucose activates the cell-surface glucose-sensing receptor and promotes its own metabolism. Glucose then enters the cells and is metabolized through already activated metabolic pathways. The glucose-sensing receptor is a key molecule regulating the action of glucose in ß-cells.


Assuntos
Glucose/metabolismo , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Modelos Biológicos , Receptores de Superfície Celular/agonistas , Animais , Sinalização do Cálcio , AMP Cíclico/metabolismo , Dimerização , Ativação Enzimática , Regulação da Expressão Gênica , Humanos , Secreção de Insulina , Células Secretoras de Insulina/enzimologia , Proteína Quinase C/química , Proteína Quinase C/metabolismo , Multimerização Proteica , Receptores de Detecção de Cálcio/agonistas , Receptores de Detecção de Cálcio/química , Receptores de Detecção de Cálcio/genética , Receptores de Detecção de Cálcio/metabolismo , Receptores de Superfície Celular/química , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores Acoplados a Proteínas-G/agonistas , Receptores Acoplados a Proteínas-G/química , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Sistemas do Segundo Mensageiro
11.
Eur J Pharmacol ; 814: 130-137, 2017 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-28823924

RESUMO

GPBA is a G protein-coupled receptor that is activated by bile acids. Because activation of GPBA leads to increased cAMP levels and secretion of incretins and insulin, GPBA has been proposed as a promising drug target for the treatment of metabolic syndrome. Previously, we have developed a ligand-screening system to identify novel agonists of GPBA by means of a fusion protein of GPBA with G protein stimulatory α subunit (Gsα) and by a [35S]GTPγS-binding assay. To express the GPBA-Gsα fusion protein, transgenic silkworms were employed in this study, and cell membrane fractions were prepared from their fat body or pupae. We applied them to the screening of a chemical library that contains 10,625 compounds from the RIKEN Natural Products Depository (NPDepo). Eventually, a unique partial agonist, GUM2, was successfully identified. Our results indicated that the GPCR-Gα fusion proteins were beneficial for ligand identification and that the transgenic silkworms were useful for large-scale production of GPCRs. In HEK293 cells transiently expressing GPBA, GUM2 showed 50% effective concentration (EC50) of 3.5 ± 2.4µM and induced GPBA internalization as effectively as did an endogenous agonist, TLC. We also confirmed that GUM2 stimulates insulin secretion in MIN6 cells. Moreover, a single 2mg/kg dose of GUM2 significantly reduced blood glucose levels in mice during an intraperitoneal glucose tolerance test even though GUM2 is only a partial agonist with a low intrinsic activity. We concluded that GUM2 is a good candidate for research on GPBA signaling under physiological conditions and for the development of GPBA-targeting therapeutic compounds.


Assuntos
Produtos Biológicos/farmacologia , Glicemia/metabolismo , Teste de Tolerância a Glucose , Receptores Acoplados a Proteínas-G/agonistas , Animais , Células HEK293 , Humanos , Insulina/metabolismo , Secreção de Insulina , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Larva/metabolismo , Camundongos , Pupa/metabolismo
13.
J Obstet Gynaecol Res ; 43(8): 1326-1334, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28585749

RESUMO

AIM: Glycosylation of proteins is altered in cancer cells and distinctive glycan structures are associated with specific cancers, but little is known about the complete glycan profile of particular tumors. In this study, glycomic analysis of squamous cell carcinoma (SCC) of the uterine cervix was performed to search for useful markers. METHODS: A lectin microarray containing 45 lectins with different binding preferences that covered N- and O-linked glycans was coupled with evanescent field-activated fluorescent detection for glycomic analysis of SCC and normal squamous epithelium (NSE) of the cervix. Formalin-fixed, paraffin-embedded tissue specimens were obtained from 16 patients with uterine cervical cancer. Sections that included both tumor and non-tumor tissues were examined to detect alterations of glycans based on the lectin-binding pattern. RESULTS: Hippeastrum hybrid lectin was found to be a sensitive marker for distinguishing SCC of the cervix from NSE. It was the best lectin for discriminating SCC from other tissues according to receiver-operator curve analysis, as it showed a high sensitivity (81.8%), a high specificity (70.1%), and a large area under the curve (0.8182). Histochemistry confirmed specific cytoplasmic staining of SCC cells by Hippeastrum hybrid lectin, while there was little staining of cervical intraepithelial neoplasia and no staining of NSE. CONCLUSION: The present lectin microarray technique could be applied for tissue-based glycomic analysis of various tumors and for discovery of glycan-related biomarkers.


Assuntos
Amaryllidaceae/química , Carcinoma de Células Escamosas/química , Lectinas de Plantas , Polissacarídeos/química , Neoplasias do Colo do Útero/química , Adulto , Biomarcadores/análise , Feminino , Glicômica , Humanos , Análise em Microsséries
14.
PLoS One ; 12(5): e0176841, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28472098

RESUMO

We previously reported that 3T3-L1 cells express a functional sweet taste receptor possibly as a T1R3 homomer that is coupled to Gs and negatively regulates adipogenesis by a Gαs-mediated but cAMP-independent mechanism. Here, we show that stimulation of this receptor with sucralose or saccharin induced disassembly of the microtubules in 3T3-L1 preadipocytes, which was attenuated by overexpression of the dominant-negative mutant of Gαs (Gαs-G226A). In contrast, overexpression of the constitutively active mutant of Gαs (Gαs-Q227L) as well as treatment with cholera toxin or isoproterenol but not with forskolin caused disassembly of the microtubules. Sweetener-induced microtubule disassembly was accompanied by activation of RhoA and Rho-associated kinase (ROCK). This was attenuated with by knockdown of GEF-H1, a microtubule-localized guanine nucleotide exchange factor for Rho GTPase. Furthermore, overexpression of the dominant-negative mutant of RhoA (RhoA-T19N) blocked sweetener-induced dephosphorylation of Akt and repression of PPARγ and C/EBPα in the early phase of adipogenic differentiation. These results suggest that the T1R3 homomeric sweet taste receptor negatively regulates adipogenesis through Gαs-mediated microtubule disassembly and consequent activation of the Rho/ROCK pathway.


Assuntos
Adipogenia/fisiologia , Cromograninas/fisiologia , Subunidades alfa Gs de Proteínas de Ligação ao GTP/fisiologia , Microtúbulos/fisiologia , Receptores Acoplados a Proteínas-G/fisiologia , Proteínas rho de Ligação ao GTP/metabolismo , Células 3T3-L1 , Animais , Transferência Ressonante de Energia de Fluorescência , Camundongos
15.
Ind Health ; 55(3): 243-251, 2017 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-28123138

RESUMO

This study investigated whether non- or occasional drinkers' changes in drinking habits during a one-year period were related to psychological distress. Overall, 2,495 non- or occasional drinking employees (2,109 men and 386 women) completed a self-administered questionnaire measuring alcohol intake, psychological distress (12-item General Health Questionnaire), and demographic characteristics at baseline and one-year follow-up. They also completed a Web-based version of the Brief Job Stress Questionnaire to assess job stressors at baseline. Participants were categorized into three groups (stable non- or occasional drinkers; new light drinkers; new moderate drinkers) according to weekly alcohol consumption at follow-up (males 0 g/wk, 1-79 g/wk, and ≥80 g/wk; females 0 g/wk, 1-39 g/wk, and ≥40 g/wk, respectively); multiple logistic regression analyses were conducted by sex. Among only male participants, both stable non- or occasional drinkers and new moderate drinkers showed significantly higher odds ratios for psychological distress at follow-up than new light drinkers after adjusting for demographic characteristics, job stressors, and psychological distress at baseline (adjusted odds ratios of 1.72 and 1.99, respectively). These findings suggest that men who started to drink 80 g or more alcohol per week during the one-year follow-up period should have been monitored for psychological distress.


Assuntos
Consumo de Bebidas Alcoólicas/epidemiologia , Consumo de Bebidas Alcoólicas/psicologia , Estresse Ocupacional/epidemiologia , Adulto , Estudos de Coortes , Feminino , Humanos , Japão/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Estresse Psicológico/epidemiologia , Inquéritos e Questionários
16.
J Biol Chem ; 291(44): 23126-23135, 2016 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-27613866

RESUMO

The calcium-sensing receptor (CaSR) is activated by various cations, cationic compounds, and amino acids. In the present study we investigated the effect of glucose on CaSR in HEK293 cells stably expressing human CaSR (HEK-CaSR cells). When glucose concentration in the buffer was raised from 3 to 25 mm, a rapid elevation of cytoplasmic Ca2+ concentration ([Ca2+]c) was observed. This elevation was immediate and transient and was followed by a sustained decrease in [Ca2+]c The effect of glucose was detected at a concentration of 4 mm and reached its maximum at 5 mm 3-O-Methylglucose, a non-metabolizable analogue of glucose, reproduced the effect of glucose. Sucrose also induced an elevation of [Ca2+]c in HEK-CaSR cells. Similarly, sucralose was nearly as effective as glucose in inducing elevation of [Ca2+]c Glucose was not able to increase [Ca2+]c in the absence of extracellular Ca2+ The effect of glucose on [Ca2+]c was inhibited by NPS-2143, an allosteric inhibitor of CaSR. In addition, NPS-2143 also inhibited the [Ca2+]c responses to sucralose and sucrose. Glucose as well as sucralose decreased cytoplasmic cAMP concentration in HEK-CaSR cells. The reduction of cAMP induced by glucose was blocked by pertussis toxin. Likewise, sucralose reduced [cAMP]c Finally, glucose increased [Ca2+]c in PT-r parathyroid cells and in Madin-Darby canine kidney cells, both of which express endogenous CaSR. These results indicate that glucose acts as a positive allosteric modulator of CaSR.


Assuntos
Glucose/metabolismo , Receptores de Detecção de Cálcio/química , Receptores de Detecção de Cálcio/metabolismo , Regulação Alostérica , Cálcio/metabolismo , Citoplasma/química , Citoplasma/genética , Citoplasma/metabolismo , Glucose/análise , Células HEK293 , Humanos , Receptores de Detecção de Cálcio/genética
17.
Endocr J ; 63(8): 715-25, 2016 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-27250218

RESUMO

Sucralose is an artificial sweetener and activates the glucose-sensing receptor expressed in pancreatic ß-cells. Although sucralose does not enter ß-cells nor acts as a substrate for glucokinase, it induces a marked elevation of intracellular ATP ([ATP]c). The present study was conducted to identify the signaling pathway responsible for the elevation of [ATP]c induced by sucralose. Previous studies have shown that sucralose elevates cyclic AMP (cAMP), activates phospholipase C (PLC) and stimulates Ca(2+) entry by a Na(+)-dependent mechanism in MIN6 cells. The addition of forskolin induced a marked elevation of cAMP, whereas it did not affect [ATP]c. Carbachol, an activator of PLC, did not increase [ATP]c. In addition, activation of protein kinase C by dioctanoylglycerol did not affect [ATP]c. In contrast, nifedipine, an inhibitor of the voltage-dependent Ca(2+) channel, significantly reduced [ATP]c response to sucralose. Removal of extracellular Na(+) nearly completely blocked sucralose-induced elevation of [ATP]c. Stimulation of Na(+) entry by adding a Na(+) ionophore monensin elevated [ATP]c. The monensin-induced elevation of [ATP]c was only partially inhibited by nifedipine and loading of BAPTA, both of which completely abolished elevation of [Ca(2+)]c. These results suggest that Na(+) entry is critical for the sucralose-induced elevation of [ATP]c. Both calcium-dependent and -independent mechanisms are involved in the action of sucralose.


Assuntos
Trifosfato de Adenosina/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Sinalização do Cálcio/fisiologia , Células Secretoras de Insulina/efeitos dos fármacos , Sacarose/análogos & derivados , Animais , Cálcio/metabolismo , Cálcio/farmacologia , Células Cultivadas , AMP Cíclico/metabolismo , Ácido Egtázico/análogos & derivados , Ácido Egtázico/farmacologia , Células Secretoras de Insulina/metabolismo , Camundongos , Nifedipino/farmacologia , Sacarose/farmacologia , Edulcorantes/farmacologia
18.
PLoS One ; 10(12): e0144053, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26630567

RESUMO

Glucose is a primary stimulator of insulin secretion in pancreatic ß-cells. High concentration of glucose has been thought to exert its action solely through its metabolism. In this regard, we have recently reported that glucose also activates a cell-surface glucose-sensing receptor and facilitates its own metabolism. In the present study, we investigated whether glucose activates the glucose-sensing receptor and elicits receptor-mediated rapid actions. In MIN6 cells and isolated mouse ß-cells, glucose induced triphasic changes in cytoplasmic Ca(2+) concentration ([Ca(2+)]c); glucose evoked an immediate elevation of [Ca(2+)]c, which was followed by a decrease in [Ca(2+)]c, and after a certain lag period it induced large oscillatory elevations of [Ca(2+)]c. Initial rapid peak and subsequent reduction of [Ca(2+)]c were independent of glucose metabolism and reproduced by a nonmetabolizable glucose analogue. These signals were also blocked by an inhibitor of T1R3, a subunit of the glucose-sensing receptor, and by deletion of the T1R3 gene. Besides Ca(2+), glucose also induced an immediate and sustained elevation of intracellular cAMP ([cAMP]c). The elevation of [cAMP]c was blocked by transduction of the dominant-negative Gs, and deletion of the T1R3 gene. These results indicate that glucose induces rapid changes in [Ca(2+)]c and [cAMP]c by activating the cell-surface glucose-sensing receptor. Hence, glucose generates rapid intracellular signals by activating the cell-surface receptor.


Assuntos
Cálcio/metabolismo , AMP Cíclico/metabolismo , Glucose/metabolismo , Células Secretoras de Insulina/metabolismo , Receptores de Superfície Celular/metabolismo , Animais , Linhagem Celular , Citoplasma/metabolismo , Insulina/metabolismo , Camundongos
19.
Carbohydr Polym ; 134: 718-25, 2015 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-26428177

RESUMO

This study examined the effects of sub- and supercritical water pretreatments on the physicochemical properties of crab shell α-chitin and its enzymatic degradation to obtain N,N'-diacetylchitobiose (GlcNAc)2. Following sub- and supercritical water pretreatments, the protein in the crab shell was removed and the residue of crab shell contained α-chitin and CaCO3. Prolonged pretreatment led to α-chitin decomposition. The reaction of pure α-chitin in sub- and supercritical water pretreatments was investigated separately; we observed lower mean molecular weight and weaker hydrogen bonds compared with untreated α-chitin. (GlcNAc)2 yields from enzymatic degradation of subcritical (350 °C, 7 min) and supercritical water (400 °C, 2.5 min) pretreated crab shell were 8% and 6%, compared with 0% without any pretreatment. This study shows that sub- and supercritical water pretreatments of crab shell provide to an alternative method to the use of acid and base for decalcification and deproteinization of crab shell required for (GlcNAc)2 production.


Assuntos
Exoesqueleto/química , Braquiúros/química , Quitina/química , Quitina/metabolismo , Enzimas/metabolismo , Água/química , Acetilglucosaminidase/química , Animais
20.
J Endocrinol ; 226(1): 57-66, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25994004

RESUMO

Glucose activates the glucose-sensing receptor T1R3 and facilitates its own metabolism in pancreatic ß-cells. An inhibitor of this receptor would be helpful in elucidating the physiological function of the glucose-sensing receptor. The present study was conducted to examine whether or not lactisole can be used as an inhibitor of the glucose-sensing receptor. In MIN6 cells, in a dose-dependent manner, lactisole inhibited insulin secretion induced by sweeteners, acesulfame-K, sucralose and glycyrrhizin. The IC50 was ∼4 mmol/l. Lactisole attenuated the elevation of cytoplasmic Ca2+ concentration ([Ca2+]c) evoked by sucralose and acesulfame-K but did not affect the elevation of intracellular cAMP concentration ([cAMP]c) induced by these sweeteners. Lactisole also inhibited the action of glucose in MIN6 cells. Thus, lactisole significantly reduced elevations of intracellular [NADH] and intracellular [ATP] induced by glucose, and also inhibited glucose-induced insulin secretion. To further examine the effect of lactisole on T1R3, we prepared HEK293 cells stably expressing mouse T1R3. In these cells, sucralose elevated both [Ca2+]c and [cAMP]c. Lactisole attenuated the sucralose-induced increase in [Ca2+]c but did not affect the elevation of [cAMP]c. Finally, lactisole inhibited insulin secretion induced by a high concentration of glucose in mouse islets. These results indicate that the mouse glucose-sensing receptor was inhibited by lactisole. Lactisole may be useful in assessing the role of the glucose-sensing receptor in mouse pancreatic ß-cells.


Assuntos
Derivados de Benzeno/farmacologia , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/metabolismo , Receptores Acoplados a Proteínas-G/antagonistas & inibidores , Edulcorantes/farmacologia , Animais , Cálcio/metabolismo , Linhagem Celular , AMP Cíclico/metabolismo , Glucose/metabolismo , Glucose/farmacologia , Ácido Glicirrízico/farmacologia , Células HEK293 , Humanos , Insulina/metabolismo , Secreção de Insulina , Camundongos , Sacarose/análogos & derivados , Sacarose/farmacologia , Tiazinas/farmacologia
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