RESUMO
Objective: To investigate the clinical significance and correlation of arginase 1 (Arg-1) and inducible nitric oxide synthase (iNOS) expression in hepatocellular carcinoma (HCC). Methods: The expression of Arg-1and iNOS in 146 cases of hepatocellular carcinoma tissues and corresponding adjacent tissues was detected by immunohistochemistry. The clinicopathological characteristics and the correlation between the expressions and prognosis were determined by chi square test, Spearman's rank correlation, Kaplan-Meier survival analysis and Cox regression analysis. Results: The positive rates of Arg-1 and iNOS were 18.7% (23/123) and 37.0% (54/146), respectively, which was significantly lower than the adjacent tissues [100%(146/146) and 93.8% (137/146)] and the difference was statistically significant (χ (2) = 212.521, P < 0.01, χ (2) = 104.276, P < 0.01). There was a positive correlation between the both expression (r = 0.331, P < 0.01). Arg-1 low expression was correlated with preoperative serum alpha-fetoprotein (AFP) level, tumor size, differentiation degree, histological types and Edmondson's grade. iNOS low expression was correlated with the differentiation degree and Edmondson's grade (P < 0.05). Kaplan Meier survival analysis showed that in patients with recurrence-free survival (RFs), Arg-1 (+) group > Arg-1 (-) group and Arg-1 (+) iNOS (+) group > Arg-1 (+) iNOS (-) group > Arg-1 (-) iNOS (-) group (P < 0.05). Cox multivariate analysis showed that age, tumor size, Edmondson's grade, vascular tumor emboli were significantly correlated with RFs (P < 0.05). Conclusion: There is a positive correlation between Arg-1 and iNOS expressions in HCC, and both may reflect the HCC malignant degree. The reduced/absent expression of both may participate in the occurrence and development of HCC. The combined detection of Arg-1 and iNOS on HCC may have certain significance for the judgment of differentiation degree and prognosis.
Assuntos
Carcinoma Hepatocelular , Neoplasias Hepáticas , Arginase , Humanos , Óxido Nítrico Sintase Tipo II/genética , PrognósticoRESUMO
Objective: To investigate the correlation between work-related musculoskeletal disorders (WMSDs) and work ability (WA) among nurses in Xinjiang. Methods: A total of 1 873 nursing staff in the Departments of Internal Medicine, Surgery, Emergency and ICU Department, Operating Room and Supply Room of 6 Grade 3 General Hospitals in Xinjiang Beijiang Region were randomly selected by means of random sampling, and revised Nordic National Musculoskeletal Disorders Standard Questionnaire and Work Ability Index Scale were used to evaluate the status of nursing WMSDs and WA. Results: (1) The symptom incidence of work-related musculoskeletal disorders among nursing staff in the past year was 77.42%; The highest symptom incidence in all parts of the body was waist, neck, shoulder and back, and the symptom incidence was 63.59%, 59.80%, 47.78% and 37.32%, respectively; (2) The total score of nursing staff's work ability was (35.91±5.143) ; The proportion of "poor" , "middle" , "good" and "excellent" were 5.23%, 47.41%, 41.43% and 5.93% respectively; (3) The total working capacity of the sick group was (35.11±5.044) , The working ability of the non-diseased group was (38.64±4.507) , The difference between the two groups was statistically significant (t=13.773, P<0.01) ; There was a significant difference in the level of work ability between the two groups (z=-12.335, P<0.01) ; The working ability of "poor" and medium in the injury group was significantly higher than that of the non injury group, while the working ability was "better" and "excellent" than that of the non injury group; (4) The working ability of nursing staff of different age in WMSDs injury group was lower than that in non injury group, the difference was statistically significant (P<0.01) ; There was a negative correlation between the working ability of the injury group and the length of service, that is, with the increase of the length of service, the working ability index decreased (P<0.01) ; There was a positive correlation between the working ability of the non injury group and the length of service, that is, with the increase of the length of service, the work ability index increased gradually, but the correlation was not statistically significant (r=0.063, P=0.195) ; (5) The working ability of nursing staff of different departments in WMSDs injury group was lower than that in non injury group, the difference was statistically significant (P<0.01) . (6) Multiple linear regression analysis showed that the factors influencing the working ability of nursing staff were: past medical history, WMSDs disease, length of service and night shift frequency; Conclusion: The symptom incidence of WMSDs was high among nurses; Work Ability at a moderate level; work-related musculoskeletal disorders is associated with reduced ability of nursing staff; work-related musculoskeletal disorders are associated with reduced working capacity of nursing staff.
Assuntos
Doenças Musculoesqueléticas/epidemiologia , Recursos Humanos de Enfermagem Hospitalar/estatística & dados numéricos , Doenças Profissionais/epidemiologia , Avaliação da Capacidade de Trabalho , China/epidemiologia , Hospitais Gerais , Humanos , Inquéritos e QuestionáriosRESUMO
Objective: To evaluate the ergonomic load of clinical nursing procedures and to provide evidence for the prevention and management of work-related musculoskeletal disorders (WMSDs) in nurses. Methods: Based on the nursing unit characteristics and the common departments involving patient-turning procedures, 552 nurses were selected from 6 clinical departments from July to September, 2016. The ergonomic load of four types of patient-turning procedures, i.e., turning the patient's body, changing the bed linen of in-bed patients, moving patients, and chest physiotherapy, was evaluated by the on-site inspectors and self-evaluated by the operators using the Quick Exposure Check. The exposure value, exposure level, and exposure rate of WMSDs were assessed based on the procedure-related physical loads on the back, shoulders/arms, wrists/hands and neck, as well as the loads from work rhythm and work pressure. Results: All surveyed subjects were females who were aged mostly between 26-30 years (49.46%) , with a mean age of 29.66±5.28 years. These nurses were mainly from the Department of Infection (28.99%) and Spine Surgery (21.56%) . There were significant differences in the back, shoulders/arms, neck, work rhythm, and work pressure scores between different nursing procedures (F=16.613, 5.884, 3.431, 3.222, and 5.085, respectively; P<0.05) . Patient-turning nursing procedures resulted in high to intermediate physical load in nurses. Procedures with high to low level of WMSDs exposure were patient turning (72.69%) , bed linen changing (67.15%) , patient transfer (65.82%) , and chest physiotherapy (58.34%) . In particular, patient turning was considered as very high-risk procedure, whereas others were considered as high-risk procedures. Conclusion: Patient-turning nursing procedures result in high ergonomic load in the operators. Therefore, more focus should be placed on the ergonomics of the caretakers and nurses.
Assuntos
Ergonomia , Dor Lombar/prevenção & controle , Movimentação e Reposicionamento de Pacientes , Doenças Musculoesqueléticas/prevenção & controle , Enfermeiras e Enfermeiros , Cuidados de Enfermagem/métodos , Suporte de Carga , Adulto , Feminino , Humanos , Remoção/efeitos adversos , Fatores de Risco , Inquéritos e Questionários , Suporte de Carga/fisiologia , Adulto JovemRESUMO
OBJECTIVE: To identify and verify proteins that interact and collaborate with ATF3 in inhibiting hepatocarcinogenesis. METHODS: Immunoprecipitation (IP), co-IP and protein spectrum analysis were used to identify the protein which interacted with ATF3 in HepG2. Immunohistochemistry (IHC) and Western blot (WB) were used to detect the expression pattern of ATF3 and its candidate interacting proteins in liver tissue. RESULTS: The protein expression differences were detected by IP in two HepG2 groups. The experimental group was infected by lentiviral vector with ATF3 over-expression and the control group was infected by mock-vehicle. Several protein bands with expression diversity were analyzed by protein spectrum, which revealed several candidate proteins that may be related with ATF3. Peptide sequences were analyzed by Mascot software and NCBI database. Combined with the existing literature and our study results, Gelsolin (GSN) was identified as a protein closely interacting with ATF3 and confirmed by co-IP, IHC and WB. CONCLUSIONS: GSN is identified and verified as an interacting protein with ATF3. ATF3 may function as a suppressor of liver cancer via protein-protein interactions with Gelsolin.
Assuntos
Fator 3 Ativador da Transcrição/metabolismo , Gelsolina/metabolismo , Neoplasias Hepáticas , Fígado/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Western Blotting , Gelsolina/análise , Humanos , Imuno-Histoquímica , Imunoprecipitação , Proteínas Supressoras de Tumor/análiseRESUMO
Objective: To investigate the prevalence of work-related musculoskeletal disorders (WMSDs) in nurses in Xinjiang, China, to analyze the influencing factors for the development of WMSDs, and to provide a reference for the prevention and treatment of WMSDs in nurses. Methods: Stratified cluster sampling was used to randomly select 8 422 nurses in 8 tertiary hospitals and 4 secondary hospitals in Xinjiang from January to October, 2015. A questionnaire survey was performed to investigate the prevalence of WMSDs in nurses from departments of internal medicine, surgery, gynecology, pediatrics, emergency, and intensive care and operating rooms and analyze related influencing factors. Results: A total of 3 000 questionnaires were distributed, and 2 851 questionnaires were returned, resulting in a recovery rate of 95.00%. In the nurses who participated in the survey, the prevalence of WMSDs was 78.58%. As for different body parts, the prevalence of WMSDs was 63.36% in the waist, 61.79% in the neck, 52.52% in the shoulder, 41.83% in the back, 35.56% in the knees, 33.35% in the ankles, 26.02% in the wrists, 23.41% in the hip, and 17.62% in the elbows. There were significant differences in the prevalence of WMSDs across the nurses with different ages and working years (χ2=28.29 and 27.73, both P<0.01) , and the prevalence of WMSDs tended to increase with the increasing age and working years (χ2=22.94 and 17.56, both P<0.01) . The results of multivariate logistic regression analysis showed that sex, working years, intensive care/emergency, outpatient service, surgical anesthesia, weekly working hours, physical condition, and fatigue in work were risk factors for WMSDs in nurses. Conclusion: In the nurses in Xinjiang, the prevalence of WMSDs in the neck, shoulder, back, and waist is high, and nurses with different ages, working years, departments, sexes, physical conditions, and weekly working hours have varying degrees of risk of WMSDs.
Assuntos
Doenças Musculoesqueléticas/epidemiologia , Enfermeiras e Enfermeiros/estatística & dados numéricos , Doenças Profissionais/epidemiologia , Dorso , China , Estudos Transversais , Hospitais , Humanos , Prevalência , Fatores de Risco , Ombro , Inquéritos e QuestionáriosRESUMO
HBV genotypes have specific geographical distributions and can serve as epidemiological markers. Accumulated data have shown that the major HBV genotypes in China are B and C. Here, the HBV genotypes were examined from 6817 blood samples, which were collected from patients with chronic HBV infection in Fujian Province during 2006-2013; genotype B was identified in 3384 patients (49·6%), while genotype C was identified in 3430 patients (50·3%). The percentage of patients infected with genotype C gradually increased with age from 39·5% (patients aged 50 years), reaching a peak of 67·3% in the 45-50 years age group. These results clearly demonstrate that the genotype distribution of HBV in Fujian Province has significantly changed in recent years with almost equal numbers of genotype B and genotype C infections existing in the entire patient population, while higher incidence of genotype C infection exists in older patients, but genotype B is no longer dominant in the Fujian area as previously reported.
Assuntos
Vírus da Hepatite B/genética , Hepatite B Crônica/epidemiologia , Hepatite B Crônica/virologia , Adolescente , Adulto , China/epidemiologia , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Recurrent Clostridium difficile infection (CDI) represents a significant burden on the healthcare system and is associated with poor outcomes in hematopoietic stem cell transplant (HSCT) patients. Data are limited evaluating recurrence rates and risk factors for recurrence in HSCT patients. METHODS: HSCT patients who developed CDI between January 2010 and December 2012 were divided into 2 groups: non-recurrent CDI (nrCDI) and recurrent CDI (rCDI). Risk factors for rCDI were compared between groups. Rate of recurrence in HSCT patients was compared to that in other hospitalized patients. RESULTS: CDI was diagnosed in 95 of 711 HSCT patients (22 rCDI and 73 nrCDI). Recurrence rates were similar in HSCT patients compared with other hospitalized patients (23.2% vs. 22.9%, P > 0.99). Patients in the rCDI group developed the index case of CDI significantly earlier than the nrCDI group (3.5 days vs. 7.0 days after transplant, P = 0.05). On univariate analysis, patients with rCDI were more likely to have prior history of CDI and neutropenia at the time of the index CDI case. Neutropenia at the time of the index CDI case was the only independent predictor of rCDI (78.8 vs. 34.8%, P = 0.006) on multivariate analysis. CONCLUSIONS: The rate of rCDI was similar between HSCT and other hospitalized patients, and the majority of patients developed the index case of CDI within a week of transplantation. Neutropenia at the index CDI case may be associated with increased rates of rCDI.
Assuntos
Clostridioides difficile , Enterocolite Pseudomembranosa/epidemiologia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Neutropenia/epidemiologia , Adulto , Idoso , Estudos de Casos e Controles , Enterocolite Pseudomembranosa/microbiologia , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Recidiva , Estudos Retrospectivos , Fatores de Risco , Fatores de Tempo , Adulto JovemRESUMO
This study evaluated the efficacy of slightly acidic electrolyzed water (SAEW) for the inactivation of Salmonella Enteritidis and Escherichia coli on shelled eggs as well as studied the shelf life and internal quality attributes (i.e., weight loss, Haugh unit, yolk index, albumen pH, and yolk pH) of eggs during storage at 25°C. The decontamination test egg samples (freshly laid) were inoculated and immersed for 1, 2, 3, and 4 min in each treatment (i.e., SAEW, acidic electrolyzed water, NaClO solution, and sterile deionized water) at available chlorine concentrations (ACCs) of 10, 18 and 26 mg/L. The storage test eggs (freshly laid) were immersed for 3 min in 4 treatments (i.e., SAEW, acidic electrolyzed water, NaClO solution, and no treatment) and stored for 30 D at 25°C. The effects of the SAEW on microbiological qualities and freshness parameters were investigated. Decontamination of eggs with SAEW showed an equivalent or higher bactericidal effect compared to other treatments. A complete inactivation of S. Enteritidis and E. coli on the surface of shelled egg samples resulted from treatment with SAEW at an ACC of 26 mg/L for 3 and 4 min, respectively. Almost all of the egg quality parameters investigated in the present study were significantly (P < 0.05) affected by the storage time. As storage time increased, the yolk index and Haugh unit value decreased, and weight loss, albumen pH, and yolk pH increased. However, SAEW treatment minimized weight loss (5.52%) and preserved the albumen and yolk quality better than no treatment at 25°C. Relative to acidic electrolyzed water and NaClO solution, the advantages of SAEW are reduced corrosion of egg surfaces and potentially less water and CO2 escaping from eggshell pores. The results highlight the promising use of SAEW to enhance the microbial safety and to extend the shelf life of shelled eggs. Future combined methods with SAEW and other treatments are also needed.
Assuntos
Desinfetantes/farmacologia , Ovos/microbiologia , Escherichia coli/efeitos dos fármacos , Óvulo/química , Salmonella enteritidis/efeitos dos fármacos , Água/farmacologia , Eletrólise , Concentração de Íons de Hidrogênio , Óvulo/efeitos dos fármacos , Água/químicaRESUMO
Differentiation of the R7 photoreceptor cell is dependent on the Sevenless receptor tyrosine kinase, which activates the RAS1/mitogen-activated protein kinase signaling cascade. Kinase suppressor of Ras (KSR) functions genetically downstream of RAS1 in this signal transduction cascade. Expression of dominant-negative KSR (KDN) in the developing eye blocks RAS pathway signaling, prevents R7 cell differentiation, and causes a rough eye phenotype. To identify genes that modulate RAS signaling, we screened for genes that alter RAS1/KSR signaling efficiency when misexpressed. In this screen, we recovered three known genes, Lk6, misshapen, and Akap200. We also identified seven previously undescribed genes; one encodes a novel rel domain member of the NFAT family, and six encode novel proteins. These genes may represent new components of the RAS pathway or components of other signaling pathways that can modulate signaling by RAS. We discuss the utility of gain-of-function screens in identifying new components of signaling pathways in Drosophila.
Assuntos
Proteínas de Drosophila , Drosophila melanogaster/genética , Proteínas de Ligação ao GTP/metabolismo , Genes de Insetos , Proteínas de Insetos/genética , Transdução de Sinais/genética , Proteínas ras , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Cruzamentos Genéticos , Drosophila melanogaster/fisiologia , Feminino , Regulação da Expressão Gênica , Genótipo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Masculino , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Dados de Sequência Molecular , Células Fotorreceptoras de Invertebrados/fisiologia , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Using the polymerase chain reaction (PCR) differential display directed to cloning transcripts related to retention performance for the inhibitory avoidance learning task, we have isolated two rat cDNA fragments homologous to the 3' untranslated region (3'UTR) of glial fibrillary acid protein (GFAP) cDNA. The GFAP upper cDNA represents a new GFAP mRNA isotype which has a 7 bp insertion in the 3'UTR. Further analyses indicated that retention performance in rats containing only the new GFAP mRNA isotype was significantly better than that of rats containing both the new GFAP mRNA isotype and the standard GFAP mRNA simultaneously. These results suggest that expression of the GFAP isotypes may be related to memory retention in rats.
Assuntos
Proteína Glial Fibrilar Ácida/metabolismo , Hipocampo/metabolismo , Retenção Psicológica/fisiologia , Animais , Aprendizagem da Esquiva , Sequência de Bases , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Transcrição GênicaRESUMO
The present study investigated the role of hippocampal nitric oxide (NO) in memory retention of an inhibitory avoidance learning task in rats. The anatomical locus was aimed at the dentate gyrus (DG). Results indicated that intra-DG administration of a NO generator, sodium nitroprusside (SNP), at moderate doses enhanced retention performance in a dose-response fashion in rats. SNP at higher doses, on the other hand, impaired memory retention. Intra-DG injection of a NO inhibitor, L-NG-monomethylarginine (L-MeArg), impaired retention performance at moderate doses. Coadministration of a NO precursor L-arginine (2.9 and 7.2 micrograms) reversed the memory-impairing effect of L-MeArg. An in vitro ADP-ribosylation experiment showed five protein bands with molecular weights around 118, 94, 54, 43, and 39 kDa that were labeled. The labeling intensity of these proteins decreased as the concentration of in vivo SNP increased. These results suggest that hippocampal NO plays a facilitatory role in the memory process of an inhibitory avoidance learning task in rats.
Assuntos
Arginina/análogos & derivados , Arginina/farmacologia , Hipocampo/fisiologia , Óxido Nítrico/fisiologia , Nitroprussiato/farmacologia , Retenção Psicológica/fisiologia , Adenosina Difosfato Ribose/metabolismo , Animais , Hipocampo/química , Hipocampo/efeitos dos fármacos , Masculino , Óxido Nítrico/antagonistas & inibidores , Ratos , Ratos Sprague-Dawley , Retenção Psicológica/efeitos dos fármacos , ômega-N-MetilargininaRESUMO
Corticotropin-releasing factor (CRF) has been found to play an important role in modulating the learning and memory processes in rats. In the present study, we examined the alteration of CRF mRNA level in rat hippocampus at different stages of the memory process. Results indicated that CRF mRNA is quite homogenously distributed within subdivisions of the hippocampus, with the dentate gyrus (DG) has a relatively lower expression level. By using the one-way inhibitory avoidance learning paradigm, we have selected the good memory (GM) rats (with a retention score of 600) and the poor memory (PM) rats (with retention scores < 80). Results indicated that CRF mRNA level in the hippocampus was significantly increased in GM rats when compared with that in the PM rats at 1 hr, 3 hr and 6 hr after training. It was not markedly altered at 24 hr and 72 hr post-training. However, it was again increased at 21 days post-training in both the hippocampus and the frontal cortex. Further, this effect was demonstrated not to be associated with the acute footshock stress. These results together suggest that enhanced hippocampal CRF gene expression is probably associated with both memory consolidation and memory storage, but is not associated with maintenance of formed memory in rats.
Assuntos
Aprendizagem da Esquiva/fisiologia , Hormônio Liberador da Corticotropina/genética , Expressão Gênica , Hipocampo/fisiologia , Memória/fisiologia , Animais , Eletrochoque , Masculino , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Estresse Fisiológico/metabolismo , Fatores de Tempo , Distribuição Tecidual , Transcrição GênicaAssuntos
Antagonistas Muscarínicos/uso terapêutico , Quinuclidinas/uso terapêutico , Tetra-Hidroisoquinolinas/uso terapêutico , Bexiga Urinária Hiperativa/tratamento farmacológico , Adulto , Relação Dose-Resposta a Droga , Método Duplo-Cego , Esquema de Medicação , Feminino , Humanos , Masculino , Antagonistas Muscarínicos/administração & dosagem , Antagonistas Muscarínicos/efeitos adversos , Quinuclidinas/administração & dosagem , Quinuclidinas/efeitos adversos , Succinato de Solifenacina , Tetra-Hidroisoquinolinas/administração & dosagem , Tetra-Hidroisoquinolinas/efeitos adversos , Resultado do Tratamento , Bexiga Urinária Hiperativa/metabolismo , Micção/efeitos dos fármacosRESUMO
Znf179 is a member of the RING finger protein family. During embryogenesis, Znf179 is expressed in a restricted manner in the brain, suggesting a potential role in nervous system development. In this report, we show that the expression of Znf179 is upregulated during P19 cell neuronal differentiation. Inhibition of Znf179 expression by RNA interference significantly attenuated neuronal differentiation of P19 cells and a primary culture of cerebellar granule cells. Using a microarray approach and subsequent functional annotation analysis, we identified differentially expressed genes in Znf179-knockdown cells and found that several genes are involved in development, cellular growth, and cell cycle control. Flow cytometric analyses revealed that the population of G0/G1 cells decreased in Znf179-knockdown cells. In agreement with the flow cytometric data, the number of BrdU-incorporated cells significantly increased in Znf179-knockdown cells. Moreover, in Znf179-knockdown cells, p35, a neuronal-specific Cdk5 activator that is known to activate Cdk5 and may affect the cell cycle, and p27, a cell cycle inhibitor, also decreased. Collectively, these results show that induction of the Znf179 gene may be associated with p35 expression and p27 protein accumulation, which lead to cell cycle arrest in the G0/G1 phase, and is critical for neuronal differentiation of P19 cells.
Assuntos
Diferenciação Celular , Proteínas de Ligação a DNA/metabolismo , Células-Tronco de Carcinoma Embrionário/citologia , Neurônios/citologia , Animais , Bromodesoxiuridina/farmacologia , Células Cultivadas , Quinase 5 Dependente de Ciclina/antagonistas & inibidores , Quinase 5 Dependente de Ciclina/genética , Quinase 5 Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/genética , Células-Tronco de Carcinoma Embrionário/metabolismo , Fase G1 , Pontos de Checagem da Fase G1 do Ciclo Celular , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurogênese/genética , Neurônios/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Fase de Repouso do Ciclo Celular , Tretinoína/farmacologiaAssuntos
Células Híbridas/enzimologia , Hipoxantina Fosforribosiltransferase/genética , Mieloma Múltiplo/enzimologia , Animais , Linhagem Celular , Regulação da Expressão Gênica , Humanos , Hipoxantina Fosforribosiltransferase/análise , Camundongos , Mieloma Múltiplo/patologia , Coelhos , Ratos , Reticulócitos/enzimologiaRESUMO
This study was to examine the effects of treadmill exercise on the expression of brain-derived neurotrophic factor (BDNF) in rat hippocampus. After 1-wk treadmill familiarization, animals in exercise groups received a 4-wk exercise training or an acute exercise. They were sacrificed 2 h or 2 d after exercise and their hippocampal BDNF mRNA and protein levels were determined. We demonstrated that 1) hippocampal BDNF mRNA and protein levels were both elevated in response to exercise training at 2 h after the last run but not after 2 d; 2) an acute moderate exercise (1 or 3 d) increased BDNF protein levels; 3) acute severe exercise increased BDNF protein and mRNA levels in animals under a familiarization regimen, while suppressed the BDNF mRNA level in rats without treadmill familiarization, paralleling the stress effect of immobilization/water exposure. We conclude that compulsive treadmill exercise with pre-familiarization acutely upregulates rat hippocampal BDNF gene expression.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Hipocampo/metabolismo , Condicionamento Físico Animal/fisiologia , Regulação para Cima/fisiologia , Animais , Fator Neurotrófico Derivado do Encéfalo/genética , Regulação para Baixo/fisiologia , Teste de Esforço , Regulação da Expressão Gênica/fisiologia , Masculino , Aprendizagem em Labirinto/fisiologia , Plasticidade Neuronal/fisiologia , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Estresse Fisiológico/genética , Estresse Fisiológico/metabolismo , Estresse Fisiológico/fisiopatologiaRESUMO
Dorsoventral (DV) patterning of the Drosophila embryo is initiated by a broad Dorsal (Dl) nuclear gradient, which is regulated by a conserved signaling pathway that includes the Toll receptor and Pelle kinase. We investigate the consequences of expressing a constitutively activated form of the Toll receptor, Toll(10b), in anterior regions of the early embryo using the bicoid 3' UTR. Localized Toll(10b) products result in the formation of an ectopic, anteroposterior (AP) Dl nuclear gradient along the length of the embryo. The analysis of both authentic dorsal target genes and defined synthetic promoters suggests that the ectopic gradient is sufficient to generate the full repertory of DV patterning responses along the AP axis of the embryo. For example, mesoderm determinants are activated in the anterior third of the embryo, whereas neurogenic genes are expressed in central regions. These results raise the possibility that Toll signaling components diffuse in the plasma membrane or syncytial cytoplasm of the early embryo. This study also provides evidence that neurogenic repressors may be important for the establishment of the sharp mesoderm/neuroectoderm boundary in the early embryo.
Assuntos
Padronização Corporal/fisiologia , Proteínas de Drosophila , Drosophila/embriologia , Proteínas de Insetos/fisiologia , Glicoproteínas de Membrana/fisiologia , Receptores de Superfície Celular/fisiologia , Transdução de Sinais/fisiologia , Animais , Animais Geneticamente Modificados , Cruzamentos Genéticos , Drosophila/genética , Ectoderma , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Insetos/análise , Proteínas de Insetos/genética , Glicoproteínas de Membrana/análise , Glicoproteínas de Membrana/genética , Mesoderma , Regiões Promotoras Genéticas/genética , RNA Mensageiro/análise , Receptores de Superfície Celular/análise , Receptores de Superfície Celular/genética , Receptores Toll-Like , Transativadores/genéticaRESUMO
Five lacY mutants with amber stop codons at known positions were each placed into 12 different suppressor strains. The 60 amino acid substitutions obtained in this manner were tested for growth on lactose-minimal medium plates and for transport of lactose, melibiose, and thiomethylgalactoside. Most of the amino acid substitutions in the regions of the putative loops (between transmembrane alpha helices) resulted in a reasonable growth rate on lactose with moderate-to-good transport activity. In one strain (glycine substituted for Trp-10), abnormal sugar recognition was found. The substitution of proline for Trp-33 (in the region of the first alpha helix) showed no activity, while four additional substitutions (lysine, leucine, cysteine, and glutamic acid) showed low activity. Altered sugar specificity was observed when Trp-33 was replaced by serine, glutamine, tyrosine, alanine, histidine, or phenylalanine. It is concluded that Trp-33 may be involved directly or indirectly in sugar recognition.
Assuntos
Aminoácidos/genética , Proteínas de Escherichia coli , Proteínas de Membrana Transportadoras/genética , Proteínas de Transporte de Monossacarídeos , Supressão Genética , Simportadores , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/metabolismo , Imuno-Histoquímica , Lactose/metabolismo , Melibiose/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Metilgalactosídeos/metabolismo , Dados de Sequência Molecular , Mutação , Tiogalactosídeos/metabolismoRESUMO
A new member of the NAP/SET gene family, named MB20, was isolated from a mouse brain cDNA library by virtue of its CAG trinucleotide repetitive sequence and a brain-specific gene expression pattern. The complementary DNA sequence predicted an open reading frame of 545 amino acids, with four copies of an 11-amino-acid direct repeat. The consensus sequence for these repeats, PKE-P--K-EE, is present in the largest subunit of murine neurofilament (NF-H). The MB20 protein sequence is homologous to nucleosome assembly proteins of several species, and its C-terminus is homologous to SET proteins. Immunoblot analysis revealed that MB20 protein is expressed in the brain. Transient transfection and immunofluorescence microscopy demonstrated that MB20 is distributed in the cytoplasm as well as in the nucleus. Deletion of the N-terminal end imparts the complete localization of MB20 protein to the nucleus. The ability of MB20 to bind histone proteins was analyzed by sucrose gradient sedimentation and by retention of histone proteins by immobilized MB20 protein. On the basis of its expression pattern, predicted sequence, and protein properties, we propose that MB20 plays a unique role in modulating nucleosome structure and gene expression during brain development.