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1.
Braz J Med Biol Res ; 51(10): e7423, 2018 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-30066727

RESUMO

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.


Assuntos
Movimento Celular/fisiologia , Escherichia coli Enteropatogênica/patogenicidade , Células Epiteliais/microbiologia , Sistemas de Secreção Tipo III/fisiologia , Fatores de Virulência/genética , Proteínas rho de Ligação ao GTP/fisiologia , Apoptose , Western Blotting , Citometria de Fluxo , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Virulência/fisiologia
2.
Sports Med Open ; 4(1): 7, 2018 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-29388074

RESUMO

ERRATUM: The original article [1] mistakenly omits a grant acknowledgement; thus, the authors would like to acknowledge that the original article was supported by FAPESP 2015/20082-7.

3.
Braz. j. med. biol. res ; 51(10): e7423, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-951708

RESUMO

Epithelial cell migration is an essential response to enteric pathogens such as enteropathogenic Escherichia coli (EPEC). This study aimed to investigate the effects of EPEC infection on intestinal epithelial cell migration in vitro, as well as the involvement of type III secretion system (T3SS) and Rho GTPases. Crypt intestinal epithelial cells (IEC-6) were infected with EPEC strains (E2348/69, ΔescF, and the LDI001 strain isolated from a malnourished Brazilian child) and commensal E. coli HS. Wound migration and cell death assays were performed at different time-points. Transcription and expression of Rho GTPases were evaluated using real-time PCR and western blotting. Overall, EPEC E2348/69 reduced migration and increased apoptosis and necrosis levels compared to EPEC LDI001 and E. coli HS strains. Moreover, EPEC LDI001 impaired cell migration at a higher level than E. coli HS and increased necrosis after 24 hours compared to the control group. The different profiles of virulence genes between the two wild-type EPEC strains, characterized by the absence of espL and nleE genes in the LDI001, might explain the phenotypic results, playing significant roles on cell migration impairment and cell death-related events. Moreover, the type III secretion system is determinant for the inhibition of intestinal epithelial cell migration by EPEC 2348/69, as its deletion prevented the effect. Active Rac1 concentrations were increased in E2348/69 and LDI001-infected cells, while the T3SS-deficient strain did not demonstrate this activation. This study contributes with valuable insight to characterize the mechanisms involved in the impairment of intestinal cell migration induced by EPEC.


Assuntos
Humanos , Movimento Celular/fisiologia , Proteínas rho de Ligação ao GTP/fisiologia , Fatores de Virulência/genética , Células Epiteliais/microbiologia , Escherichia coli Enteropatogênica/patogenicidade , Sistemas de Secreção Tipo III/fisiologia , Western Blotting , Apoptose , Fatores de Virulência/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Citometria de Fluxo
4.
Braz J Med Biol Res ; 49(10): e5340, 2016 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-27737316

RESUMO

Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Crescimento/fisiologia , Íleo/anatomia & histologia , Íleo/metabolismo , Desnutrição/metabolismo , Desnutrição/fisiopatologia , Doença Aguda , Animais , Peso Corporal , Modelos Animais de Doenças , Ingestão de Energia/fisiologia , Immunoblotting , Absorção Intestinal/fisiologia , Transporte de Íons/fisiologia , Masculino , Desnutrição/complicações , Proteínas de Membrana Transportadoras/análise , Camundongos , RNA Mensageiro , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas de Junções Íntimas/análise , Proteínas de Junções Íntimas/metabolismo , Fatores de Tempo
5.
Braz. j. med. biol. res ; 49(10): e5340, 2016. tab, graf
Artigo em Inglês | LILACS | ID: biblio-951651

RESUMO

Undernutrition represents a major public health challenge for middle- and low-income countries. This study aimed to evaluate whether a multideficient Northeast Brazil regional basic diet (RBD) induces acute morphological and functional changes in the ileum of mice. Swiss mice (∼25 g) were allocated into two groups: i) control mice were fed a standard diet and II) undernourished mice were fed the RBD. After 7 days, mice were killed and the ileum collected for evaluation of electrophysiological parameters (Ussing chambers), transcription (RT-qPCR) and protein expression (western blotting) of intestinal transporters and tight junctions. Body weight gain was significantly decreased in the undernourished group, which also showed decreased crypt depth but no alterations in villus height. Electrophysiology measurements showed a reduced basal short circuit current (Isc) in the undernourished group, with no differences in transepithelial resistance. Specific substrate-evoked Isc related to affinity and efficacy (glutamine and alanyl-glutamine) were not different between groups, except for the maximum Isc (efficacy) induced by glucose. Transcription of Sglt1 and Pept1 was significantly higher in the undernourished group, while SN-2 transcription was decreased. No changes were found in transcription of CAT-1 and CFTR, while claudin-2 and occludin transcriptions were significantly increased in the undernourished group. Despite mRNA changes, SGLT-1, PEPT-1, claudin-2 and occludin protein expression showed no difference between groups. These results demonstrate early effects of the RBD on mice, which include reduced body weight and crypt depth in the absence of significant alterations to villus morphology, intestinal transporters and tight junction expression.


Assuntos
Animais , Masculino , Coelhos , Desnutrição/fisiopatologia , Desnutrição/metabolismo , Crescimento/fisiologia , Íleo/anatomia & histologia , Íleo/metabolismo , Fenômenos Fisiológicos da Nutrição Animal , Fatores de Tempo , Peso Corporal , Ingestão de Energia/fisiologia , RNA Mensageiro , Immunoblotting , Doença Aguda , Transporte de Íons/fisiologia , Desnutrição/complicações , Modelos Animais de Doenças , Absorção Intestinal/fisiologia
6.
Artigo em Inglês | MEDLINE | ID: mdl-23366321

RESUMO

This paper presents a medical remote monitoring application which aims at detecting falls. The detection system is based on three modalities: a wearable sensor, infrared sensors and a sound analysis module. The sound analysis is presented briefly. The multimodal fusion is made using the Dempster Schaffer theory through Evidential Network. A first evaluation of the use of data mining techniques in order to extract blindly data representatives is proposed. These representatives are used to continuously increase the system performances. The system is evaluated on a local recorded data base.


Assuntos
Acidentes por Quedas/prevenção & controle , Actigrafia/métodos , Algoritmos , Inteligência Artificial , Mineração de Dados/métodos , Monitorização Ambulatorial/métodos , Telemedicina/métodos , Acidentes por Quedas/estatística & dados numéricos , Sistemas de Apoio a Decisões Clínicas , Humanos
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