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1.
Braz J Med Biol Res ; 57: e13309, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38656073

RESUMO

Diabetic-metabolic syndrome (MetS-D) has a high prevalence worldwide, in which an association with the rupture of the intestinal epithelium barrier function (IEBF) has been pointed out, but the functional and morphological properties are still not well understood. This study aimed to evaluate the impact of acute hyperglycemia diabetes on intestinal tight junction proteins, metabolic failure, intestinal ion and water transports, and IEBF parameters. Diabetes was induced in male Rattus norvegicus (200-310 g) with 0.5 mL of streptozotocin (70 mg/kg). Glycemic and clinical parameters were evaluated every 7 days, and intestinal parameters were evaluated on the 14th day. The MetS-D animals showed a clinical pattern of hyperglycemia, with increases in the area of villi and crypts, lactulose:mannitol ratio, myeloperoxidase (MPO) activity, and intestinal tissue concentrations of malondialdehyde (MDA), but showed a reduction in reduced glutathione (GSH) when these parameters were compared to the control. The MetS-D group had increased secretion of Na+, K+, Cl-, and water compared to the control group in ileal tissue. Furthermore, we observed a reduction in mRNA transcript of claudin-2, claudin-15, and NHE3 and increases of SGLT-1 and ZO-1 in the MetS-D group. These results showed that MetS-D triggered intestinal tissue inflammation, oxidative stress, complex alterations in gene regulatory protein transcriptions of intestinal transporters and tight junctions, damaging the IEBF and causing hydroelectrolyte secretion.


Assuntos
Diabetes Mellitus Experimental , Hiperglicemia , Mucosa Intestinal , Junções Íntimas , Animais , Masculino , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Diabetes Mellitus Experimental/metabolismo , Hiperglicemia/metabolismo , Junções Íntimas/metabolismo , Ratos , Inflamação/metabolismo , Modelos Animais de Doenças , Ratos Wistar , Síndrome Metabólica/metabolismo , Síndrome Metabólica/fisiopatologia
2.
Braz J Med Biol Res ; 56: e12946, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37909497

RESUMO

The role of cyclooxygenase (COXs) isoforms in maintaining colonic mucosal integrity is not fully understood. This study aimed to evaluate the role of COX-1 and -2 on colonic mucosal integrity in an experimental colitis model. Colitis was induced in Wistar rats by intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (20 mg + 50% ethanol). The control group (sham group) received saline only. After 7, 14, or 28 days, colonic samples were removed, and macroscopic lesion scores, wet weight, myeloperoxidase activity, and transepithelial electrical resistance (TER) were determined. In other rat groups, colonic samples from the sham group and a 7th day post-colitis group were mounted in Üssing chambers with the luminal side exposed to a buffer solution (control), acetylsalicylic acid (ASA), SC-560 (COX-1 inhibitor), or celecoxib (COX-2 inhibitor). TER and epithelial permeability to fluorescein were measured. The 7th day colitis group had higher macroscopic damage scores, wet weight, and myeloperoxidase activity and lower basal TER than the sham, 14th day colitis, and 28th day colitis groups. Inhibition of COX-1 but not COX-2 significantly decreased TER and increased permeability to fluorescein in the 7th day post-colitis group compared to the sham group. Additionally, ASA decreased the colonic mucosal integrity on day seven post-colitis compared to the sham group. A decrease in the colonic mucosa integrity in the experimental colitis model can be aggravated only by the inhibition of COX-1, which demonstrated the importance of this enzyme in the maintenance of colonic mucosal integrity.


Assuntos
Colite , Peroxidase , Ratos , Animais , Ratos Wistar , Colite/induzido quimicamente , Colite/patologia , Mucosa Intestinal , Aspirina , Ciclo-Oxigenase 2 , Fluoresceínas
3.
Braz. j. med. biol. res ; 56: e12946, 2023. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1520470

RESUMO

The role of cyclooxygenase (COXs) isoforms in maintaining colonic mucosal integrity is not fully understood. This study aimed to evaluate the role of COX-1 and -2 on colonic mucosal integrity in an experimental colitis model. Colitis was induced in Wistar rats by intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (20 mg + 50% ethanol). The control group (sham group) received saline only. After 7, 14, or 28 days, colonic samples were removed, and macroscopic lesion scores, wet weight, myeloperoxidase activity, and transepithelial electrical resistance (TER) were determined. In other rat groups, colonic samples from the sham group and a 7th day post-colitis group were mounted in Üssing chambers with the luminal side exposed to a buffer solution (control), acetylsalicylic acid (ASA), SC-560 (COX-1 inhibitor), or celecoxib (COX-2 inhibitor). TER and epithelial permeability to fluorescein were measured. The 7th day colitis group had higher macroscopic damage scores, wet weight, and myeloperoxidase activity and lower basal TER than the sham, 14th day colitis, and 28th day colitis groups. Inhibition of COX-1 but not COX-2 significantly decreased TER and increased permeability to fluorescein in the 7th day post-colitis group compared to the sham group. Additionally, ASA decreased the colonic mucosal integrity on day seven post-colitis compared to the sham group. A decrease in the colonic mucosa integrity in the experimental colitis model can be aggravated only by the inhibition of COX-1, which demonstrated the importance of this enzyme in the maintenance of colonic mucosal integrity.

4.
Braz J Med Biol Res ; 53(5): e9211, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32321150

RESUMO

Strenuous exercise triggers deleterious effects on the intestinal epithelium, but their mechanisms are still uncertain. Here, we investigated whether a prolonged training and an additional exhaustive training protocol alter intestinal permeability and the putative effect of alanyl-glutamine (AG) pretreatment in this condition. Rats were allocated into 5 different groups: 1) sedentary; 2 and 3) trained (50 min per day, 5 days per week for 12 weeks) with or without 6 weeks oral (1.5 g/kg) AG supplementation; 4 and 5) trained and subjected to an additional exhaustive test protocol with or without oral AG supplementation. Venous blood samples were collected to determine gasometrical indices at the end of the 12-week protocol or after exhaustive test. Lactate and glucose levels were determined before, during, and after the exhaustive test. Ileum tissue collected after all experimental procedures was used for gene expression analysis of Zonula occludens 1 (ZO-1), occludin, claudin-2, and oligopeptide transporter 1 (PepT-1). Intestinal permeability was assessed by urinary lactulose/mannitol test collected after the 12-week protocol or the exhaustive test. The exhaustive test decreased pH and base excess and increased pCO2. Training sessions delayed exhaustion time and reduced the changes in blood glucose and lactate levels. Trained rats exhibited upregulation of PEPT-1, ZO-1, and occludin mRNA, which were partially protected by AG. Exhaustive exercise induced intestinal paracellular leakage associated with the upregulation of claudin-2, a phenomenon protected by AG treatment. Thus, AG partially prevented intestinal training adaptations but also blocked paracellular leakage during exhaustive exercise involving claudin-2 and occludin gene expression.


Assuntos
Dipeptídeos/administração & dosagem , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/fisiopatologia , Permeabilidade/efeitos dos fármacos , Condicionamento Físico Animal/fisiologia , Animais , Masculino , Modelos Animais , Ratos , Ratos Wistar
5.
Braz. j. med. biol. res ; 53(5): e9211, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1098114

RESUMO

Strenuous exercise triggers deleterious effects on the intestinal epithelium, but their mechanisms are still uncertain. Here, we investigated whether a prolonged training and an additional exhaustive training protocol alter intestinal permeability and the putative effect of alanyl-glutamine (AG) pretreatment in this condition. Rats were allocated into 5 different groups: 1) sedentary; 2 and 3) trained (50 min per day, 5 days per week for 12 weeks) with or without 6 weeks oral (1.5 g/kg) AG supplementation; 4 and 5) trained and subjected to an additional exhaustive test protocol with or without oral AG supplementation. Venous blood samples were collected to determine gasometrical indices at the end of the 12-week protocol or after exhaustive test. Lactate and glucose levels were determined before, during, and after the exhaustive test. Ileum tissue collected after all experimental procedures was used for gene expression analysis of Zonula occludens 1 (ZO-1), occludin, claudin-2, and oligopeptide transporter 1 (PepT-1). Intestinal permeability was assessed by urinary lactulose/mannitol test collected after the 12-week protocol or the exhaustive test. The exhaustive test decreased pH and base excess and increased pCO2. Training sessions delayed exhaustion time and reduced the changes in blood glucose and lactate levels. Trained rats exhibited upregulation of PEPT-1, ZO-1, and occludin mRNA, which were partially protected by AG. Exhaustive exercise induced intestinal paracellular leakage associated with the upregulation of claudin-2, a phenomenon protected by AG treatment. Thus, AG partially prevented intestinal training adaptations but also blocked paracellular leakage during exhaustive exercise involving claudin-2 and occludin gene expression.


Assuntos
Animais , Masculino , Ratos , Permeabilidade/efeitos dos fármacos , Condicionamento Físico Animal/fisiologia , Dipeptídeos/administração & dosagem , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/fisiopatologia , Ratos Wistar , Modelos Animais
6.
Arq. bras. med. vet. zootec. (Online) ; 71(5): 1488-1496, set.-out. 2019. tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-1038649

RESUMO

A ordem dos Passeriformes é uma das mais pressionadas pelas ações antrópicas, especialmente as relativas ao tráfico de animais, que, devido às más condições de manejo e higiênico-sanitárias, favorecem a infecção dos espécimes por patógenos virulentos e zoonóticos, como cepas de Escherichia coli e Salmonella spp., cujo isolamento em suabes cloacais, bem como a análise dos genes de virulência das cepas de E. coli foram objetivos do estudo. Para isso, 120 Passeriformes silvestres nativos, recebidos pelo Cetas/CE, foram avaliados individualmente. As cepas isoladas foram submetidas a teste de disco difusão para determinação da sensibilidade aos antimicrobianos. Em etapa posterior, foi realizada PCR para a detecção de oito genes de virulência dos principais patotipos diarreiogênicos de E. coli. Quanto aos resultados, nenhuma cepa de Salmonella spp. foi isolada, no entanto a ocorrência de E. coli foi de 40,8%. Foi observada elevada resistência, principalmente aos antimicrobianos tetraciclina, ampicilina e sulfazotrim, ocorrendo multirresistência em 42,8% das cepas. Pela análise molecular, foram diagnosticados quatro entre os nove genes pesquisados, com a identificação de EPEC típicas, EPEC atípicas, ETEC, EHEC e EAEC. Os resultados apontam para a importância de Passeriformes como possíveis disseminadores de zoonoses.(AU)


The order Passeriformes is one of the most pressured by anthropic actions, especially those related to animal trafficking. Due to poor sanitary and hygienic conditions, the infection of the specimens is favored by virulent and zoonotic pathogens such as strains of Escherichia coli and Salmonella spp., whose isolation in cloacal swabs as well as the analysis of the virulence genes of E. coli strains were the objectives of the study. For this, 120 native wild Passeriformes, received by CETAS/CE were individually evaluated. The isolated strains were submitted to diffusion disc test to determine sensitivity to antimicrobials. In a later stage, PCR was performed for the detection of eight virulence genes from the main E. coli diarrhoeagenic pathogens. Regarding the results, no strain of Salmonella spp. was isolated; however, the occurrence of E. coli was 40.8%. High resistance was observed, mainly to the antimicrobials Tetracycline, Ampicillin and Sulfazotrim, with multi-resistance in 42.8% of the strains. By molecular analysis, four of the nine genes were diagnosed, identifying typical EPEC, atypical EPEC, ETEC, EHEC and EAEC. The results point to the importance of Passeriformes as possible disseminators of zoonoses.(AU)


Assuntos
Salmonella/isolamento & purificação , Salmonella/patogenicidade , Passeriformes/parasitologia , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Animais Selvagens/parasitologia
7.
Curr Top Med Chem ; 19(22): 2049-2057, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31364515

RESUMO

BACKGROUND: Scorpion venom causes renal injury and affects vascular ion-channels function. Centruroides margaritatus scorpion is found in Colombia and is frequently the cause of envenomation accidents; however, its renal impact has never been investigated. OBJECTIVE: To evaluate the effects of C. margaritatus venom (CmV) on renal parameters using isolated rat kidney and renal cell culture models. METHODS: Wistar rats (n = 5, weighing 240-300 g) were first perfused with Krebs-Henseleit solution containing 6 g 100 mL-1 bovine serum albumin. After 30 minutes, the kidneys were perfused with CmV to a final concentration of 10 µgmL-1; evaluation was performed by measuring Perfusion Pressure (PP), Renal Vascular Resistance (RVR), Urinary Flow (UF), Glomerular Filtration Rate (GFR), and percentage of electrolyte tubular transport. Moreover, kidney histological analyses and cell cytotoxicity in renal tubule epithelial cells (MDCK) and proximal tubular cells (LLC-MK2) were assessed. RESULTS: CmV increased PP and RVR 60 min after perfusion. On the other hand, UF, GFR, and the percentages of sodium, potassium and chloride tubular transport decreased after experimental envenomation. UF dropped after 120 min, while GFR and percentage of electrolyte tubular transport diminished after 60, 90 and 120 min. CmV was not toxic to MDCK cell line but reduced the viability of LLC-MK2 cells at concentrations ranging from 6.25 to 200 µgmL-1. Histological analyses disclosed hydropic degeneration, edema, and protein deposits. Flow cytometry disclosed that cell death occurred predominantly by necrosis. CONCLUSION: Our results suggest that C. margaritatus venom can trigger renal impairment, mainly in the proximal kidney tubule.


Assuntos
Rim/efeitos dos fármacos , Venenos de Escorpião/farmacologia , Animais , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colômbia , Cães , Relação Dose-Resposta a Droga , Rim/patologia , Células Madin Darby de Rim Canino/efeitos dos fármacos , Células Madin Darby de Rim Canino/patologia , Masculino , Ratos , Ratos Wistar , Escorpiões , Relação Estrutura-Atividade
8.
Braz J Med Biol Res ; 52(6): e8589, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31166385

RESUMO

The transport of myo-inositol is the main mechanism for the maintenance of its high intracellular levels. We aimed to measure the mRNA and protein levels of myo-inositol cotransporters in the sciatic nerve (SN) and dorsal root ganglia (DRG) during experimental diabetes. Streptozotocin-induced (STZ; 4, 8, and 12 weeks; 65 mg/kg; ip) diabetic rats (DB) and age-matched euglycemic (E) rats were used for the analysis of mRNA and protein levels of sodium myo-inositol cotransporters 1, 2 (SMIT1, SMIT2) or H+/myo-inositol cotransporter (HMIT). There was a significant reduction in the mRNA levels for SMIT1 in the SN and DRG (by 36.9 and 31.0%) in the 4-week DB (DB4) group compared to the E group. SMIT2 was not expressed in SN. The mRNA level for SMIT2 was up-regulated only in the DRG in the DB4 group. On the other hand, the protein level of SMIT1 decreased by 42.5, 41.3, and 44.8% in the SN after 4, 8, and 12 weeks of diabetes, respectively. In addition, there was a decrease of 64.3 and 58.0% of HMIT in membrane and cytosolic fractions, respectively, in the SN of the DB4 group. In the DRG, there was an increase of 230 and 86.3% for SMIT1 and HMIT, respectively, in the DB12 group. The levels of the main inositol transporters, SMIT1 and HMIT, were greatly reduced in the SN but not in the DRG. SMIT-1 was selectively reduced in the sciatic nerve during experimental STZ-induced diabetes.


Assuntos
Transporte Biológico Ativo/fisiologia , Diabetes Mellitus Experimental/metabolismo , Gânglios Espinais/metabolismo , Inositol/metabolismo , RNA Mensageiro/metabolismo , Nervo Isquiático/metabolismo , Animais , Western Blotting , Masculino , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estreptozocina , Regulação para Cima
9.
Braz. j. med. biol. res ; 52(6): e8589, 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1011585

RESUMO

The transport of myo-inositol is the main mechanism for the maintenance of its high intracellular levels. We aimed to measure the mRNA and protein levels of myo-inositol cotransporters in the sciatic nerve (SN) and dorsal root ganglia (DRG) during experimental diabetes. Streptozotocin-induced (STZ; 4, 8, and 12 weeks; 65 mg/kg; ip) diabetic rats (DB) and age-matched euglycemic (E) rats were used for the analysis of mRNA and protein levels of sodium myo-inositol cotransporters 1, 2 (SMIT1, SMIT2) or H+/myo-inositol cotransporter (HMIT). There was a significant reduction in the mRNA levels for SMIT1 in the SN and DRG (by 36.9 and 31.0%) in the 4-week DB (DB4) group compared to the E group. SMIT2 was not expressed in SN. The mRNA level for SMIT2 was up-regulated only in the DRG in the DB4 group. On the other hand, the protein level of SMIT1 decreased by 42.5, 41.3, and 44.8% in the SN after 4, 8, and 12 weeks of diabetes, respectively. In addition, there was a decrease of 64.3 and 58.0% of HMIT in membrane and cytosolic fractions, respectively, in the SN of the DB4 group. In the DRG, there was an increase of 230 and 86.3% for SMIT1 and HMIT, respectively, in the DB12 group. The levels of the main inositol transporters, SMIT1 and HMIT, were greatly reduced in the SN but not in the DRG. SMIT-1 was selectively reduced in the sciatic nerve during experimental STZ-induced diabetes.


Assuntos
Animais , Masculino , Ratos , Nervo Isquiático/metabolismo , Transporte Biológico Ativo/fisiologia , RNA Mensageiro/metabolismo , Diabetes Mellitus Experimental/metabolismo , Gânglios Espinais/metabolismo , Inositol/metabolismo , Regulação para Cima , Western Blotting , Estreptozocina , Reação em Cadeia da Polimerase Via Transcriptase Reversa
10.
Braz. j. med. biol. res ; 52(1): e7581, 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-974275

RESUMO

Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3′,4′-OH flavonoids are known inhibitors of phospholipases A2.


Assuntos
Animais , Masculino , Ratos , Antivenenos/farmacologia , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Venenos de Crotalídeos/antagonistas & inibidores , Edema/tratamento farmacológico , Hemorragia/etiologia , Antivenenos/isolamento & purificação , Bothrops , Venenos de Crotalídeos/toxicidade , Polygalaceae/química , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Edema/etiologia , Hemorragia/tratamento farmacológico
11.
Braz J Med Biol Res ; 52(1): e7581, 2018 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-30517287

RESUMO

Bredemeyera floribunda roots are popularly used to treat snakebites in the semiarid region of Northeast Brazil, and previous studies indicate the anti-ophidian actions of triterpenoid saponins found in its roots. To assess B. floribunda root extract (BFRE) activity against the effects of Bothrops jararacussu venom (BjuV), antiphospholipasic, antiproteolytic, antihemorrhagic, antinecrotic, and anti-edematogenic activities were investigated in mice. Phytochemical analysis revealed the presence of saponins, flavonoids, and sugars, with rutin and saccharose being the major constituents of BFRE. Acute toxicity was determined and BFRE was nontoxic to mice. Phospholipase A2 and proteolytic activities induced by BjuV were inhibited in vitro by BFRE at all concentrations tested herein. BFRE (150 mg/kg) inhibited paw edema induced by BjuV (50 µg/animal), reducing total edema calculated by area under the curve, but carrageenan-induced paw edema was unchanged. Hemorrhagic and necrotizing actions of BjuV (50 µg/animal) were considerably decreased by BFRE treatment. Thus, BFRE blocked the toxic actions of B. jararacussu venom despite having no anti-inflammatory activity, which points to a direct inhibition of venom's toxins, as demonstrated in the in vitro assays. The larger amounts of rutin found in BFRE may play a role in this inhibition, since 3',4'-OH flavonoids are known inhibitors of phospholipases A2.


Assuntos
Antivenenos/farmacologia , Venenos de Crotalídeos/antagonistas & inibidores , Edema/tratamento farmacológico , Hemorragia/etiologia , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Polygalaceae/química , Animais , Antivenenos/isolamento & purificação , Bothrops , Venenos de Crotalídeos/toxicidade , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Edema/etiologia , Hemorragia/tratamento farmacológico , Masculino , Ratos
12.
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
13.
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
14.
Arq. bras. med. vet. zootec. (Online) ; 69(5): 1236-1242, set.-out. 2017. ilus, tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-878737

RESUMO

This study reports a co-infection of Escherichia coli and Salmonella in a free-living ruddy ground dove (Columbina talpacoti) received at the Laboratory of Ornithological Studies of the State University of Ceará, Brazil. The bird presented diarrhea, leg paralysis and anorexia, and died shortly after. Necropsy was then performed and samples from lung, kidney, liver and intestine were collected for microbiological and histopathological analyses. Escherichia coli was isolated from cloacal swab, lung and kidney samples. Salmonella ser. Saintpaul was identified in liver and spleen samples. Escherichia coli isolates were tested for the presence of eight diagnostic genes for diarrheagenic pathotypes (STEC, ETEC, EPEC, EIEC, EAEC) with conventional polymerase chain reaction (PCR). EAEC was detected in the lung and kidney, and STEC in the intestine. In conclusion, Columbina talpacoti is susceptible to enteroaggregative Escherichia coli and Salmonella ser. Saintpaul infection, which may have public health implications.(AU)


Este estudo relata um caso de coinfecção por Escherichia coli e Salmonella ser. Saintpaul em uma rolinha-roxa (Columbina talpacoti) recebida pelo Laboratório de Estudos Ornitológicos da Universidade Estadual do Ceará, Brasil. A ave apresentava diarreia, paralisia nas pernas e anorexia, indo a óbito rapidamente. A necropsia foi realizada e amostras de pulmão, rim, fígado e intestino foram coletados para isolamento microbiológico e análise histopatológica. Escherichia coli foi identificada em amostras de suabe cloacal, pulmão e rim. Salmonella ser. Saintpaul foi identificada no fígado e baço. Isolados de E. coli foram testados para a presença de oito genes de diagnóstico para patotipos diarreiogênicos (STEC, ETEC, EPEC, EIEC, EAEC) através de reação em cadeia de polimerase (PCR) convencional. EAEC foi detectada no pulmão e rim, e STEC foi identificada no intestino. Em conclusão, Columbina talpacoti é suscetível a infecção por Escherichia coli enteroagregativa e Salmonella ser. Saintpaul, o que pode implicar em risco para a saúde pública.(AU)


Assuntos
Animais , Coinfecção/veterinária , Columbidae , Farmacorresistência Bacteriana , Infecções por Escherichia coli/veterinária , Salmonelose Animal/diagnóstico
15.
J Transl Sci ; 2(2): 134-139, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27746954

RESUMO

Fecal biomarkers have emerged as important tools to assess intestinal inflammation and enteropathy. The aim of this study was to investigate the correlations between the fecal markers, myeloperoxidase (MPO), lactoferrin (FL), calprotectin (FC) and lipocalin-2 (Lcn-2), and to compare differences by breastfeeding status as well as normalization by fecal protein or by fecal weight. Simultaneous, quantitative MPO, FL, FC and Lcn-2, levels were determined in frozen fecal specimens collected from 78 children (mean age 15.2 ± 5.3 months) in a case-control study of childhood malnutrition in Brazil. The biomarker concentrations were measured by enzymelinked immunosorbent assay. The correlations among all biomarkers were significant (P<0.01). There were stronger correlations of fecal MPO with fecal lactoferrin and calprotectin, with lower, but still highly significant correlations of all 3 inflammatory biomarkers with Lcn-2 likely because the latter may also reflect enterocyte damage as well as neutrophil presence. Furthermore, the biomarker results with protein normalized compared to simple fecal weight normalized values showed only a slightly better correlation suggesting that the added cost and time for protein normalization added little to carefully measured fecal weights as denominators. In conclusion, fecal MPO correlates tightly with fecal lactoferrin and calprotectin irrespective of breastfeeding status and provides a common, available biomarker for comparison of human and animal model studies.

16.
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
17.
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
18.
Toxicon ; 90: 134-47, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25127849

RESUMO

Local tissue reactions provoked by Bothrops venoms are characterized by edema, hemorrhage, pain, and inflammation; however, the mechanisms of tissue damage vary depending upon the species of snake. Here, we investigated the mechanisms involved in the local inflammatory response induced by the Bothrops jararacussu venom (BjcuV). Female Swiss mice were injected with either saline, BjcuV (0.125-8 µg/paw) or loratadine (an H1 receptor antagonist), compound 48/80 (for mast cell depletion), capsaicin (for C-fiber desensitization), infliximab (an anti-TNF-α antibody), indomethacin (a non-specific COX inhibitor), celecoxib (a selective COX-2 inhibitor) or fucoidan (a P- and L-selectins modulator) given before BjcuV injection. Paw edema was measured by plethysmography. In addition, paw tissues were collected for the measurement of myeloperoxidase activity, TNF-α and IL-1 levels, and COX-2 immunoexpression. The direct chemotactic effect of BjcuV and the in vitro calcium dynamic in neutrophils were also investigated. BjcuV caused an edematogenic response with increased local production of TNF-α and IL-1ß as well as COX-2 expression. Both edema and neutrophil migration were prevented by pretreatment with indomethacin, celecoxib or fucoidan. Furthermore, BjcuV induced a direct in vitro neutrophil chemotaxis by increasing intracellular calcium. Therefore, BjcuV induces an early onset edema dependent upon prostanoid production and neutrophil migration.


Assuntos
Venenos de Crotalídeos/farmacologia , Inflamação/induzido quimicamente , Neutrófilos/efeitos dos fármacos , Prostaglandinas/metabolismo , Animais , Bothrops , Quimiotaxia de Leucócito/efeitos dos fármacos , Ciclo-Oxigenase 2/metabolismo , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-1beta/metabolismo , Camundongos , Neutrófilos/imunologia , Fator de Necrose Tumoral alfa/metabolismo
19.
J Appl Microbiol ; 117(2): 390-6, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24848589

RESUMO

AIMS: Dinoponera quadriceps venom (DqV) was examined to evaluate the antibacterial activity and its bactericidal action mechanism against Staphylococcus aureus. METHODS AND RESULTS: DqV was tested against a standard strain of methicillin-sensitive Staphylococcus aureus (MSSA), Staph. aureus ATCC 6538P and two standard strains of methicillin-resistant Staphylococcus aureus (MRSA), Staph. aureus ATCC 33591 and Staph. aureus CCBH 5330. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the rate of kill and pH sensitivity of the DqV were determined by microdilution tests. Bactericidal and inhibitory concentrations of DqV were tested to check its action on Staph. aureus membrane permeability and cell morphology. The MIC and MBC of DqV were 6·25 and 12·5 µg ml(-1) for Staph. aureus ATCC 6538P, 12·5 and 50 µg ml(-1) for Staph. aureus CCBH 5330 and 100 and 100 µg ml(-1) for Staph. aureus ATCC 33591, respectively. Complete bacterial growth inhibition was observed after 4 h of incubation with the MBC of DqV. A lowest MIC was observed in alkaline pH. Alteration in membrane permeability was observed through the increase in crystal violet uptake, genetic material release and morphology in atomic force microscopy. CONCLUSIONS: The results suggest antibacterial activity of DqV against Staph. aureus and that the venom acts in the cell membrane. SIGNIFICANCE AND IMPACT OF THE STUDY: Alteration in membrane permeability may be associated with the antimicrobial activity of hymenopteran venoms.


Assuntos
Venenos de Formiga/farmacologia , Antibacterianos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Animais , Formigas
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