RESUMO
The Continuous Visual Attention Test (CVAT) is a test that detects visuomotor reaction time (RT, alertness), variability of reaction time (VRT, sustained attention), omission errors (OE, focused attention), and commission errors (CE, response inhibition). The standard test takes 15 min, while the ultrafast version only 90 s. Besides overall task length, the two versions differ by target probability (20% and 80% in the 15-min vs. only 80% in the 90-s test) and stimulus-onset asynchrony (SOA) (1, 2, and 4 s in the 15-min vs. only 1 s in the 90-s test. We aimed to analyze the effect of target probability, SOA, and time length on the CVAT variables across the 15-min task and to verify correlations and agreements between the 15-min and the 90-s CVATs. 205 healthy participants performed the two CVATs on the same day. Considering the 15-min task, RT and CE were strongly affected by target probability. Conversely, VRT was not affected. When the 15-min task was compared to the 90-s task, we found no significant difference in the VRT variable. Additionally, a significant agreement between the two tasks was found for the VRT variable. We concluded that sustained attention can be measured with the 90-s CVAT.
Assuntos
Atenção , Tempo de Reação , Humanos , Atenção/fisiologia , Tempo de Reação/fisiologia , Masculino , Feminino , Adulto , Adulto Jovem , Desempenho Psicomotor/fisiologia , Testes Neuropsicológicos , Adolescente , Pessoa de Meia-Idade , Percepção Visual/fisiologia , Estimulação LuminosaRESUMO
Introdução: A lipoenxertia é um enxerto autólogo de células do tecido celular subcutâneo, que pode ser utilizada como técnica complementar na reconstrução mamária. Diante disso, a criopreservação de células-tronco mesenquimais provenientes de tecido adiposo (CTDAs) poderia ser uma maneira de realizar a coleta em um tempo cirúrgico e após realizar a lipoenxertia de forma fracionada. O dimetilsulfóxido (DMSO) é um criopreservante utilizado em pesquisas com células, porém é potencialmente tóxico, o que impossibilitaria a utilização de CTDAs criopreservadas na prática clínica. Novos criopreservantes celulares, sem toxicidade, vêm sendo descritos na literatura científica experimental, como as substâncias L-prolina e trealose. Com isso, esse trabalho teve como objetivo avaliar a viabilidade de CTDAs criopreservadas com a combinação de L-prolina e trealose, em um período de até 90 dias. Método: Estudo experimental, no qual foram obtidas amostras de lipoaspirado provenientes de 9 pacientes. A fração celular foi processada e congelada com L-prolina (1,5M) + trealose (0,2M), ou com DMSO + soro fetal bovino (SFB), como controle. Após 30 e 90 dias, as amostras foram descongeladas e a viabilidade celular foi avaliada pela técnica de MTT. Resultados: A análise das CTDAs, após 1 e 3 meses de congelamento, indicou que as amostras tratadas com L-prolina + trealose apresentaram viabilidade semelhante àquelas preservadas com DMSO e SFB (p=0,444). Conclusão: A associação de L-prolina e trealose manteve CTDA viáveis por 30 e 90 dias de congelamento, podendo ser uma alternativa como criopreservante celular sem toxicidade e viabilizando o uso de lipoenxertia seriada.
Introduction: Fat grafting is an autologous graft of cells from subcutaneous tissue, which can be used as a complementary technique in breast reconstruction. Given this, the cryopreservation of adipose tissue-derived mesenchymal stem cells (ADMSCs) could be a way to collect them in one surgical procedure and after performing fractional fat grafting. Dimethyl sulfoxide (DMSO) is a cryopreservative used in cell research, but it is potentially toxic, which would make it impossible to use cryopreserved ADMSCs in clinical practice. New cellular cryopreservatives, without toxicity, have been described in the experimental scientific literature, such as the substances L-proline and trehalose. Therefore, this work aimed to evaluate the viability of ADMSCs cryopreserved with the combination of L-proline and trehalose over up to 90 days. Method: Experimental study in which lipoaspirate samples were obtained from 9 patients. The cellular fraction was processed and frozen with L-proline (1.5M) + trehalose (0.2M) or with DMSO + fetal bovine serum (FBS) as control. After 30 and 90 days, the samples were thawed, and cell viability was assessed using the MTT technique. Results: The analysis of ADMSCs, after 1 and 3 months of freezing, indicated that samples treated with L-proline + trehalose showed similar viability to those preserved with DMSO and SFB (p=0.444). Conclusion: The association of L-proline and trehalose kept ADMSC viable for 30 and 90 days of freezing, and could be an alternative as a cellular cryopreservative without toxicity and enabling the use of serial fat grafting.
RESUMO
Previous studies have shown that COVID-19 inpatients exhibited significant attentional deficits on the day of discharge. However, the presence of gastrointestinal symptoms (GIS) has not been evaluated. Here, we aimed to verify: (1) whether COVID-19 patients with GIS exhibited specific attention deficits; (2) which attention subdomain deficits discriminated patients with GIS and without gastrointestinal symptoms (NGIS) from healthy controls. On admission, the presence of GIS was recorded. Seventy-four physically functional COVID-19 inpatients at discharge and sixty-eight controls underwent a Go/No-go computerized visual attentional test (CVAT). A Multivariate Analysis of Covariance (MANCOVA) was performed to examine group differences in attentional performance. To discriminate which attention subdomain deficits discriminated GIS and NGIS COVID-19 patients from healthy controls, a discriminant analysis was applied using the CVAT variables. The MANCOVA showed a significant overall effect of COVID-19 with GIS on attention performance. The discriminant analysis indicated that the GIS group could be differentiated from the controls by variability of reaction time and omissions errors. The NGIS group could be differentiated from controls by reaction time. Late attention deficits in COVID-19 patients with GIS may reflect a primary problem in the sustained and focused attention subsystems, whereas in NGIS patients the attention problems are related to the intrinsic-alertness subsystem.
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Cerebellar manifestations have been described in patients with gluten sensitivity (GS)-related disorders. A better understanding of the neurological manifestations of GS requires the use of neuroimaging techniques. We performed a systematic review on neuroimaging findings in GS patients with cerebellar symptoms. We also included a specific search on neuroimaging findings in GS patients with cerebellar manifestations on a gluten-free diet (GFD). PubMed, Embase, and Bireme were systematically searched to identify studies assessing neuroimaging features of adults with cerebellar manifestations and GS with or without enteropathy on a GFD. Ten studies with a total of 222 adult-GS patients were included. Magnetic resonance imaging was used in 100% of the studies. Cerebellar atrophy was evaluated in 7 studies and observed in 63% of the patients. White matter abnormalities were described in 2 studies. Single-photon emission computed tomography was used in 2 studies, and decreased cerebellar perfusion was detected in 92% of the included patients. No study employed nuclear medicine after the start of GFD. Magnetic resonance spectroscopy (MRS) was performed in 2 studies before and after GFD. An increase in the Naa/Cr ratio in cerebellar vermis was seen in 98% of the cases on a strict GFD. Cerebellar atrophy was found to be a prevalent condition in GS patients. MRS demonstrated to be useful in the follow-up of GS patients with cerebellar manifestations on a GFD. Prospective studies using nuclear medicine imaging are needed to study brain changes in GS patients on a GFD.
Assuntos
Doença Celíaca , Dieta Livre de Glúten , Adulto , Humanos , Doença Celíaca/diagnóstico por imagem , Estudos Prospectivos , Cerebelo/diagnóstico por imagem , Atrofia , Neuroimagem , Glutens/efeitos adversosRESUMO
Neste estudo foram identificadas espécies de Candida em isolados de secreção vaginal, avaliados os perfis de suscetibilidade in vitro a antifúngicos e correlacionados com os antifúngicos prescritos para pacientes em um serviço de atenção primária. A identificação das espécies pela Reação em Cadeia da Polimerase mostrou que 36,5% dos isolados foram caracterizados como espécie não-C.albicans. Nos testes de sensibilidade a maioria dos isolados foi suscetível a cetoconazol, fluconazol e itraconazol, contudo cerca de 40% e 50% apresentaram resistência ou sensibilidade dose-dependente a miconazol e nistatina, respectivamente. A análise dos fármacos prescritos para as pacientes revelou que 34,2% dos isolados foram considerados resistentes aos agentes utilizados no tratamento. Diversas espécies de Candida podem causar vulvovaginite com variados perfis de suscetibilidade aos antifúngicos comumente utilizados no tratamento. A identificação das espécies de Candida é relevante para o gerenciamento epidemiológico das infecções, além de ser útil, assim como os testes de suscetibilidade, na escolha do tratamento farmacológico mais eficaz para a paciente.
In this study were identified Candida species from vaginal secretion isolates, evaluated their in vitro antifungal susceptibilities, and correlated these features with antifungal agents prescribed for patients assisted in a primary care service. Species identification by Polymerase Chain Reaction showed that 36.5% of isolates were characterized as non-C. albicans species. In antifungal susceptibility tests most isolates were susceptible to ketoconazole, fluconazole, and itraconazole, although between 40% and 50% of isolates show resistance or dose-dependent susceptibility to miconazole and nystatin, respectively. Analysis of drugs prescribed to patients revealed that 34.2% of the isolates were considered resistant to agents used in treatment. Several Candida species can cause vulvovaginitis and exhibit different susceptibility profiles to antifungal drugs used in treatment. The identification of Candida species is relevant and useful to the epidemiological management of infections. The antifungal susceptibility test may also be useful for choosing most effective drug treatment for each patient.
RESUMO
BACKGROUND: The Clinical Dementia Rating (CDR) scale is commonly used to stage cognitive impairment, despite having educational limitations. In elderly with low education, a previous study has shown that intraindividual variability of reaction time (CV) and commission errors (CE), measured using a culture-free Go/No-Go task, can reliably distinguish early Alzheimer's disease (AD) from mild cognitive impairment (MCI) and healthy controls. OBJECTIVE: We aimed to extend the clinical utility of this culture-free Go/No-Go task in a sample with high educational disparity. METHODS: One hundred and ten participants with a wide range of years of formal education (0-14 years) were randomly selected from a geriatric unit and divided based on their CDR scores into cognitively unimpaired (CDRâ=â0), MCI (CDRâ=â0.5), and early AD (CDRâ=â1). All underwent a 90-s reaction-time test that measured the variables previously found to predict CDR in low educated elderly. Here we added years of formal education (educational level) to the model. Multivariate analyses compared differences in group means using educational level as confounding factor. A confirmatory discriminant analyses was performed, to assess if CDR scores could be predicted by the two Go/No-Go variables in a sample with high educational disparity. RESULTS: Over all three groups, differences in both CE and CV reached statistical significance (pâ<â0.05). The discriminant analysis demonstrated that CV and CE discriminated cognitively impaired from cognitively normal elderly. These results remained similar when discriminating MCI from cognitively unimpaired elderly. CONCLUSION: The Go/No-Go task reliably discriminates elderly with MCI from elderly without cognitive impairment independent of educational disparity.
Assuntos
Doença de Alzheimer/fisiopatologia , Atenção/fisiologia , Transtornos Cognitivos/fisiopatologia , Disfunção Cognitiva/fisiopatologia , Análise e Desempenho de Tarefas , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/diagnóstico , Cognição/fisiologia , Transtornos Cognitivos/diagnóstico , Disfunção Cognitiva/diagnóstico , Progressão da Doença , Escolaridade , Feminino , Humanos , Masculino , Testes de Estado Mental e Demência , Índice de Gravidade de DoençaRESUMO
Introdução: A lipoenxertia é uma alternativa com importante aplicabilidade para reconstrução de mama e/ou correções de assimetrias decorrentes do tratamento oncológico. Esta técnica consiste na transferência de gordura autóloga, cujo estroma contém células-tronco derivadas do tecido adiposo que tem capacidade de diferenciar-se em toda a linhagem mesodermal. Para o preparo do tecido adiposo, Coleman fundamentou a centrifugação, de material aspirado por seringa, em 3000 rotações por minuto (rpm) durante 3 minutos. Contudo, estudos questionam se velocidades menores de centrifugação poderiam ser menos deletérias para viabilidade celular. Métodos: Foi realizado um estudo experimental, onde foram avaliadas as células adiposas de seis pacientes; a partir de 60mL de lipoaspirado de cada um. A amostra coletada foi fracionada em quatro tubos, e submetidos a diferentes protocolos, decantação e centrifugação nas velocidades 500, 1000 e 3000rpm por 3 minutos. Após as amostras foram processadas com colagenase IA por 30 min, submetidas ao cultivo celular por 24 horas e realizado a análise da viabilidade celular. Os resultados foram tabulados e analisados pelo teste ANOVA utilizando os programas Graphpad Prism 6.0® e SAS®. Resultados: A viabilidade celular foi maior na amostra celular centrifugada a 3000rpm e menor na amostra decantada. A coloração com Giemsa indicou manutenção da morfologia celular entre as amostras. Conclusão: As células centrifugadas na velocidade de 3000rpm apresentaram maior viabilidade celular. A centrifugação foi efetiva na compactação do tecido e eliminação de resíduos indesejados (sangue e óleo residual).
Introduction: Lipografting is an alternative with important applicability for breast reconstruction and/or corrections of asymmetries resulting from cancer treatment. This technique consists of autologous fat transfer, whose stroma contains stem cells derived from adipose tissue that can differentiate itself throughout the mesodermal lineage. For adipose tissue preparation, Coleman-based centrifugation of syringe-aspirated material at 3000 revolutions per minute (rpm) for 3 minutes. However, studies question whether lower centrifugation speeds could be less harmful to cell viability. Methods: An experimental study was conducted to evaluate the adipose cells of six patients; from 60mL of liposuction of each one. The sample collected was fractionated into four tubes and submitted to different protocols, decanting and centrifugation at speeds 500, 1000, and 3000rpm for 3 minutes. Afterward, the samples were processed with collagenase IA for 30 min, submitted to cell culture for 24 hours, and a cell viability analysis. The results were tabulated and analyzed by the ANOVA test using the Graphpad Prism 6.0® and SAS®. Results: Cell viability was higher in the cell sample centrifuged at 3000rpm and lower in the decanted sample. Giemsa staining indicated maintenance of cell morphology on the samples. Conclusion: Centrifuged cells at a speed of 3000rpm showed higher cell viability. Centrifugation was effective in compacting tissue and eliminating unwanted waste (blood and residual oil).
RESUMO
Celiac disease (CD) is an immune-mediated systemic disorder triggered by gluten and related prolamins in genetically predisposed individuals. Here, we described a case of a 31-year-old Caucasian woman who exhibited cerebellar and psychiatric dysfunctions. The patient underwent single-photon emission computed tomography (SPECT-CT) before and after a gluten-free diet (GFD). There was an improvement in cerebellar perfusion accompanied by a remission of cerebellar manifestations. The maintenance of the psychiatric manifestations was related to the persistence of the hypoperfusion in the frontal lobes. The patient's psychiatric symptoms did not change after 4 months under a GFD in the hospital. To our knowledge, this is the first case that shows the relationship between improvement in cerebellar perfusion and remission of cerebellar clinical manifestations in a CD patient under a GFD.
Assuntos
Doença Celíaca , Ataxia Cerebelar , Adulto , Doença Celíaca/complicações , Doença Celíaca/diagnóstico por imagem , Ataxia Cerebelar/complicações , Ataxia Cerebelar/diagnóstico por imagem , Dieta Livre de Glúten , Feminino , Humanos , Tomografia Computadorizada de Emissão de Fóton Único , Tomografia Computadorizada por Raios XRESUMO
BACKGROUND: Scales for cognitive deterioration usually depend on education level. OBJECTIVE: We aimed to study the clinical utility of a culture-free Go/No-Go task in a multi-ethnic cohort with low education level. METHODS: Sixty-four participants with less than 4 years of formal education were included and divided on the basis of their Clinical-Dementia-Rate scores (CDR) into cognitively unimpaired (CDRâ=â0), mild cognitive impairment (MCI; CDRâ=â0.5), and early Alzheimer's disease (AD, CDRâ=â1). All underwent a 90-s Continuous Visual Attention Test. This test consisted of a 90-s Go/No-go task with 72 (80%) targets and 18 (20%) non-targets. For each participant, reaction times and intraindividual variability of reaction times of all correct target responses, as well as the number of omission and commission errors were evaluated. Coefficient of variability was calculated for each participant by dividing the standard deviation of the reaction times by the mean reaction time. A MANCOVA was performed to examine between-group differences using age and sex as covariates. Discriminate analysis was performed to find the most reliable test-variable to discriminate the three groups. RESULTS: Commission error, intraindividual variability of reaction time, and coefficient of variability progressively worsened with increasing CDR level. Discriminant analysis demonstrated that coefficient of variability was the best discriminant factor, followed by intraindividual variability of reaction time and commission error. CONCLUSION: The Go/No-Go task was able to discriminate people with MCI or early AD from controls in the setting of illiteracy.
Assuntos
Doença de Alzheimer/diagnóstico , Atenção , Disfunção Cognitiva/diagnóstico , Alfabetização , Testes Neuropsicológicos , Tempo de Reação , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/fisiopatologia , Disfunção Cognitiva/fisiopatologia , Escolaridade , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: In the early phases of multiple sclerosis (MS), patients exhibit slight neuropsychiatric deficits that can only be detected using reliable tools. AIM: The present investigation aimed to examine neuropsychological performance in 35 patients with incipient MS. PATIENTS AND METHODS: For the MS group, the inclusion criteria included time of disease <3 years and low disability. The neuropsychological battery consisted of Rey Auditory Learning Test, Controlled Oral Word Association Test, Hooper Visual Organization Test, and Symbol Digit Modalities Test (SDMT). RESULTS: After correction for the educational level, no significant effect of MS on performance was found for all the tests except for the number of errors of the SDMT (NE-SDMT). Higher levels of education were associated with better performances in all tests, except for the NE-SDMT. MS patients made more errors than the controls. CONCLUSION: The effect on the NE-SDMT may reflect difficulties in the ability to inhibit inadequate responses. Patients may exhibit impulsive control disorders in incipient MS, independent of their educational level.
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The protozoan Trypanosoma cruzi is a parasite exposed to several environmental stressors inside its invertebrate and vertebrate hosts. Although stress conditions are involved in its differentiation processes, little information is available about the stress response proteins engaged in these activities. This work reports the first known association of the stress-inducible protein 1 (STI1) with the cellular differentiation process in a unicellular eukaryote. Albeit STI1 expression is constitutive in epimastigotes and metacyclic trypomastigotes, higher protein levels were observed in late growth phase epimastigotes subjected to nutritional stress. Analysis by indirect immunofluorescence revealed that T. cruzi STI1 (TcSTI1) is located throughout the cell cytoplasm, with some cytoplasmic granules appearing in greater numbers in late growing epimastigotes and late growing epimastigotes subjected to nutritional stress. We observed that part of the fluorescence signal from both TcSTI1 and TcHSP70 colocalized around the nucleus. Gene silencing of sti1 in Trypanosoma brucei did not affect cell growth. Similarly, the growth of T. cruzi mutant parasites with a single allele sti1 gene knockout was not affected. However, the differentiation of epimastigotes in metacyclic trypomastigotes (metacyclogenesis) was compromised. Lower production rates and numbers of metacyclic trypomastigotes were obtained from the mutant parasites compared with the wild-type parasites. These data indicate that reduced levels of TcSTI1 decrease the rate of in vitro metacyclogenesis, suggesting that this protein may participate in the differentiation process of T. cruzi.
Assuntos
Proteínas de Choque Térmico/metabolismo , Proteínas de Protozoários/genética , Trypanosoma cruzi/crescimento & desenvolvimento , Trypanosoma cruzi/fisiologia , Técnica Indireta de Fluorescência para Anticorpo , Técnicas de Inativação de Genes , Inativação Gênica , Proteínas de Choque Térmico/genética , Estágios do Ciclo de Vida/genética , Estágios do Ciclo de Vida/fisiologia , Mutação , Proteínas de Protozoários/metabolismo , Estresse Fisiológico , Trypanosoma cruzi/química , Trypanosoma cruzi/ultraestruturaRESUMO
Mental stress-induced myocardial ischemia (MSIMI) has been described in patients with acute coronary syndrome (ACS). We reported the case of a patient admitted with ACS and MSIMI related to generalized anxiety disorder (GAD). Myocardial perfusion imaging (MPI) after exercise stress and computed tomography coronary angiography showed normal results. In contrast, MPI with mental stress documented MSIMI. He began to improve 7 days after starting escitalopram and remained without angina at 1-year follow-up. These data suggest the applicability of mental stress radionuclide imaging for the detection of MSIMI in patients with ACS and GAD, even in normal coronary arteries.
Assuntos
Síndrome Coronariana Aguda/diagnóstico por imagem , Transtornos de Ansiedade/psicologia , Vasos Coronários/diagnóstico por imagem , Isquemia Miocárdica/diagnóstico por imagem , Estresse Psicológico/psicologia , Síndrome Coronariana Aguda/etiologia , Síndrome Coronariana Aguda/psicologia , Transtornos de Ansiedade/complicações , Transtornos de Ansiedade/tratamento farmacológico , Citalopram/uso terapêutico , Angiografia por Tomografia Computadorizada , Angiografia Coronária , Teste de Esforço , Humanos , Masculino , Isquemia Miocárdica/etiologia , Isquemia Miocárdica/psicologia , Imagem de Perfusão do Miocárdio , Inibidores Seletivos de Recaptação de Serotonina/uso terapêutico , Estresse Psicológico/complicações , Estresse Psicológico/tratamento farmacológico , Adulto JovemRESUMO
BACKGROUND: Limbic encephalitis was originally described as a rare clinical neuropathological entity involving seizures and neuropsychological disturbances. In this report, we describe cerebral patterns visualized by positron emission tomography in a patient with limbic encephalitis and cholangiocarcinoma. To our knowledge, there is no other description in the literature of cerebral positron emission tomography findings in the setting of limbic encephalitis and subsequent diagnosis of cholangiocarcinoma. CASE PRESENTATION: We describe a case of a 77-year-old Caucasian man who exhibited persistent cognitive changes 2 years before his death. A cerebral scan obtained at that time by 2-deoxy-2-[fluorine-18]fluoro- D -glucose integrated with computed tomography-positron emission tomography showed low radiotracer uptake in the frontal and temporal lobes. Cerebrospinal fluid analysis indicated the presence of voltage-gated potassium channel antibodies. Three months before the patient's death, a lymph node biopsy indicated a cholangiocarcinoma, and a new cerebral scan obtained by 2-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography-positron emission tomography showed an increment in the severity of metabolic deficit in the frontal and parietal lobes, as well as hypometabolism involving the temporal lobes. Two months before the patient's death, cerebral metastases were detected on a contrast-enhanced computed tomographic scan. Postmortem examination revealed a cholangiocarcinoma with multiple metastases including the lungs and lymph nodes. The patient's brain weighed 1300 g, and mild cortical atrophy, ex vacuo dilation of the ventricles, and mild focal thickening of the cerebellar leptomeninges, which were infiltrated by neoplastic epithelial cells, were observed. CONCLUSIONS: These findings support the need for continued vigilance in malignancy surveillance in patients with limbic encephalitis and early cerebral positron emission tomographic scan abnormalities. The difficulty in early diagnosis of small tumors, such as a cholangiocarcinoma, is discussed in the context of the clinical utility of early cerebral hypometabolism detected by 2-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography-positron emission tomography in patients with rapidly progressive dementia.
Assuntos
Neoplasias dos Ductos Biliares/complicações , Encefalopatias/complicações , Encefalopatias/diagnóstico por imagem , Colangiocarcinoma/complicações , Fluordesoxiglucose F18 , Encefalite Límbica/complicações , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Idoso , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Evolução Fatal , Humanos , Masculino , Compostos RadiofarmacêuticosRESUMO
The life cycle of the protozoan Trypanosoma cruzi exposes it to several environmental stresses in its invertebrate and vertebrate hosts. Stress conditions are involved in parasite differentiation, but little is known about the stress response proteins involved. We report here the first characterization of stress-induced protein-1 (STI-1) in T. cruzi (TcSTI-1). This co-chaperone is produced in response to stress and mediates the formation of a complex between the stress proteins HSP70 and HSP90 in other organisms. Despite the similarity of TcSTI-1 to STI-1 proteins in other organisms, its expression profile in response to various stress conditions, such as heat shock, acidic pH or nutrient starvation, is quite different. Neither polysomal mRNA nor protein levels changed in exponentially growing epimastigotes cultured under any of the stress conditions studied. Increased levels of TcSTI-1 were observed in epimastigotes subjected to nutritional stress in the late growth phase. Co-immunoprecipitation assays revealed an association between TcSTI-1 and TcHSP70 in T. cruzi epimastigotes. Immunolocalization demonstrated that TcSTI-1 was distributed throughout the cytoplasm and there was some colocalization of TcSTI-1 and TcHSP70 around the nucleus. Thus, TcSTI-1 associates with TcHSP70 and TcSTI-1 expression is induced when the parasites are subjected to stress conditions during specific growth phase.
Assuntos
Proteínas de Choque Térmico/metabolismo , Trypanosoma cruzi/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Imunofluorescência , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico HSP90/genética , Proteínas de Choque Térmico HSP90/metabolismo , Proteínas de Choque Térmico/genética , ImunoprecipitaçãoRESUMO
The life cycle of the protozoan Trypanosoma cruzi exposes it to several environmental stresses in its invertebrate and vertebrate hosts. Stress conditions are involved in parasite differentiation, but little is known about the stress response proteins involved. We report here the first characterization of stress-induced protein-1 (STI-1) in T. cruzi (TcSTI-1). This co-chaperone is produced in response to stress and mediates the formation of a complex between the stress proteins HSP70 and HSP90 in other organisms. Despite the similarity of TcSTI-1 to STI-1 proteins in other organisms, its expression profile in response to various stress conditions, such as heat shock, acidic pH or nutrient starvation, is quite different. Neither polysomal mRNA nor protein levels changed in exponentially growing epimastigotes cultured under any of the stress conditions studied. Increased levels of TcSTI-1 were observed in epimastigotes subjected to nutritional stress in the late growth phase. Co-immunoprecipitation assays revealed an association between TcSTI-1 and TcHSP70 in T. cruzi epimastigotes. Immunolocalization demonstrated that TcSTI-1 was distributed throughout the cytoplasm and there was some colocalization of TcSTI-1 and TcHSP70 around the nucleus. Thus, TcSTI-1 associates with TcHSP70 and TcSTI-1 expression is induced when the parasites are subjected to stress conditions during specific growth phase.