O exerc?cio intervalado de alta intensidade induz desequil?brio redox em c?lulas mononucleares do sangue perif?rico e reduz a resposta proliferativa de linf?citos ao est?mulo superantig?nico por altera??o da propor??o de subpopula??es linfocit?rias

O exerc?cio intervalado de alta intensidade (HIIE) ? caracterizado por breves e repetidas sess?es de exerc?cio intenso, intercaladas por per?odos de repouso ou exerc?cio de baixa intensidade. ? uma modalidade de exerc?cio que tem ganhado muito destaque nos ?ltimos anos por ser uma modalidade de treinamento de baixo volume, embora pouco se saiba a respeito dos seus efeitos na fun??o imune e no estado redox celular. Assim, este estudo avaliou o efeito de uma sess?o de HIIE sobre o estado redox e a fun??o de linf?citos em homens jovens sedent?rios. Esse trabalho foi dividido em dois estudos. No estudo 1 avaliou-se o efeito do HIIE sobre a prolifera??o e produ??o de citocinas e o estado redox de c?lulas mononucleares do sangue perif?rico (PBMC). No estudo 2 avaliou-se o efeito do HIIE sobre a viabilidade e express?o de marcadores de ativa??o em linf?citos. As sess?es de HIIE foram realizadas em bicicleta ergom?trica, e consistiram em oito s?ries de 1 min a 90-100% de pot?ncia pico, com 75 segundos de recupera??o ativa, a 30W, entre as s?ries. O sangue venoso foi colhido antes, imediatamente ap?s e 30 minutos ap?s a sess?o de HIIE. Para avalia??o da prolifera??o celular, por citometria de fluxo, as PBMC foram coradas com Carboxifluoresce?na Succinimidil Ester (CFSE) (10 ?M) e estimuladas com o superant?geno SEB (100 ng/mL), durante 5 dias a 37? C, 5% de CO2. A produ??o de IL-2 e IFN-? em resposta a estimula??o por SEB durante 18 horas, foi avaliada por ELISA. O estado redox celular foi avaliado pela mensura??o da atividade das enzimas catalase (CAT) e super?xido dismutase (SOD), a concentra??o de subst?ncias que reagem ao ?cido tiobarbit?rico (TBARS) e conte?do de glutationa reduzida (GSH). Para avalia??o da viabilidade celular, por citometria de fluxo, as c?lulas foram marcadas com anticorpos anti-Anexina V FITC e iodeto de prop?deo. Para an?lise da ativa??o dos linf?citos, por citometria de fluxo, as PBMC foram estimuladas com SEB por 18 horas, e em seguida marcadas com anticorpos fluorescentes dirigidos contra CD4, CD8, CD19, CD25 e CD69. Os dados foram analisados utitlizando os testes one-way ou two-way ANOVA, considerando p ? 0,05. O HIIE promoveu redu??o na prolifera??o de linf?citos (p = 0,01), aumento na concentra??o de IL-2 (p = 0,02), e desequil?brio redox nas PBMC, marcado por aumento nas concentra??es de TBARS (p = 0,02) e diminui??o na atividade da CAT (p = 0,04). O HIIE n?o alterou a viabilidade das PBMC, mas a frequ?ncia de c?lulas CD4 e CD19, positivas para os marcadores CD25 e CD69, foi menor ap?s o exerc?cio. Contudo, como foi observada redu??o na frequ?ncia de c?lulas CD4+ e CD19+, a redu??o da frequ?ncia de express?o de marcadores de ativa??o refletiu a redu??o do n?mero de c?lulas, e n?o da resposta ao est?mulo superantig?nico. Nossos resultados mostram portanto, que apesar do HIIE promover desequil?brio redox nas PBMC a resposta dos linf?citos ao est?mulo superantig?nico n?o ? alterada. A redu??o da resposta proliferativa ?, provavelmente, reflexo da altera??o da distribui??o das subpopula??es linfocit?rias em decorr?ncia do HIIE.Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015.ABSTRACT High-intensity interval exercise (HIIE) is characterized by brief and repeated intense exercise sessions, interspersed with periods of rest or low intensity exercise. This exercise modality has gained much attention in recent years, although little is known about its effects on immune function and cellular redox state. This study evaluated the effect of HIIE on the redox state and lymphocyte function in sedentary young males. This work was divided in two studies. The firsty study evaluated the effect of HIIE on peripheral blood mononuclear cells (PBMC) proliferation, cytokine production and redox status. The second study evaluated the effect of HIIE on lymphocyte viability and activation markers expression. HIIE was performed on cycloergometer, and consisted of eight series of 1 min at 90-100% of peak power, with 75 seconds of active recovery, at 30W, between sets. Venous blood was collected before, immediately after and 30 minutes after HIIE. For cell proliferation evaluation by flow cytometry, PBMC were stained with Carboxyfluorescein Succinimidyl Ester (CFSE) (10 mM) and stimulated with the superantigen SEB (100 ng/ml) for 5 days at 37? C, 5% CO2. IL-2 and IFN-? secretion in response to SEB stimulation for 18 hours was assessed by ELISA. The cellular redox status was assessed by measuring the activity of catalase (CAT) and superoxide dismutase (SOD), the concentration of thiobarbituric acid reactive substances (TBARS) and reduced glutathione content (GSH). To assess cellular viability, using flow cytometry, cells were labeled with anti-Annexin V-FITC and propidium iodide. To analyze lymphocyte activation, by flow cytometry, PBMC were stimulated with SEB for 18 hours, and then stained with fluorescent antibodies directed against CD4, CD8, CD19, CD25 and CD69. One-way or two-way ANOVA was employed for statistical analyzis, with ? ? 0.05. Lymphocyte proliferation was reduced (p = 0.01) after HIIE, despite increased IL-2 concentration (p = 0.02), and HIIE also induced PBMC redox imbalance characterized by increased TBARS concentration (p = 0.02) and decreased CAT activity (p = 0.04). PBMC viability was not affected by HIIE, but the frequency of CD4+CD25+/CD69+ and CD19+CD25+/CD69+ cells in response to SEB stimulation was lower after exercise. However, as CD4+ and CD19+ frequencies were reduced, reduced activation markers expression was a consequence of cell number reduction. Our results therefore show that, although HIIE induced redox imbalance, lymphocyte response to superantigen stimulation was not affected. Reduced lymphocyte proliferative response after HIIE is probably due to modifications in lymphocyte subpopulations distribution

Meios não farmacológicos utilizados por fisioterapeutas para conforto e alívio da dor em unidades de terapia intensiva neonatal no estado de Minas Gerais

ResumoIntrodução: o neonato tem baixa sensibilidade de processar o estímulo nociceptivo da dor, logo, os procedimentos dolorosos repetidos que ocorrem dentro das Unidades de Terapia Intensiva Neonatais podem ocasionar prejuízos e levar à piora do seu quadro clínico. Dessa forma, recursos e técnicas não farmacológicas são indicados para amenizar possíveis prejuízos ao bebê na Unidade de Terapia Intensiva Neonatal. Objetivo: avaliar a utilização dos meios não farmacológicos para conforto e alívio da dor aplicados pelos fisioterapeutas dentro das Unidades de Terapia Intensiva Neonatais do estado de Minas Gerais. Métodos: trata-se de um estudo descritivo transversal, aprovado pelo Comitê de Ética em Pesquisa da Universidade Vale do Rio Doce, no qual fisioterapeutas que trabalham em Unidades de Terapia Intensiva Neonatais de Minas Gerais responderam a um questionário online acerca dos principais métodos não farmacológicos por eles utilizados. Resultados: participaram do presente estudo, 61 fisioterapeutas divididos nas mesorregiões do estado de Minas Gerais. As intervenções mais utilizadas dentro das Unidades de Terapia Intensiva Neonatais foram posicionamento e método canguru, seguido por sucção não nutritiva, massagem terapêutica, banho de ofurô e redeterapia. Os principais benefícios relatados pelos fisioterapeutas foram estabilidade clínica e hemodinâmica, conforto e alívio da dor, melhora do desenvolvimento neuropsicomotor, ganho de peso e diminuição do tempo de internação. Em relação aos critérios considerados para a realização dos métodos, foram citados instabilidade hemodinâmica, extremo baixo peso e entubação, e os empecilhos foram prematuridade extrema, alterações de pele e falta de capacitação da equipe. Conclusão: todos os métodos utilizados apresentaram diversos benefícios quanto ao conforto e alívio da dor, além de proporcionar calma e influenciar no desenvolvimento neuropsicomotor dos neonatos. Implicações clínicas: a prática desses métodos traz conforto e alívio da dor, o que implica em diversos benefícios ao neonato, fazendo com que o tempo de internação seja voltado para um cuidado humanizado

Lymphocyte Redox Imbalance and Reduced Proliferation after a Single Session of High Intensity Interval Exercise

<div><p>This study investigated whether an acute session of high-intensity interval training (HIIT) is sufficient to alter lymphocyte function and redox status. Sixteen young healthy men underwent a HIIT session on a cycloergometer, consisting of eight bouts of 1 min at 90–100% of peak power, with 75 seconds of active recovery at 30 W between bouts. Venous blood was collected before, immediately after, and 30 minutes after the HIIT session. In response to <i>Staphylococcus aureus</i> superantigen B (SEB) stimulation, lymphocyte proliferation decreased and the IL-2 concentration increased after the HIIT session. However, the HIIT session had no effect on lymphocyte proliferation or IL-2 response to phytohemagglutinin stimulation. The HIIT session also induced lymphocyte redox imbalance, characterized by an increase in the concentration of thiobarbituric acid reactive substances and a decrease in the activity of the antioxidant enzyme catalase. Lymphocyte viability was not affected by the HIIT session. The frequencies of CD25⁺ and CD69⁺ T helper and B lymphocytes in response to superantigen stimulation were lower after exercise, suggesting that superantigen-induced lymphocyte activation was reduced by HIIT. However, HIIT also led to a reduction in the frequency of CD4⁺ and CD19⁺ cells, so the frequencies of CD25⁺ and CD69⁺ cells within the CD4 and CD19 cell populations were not affected by HIIT. These data indicate that the reduced lymphocyte proliferation observed after HIIT is not due to reduced early lymphocyte activation by superantigen. Our findings show that an acute HIIT session promotes lymphocyte redox imbalance and reduces lymphocyte proliferation in response to superantigenic, but not to mitogenic stimulation. This observation cannot be explained by alteration of the early lymphocyte activation response to superantigen. The manner in which lymphocyte function modulation by an acute HIIT session can affect individual immunity and susceptibility to infection is important and requires further investigation.</p></div

HIIT effect on cytotoxic T CD8 cell activation by SEB.

<p><b>(A)</b> Frequency of CD8^highCD25⁺ lymphocytes. <b>(B)</b> Frequency of CD8^highCD69⁺ lymphocytes. Data shown as mean ± SE. pre-ex = before exercise, post-ex = immediately after exercise, 30 min post-ex = 30 min after exercise. N = 6.</p

HIIT effect on TCD4 activation by SEB.

<p><b>(A)</b> Frequency of CD4⁺CD25⁺ lymphocytes. <b>(B)</b> Frequency of CD4⁺CD69⁺ lymphocytes. <b>(C)</b> Frequency of CD4⁺CD25⁺ cells within the T helper (CD4⁺) cell subpopulation. <b>(D)</b> Frequency of CD4⁺CD69⁺ cells within the T helper (CD4⁺) cell subpopulation. Data shown as mean ± SE. # P <0.05, compared to non-stimulated cells in all moments. *P < 0.05, compared to pre-ex. ^§P < 0.05, compared to post-ex. Two way Anova, Tukey <i>post-hoc</i>. pre-ex = before exercise, post-ex = immediately after exercise, 30 min post-ex = 30 min after exercise. N = 6.</p

HIIT effect on lymphocyte proliferative response.

<p><b>(A)</b> SEB-stimulated cells. <b>(B)</b> PHA-stimulated cells. Data shown as mean ± SE. ^#P < 0.05, compared to non-stimulated cells, in all moments. *P < 0.05, compared to pre-ex, Anova-two way, <i>Tukey post-hoc</i>. pre-ex = before exercise, post-ex = immediately after exercise, 30 min post-ex = 30 min after exercise. N = 10.</p

Analytic strategy used to evaluate lymphocyte GSH content by flow cytometry.

<p>Profile of size (FSC) and granularity (SSC) (A). Fluorescence intensity histograms of non-labeled (control) (B) and Thiol Tracker-labeled cells (C).</p

Heart rate and perceived exertion during the HIIT sessions.

<p>Heart rate and perceived exertion during the HIIT sessions.</p

HIIT effect on IL-2 concentration in culture supernatant.

<p><b>(A)</b> SEB-stimulated cells. <b>(B)</b> PHA-stimulated cells. Data shown as mean ± SE. #P < 0.05, compared to non-stimulated cells, in all moments. *P < 0.05, compared to pre-ex. Anova-two way, Tukey <i>post-hoc</i>. pre-ex = before exercise, post-ex = immediately after exercise, 30 min post-ex = 30 min after exercise. N = 10.</p

Physical and physiological characteristics of volunteers.

<p>Physical and physiological characteristics of volunteers.</p