Smoking history: relationships with inflammatory markers, metabolic markers, body composition, muscle strength, and cardiopulmonary capacity in current smokers / Relação da carga tabágica com marcadores inflamatórios, marcadores metabólicos, composição corporal, força muscular e capacidade cardiorrespiratória em tabagistas

ABSTRACT Objective: To determine the relationships that smoking history has with inflammatory markers, metabolic markers, body composition, muscle strength, and cardiopulmonary capacity in current smokers. Methods: This was a cross-sectional study involving 65 smokers (age range: 18-60 years). On three non-consecutive days, each participant was evaluated in terms of smoking history, pre-existing comorbidities, lung function (by spirometry), peripheral muscle strength (by dynamometry), body composition (by bioelectrical impedance analysis), levels of metabolic/inflammatory markers, and maximum cardiopulmonary capacity (by treadmill exercise test). We evaluated the relationships that smoking history has with inflammatory markers, metabolic markers, body composition, muscle strength, and cardiopulmonary capacity, using logarithmic transformation of the data and calculating Pearson's correlation coefficient and for partial correlations adjusted for age, gender, body mass index (BMI), and comorbidities. To identify the influence of smoking history on pre-existing comorbidities, we used a logistic regression model adjusted for age, BMI, and duration of smoking. Results: Smoking history correlated significantly, albeit weakly, with triglyceride level (r = 0.317; p = 0.005), monocyte count (r = 0.308; p = 0.013), and waist circumference (r = 0.299; p = 0.017). However, those correlations did not retain their significance in the adjusted analysis. In the logistic regression model, smoking more than 20 cigarettes/day correlated significantly with the presence of metabolic diseases (OR = 0.31; 95% CI: 1.009-1.701; p = 0.043). Conclusions: In this sample of smokers, smoking history correlated positively with the triglyceride level, the monocyte count, and waist circumference. The prevalence of metabolic disease was highest in those who smoked more than 20 cigarettes/day.

RESUMO Objetivo: Verificar a relação da carga tabágica com marcadores inflamatórios, marcadores metabólicos, composição corporal, força muscular e capacidade cardiorrespiratória em tabagistas. Métodos: Estudo transversal com 65 tabagistas de ambos os sexos (idade: 18-60 anos). Todos os participantes foram avaliados em três dias não consecutivos quanto ao histórico de tabagismo, comorbidades pré-existentes, função pulmonar (espirometria), força muscular periférica (dinamometria), composição corporal (bioimpedância), dosagem de marcadores metabólicos e inflamatórios e teste cardiopulmonar em esteira para avaliar a capacidade cardiorrespiratória máxima. Avaliou-se a relação da carga tabágica com marcadores inflamatórios, marcadores metabólicos, composição corporal, força muscular e capacidade cardiorrespiratória com transformação logarítmica através da correlação de Pearson e correlações parciais ajustadas para idade, sexo, índice de massa corpórea (IMC) e comorbidades. A regressão logística com modelo ajustado para idade, IMC e tempo de tabagismo foi utilizada para identificar a influência do histórico de tabagismo sobre as comorbidades pré-existentes. Resultados: Observaram-se correlações positivas fracas somente para dados não ajustados da carga tabágica com nível de triacilglicerol (r = 0,317; p = 0,005), contagem de monócitos (r = 0,308; p = 0,013) e circunferência abdominal (r = 0,299; p = 0,017). No modelo de regressão logística, fumar mais de 20 cigarros/dia correlacionou-se significativamente com a presença de doenças metabólicas (OR = 0,31; IC95%: 1,009-1,701; p = 0,043). Conclusões: Nesta amostra de tabagistas, a carga tabágica se correlacionou positivamente com nível de triacilglicerol, contagem de monócitos e circunferência abdominal. A prevalência de doenças metabólicas foi maior em tabagistas que fumam mais de 20 cigarros/dia.

Lipopolysaccharide-stimulated intracellular cytokines and depressive symptoms in community-dwelling older adults

Abstract Background Inflammation is involved in the pathophysiology of depression, and circulating inflammatory cytokines have been associated with depressive symptoms. However, measuring circulating cytokines have inherent methodological limitations. In vitro lipopolysaccharide (LPS)-stimulated intracellular cytokines (ICCs) overcome these limitations. Furthermore, because psychosocial and physiological stressors activate inflammatory responses and LPS-stimulated ICCs reflect the inflammatory responsivity of monocytes to such stressors, ICCs may reflect individual stress responsivity. Methods This cross-sectional study examined whether LPS-stimulated expression of ICCs in peripheral blood mononuclear cells (PBMCs) is a sensitive inflammation measure correlated with depressive symptoms in 180 community-dwelling older adults. We tested correlations of not only intracellular but also circulating inflammatory markers with depressive symptoms assessed using the 10-item Center for Epidemiological Studies Depression Scale (CES-D). Intracellular markers included expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and both in PBMCs. Circulating markers included IL-6, TNF-α, and C-reactive protein (CRP) in plasma. Results None of the correlations were statistically significant. However, in contrast to circulating markers, the correlations of ICCs were consistently in the expected direction, i.e., higher ICC expression correlating with higher depression severity. Discussion Despite the non-significant findings, further research is required for the evaluation of LPS-stimulated ICC expression as biomarkers of depressive symptoms.

Androgen receptor deficiency in monocytes/macrophages does not alter adiposity or glucose homeostasis in male mice / 亚洲男科学杂志(英文版)

Androgen deprivation in men leads to increased adiposity, but the mechanisms underlying androgen regulation of fat mass have not been fully defined. Androgen receptor (AR) is expressed in monocytes/macrophages, which are resident in key metabolic tissues and influence energy metabolism in surrounding cells. Male mice bearing a cell-specific knockout of the AR in monocytes/macrophages (M-ARKO) were generated to determine whether selective loss of androgen signaling in these cells would lead to altered body composition. Wild-type (WT) and M-ARKO mice (12-22 weeks of age, n = 12 per group) were maintained on a regular chow diet for 8 weeks and then switched to a high-fat diet for 8 additional weeks. At baseline and on both the regular chow and high-fat diets, no differences in lean mass or fat mass were observed between groups. Consistent with the absence of differential body weight or adiposity, no differences in food intake (3.0 ± 0.5 g per day for WT mice vs 2.8 ± 0.4 g per day for M-ARKO mice) or total energy expenditure (0.6 ± 0.1 Kcal h-1 for WT mice vs 0.5 ± 0.1 Kcal h-1 for M-ARKO mice) were evident between groups during high-fat feeding. Liver weight was greater in M-ARKO than that in WT mice (1.5 ± 0.1 g vs 1.3 ± 0.0 g, respectively, P = 0.02). Finally, M-ARKO mice did not exhibit impairments in glucose tolerance or insulin sensitivity relative to WT mice at any study time point. In aggregate, these findings suggest that AR signaling specifically in monocytes/macrophages does not contribute to the regulation of systemic energy balance, adiposity, or insulin sensitivity in male mice.

Circulatory and Ventilatory Power: Characterization in Patients with Coronary Artery Disease / Potência Circulatória e Ventilatória: Caracterização em Pacientes com Doença Arterial Coronariana

Background: Circulatory power (CP) and ventilatory power (VP) are indices that have been used for the clinical evaluation of patients with heart failure; however, no study has evaluated these indices in patients with coronary artery disease (CAD) without heart failure. Objective: To characterize both indices in patients with CAD compared with healthy controls. Methods: Eighty-seven men [CAD group = 42 subjects and healthy control group (CG) = 45 subjects] aged 40–65 years were included. Cardiopulmonary exercise testing was performed on a treadmill and the following parameters were measured: 1) peak oxygen consumption (VO2), 2) peak heart rate (HR), 3) peak blood pressure (BP), 4) peak rate-pressure product (peak systolic HR x peak BP), 5) peak oxygen pulse (peak VO2/peak HR), 6) oxygen uptake efficiency (OUES), 7) carbon dioxide production efficiency (minute ventilation/carbon dioxide production slope), 8) CP (peak VO2 x peak systolic BP) and 9) VP (peak systolic BP/carbon dioxide production efficiency). Results: The CAD group had significantly lower values for peak VO2 (p < 0.001), peak HR (p < 0.001), peak systolic BP (p < 0.001), peak rate-pressure product (p < 0.001), peak oxygen pulse (p = 0.008), OUES (p < 0.001), CP (p < 0.001), and VP (p < 0.001) and significantly higher values for peak diastolic BP (p = 0.004) and carbon dioxide production efficiency (p < 0.001) compared with CG. Stepwise regression analysis showed that CP was influenced by group (R2 = 0.44, p < 0.001) and VP was influenced by both group and number of vessels with stenosis after treatment (interaction effects: R2 = 0.46, p < 0.001). Conclusion: The indices CP and VP were lower in men with CAD than healthy controls. .

Fundamento: Os índices da Potência Circulatória (PC) e Potência Ventilatória (PV) têm sido utilizados para avaliação clínica de pacientes com insuficiência cardíaca, mas nenhum estudo avaliou esses índices em pacientes com Doença Arterial Coronariana (DAC). Objetivo: Caracterizar ambos os índices em pacientes com DAC comparados a indivíduos saudáveis. Métodos: Oitenta e sete homens [grupo DAC = 42 sujeitos e, grupo controle (GC) = 45 sujeitos] com idade entre 45 e 65 anos foram incluídos. Um Teste de Exercício Cardiopulmonar (TECP) foi realizado em esteira e as seguintes variáveis foram obtidas: 1) consumo de oxigênio (VO2) pico; 2) Frequência Cardíaca (FC) pico; 3) Pressão Arterial (PA) pico; 4) duplo produto pico (PA sistólica pico x FC pico); 5) pulso de oxigênio pico (VO2 pico dividido pela FC pico); 6) eficiência ventilatória para o consumo de oxigênio (OUES); 7) eficiência ventilatória para a produção de dióxido de carbono (VE/VCO2 slope); 8) PC (VO2 pico x PA sistólica pico); e 9) PV (PA sistólica pico dividido pelo VE/VCO2 slope). Resultados: O grupo DAC apresentou valores significativamente menores das seguintes variáveis no pico do exercício: VO2 (p < 0,001), FC (p < 0,001), PA sistólica (p < 0,001), duplo produto (p < 0,001), pulso de oxigênio (p = 0,008), OUES (p < 0,001), PC (p < 0,001) e PV (p < 0,001), e valores significativamente maiores de PA diastólica (p = 0,004) e VE/VCO2 slope (p < 0,001) em relação ao GC. Uma análise de regressão pelo método stepwise mostrou que a PC foi influenciada pelo grupo (R2 = 0,44, p < 0,001) e a PV tanto pelo grupo quanto pelo número de vasos com estenose pós tratamento (efeito de interação: R2 = 0,46, p < 0,001). Conclusion: Os índices da PC e PV foram menores em homens com DAC comparados ao GC, podendo dessa forma ser utilizados na caracterização dessa população. .

Percepção dos graduandos de enfermagem sobre o seu envelhecimento / Percepción de los estudiantes de enfermería sobre lo proprio envejecimiento / Perception of nursing undergraduate students on self-aging

RESUMO Objetivo: analisar a percepção dos graduandos de enfermagem sobre o próprio envelhecimento. Método: pesquisa de abordagem qualitativa, realizada em agosto e setembro de 2011, com 18 graduandos de enfermagem de uma Universidade pública de Salvador (Bahia). Os depoimentos foram analisados por meio da Análise de Conteúdo. Resultados: apreendeu-se o núcleo temático: Percepção do graduando de enfermagem sobre o próprio envelhecimento e, a partir deste, emergiram duas subcategorias: A) O Não Pensar; B) O contexto influenciando no processo. Conclusão: os graduandos revelam que o envelhecimento está intrínseco ao desenvolvimento humano, e possui o vínculo familiar, a espiritualidade e atividade física como ferramentas fundamentais para um envelhecimento ativo. Entretanto, os mesmos relatam que, o modo de vida acelerado e estressante vivido na sociedade possibilita inserir hábitos considerados inadequados, como o consumo de “fast food” e álcool, que trazem influências negativas para o próprio processo de envelhecimento. .

RESUMEN Objetivo: analizar la percepción de los estudiantes de enfermería sobre su proprio envejecimiento. Método: estudio cualitativo, realizado en agosto y septiembre de 2011, con 18 estudiantes de enfermería de una universidad pública en Salvador/Bahia. Los datos fueron analizados através de análisis de contenido. Resultado: incautados el tema central: Percepción de alumnos de enfermería sobre su propio envejecimiento y de esto surgieron dos subcategorías: A) No creo; B) El contexto influye en el proceso. Conclusión: los estudiantes revelan que el envejecimiento es intrínseco al desarrollo humano, y tiene los vínculos familiares, la espiritualidad y la actividad física como herramienta clave para el envejecimiento activo. Sin embargo, el mismo informe que, debido a la forma de vida que se vive en la sociedad de ritmo rápido y estresante permite insertar hábitos considerados inadecuados, como el consumo de “comida rápida” y el alcohol y convertirse en influencias negativas para su propio proceso tuvo como objetivo analizar de los estudiantes de enfermería su propio envejecimiento. .

ABSTRACT Objective: to analyze the perceptions of nursing undergraduate students on their self-aging process. Method: qualitative study carried out between August and September, 2011 with 18 nursing undergraduate students of a public university in Salvador, Bahia. The interviews were analyzed by means of the Content Analysis method. Results: the following thematic concept was apprehended: Perceptions of nursing undergraduates on their self-aging, which generated two subcategories: A) The “don’t think about it” process; B) The context infl uencing the process. Conclusion: undergraduates reveal that the aging process is an intrinsic factor to human development. Family ties, spirituality and physical activity would be key mechanisms toward active aging. However, students also reported that their accelerated and stressed social lifestyles led to inadequate habits, such as the consumption of fast food and alcohol, which become negative infl uences in their aging process. .

Asociación entre los niveles de zinc intracelular y el estado nutricional de los niños infectados por el virus de la inmunodeficiencia humana (VIH) y los niños expuestos al virus no infectados / Association between Intracellular zinc levels and nutritional status in HIV-infected and uninfected children exposed to the virus

Introducción: Desnutrición, retardo en el crecimiento e infecciones oportunistas sobrevienen a alteraciones metabólicas, inmunológicas y gastrointestinales que produce el virus de la inmunodeficiencia humana (VIH). La deficiencia de zinc se ha asociado con deterioro nutricional, falla en el crecimiento y riesgo de infecciones. El objetivo de este estudio fue asociar los niveles de zinc en células mononucleares de sangre periférica (PBMC) con el estado nutricional en niños infectados por el VIH y en niños no infectados expuestos al virus. Pacientes y Método: Estudio analítico observacional, transversal, en 17 niños infectados y 17 expuestos, entre 2 y 10 años de edad. Se realizó valoración antropométrica, historia clínica-nutricional, recordatorio de 24 horas, medición de actividad física y determinación de zinc en PBMC por citometría de fiujo. Resultados: La talla para la edad, el consumo de energía, y la adecuación de energía, proteínas y zinc alimentario fueron significativamente mayores en los niños expuestos comparados con los niños infectados (p < 0,05). No se hallaron diferencias significativas en el índice de masa corporal, los niveles de zinc en monocitos, linfocitos CD4+ y CD4- entre los dos grupos de estudio (p > 0,05); sin embargo, la mediana de los niveles de zinc en monocitos de pacientes infectados fue mayor (218,6) comparado con el grupo control (217,0). No se encontró asociación entre consumo de zinc y niveles de zinc intracelular. Conclusiones: El deterioro del estado nutricional y el retardo en el crecimiento en niños estuvo asociado al VIH, pero no a los niveles de zinc intracelular. El consumo alimentario de este nutriente no se asoció a niveles de zinc en monocitos y linfocitos CD4+ y CD4-.

Introduction: Malnutrition, growth retardation and opportunistic infections outlast the metabolic, immune and gastrointestinal disorders produced by HIV. Zinc deficiency has been associated with deteriorating nutritional status, growth failure, and risk of infection. The aim of this study is to determine the association between zinc levels in peripheral blood mononuclear cells (PBMC) and the nutritional status of HIV-infected and uninfected children exposed to the virus. Patients and Methods: An analytical, observational, cross-sectional study was conducted on 17 infected and 17 exposed children, aged 2-10 years. Anthropometric measurements, clinical and nutritional history, 24 h recall, measurement of physical activity, and zinc in PBMC by fiow cytometry analysis were recorded. Results: Height according to age, energy consumption and adequacy of energy, protein and dietary zinc were significantly higher in children exposed to the virus compared to those infected with HIV (P < 0.05). No significant differences were found in BMI, levels of zinc in monocytes, CD4+ and CD4- lymphocytes between the two study groups (P > 0.05). However, the median levels of zinc in monocytes of infected patients was higher (218.6) compared to the control group (217.0). No association was found between zinc intake and levels of intracellular zinc. Conclusions: The deterioration of nutritional status and growth retardation in children were associated with HIV, but not with the levels of intracellular zinc. The dietary intake of this nutrient was not associated with levels of zinc in monocytes or CD4+ and CD4- lymphocytes.

Role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism

Our aim was to investigate the role of chemokines in promoting instability of coronary atherosclerotic plaques and the underlying molecular mechanism. Coronary angiography and intravascular ultrasound (IVUS) were performed in 60 stable angina pectoris (SAP) patients and 60 unstable angina pectoris (UAP) patients. The chemotactic activity of monocytes in the 2 groups of patients was examined in Transwell chambers. High-sensitivity C-reactive protein (hs-CRP), monocyte chemoattractant protein-1 (MCP-1), regulated on activation in normal T-cell expressed and secreted (RANTES), and fractalkine in serum were examined with ELISA kits, and expression of MCP-1, RANTES, and fractalkine mRNA was examined with real-time PCR. In the SAP group, 92 plaques were detected with IVUS. In the UAP group, 96 plaques were detected with IVUS. The plaques in the UAP group were mainly lipid 51.04% (49/96) and the plaques in the SAP group were mainly fibrous 52.17% (48/92). Compared with the SAP group, the plaque burden and vascular remodeling index in the UAP group were significantly greater than in the SAP group (P<0.01). Chemotactic activity and the number of mobile monocytes in the UAP group were significantly greater than in the SAP group (P<0.01). Concentrations of hs-CRP, MCP-1, RANTES, and fractalkine in the serum of the UAP group were significantly higher than in the serum of the SAP group (P<0.05 or P<0.01), and expression of MCP-1, RANTES, and fractalkine mRNA was significantly higher than in the SAP group (P<0.05). MCP-1, RANTES, and fractalkine probably promote instability of coronary atherosclerotic plaque.

Effects of Titanium Surfaces on the Developmental Profile of Monocytes/Macrophages

Due to the critical role of monocytes/macrophages (Mϕ) in bone healing, this study evaluated the effects of bio-anodized, acid-etched, and machined titanium surfaces (Ti) on Mϕ behavior. Cells were separated from whole human blood from 10 patients, plated on Ti or polystyrene (control) surfaces, and cultured for 72 h. At 24, 48 and 72 h, cell viability, levels of IL1β, IL10, TNFα, TGFβ1 inflammatory mediators, and nitric oxide (NO) release were analyzed by mitochondrial colorimetric assay (MTT assay) and immunoenzymatic assays, respectively. Real-time PCR was used to verify the expression of TNFα and IL10 at 72 h. The data were subjected to a Kruskal-Wallis analysis. IL1β, TNFα and TGFβ1 release were not significantly different between the Ti surfaces (p>0.05). The presence of NO and IL10 was not detected in the samples. Cell viability did not differ between the samples cultivated on Ti and those cultivated on control surfaces, except at 24 h (p=0.0033). With respect to the mediators evaluated, the surface characteristics did not induce a typical Th1 or Th2 cytokine profile, although the cell morphology and topography were influenced by the Ti surface during the initial period.

Devido ao papel crítico dos monócitos / macrófagos (Mϕ) na cicatrização óssea, este estudo avaliou os efeitos de superficies de titânio (Ti) bio-anodizada, ataque ácido e usinada sobre o comportamento Mϕ. As células foram separadas a partir de sangue humano de 10 pacientes, plaqueadas em Ti ou superfícies de poliestireno (controle), e cultivadas durante 72 h. Às 24, 48 e 72 h, a viabilidade celular e IL1β, IL10, TNFα, TGFβ1 e liberação de óxido nítrico foram analisados por ensaio colorimétrico mitocondrial (MTT) e ensaios imunoenzimáticos, respectivamente. PCR em tempo real foi utilizado para verificar a expressão de TNFα e IL-10 às 72 h. Os dados foram submetidos a uma análise de Kruskal-Wallis. IL1β autorização, TNFα e TGFβ1, não foram significativamente diferentes entre as superfícies de Ti (p>0,05). A presença de NO e de IL-10 não foi detectada nas amostras. A viabilidade celular não diferiu entre as amostras cultivadas em Ti e aquelas cultivadas em superfícies de controle, exceto às 24 h (p=0,0033). No que diz respeito aos mediadores avaliados, as características da superfície, não induziu resposta típica de citocinas Th1 ou Th2, embora a morfologia da célula e a topografia foram influenciadas pela superfície de Ti, durante o período inicial.

The presence of high level soluble herpes virus entry mediator in sera of gastric cancer patients

The development of gastric cancer (GC) is closely related to chronic inflammation caused by Helicobacter pylori infection, and herpes virus entry mediator (HVEM) is a receptor expressed on the surface of leukocytes that mediates potent inflammatory responses in animal models. However, the role of HVEM in human GC has not been studied. Previously, we showed that the interaction of HVEM on human leukocytes with its ligand LIGHT induces intracellular calcium mobilization, which results in inflammatory responses including induction of proinflammatory cytokine production and anti-bacterial activities. In this study, we report that leukocytes from GC patients express lower levels of membrane HVEM (mHVEM) and have lower LIGHT-induced bactericidal activities than those from healthy controls (HC). In contrast, levels of soluble HVEM (sHVEM) in the sera of GC patients were significantly higher than in those of HC. We found that monocyte membrane-bound HVEM is released into the medium when cells are activated by proinflammatory cytokines such as TNF-alpha and IL-8, which are elevated in the sera of GC patients. mHVEM level dropped in parallel with the release of sHVEM, and release was completely blocked by the metalloprotease inhibitor, GM6001. We also found that the low level of mHVEM on GC patient leukocytes was correlated with low LIGHT-induced bactericidal activities against H. pylori and S. aureus and production of reactive oxygen species. Our results indicate that mHVEM on leukocytes and sHVEM in sera may contribute to the development and/or progression of GC.

High-density lipoprotein prevents SAA-induced production of TNF-α in THP-1 monocytic cells and peripheral blood mononuclear cells

In this study, we evaluated whether human serum and lipoproteins, especially high-density lipoprotein (HDL), affected serum amyloid A (SAA)-induced cytokine release. We verified the effects of SAA on THP-1 cells in serum-free medium compared to medium containing human serum or lipoprotein-deficient serum. SAA-induced tumour necrosis factor-alpha (TNF-α) production was higher in the medium containing lipoprotein-deficient serum than in the medium containing normal human serum. The addition of HDL inhibited the SAA-induced TNF-α release in a dose-dependent manner. This inhibitory effect was specific for HDL and was not affected by low-density lipoprotein or very low-density lipoprotein. In human peripheral blood mononuclear cells, the inhibitory effect of HDL on TNF-α production induced by SAA was less pronounced. However, this effect was significant when HDL was added to lipoprotein-deficient medium. In addition, a similar inhibitory effect was observed for interleukin-1 beta release. These findings confirm the important role of HDL and support our previous hypothesis that HDL inhibits the effects of SAA during SAA transport in the bloodstream. Moreover, the HDL-induced reduction in the proinflammatory activity of SAA emphasizes the involvement of SAA in diseases, such as atherosclerosis, that are characterized by low levels of HDL.

Angiotensin II induces NF-κB, JNK and p38 MAPK activation in monocytic cells and increases matrix metalloproteinase-9 expression in a PKC- andRho kinase-dependent manner

Angiotensin II (ANG II), the main effector of the renin-angiotensin system, is implicated in endothelial permeability, recruitment and activation of the immune cells, and also vascular remodeling through induction of inflammatory genes. Matrix metalloproteinases (MMPs) are considered to be important inflammatory factors. Elucidation of ANG II signaling pathways and of possible cross-talks between their components is essential for the development of efficient inhibitory medications. The current study investigates the inflammatory signaling pathways activated by ANG II in cultures of human monocytic U-937 cells, and the effects of specific pharmacological inhibitors of signaling intermediates on MMP-9 gene (MMP-9) expression and activity. MMP-9 expression was determined by real-time PCR and supernatants were analyzed for MMP-9 activity by ELISA and zymography methods. A multi-target ELISA kit was employed to evaluate IκB, NF-κB, JNK, p38, and STAT3 activation following treatments. Stimulation with ANG II (100 nM) significantly increased MMP-9 expression and activity, and also activated NF-κB, JNK, and p38 by 3.8-, 2.8- and 2.2-fold, respectively (P < 0.01). ANG II-induced MMP-9 expression was significantly reduced by 75 and 67 percent, respectively, by co-incubation of the cells with a selective inhibitor of protein kinase C (GF109203X, 5 µM) or of rho kinase (Y-27632, 15 µM), but not with inhibitors of phosphoinositide 3-kinase (wortmannin, 200 nM), tyrosine kinases (genistein, 100 µM) or of reactive oxygen species (α-tocopherol, 100 µM). Thus, protein kinase C and Rho kinase are important components of the inflammatory signaling pathways activated by ANG II to increase MMP-9 expression in monocytic cells. Both signaling molecules may constitute potential targets for effective management of inflammation.

Produção de citocinas pelos monócitos de idosos com estomatite protética associada a Candida / Cytokine production by monocytes from elderly patients with Candida-associated denture stomatitis

Estomatite protética associada a Cândida (EPC), a lesão mais frequente em usuários de próteses removíveis, principalmente os idosos, caracteriza-se por uma inflamação da mucosa bucal que suporta a prótese. Está fortemente associada com Candida Albicans, bem como com fatores locais e sistêmicos, como a deficiência da resposta imune. Os monócitos são importantes na resposta protetora contra o fungo, produzindo citocinas que recrutam e ativam leucócitos. Existem alterações funcionais dessas células com o avanço da idade. Não foi possível encontrar na literatura dados referentes à função imunomodulatória dos monócitos de idosos com EPC. O presente trabalho pretendeu avaliar a produção de citocinas por essas células, estimuladas in vitro com C. albicans, obtidas do sangue periférico de idosos usuários de prótese total superior (PTS) com EPC, comparando-se com idosos usuários de PTS sem EPC, e com idosos e jovens não usuários de PTS. Os monócitos isolados foram cultivados em placas de cultura de 24 poços, na ausência ou presença de lipopolissacarídeo (LPS) ou C. albicans ATCC 90028 morta pelo calor. Após 18 horas, o sobrenadante foi coletado e submetido ao ensaio de imunoabsorção por ligação enzimática (ELISA) para dosagem das citocinas pró- inflamatórias fator de necrose tumoral- (TNF-), interleucina-6 (IL-6), IL-1, CXCL8 e proteína quimiotática de monócito (MCP-1), e anti-inflamatórias IL-10 e fator transformador de crescimento- (TGF-). Os resultados estão expressos como média ± desvio padrão dos valores obtidos para cada grupo, e foram analisados por meio de testes estatísticos não-paramétricos. Valores de p<0,05 foram considerados estatisticamente significativos. Os resultados demonstraram, de uma forma geral, alterações nos monócitos oriundos dos idosos com EPC, em comparação aos outros grupos: menor produção espontânea de CXCL8 e de MCP-1; menores níveis de TNF-, de IL-6, de IL-1, de CXCL8, de MCP-1 e de IL-10, após estímulo com LPS; e menor...

Candida-associated denture stomatitis (DS), the most frequent lesion among denture wearers, especially the elderly, is characterized by inflammation of the denture-bearing mucosa. It is strongly associated with Candida albicans, as well as with local and systemic factors, such as impaired immune response. Monocytes are important in the protective immune response against the fungus, by the production of cytokines that recruit and activate leukocytes. There are functional changes of these cells with advancing age. No data were found in the literature concerning the immunomodulatory function of these phagocytes in elderly patients with DS. This study aimed to evaluate the cytokine production by monocytes, challenged in vitro with C. albicans, obtained from peripheral blood of elderly denture wearers with DS, compared with elderly denture wearers without DS and elderly and young non-denture wearers. The isolated monocytes were cultivated in 24-well flat-bottomed culture plates, in the absence or presence of lipopolysaccharide (LPS) or heat-killed C.albicans ATCC 90028. After 18 hours, the supernatant was collected and submitted to the enzyme-linked immunosorbent assay (ELISA) for determination of the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-), interleukin-6 (IL-6), CXCL8, IL-1, monocyte chemotactic protein-1 (MCP-1) and anti-inflammatory cytokines IL-10 and transforming growth factor- (TGF-). The results are expressed as mean ± standard deviation of the values obtained for each group, and were analyzed using nonparametric statistical tests; p values <0.05 were considered statistically significant. The results demonstrated, in general, changes in monocytes from the elderly with DS, as compared to other groups: lower spontaneous production of CXCL8 and MCP-1; lower levels of TNF-, IL-6, IL-1, CXCL8, MCP-1 and IL-10 after stimulation with LPS; and reduced production of TNF- and IL-6 after stimulation with C. albicans. Comparing...

Lipopolysaccharide-induced expression of cell surface receptors and cell activation of neutrophils and monocytes in whole human blood

Lipopolysaccharide (LPS) activates neutrophils and monocytes, inducing a wide array of biological activities. LPS rough (R) and smooth (S) forms signal through Toll-like receptor 4 (TLR4), but differ in their requirement for CD14. Since the R-form LPS can interact with TLR4 independent of CD14 and the differential expression of CD14 on neutrophils and monocytes, we used the S-form LPS from Salmonella abortus equi and the R-form LPS from Salmonella minnesota mutants to evaluate LPS-induced activation of human neutrophils and monocytes in whole blood from healthy volunteers. Expression of cell surface receptors and reactive oxygen species (ROS) and nitric oxide (NO) generation were measured by flow cytometry in whole blood monocytes and neutrophils. The oxidative burst was quantified by measuring the oxidation of 2',7'-dichlorofluorescein diacetate and the NO production was quantified by measuring the oxidation of 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate. A small increase of TLR4 expression by monocytes was observed after 6 h of LPS stimulation. Monocyte CD14 modulation by LPS was biphasic, with an initial 30 percent increase followed by a 40 percent decrease in expression after 6 h of incubation. Expression of CD11b was rapidly up-regulated, doubling after 5 min on monocytes, while down-regulation of CXCR2 was observed on neutrophils, reaching a 50 percent reduction after 6 h. LPS induced low production of ROS and NO. This study shows a complex LPS-induced cell surface receptor modulation on human monocytes and neutrophils, with up- and down-regulation depending on the receptor. R- and S-form LPS activate human neutrophils similarly, despite the low CD14 expression, if the stimulation occurs in whole blood.

Human monocytes tolerant to LPS retain the ability to phagocytose bacteria and generate reactive oxygen species

Tolerance to lipopolysaccharide (LPS) occurs when animals or cells exposed to LPS become hyporesponsive to a subsequent challenge with LPS. This mechanism is believed to be involved in the down-regulation of cellular responses observed in septic patients. The aim of this investigation was to evaluate LPS-induced monocyte tolerance of healthy volunteers using whole blood. The detection of intracellular IL-6, bacterial phagocytosis and reactive oxygen species (ROS) was determined by flow cytometry, using anti-IL-6-PE, heat-killed Staphylococcus aureus stained with propidium iodide and 2',7'-dichlorofluorescein diacetate, respectively. Monocytes were gated in whole blood by combining FSC and SSC parameters and CD14-positive staining. The exposure to increasing LPS concentrations resulted in lower intracellular concentration of IL-6 in monocytes after challenge. A similar effect was observed with challenge with MALP-2 (a Toll-like receptor (TLR)2/6 agonist) and killed Pseudomonas aeruginosa and S. aureus, but not with flagellin (a TLR5 agonist). LPS conditioning with 15 ng/mL resulted in a 40 percent reduction of IL-6 in monocytes. In contrast, phagocytosis of P. aeruginosa and S. aureus and induced ROS generation were preserved or increased in tolerant cells. The phenomenon of tolerance involves a complex regulation in which the production of IL-6 was diminished, whereas the bacterial phagocytosis and production of ROS was preserved. Decreased production of proinflammatory cytokines and preserved or increased production of ROS may be an adaptation to control the deleterious effects of inflammation while preserving antimicrobial activity.

Expression of cannabinoid receptor I during mice skin incised wound healing course / 法医学杂志

OBJECTIVE@#To investigate the expression of cannabinoid receptor I (CB1R) during mice skin incised wound healing course and time-dependent changes of CB1R in wound age determination.@*METHODS@#The changes of CBIR expression in skin incised wound were detected by immunohistochemistry and Western blotting.@*RESULTS@#The control group showed a low expression of CB1R detected mainly in epidermis, hair follicles, sebaceous gland and dermomuscular layer. CB1R expression was undetectable in neutrophils in the wound specimens from 6h to 12h post-injury. CB1R positive cells were mostly mononuclear cells (MNCs) and fibroblastic cells (FBCs) from 1 d to 5 d post-injury. CB1R positive cells were mostly FBCs from 7 d to 14d post-injury. The ratio of the CB1R positive cells increased gradually in the wound specimens from 6 h to 3 d post-injury, reached peak level at 5 d, and then decreased gradually from 7d to 14 d post-injury. The positive bands of CB1R were observed in all time points of the wound healing course by Western blotting. The expression peak showed at 5 d post-injury.@*CONCLUSION@#CB1R is activated during the wound healing course. The expression of CB1R is found in mononuclear cells, which could be involved in inflammation reaction. CBIR is observed in fibroblastic cells, which could participate in the wound healing. CB1R may be a potentially useful marker for determination of wound healing age.

Expression of M3 subtype of muscarinic receptor during the skin incised wound healing in mice / 法医学杂志

OBJECTIVE@#To investigate the expression of M3 subtype of muscarinic receptors (M3R) during the incised wound healing of the skin in mice and the characteristics of its time-dependent.@*METHODS@#The change of M3R in skin incised wound was detected by immunohistochemical staining and Western blot.@*RESULTS@#M3R-positive cells were detected in epidermis, hair follicle, sebaceous glands, sweat glands, dermomuscular layer in normal mouse skin. Expression of M3R was mainly detectable in polymorphonuclear cells (PMNs) in the wound specimens aged from 6h to 12h after injury. Afterwards, the M3R-positive cells were mostly mononuclear cells (MNCs) and fibroblastic cells (FBCs) at 1 d to 3d post-injury, whereas the M3R-positive cells were mostly FBCs aged from 5 d to 14d. Morphometrically, the ratio of the M3R-positive cells increased aged from 6h to 12h after injury, with a peak at 12h. The ratios kept a high relatively level aged from 1 d to 5 d, but significantly that lowered as compared with aged 12h after injury. The ratio reached the peak at 7 d again after injury, and then decreased gradually. The M3R protein also revealed a time-dependent tendency with double peaks at 12h and 7 d after injury as detected by Western blotting.@*CONCLUSION@#M3R is time-dependently expression in PMNs, MNCs and FBCs suggesting that it may play roles during the skin incised wound healing, and M3R may be used as a marker for wound age determination.

Annexin A6 is highly abundant in monocytes of obese and type 2 diabetic individuals and is downregulated by adiponectin in vitro

Adiponectin stimulates cholesterol efflux in macrophages and low adiponectin may in part contribute to disturbed reverse cholesterol transport in type 2 diabetes. Monocytes express high levels of annexin A6 that could inhibit cholesterol efflux and it was investigated whether the atheroprotective effects of adiponectin are accompanied by changes in annexin A6 levels. Adiponectin reduces annexin A6 protein whereas mRNA levels are not affected. Adiponectin-mediated activation of peroxisome proliferator-activated receptor alpha (PPARalpha) and AMP-activated protein kinase (AMPK) does not account for reduced annexin A6 expression. Further, fatty acids and lipopolysaccharide that are elevated in obesity do not influence annexin A6 protein levels. Annexin A6 in monocytes from overweight probands or type 2 diabetic patients is significantly elevated compared to monocytes of normal-weight controls. Monocytic annexin A6 positively correlates with body mass index and negatively with systemic adiponectin of the blood donors. Therefore, the current study demonstrates that adiponectin reduces annexin A6 in monocytes and thereby may enhance cholesterol efflux. In agreement with these in vitro finding an increase of monocytic annexin A6 in type 2 diabetes monocytes was observed.

Expression of Rap1GAP in human myeloid disease following microarray selection

To find the underlying causes of primary myelodysplastic syndrome (MDS), the gene expression profiling of both CD34+ cells and bone marrow mononuclear cells from MDS patients was performed using oligonucleotide microarray and cDNA microarrays, respectively. Several candidate genes which were differentially expressed in MDS patients versus normal controls were selected and confirmed in expanding samples by quantitative real-time reverse transcription-polymerase chain reaction after clustering and bioinformatics analysis. one of these genes, RAP1GAP, was found to be expressed at a significantly higher level in patients with MDS in comparison with those suffering from other hematopoietic diseases including leukemia (P < 0.01). We propose that over-expression of RAP1GAP gene may play a role in the pathogenesis of MDS.

A study on the time-dependent expression of caspase-6 during repair of skin contusion in rats / 法医学杂志

OBJECTIVE@#To investigate the expression of caspase-6 in rat skin contusion and its surrounding areas during repairment.@*METHODS@#Immunohistochemical SP method and Western blot technique were used to study the expression and activation of caspase-6 in rat skin contusion and its surrounding areas.@*RESULTS@#Weak expression of caspase-6 was detected in cytoplasms of polymorphonuchear cells (PMNs) infiltrated in the injured area at 3 hours post-contusion. The ratio of the caspase-6 positive cells was low (25.78 +/- 1.38)%. The expression of caspase-6 was increased prominently (47.70 +/- 5.14)% at 12 hours post-contusion. Almost all of the PMNs, mononuclear cells (MNCs) and fibroblastic cells (FBCs) were caspase-6 positive with both cytoplasm and nucleus staining (54.58 +/- 5.64)% on post-contusion day 3. The expression of caspase-6 decreased gradually thereafter. The expression of the 34-kDa pro-caspase-6 was detected by Western blot in both control and the post-contusion groups with time dependent dynamics.@*CONCLUSION@#These results suggest that caspase-6 may play a major role in trauma-induced inflammatory response. Since caspase-6 shows a timely dependent expression in PMNs, MNCs and FBCs during skin injury repair in rat, it may be used as a marker for the contusion age determination,

A study on the time-dependent expression of phospho-p38MAPK during the skin incised wound healing in mice / 法医学杂志

OBJECTIVE@#To investigate the expressions of phospho-p38MAPK(p-p38MAPK) during the incised wound healing of the skin in mice and to explore the applicability of the time-dependent expressions of p-p38MAPK in the determination of wound age.@*METHODS@#The expression of p-p38MAPK in incised skin wound was detected by immunohistochemistry and Western blot.@*RESULTS@#There was a minimal baseline expression of p-p38MAPK in control mouse skin. Expression of p-p38MAPK was mainly detectable in neutrophils in the wound specimens from 3hours to 12 hours after injury. Afterwards, the p-p38MAPK positive cells were mostly mononuclear cells and fibroblasts between post-injury day 1 and day 5, whereas the p-p38MAPK positive cells were mostly fibroblasts between post-injury day 7 and day 14. Morphometrically, the ratio of the p-p38MAPK positive cells to total number of the cells increased gradually in the wound specimens aged from 3 hours to 12 hours, and maximized in the wound specimens aged 12 hours with a slight decrease at 24 hours after injury. The ratio maintained at high level from post-injury day 3 to day 5, and then decreased gradually from post-injury day 7 to day 14. The expression of p-p38MAPK was observed throughout the wound healing stages by Western blot as well with a peak expression occurring between 12hour and day 3 after injury.@*CONCLUSION@#Our data suggest that p-p38MAPK may play an important role in inducing apoptosis of neutrophils, mononuclear cells, and fibroblasts during skin wound healing and meanwhile, p-p38MAPK may be a potentially useful marker for wound age determination.