Long-term effects of a neonatal low-protein diet in rats on the number of macrophages in culture and the expression/production of fusion proteins.

AIM: To investigate the effects of a neonatal low-protein diet on the number of macrophages in culture and the expression/production of proteins that regulate macrophage fusion in young and adult rats. METHODS: Male Wistar rats (n = 18) were suckled by mothers fed diets containing 17 % protein (controls, C) or 8 % protein (undernourished, UN). All rats were fed a normal protein diet after weaning. Bronchoalveolar lavage was collected from 42-, 60- and 90-day-old rats. Alveolar macrophages were cultured for 4 days to assess the number of cells and the expression of cadherins, key proteins involved in macrophage fusion, by western blotting. IL-4 and IFN-γ levels in culture supernatants were measured by ELISA. RESULTS: Offspring from mothers fed a low-protein diet showed a lower body weight gain. The number of cells in cultured macrophages from UN was reduced at 42 and 60 days and increased at 90 days. IL-4 production was increased in the supernatants from UN group at 60 days but did not affect the expression of cadherins. IFN-γ production was increased in the supernatants from UN group at 42 and 60 days and reduced at 90 days. CONCLUSIONS: This study thus demonstrated that dietary restriction during lactation altered the number of alveolar macrophages in culture and the production of fusion proteins of offspring aged 42, 60 or 90 days but did not modify the expression of adhesion molecules important for the fusion of these cells.

Depressive symptoms, inflammatory markers and body composition in elderly with and without chronic obstructive pulmonary disease (COPD).

UNLABELLED: Our aim was to assess the relationships between cortisol, interleukin-2 (Il-2) and tumor necrosis factor-α (TNF-α) levels in elderly with and without COPD presenting with or without depressive symptoms. Forty COPD patients and 53 elderly individuals with no COPD took part in the study. Depressive symptoms (Geriatric Depression Scale=GDS-15), IL-2 and TNF-α, serum cortisol, number of comorbidities, smoking habits and body composition were evaluated. The prevalence of depressive symptoms was higher in COPD group. The number of comorbidities was higher in patients with depressive symptoms. No differences were found between IL-2, TNF-α and cortisol levels, years of smoking and smoked pack-years in the groups. The COPD group obtained lower body mass index (BMI) and fat content and higher fat free mass index as well as greater nutritional depletion. CONCLUSIONS: Depressive symptoms as well as fat and lean body composition, due to preserved BMI in those with nutritional depletion, must be investigated.

Cytotoxicity of current adhesive systems: in vitro testing on cell cultures of primary murine macrophages.

OBJECTIVES: The aim of this study was to evaluate, in vitro, the potential cytotoxicity of dentinal adhesives on alveolar macrophages of Wistar rats, after diffusion through dentin. METHODS: The cytotoxicity of adhesives [single bond plus (SB), clearfil SE bond (CF) and Xeno V (XE)] applied to the occlusal surface of human dentin disks adapted to a dentin barrier test device were analyzed. The sets placed on a monolayer of cells were incubated for 24, 48 and 72h. Culture medium and Escherichia coli lipopolysaccharides (LPS) were used as negative and positive controls, respectively. Cellular cytotoxicity was evaluated by observing the cell survival rate (MTT assay) and nitric oxide production (NO). The data were analyzed by one-way factorial ANOVA and Tukey's and Tamhane's paired comparisons T2 (α=0.05). RESULTS: All the adhesive systems reduced the percentage of live cells by over 50%, compared with the control group. Within the same period of time, there was a statistically significant difference between the adhesives and LPS compared with the negative control group. SB presented a statistically significant difference between 24h and 72h, and XE between 48h and 72h. The quantity of NO produced in 24h did not differ statistically between the NC and adhesive groups. After 48h there was a significant difference between SB/CF and XE/NC. At 72h only CF showed a significant difference from each of the other groups. LPS differed statistically from all the other groups at all the evaluation times. SIGNIFICANCE: Components of the adhesives tested may permeate the dentin in sufficient concentrations to cause death and damage to cell metabolism in the alveolar macrophages of rats, which indicates potential cytotoxicity to pulpal cells.

Stress-induced downregulation of macrophage phagocytic function is attenuated by exercise training in rats.

BACKGROUND/AIMS: Acute restraint stress may induce impaired macrophage phagocytic function. Moderate physical training is associated with beneficial effects on immunological functions. We investigated the effects of moderate physical training on phagocytic function of alveolar macrophages in rats submitted to acute restraint stress. METHODS: Thirty male Wistar rats weighing 210-226 g were randomly divided into 4 groups: nontrained rats (n = 7), nontrained rats submitted to stress (n = 8), trained rats (n = 7) and trained rats submitted to stress (n = 8). Trained rats were submitted to a program of moderate running training over a period of 8 weeks. Rats subjected to restraint stress were kept immobilized in glass cylinders (8 cm in diameter and 24 cm long) during 60 min. Phagocytosis capacity of macrophages was evaluated by either Escherichia coli orzymosan stimuli. RESULTS: Restraint stress induced a decrease in phagocytosis of E. coli and zymosan particle stimulation by macrophages. Neither of these alterations was observed in trained animals submitted to acute restraint stress. CONCLUSIONS: Our data confirm that acute restraint stress is associated with impaired function of macrophages. Moreover, moderate physical training attenuates the effects of acute stress by a mechanism that involves an increase in tolerance of macrophages.

Physical training attenuates the stress-induced changes in rat T-lymphocyte function.

BACKGROUND/AIMS: Modulations in the immune function by stress are a well-known phenomenon. Acute restraint stress may induce impaired T-lymphocyte responses. Moderate physical training is associated with beneficial effects on immunological functions. We investigated the effects of a moderate physical training on T-lymphocyte function in rats submitted to acute restraint stress. METHODS: Thirty male Wistar rats weighing 210-226 g were randomly divided into four groups: non-trained rats (NT, n = 7), and non-trained rats submitted to stress (NT + S, n = 8); trained rats (T, n = 7), and trained rats submitted to stress (T + S, n = 8). Trained rats were submitted to a program of moderate running over a period of 8 weeks. Rats subjected to restraint stress were kept immobilized in glass cylinders (8 cm in diameter and 24 cm long) during 60 min. Plasma corticosterone concentration, peripheral blood leukocyte number, indicators of apoptosis of T lymphocytes in blood and lymphoid organs, and mitogen-induced proliferation of T lymphocytes in lymphoid organs were evaluated. RESULTS: Acute stress exposure raised plasma corticosterone concentration (p < 0.001), but not in previously trained animals. Restraint stress induced an increase in the percentage of lymphocytes in apoptosis, and a decrease in the concanavalin-A-induced proliferation of lymphocytes from the thymus and lymph nodes, and an increase in lymphocytes of the spleen. Neither of these alterations was observed in trained animals submitted to acute restraint stress. CONCLUSIONS: Our data confirm that acute restraint stress is associated with changes in T-lymphocyte function. Moreover, moderate physical training attenuates the effects of acute stress by a mechanism that involves the hypothalamic-pituitary-adrenal axis and an increase in tolerance of leukocytes.

[Therapeutical and inflammatory response in rats with secondary peritonitis and topical use of ampicilin/sulbactam]. / Resposta terapêutica e inflamatória de ratos com peritonite secundária submetidos ao uso tópico de ampicilina/sulbactam.

OBJECTIVES: The acute peritonitis is an important cause of sepsis and death on intensive care units and surgery. The treatment must include: systemical use of antibiotics, drainage of abscess and restauration of gastrointestinal integrity. The topical use of antibiotics in the peritoneal cavity is controversial. The aim of this study was to evaluate the use of topical use of ampicilin/ sulbactam in the treatment of peritonitis. METHODS: We measured the plasmatic levels of nitric oxide, count of eosinophils, lymphocytes, monocytes, and neutrophils in blood and peritoneal cavity, using a model of peritonitis in rats (transfixation and ligature of cecum). Twenty four Wistar rats were divided in 4 groups (n = 6 each). group A: induction of peritonitis with ligature of cecum and topical treatment with saline; group B: induction of peritonitis with ligature of cecum and topical treatment with ampicilin/sulbactam; group C: transfixation of cecum; group D: laparotomy and peritoneal exsudate + blood sample. The transfixation-ligture of cecum remained for 24 hs before treatment. A relaparotomy was performed in 18 rats and peritoneal exsudate/blood were collected. Dosage of Nitric oxide, count of eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudte were done. RESULTS: The difference was not significant in the levels of nitric oxide, eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudate (p > 0,05) among the studied groups. CONCLUSION: The use of ampicilin associated to sulbactam via intraperitoneal in rats with fecal peritonitis did not change survival.; the levels of plama nitric oxide, count of eosinophil, lymphocytes, monocytes, and neutrophils in blood and peritoneal exsudate were not affected.

Interferência de ondas curtas pulsadas na adesividade e na fagocitose de macrófagos em ratos com desnutrição neonatal / Interference of pulse short waves on macrophage adhesion and phagocytosis in rats with neonatal malnutrition

Tendo em vista a relação estabelecida entre desnutrição neonatal e atividade celular ligada ao sistema imune, macrófagos peritoniais de ratos Wistar machos, desnutridos e recuperados, foram expostos à diatermia de ondas curtas pulsadas (DOCP) e a campo magnético (CM) alternado em frequência ultrabaixa para verificar in vitro as possíveis alterações no índice de aderência.

In view of the established relationships between neonatal malnutrition and the immuno system, peritoneal macrophages of male Wistar rats were exposed to diatermic pulsate short wave (DPSW) and to alterned magnetic field on extreme low frequency, in order to verify in vitro adhesion index and phagocytic activity...

Resposta terapêutica e inflamatória de ratos com peritonite secundária submetidos ao uso tópico de ampicilina/sulbactam / Therapeutical and inflammatory response in rats with secondary peritonitis and topical use of ampicilin/sulbactam

OBJETIVOS: A peritonite aguda representa uma importante causa de sepsis e óbito nas unidades de terapia intensiva e cirurgia. Classicamente o seu tratamento deve incluir: a administração sistêmica de antibióticos, a remoção mecânica dos contaminantes e a restauração da integridade gastrintestinal. A utilização de antibióticos diretamente na cavidade peritoneal é controversa. Estudo com o objetivo de avaliar o uso terapêutico, intraperitoneal da ampicilina associada ao sulbactam. MÉTODOS: foram mensurados os níveis plasmáticos do óxido nítrico, bem como a contagem de eosinófilos, linfócitos, monócitos e neutrófilos no sangue e no lavado peritoneal, utilizando-se modelo de peritonite em ratos (ligadura-transfixação cecal). Vinte quatro ratos Wistar, machos, foram divididos em quatro grupos de seis animais, assim distribuídos: grupo A: método de indução de peritonite - soltura da ligadura + tratamento com soro fisiológico; grupo B: método de indução de peritonite + soltura da ligadura + tratamento com soro fisiológico acrescido de ampicilina / sulbactam; grupo C: método de indução de peritonite + soltura da ligadura-transfixação cecal; e grupo D: laparatomia para realização de lavado peritoneal mais coleta de sangue. A ligadura-transfixação do cecum permaneceu por 24 horas, antes do tratamento instaurado. Foi realizada uma relaparotomia nos 18 ratos com coleta de líquido de lavado peritoneal e sangue. Foram dosados os níveis plasmáticos de óxido nítrico e determinado o número de eosinófilos, linfócitos, monócitos e neutrófilos no sangue e no lavado peritoneal. RESULTADOS: Não ocorreu diferença estatisticamente significante (p > 0,05) nos níveis de óxido nítrico, bem como no número de eosinófilos, linfócitos, monócitos e neutrófilos no sangue e no lavado peritoneal, entre os grupos. CONCLUSÃO: Neste estudo, concluiu-se que: a utilização de ampicilina associada a sulbactam por via intraperitoneal nos ratos com peritonite fecal: não modificou a sobrevida; não alterou os níveis plasmáticos de óxido nítrico; não alterou a contagem de eosinófilos, linfócitos, monócitos e neutrófilos tanto no sangue como no lavado peritoneal.

Esquitossomose mansônica hepatoesplênica humana: produção de TNF-alfa em monócitos / Hepatosplenic human schistosomiasis: TNF-alpha production in monocytes

Objetivo: Investigar os níveis de liberação de TNF-a?em cultura de monócitos em portadores humanos da forma hepatoesplênica de esquistossomose mansônica. Método: Foram incluídos aleatoriamente, no estudo, 39 voluntários de idades variando entre 15 e 31 anos, 19 homens e 20 mulheres, divididos em três grupos. Grupo 1 (GC) 12 indivíduos sadios, sem esquistossomose. Grupo 2 (AI) 18 indivíduos portadores de esquistossomose mansônica na forma hepatoesplênica, que tinham se submetido a esplenectomia, ligadurada veia gástrica esquerda e auto-implante de tecido esplênico no omento maior, quando tinham idades entre 7 e 16 anos. Esses pacientes receberam oxaminiquine na dose de 20mg/kg 30 dias antes do procedimento cirúrgico. O seguimento médio atual é de cerca de 8 anos. Grupo 3 - pacientes esplenectomizados sem autoimplanteesplênico (ESAI) constituído de nove adultos jovens que tinham se submetido à esplenectomia sem auto-implante esplênico e desconexão ázigo-porta. Os pacientes esquistossomóticos dos grupos 2 e 3 tiveram confirmação dessa doença pela presença de fibrose de Symmers nas biópsias hepáticas realizadas durante o ato cirúrgico. Foram colhidos 6 ml de sangue periférico de cada um dos voluntários incluídos no presente estudo, cujos monócitos foram separados por centrifugação e cultivados no meio de cultura CultilabÒ). Amostras de 100μl do sobrenadante da cultura de monócitos (106 células/ml), de cada indivíduo dos três grupos, eram colhidos para determinação das concentrações de TNF-a. Essa concentração era mensurada pelo estudo colorimétrico de ELISA para citocinas (QuanticininasTM – Sistema R&D), após 4 horas de estimulação com PMA e incubação, em uma atmosfera úmida com 5% de CO2 a 37oC. Resultados: As concentrações de TNF-a? não diferiram significantemente nos três grupos estudados [(GC 135,0 ± 51,6 pg/ml; AI 97,0 ± 25,4 pg/ml e ESAI 107,0 ±. 52,1 pg/ml) - ANOVA, F = 0,210; p = 0,813]...

Background: To investigate TNF-a levels released by monocyte culture in bearers of human hepatosplenic schistosomiasis mansoni. Methods: In this study we randomly included 39 volunteers with ages from 15 to 31 years, 19 men and 20 women, divided into three groups. Control Group 1 (CG) 12 healthy peoplewithout schistosomiasis. Auto implanted spleen group 2 (AIS) 18 young adults with hepatosplenic schistosomiasis mansoni who underwent splenectomy, ligature of the left gastric vein and auto implantation of spleen tissue into the major omentum, when the age varied between 7 and 16 years. These patientsreceived oxaminiquine in a single dose of 20mg/kg 30 days before the surgical treatment. The mean followup was about 8 years. Group 3 – splenectomyzed patients without auto implantation of spleen tissue (SP).Nine young adults were included and underwent total splenectomy without auto implantation of spleen tissue and azigo portal disconnection. The patients with schistosomiasis from groups I and III had the confirmation of their disease by the presence of Symmers fibrosis in the wedge liver biopsies during thesurgical procedure. Six 6ml of blood from all volunteers we collected. The monocytes were separated by centrifugation and cultivated in CultilabÒ medium. Supernatant samples of 100μl from the monocyte culture (106cells/ml), of each person were taken for the measure of TNF-a concentrations. This measure was performed by colorimetric ELISA (QuantikineTM – R&D systems), after 4 hours of stimulation with PMA and incubation with 37oC humid atmosphere of 5% CO2. Results: The TNF-a concentrations did not differ significantly among the three groups [(GC 135.0 ± 51.6 pg/ml; AIS 97.0 ± 25.4 pg/ml and SP 107.0 ±. 52.1pg/ml) - ANOVA F = 0.210; p = 0.813]...

Campos eletromagnéticos näo-ionizantes näo alteram o desenvolvimento sensório-motor em ratos / The sensory-motor development in rat is not altered by nonionizing electromagnetic fields

No presente trabalho, foi investigado o efeito dos campos eletromagnéticos (CEM) näo ionizantes de extrema baixa frequência (ELF) e rádio-frequência (RF) sobre o desenvolvimento sensorial e motor em ratos neonatos. Ratos wistar foram expostos aos campos eletromagnéticos näo-ionizantes ELF, alternado,60Hz (n=24) e de RF,30MHz, burst de 55 Hz(n=30), no período de aleitamento (1§ ao 21§ dias pós-natais durante 30 minutos. O peso dos filhotes foram aferidos diariamente. foram diariamente observadas a maturaçäo dos reflexos (preensäo palmar, recuperaçäo de decúbito, colocaçäo pelas vibrissas, aversäo ao principío, resposta ao susto, geotaxia negativa, reaçäo de aceleraçäo) e a atividade locomotora no campo aberto nos 3§, 7§, 14§, 21§ dias pós-natais, quantificando o número de quadrados invadidos (NQI). A evoluçäo ponderal, a maturaçäo dos reflexos e o NQI dos grupos expostos aos CEM, comparados aos controles, näo sofreram alteraçöes. Estes resultados indicam que sob as condiçöes experimentais utilizadas, os campos eletromagnéticos näo alteram a expressäo comportamental de desenvolvimento sensório-motor

Serotonina: uma história permeada de avanços tecnológicos em neurociência / Serononin: a permeated history of technological progresses in neurosciences

Os autores descrevem uma breve história da 5-Hidroxitriptamina (5-HT), abordando os primeiros relatos datados do início da descoberta do neurotransmissor e os avanços metodológicos empregados em seu estudo até os dias atuais. É ressaltado, em particular, o papel da farmacologia experimental e clínica como instrumento de identificação dos mecanismos fisiológicos implicados com o sistema serotoninérgico

Estresse: interações neuroendócrinas e imunológicas / Stress: Neuroendocrine and immune interactions

O estresse é fenômeno de crescente interesse científico no qual se evidência a inter-relação entre os sistemas fisiológicos na manutenção da hemeostase. Muitos dados demonstram que informações (na forma de citocinas, neurotransmissores ou mesmo impulsos nervosos) são permutados entre os diveros órgãos durante o estresse. O estresse assim como a resposta imune são mecanismos de adaptação que apresentam elementos comuns na filogênese. são discutidos alguns pontos polêmicos do estresse e a importante interação, observada durante o fenômeno, entre o sistema nervoso, o sistema endócrino e, em particular, o sistema imune. Procura-se demonstrar que além da intuição e do empirismo a relação do estresse com a resposta imune e baseada em evidências sólidas das ciências contemporâneas