ABSTRACT Purpose: To evaluate inflammatory response in critical bone injuries after implantation of the biomaterial composed of hydroxyapatite (HA)/poly (lactic-coglycolic acid) (PLGA)/BLEED. Methods: Forty-eight male Wistar rats (280 ± 20 grams) were divided into two groups: control group (CG), in which the animals do not receive any type of treatment; and biomaterial group (BG), in which the animals received the HA/PLGA/BLEED scaffold. Critical bone injury was induced in the medial region of the skull calotte with the aid of a trephine drill 8 mm in diameter. The biomaterial was implanted in the form of 1.5-mm thick scaffolds. Serum and calotte were collected at one, three and seven days. Results: Biomaterial had a significant effect on the morphological structure of the bone, accelerating osteoblast activation within three days, without causing exacerbated systemic inflammation. In addition, quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that BG induced upregulation of osteogenic genes such as runt-related transcription factor 2, and stimulated genes of inflammatory pathways such as tumor necrosis factor-α, on the first day without overexpressing genes related to bone matrix degradation, such as tissue inhibitor of metalloproteinases-1 and matrix metalloproteinase-9. Conclusions: The HA/PLGA/BLEED® association can be used as a bone graft to aid bone repair, as it is capable of modulating expression of important genes at this stage of the repair process.

Resumo Fundamento A formação de células espumosas ocorre devido ao aumento em lipoproteína plasmática de baixa densidade (LDL) e desregulação da inflamação, sendo importante para o desenvolvimento da aterosclerose. Objetivo Avaliar o perfil do fator de necrose tumoral alfa (TNF-α) e da interleucina-6 (IL-6) no método de formação da célula espumosa existente, otimizando esse protocolo. Métodos A LDL foi isolada, oxidada e marcada com sonda de isotiocianato de fluoresceína (FITC). As células espumosas foram geradas de célula derivada de monócitos humanos THP-1 e incubadas na ausência (controle) ou presença de FITC-ox-LDL (10, 50, 100, 150 ou 200 μg/mL), por 12, 24, 48 ou 72 horas. A FITC-ox-LDL na célula foi quantificada por microscopia. O ensaio de imunoabsorção enzimática foi avaliado para quantificar a IL-6 e o TNF-α, com um p <0,05 considerado significativo. Resultados Todas as concentrações de FITC-ox-LDL testadas apresentaram fluorescência mais alta em comparação com o controle, demonstrando maior acúmulo de lipoproteínas nas células. Quanto mais alta a concentração de FITC-ox-LDL, maior a produção de TNF-α e IL-6. A produção de IL-6 pelas células espumosas foi detectada até o valor de 150 µg/mL da LDL máxima de estímulo. Concentrações acima de 50 μg/mL de LDL estimularam maior liberação de TNF-α comparado ao controle. Conclusões Nosso modelo contribui para o entendimento da liberação de IL-6 e TNF-α em resposta a várias concentrações de ox-LDL usando o método otimizado para a formação de células espumosas.

Abstract Background The formation of foam cells occurs due to the increase in low-density plasma lipoprotein (LDL) and dysregulation of inflammation, which is important for the development of atherosclerosis. Objective To evaluate the profile of tumor necrosis factor-alpha (TNF-α) and Interleukin-6 (IL-6) in the existing foam cell formation method, optimizing this protocol. Methods The LDL was isolated, oxidized, and labeled with a Fluorescein isothiocyanate (FITC) probe. Foam cells were generated from THP-1 human monocyte-derived cells and incubated in the absence (control) or presence of FITC-ox-LDL (10, 50, 100, 150, or 200 μg/mL), for 12, 24, 48, or 72 hours. The accumulated FITC-ox-LDL in the cell was quantified by microscopy. The enzyme-linked immunosorbent assay was evaluated to quantify the IL-6 and TNF-α, with p < 0.05 considered significant. Results All the FITC-ox-LDL concentrations tested showed a higher fluorescence when compared to the control, showing a greater accumulation of lipoprotein in cells. The higher the concentration of FITC-ox-LDL, the greater the production of TNF-α and IL-6. The production of IL-6 by foam cells was detected up to the value of 150 µg/mL of the maximum stimulus for LDL. Concentrations above 50 μg/mL LDL stimulated greater release of TNF-α compared to control. Conclusions Our model contributes to the understanding of the release of IL-6 and TNF-α in response to different concentrations of ox-LDL, using an optimized method for the formation of foam cells.

Resumo Fundamento: A obesidade leva a um estado de inflamação crônica, disfunção endotelial e hipertensão. Objetivo: Estabelecer a sequência de eventos relacionados a marcadores inflamatórios, disfunção endotelial e pressão arterial sistólica (PAS) na obesidade em um modelo experimental. Métodos: Ratos Wistar machos (8 semanas de idade) receberam dieta padrão (Controle - CT, n = 35) ou uma dieta palatável hiperlipídica (DHL, n = 35) por 24 semanas. A cada seis semanas, 7 animais de cada grupo foram aleatoriamente selecionados para eutanásia. Foram determinados a PAS, e níveis séricos de interleucina-6, fator de necrose tumoral-a, proteína C reativa, adiponectina e óxido nítrico. As funções do músculo liso endotelial e vascular foram determinadas na aorta dissecada, e medida a peroxidação lipídica. A significância estatística foi estabelecida em p < 0,05. Resultados: os níveis das citocinas pró-inflamatórias começaram a aumentar após seis semanas de dieta hiperlipídica, enquanto os níveis da citocina anti-inflamatória adiponectina diminuíram. Um resultado interessante foi a redução da função endotelial e do óxido nítrico após seis semanas no grupo DHL. Além disso, mostramos que a massa de tecido adiposo visceral total esteve negativamente correlacionada com função endotelial e positivamente correlacionada com a PAS. Conclusão: Nossos resultados demonstram a progressão temporal dos efeitos deletérios e sua correlação com a obesidade.

Abstract Background: Obesity leads to a chronic inflammatory state, endothelial dysfunction and hypertension. Objective: To establish the time-course of events regarding inflammatory markers, endothelial dysfunction, systolic blood pressure (SBP) in obesity in only one experimental model. Methods: We fed male Wistar rats (eight-week age) with a standard diet (Control - CT, n = 35), or palatable high-fat diet (HFD, n = 35) for 24 weeks. Every six weeks, 7 animals from each group were randomly selected for euthanasia. SBP and serum levels of interleukin-6, tumor necrosis factor-α, C-reactive protein, adiponectin and nitric oxide were determined. Endothelial and vascular smooth muscle functions were determined in dissected aorta and lipid peroxidation was measured. Statistical significance was set at p < 0.05. Results: Levels of pro-inflammatory cytokines began to increase after six weeks of a high-fat diet, while those of the anti-inflammatory cytokine adiponectin decreased. Interestingly, the endothelial function and serum nitric oxide began to decrease after six weeks in HFD group. The SBP and lipid peroxidation began to increase at 12 weeks in HFD group. In addition, we showed that total visceral fat mass was negatively correlated with endothelial function and positively correlated with SBP. Conclusion: Our results show the time-course of deleterious effects and their correlation with obesity.

Resumo Objetivo: Avaliar o papel da sequência em difusão na avaliação de lesões mamárias suspeitas na ressonância magnética (RM), correlacionando seus achados com os resultados histológicos. Materiais e Métodos: Foi realizado estudo retrospectivo, descritivo, baseado na análise de prontuários médicos de 215 pacientes que realizaram RM com sequência em difusão e que foram submetidas a biópsia em um centro de referência oncológico. Foi calculado o valor do coeficiente de difusão aparente (ADC – apparent diffusion coefficient) para cada lesão e o resultado histológico foi considerado como padrão ouro. Resultados: A idade média das pacientes foi 49 anos. Foram identificadas 252 lesões, e destas, 161 (63,9%) eram lesões malignas na avaliação histológica. A média obtida do valor do ADC nas lesões benignas (1,50 × 10–3 mm2/s) foi superior à média das lesões malignas (0,97 × 10–3 mm2/s), com significância estatística (p < 0,001). O ponto de corte com maior sensibilidade e especificidade pela curva receiver operating characteristic foi 1,03 × 10–3 mm2/s. Com a combinação da difusão com os achados da RM, a acurácia chegou a 95,9%, com sensibilidade de 95,7% e especificidade de 96,4%. Conclusão: O uso da sequência em difusão pode auxiliar na caracterização das lesões mamárias, principalmente daquelas classificadas como BI-RADS 4, aumentando a especificidade e a acurácia diagnóstica da RM.

Abstract Objective: To assess the role of diffusion-weighted imaging (DWI) in the evaluation of breast lesions classified as suspicious on magnetic resonance imaging (MRI), correlating the findings with the results of the histological analysis. Materials and Methods: This was a retrospective, descriptive study based on a review of the medical records of 215 patients who were submitted to MRI with DWI before undergoing biopsy at a cancer center. Apparent diffusion coefficient (ADC) values were calculated for each lesion, and the result of the histological analysis was considered the gold standard. Results: The mean age was 49 years. We identified 252 lesions, 161 (63.9%) of which were found to be malignant in the histological analysis. The mean ADC value was higher for the benign lesions than for the malignant lesions (1.50 × 10–3 mm2/s vs. 0.97 × 10−3 mm2/s), the difference being statistically significant (p < 0.001). The ADC cut-off point with the greatest sensitivity and specificity on the receiver operating characteristic curve was 1.03 × 10−3 mm2/s. When the DWI and conventional MRI findings were combined, the accuracy reached 95.9%, with a sensitivity of 95.7% and a specificity of 96.4%. Conclusion: The use of DWI could facilitate the characterization of breast lesions, especially those classified as BI-RADS 4, increasing the specificity and diagnostic accuracy of MRI.

The protective effect of infectious agents against allergic reactions has been thoroughly investigated. Current studies have demonstrated the ability of some helminths to modulate the immune response of infected hosts. The objective of the present study was to investigate the relationship between Toxocara canis infection and the development of an allergic response in mice immunised with ovalbumin (OVA). We determined the total and differential blood and bronchoalveolar lavage fluid cells using BALB/c mice as a model. To this end, the levels of interleukin (IL)-4, IL-5 and IL-10 and anti-OVA-IgE were measured using an ELISA. The inflammatory process in the lungs was observed using histology slides stained with haematoxylin and eosin. The results showed an increase in the total number of leukocytes and eosinophils in the blood of infected and immunised animals at 18 days after infection. We observed a slight lymphocytic inflammatory infiltrate in the portal space in all infected mice. Anti-OVA-IgE levels were detected in smaller proportions in the plasma of immunised and infected mice compared with mice that were only infected. Therefore, we concluded that T. canis potentiates inflammation in the lungs in response to OVA, although anti-OVA-IgE levels suggest a potential reduction of the inflammatory process through this mechanism.

The significance of polyphenol intake for the prevention of chronic diseases is controversial. Objective: this study investigated the chemical composition and antioxidant potential of an anthocyanin-rich extract from Euterpe edulis fruits (LPEF) and its effects on liver steatosis in dyslipidemic apoE-/- knockout mice. Materials and methods: mice were divided into G1 (C57BL/6) standard diet; G2 (apoE-/-) standard diet, G3 (apoE-/-) 2% LPEF, G4 (apoE-/-) 6% LPEF, G5 (apoE-/-) 10% LPEF, G6 (apoE-/-) 2% α-tocopherol acetate. After 75 days of treatment, the animals were euthanized. The LPEF contained a high level of monomeric anthocyanins (301.4 mg/100g) and marked antioxidant activity. Results: Catalase activity was reduced in G3, G4, G5 and G6 compared to G2. Superoxide dismutase was reduced only in G4. The animals in G4, G5, and G6 showed low HDL and triglycerides levels compared to G2. The proportion of lipid droplets in liver tissue was reduced in G4 and G5 compared to G2, G3, and G6. Conclusion: The results indicated that E. edulis pulp is rich in anthocyanins and the LPEF dietary consumption can reduce the severity of liver steatosis in apoE-/- mice, an effect that is potentially mediated by the antioxidant activity of this extract and modulation of triglyceride serum levels.

El papel de los polifenoles en la prevencion de enfermedades cronicas es controvertido. Objetivo: este estudio investigo la composicion quimica y el potencial antioxidante de un extracto del fruto de Euterpe edulis rico en antocianinas (LPEF) y sus efectos en la esteatosis hepatica en ratones apoE-/- knockout con dislipidemia. Material y metodos: los ratones fueron divididos en los siguientes grupos; G1 (C57BL/6) con una dieta estandar; G2 (apoE-/-) con dieta estandar; G3 G3 (apoE-/-) con 2% de LPEF; G4 (apoE-/-) con 6% de LPEF; G5 (apoE-/-) con 10% de LPEF y G6 (apoE-/-) con 2% acetato α-tocoferol (α-tocopherol acetate). Despues de 75 dias de tratamiento, los animales fueron eutanizados. El LPEF contenia un alto nivel de antocianinas monomericas (301,4 mg/100 g) con notable actividad antioxidante. Resultados: la actividad catalasa fue reducida en los grupos G3, G4, G5 y G6 comparada con G2. La superoxidasa dismutasa solo se redujo en el grupo G4. Los animales de G4, G5 y G6 mostraron bajos niveles de HDL trigliceridos, comparados con G2. La proporcion de lipidos en el tejido hepatico fue reducida en G4 y G5, comparado con G2, G3 y G6. Conclusion: los resultados indicaron que la pulpa de E. edulis es rica en antocianinas, y que el consumo de LPEF en la dieta puede reducir la severidad de la esteatosis hepatica en ratones apoE-/-, un efecto que es potencialmente mediado por la actividad antioxidante de este extracto y la modulacion en los niveles sericos de trigliceridos.

FUNDAMENTO: A capacidade aeróbica é fundamental para o desempenho físico, e a baixa capacidade aeróbica está relacionada ao desencadeamento de diversas doenças cardiovasculares. OBJETIVO: Comparar a contratilidade e a morfologia de cardiomiócitos isolados de ratos com baixo desempenho e desempenho padrão para o exercício físico. MÉTODOS: Ratos Wistar, com 10 semanas de idade, foram submetidos a um protocolo de corrida em esteira até a fadiga, e foram divididos em dois grupos: Baixo Desempenho (BD) e Desempenho Padrão (DP). Em seguida, após eutanásia, o coração foi removido rapidamente e, por meio de dissociação enzimática, os cardiomiócitos do ventrículo esquerdo foram isolados. O comprimento celular e dos sarcômeros e a largura dos cardiomiócitos foram medidos usando-se um sistema de detecção de bordas. Os cardiomiócitos isolados foram estimulados eletricamente a 1 e 3 Hz e a contração celular foi medida registrando-se a alteração do seu comprimento. RESULTADOS: O comprimento celular foi menor no grupo BD (157,2 ± 1,3µm; p < 0,05) em relação ao DP (161,4 ± 1,3 µm), sendo o mesmo resultado observado para o volume dos cardiomiócitos (BD, 25,5 ± 0,4 vs. DP, 26,8 ± 0,4 pL; p < 0,05). Os tempos para o pico de contração (BD, 116 ± 1 vs. DP, 111 ± 2ms) e para o relaxamento total (BD, 143 ± 3 vs. DP, 232 ± 3 ms) foram maiores no grupo BD. CONCLUSÃO: Conclui-se que os miócitos do ventrículo esquerdo dos animais de baixo desempenho para o exercício físico apresentam menores dimensões que os dos animais de desempenho padrão, além de apresentarem perdas na capacidade contrátil.

BACKGROUND: Aerobic capacity is essential to physical performance, and low aerobic capacity is related to the triggering of various cardiovascular diseases. OBJECTIVE: To compare the morphology and contractility of isolated rat cardiomyocytes with low performance and standard performance for exercise. METHODS: Wistar rats with 10 weeks of age underwent a protocol of treadmill running to fatigue, and were divided into two groups: Low Performance (LP) and Standard Performance (SP). Then, the animals were sacrificed, the heart was quickly removed and, by means of enzymatic dissociation, left ventricular cardiomyocytes were isolated. The cell and sarcomeres length and width of cardiomyocytes were measured using an edge detection system. The isolated cardiomyocytes were electrically stimulated at 1 and 3 Hz and cell contraction was measured by registering the change of their length. RESULTS: The cell length was shorter in the LP group (157.2 1.3µm; p < 0.05) compared to SP (161.4 1.3µm), and the same result was observed for the volume of cardiomyocytes (LP, 25.5 0.4. vs. SP, 26.8 ± 0.4 pL; p < 0.05). The time to peak contraction (LP, 116 1 vs. SP 111 2ms) and total relaxation (LP, 143 3 vs. SP 232 3ms) were higher in the LP group. CONCLUSION: We conclude that left ventricular myocytes of animals with low performance for exercise are smaller than animals with standard performance. In addition to that, they present losses in contractile capacity.

Enquanto o exercício aeróbico regular promove adaptações benéficas ao músculo esquelético, o exercício físico exaustivo induz lesões musculares. O objetivo deste estudo foi verificar se um programa de natação com baixa intensidade é capaz de proteger músculos esqueléticos contra lesões induzidas por exercício exaustivo. Ratos Wistar (peso: 376,50 ± 4,36g; idade: 90 dias) foram divididos aleatoriamente em quatro grupos: controle sedentário (CS); sedentário submetido a teste de exaustão (SE); treinado em natação (TN); treinado em natação submetido a teste de exaustão (TNE). Animais dos grupos TN e TNE foram submetidos a um programa de natação sem sobrecarga por 90 minutos/dia, cinco dias/semana, durante 17 semanas. Após este período, os grupos SE e TNE foram submetidos a um teste de exaustão em natação. Após eutanásia, fragmentos dos músculos sóleo e reto femoral foram coletados e submetidos à análise histológica e de proteínas de choque térmico (HSP70). Os resultados mostraram que o tempo até a exaustão foi maior no grupo TNE que no SE (125,0 ± 6,0 vs. 90,0 ± 8,5min, respectivamente, P < 0,05). Os níveis de lactato sanguíneo durante o teste e exaustão foram menores no grupo TNE que no SE (5,31 ± 0,22 vs. 8,76 ± 0,59mmol/L, respectivamente, P < 0,05). A frequência de fibras lesadas nos músculos foi maior nos grupos SE (sóleo: 34,86 ± 0,04; reto femoral: 37,57 ± 0,07) e TNE (sóleo: 41,57 ± 0,08; reto femoral: 39,57 ± 0,05), comparada aos grupos CS (sóleo: 13,88 ± 0,81; reto femoral: 16,75 ± 0,79) e TN (sóleo: 24,14 ± 0,06; reto femoral: 24,0 ± 0,05), respectivamente (P < 0,05). Não houve diferença significativa nos níveis de HSP70 dos músculos analisados entre os quatro grupos. Concluimos que apesar do treinamento em natação melhorar o desempenho dos animais no teste de exaustão, não promoveu proteção aos seus músculos esqueléticos contra as lesões induzidas pelo exercício exaustivo

While regular aerobic exercise promotes beneficial adaptations to the skeletal muscle, acute exhaustive exercise causes structural damage to the skeletal muscle cells. The aim of this study was to verify whether a low-intensity swimming program protects the skeletal muscles against damage induced by exhaustive exercise. Male Wistar rats (weight: 376.50 4.36g; age: 90 days) were randomly divided into four groups: sedentary control (SC, N=8); sedentary submitted to exhaustive test (SE, N=7); swimming trained (TN, N=7); swimming trained submitted to exhaustive test (TNE, N=7). Animals of TN and TNE groups were submitted to a swimming regimen without overload for 90 min/day, 5 days/wk, during 17 weeks. Forty-eight hours after the last training session, animals from SE and TNE groups were submitted to an exhaustive exercise protocol. At sacrifice, fragments of soleus and rectus femoris muscles were collected and submitted to histological analysis and heat shock protein (HSP70) expression measurement. The results showed that the time until exhaustion was greater in the STE than in SE group (125.0 6.00 vs. 90.0 8.48 min, respectively, P<0.05). The levels of blood lactate during exhaustive exercise were lower in animals from TNE than SE (5.31 ± 0.22 vs. 876 ± 0.59 mmol/L, respectively, P<0.05)The frequency of damaged fibers in the muscles was greater in SE (soleus: 34.86±0.04; rectus femoris: 37.57 ± 0.07) and STE (soleus: 41.57±0.08; rectus femoris: 39.57 ± 0.05), compared to groups SC (soleus: 13.88±0.81; rectus femoris: 16.75 ± 0.79) and ST (soleus: 24.14±0.06; rectus femoris: 24.0 ± 0.05), respectively (P<0.05). There was no significant difference at the HSP70 levels of the analyzed muscles among the four groups (P>0.05). In conclusion, although a low-intensity swimming training increased the animals' performance in the exhaustive exercise test, it did not protect their skeletal muscles against damage induced by exhaustive exercise