Effect of combined red and infrared wavelengths on inflammation, hemorrhage, and muscle damage caused by Bothrops leucurus snake venom.

To evaluate the effects of red and infrared wavelengths, separately and combined, on the inflammatory process and collagen deposition in muscle damage caused by B. leucurus venom. 112 mice were inoculated with diluted venom (0.6mg/kg) in the gastrocnemius muscle. The animals were divided into four groups: one control (CG) and three treatments, namely: 1) red laser (λ=660 nm) (RG), 2) infrared laser (λ=808 nm) (IG) and 3) red laser (λ=660 nm) + infrared (λ=808 nm) (RIG). Each group was subdivided into four subgroups, according to the duration of treatment application (applications every 24 hours over evaluation times of up to 144 hours). A diode laser was used (0.1 W, CW, 1J/point, ED: 10 J/cm2). Both wavelengths reduced the intensity of inflammation and the combination between them significantly intensified the anti-inflammatory response. Photobiomodulation also changed the type of inflammatory infiltrate observed and RIG had the highest percentage of mononuclear cells in relation to the other groups. Hemorrhage intensity was significantly lower in treated animals and RIG had the highest number of individuals in which this variable was classified as mild. As for collagen deposition, there was a significant increase in RG in relation to CG, in RIG in relation to CG and in RIG in relation to IG. Photobiomodulation proved to be effective in the treatment of inflammation and hemorrhage caused by B. leucurus venom and stimulated collagen deposition. Better results were obtained with the combined wavelengths.

Hepatic expression of tumor necrosis factor-α and evaluation of MAPK-p38 and NFκB signaling pathways in autoimmune hepatitis.

OBJECTIVES: Autoimmune hepatitis (AIH) is a necroinflammatory disease that occurs when genetically susceptible individuals are exposed to an environmental trigger. It is a rare disease, and its epidemiological aspects are nearly unknown in Northeast Brazil. In the literature, the activation of components of the inflammatory cascade pathways, including interleukins such as TNF-α and signaling factors like MAPK-p38 and NFκB, in the pathogenesis of AIH is well described in animal models. This study evaluated, for the first time, the immunostaining of TNF-α, MAPK-p38, and NFκB in immunohistochemical analysis of liver biopsies from AIH patients. The activation of the MAPK-p38 pathway was also studied through immunoassay analysis in the peripheral blood of AIH patients. METHODS AND RESULTS: Data from medical records of 25 AIH patients were analyzed. Histological and immunohistochemical analysis of liver tissue obtained from biopsies was performed to detect NFκB, MAPK-p38, and TNF-α. Immunoassay analysis of the MAPK-p38 pathway was performed in peripheral blood from 18 AIH patients and 8 healthy volunteers. Medical record analysis showed an average age of 33.3 years, with a female predominance in a ratio of 7.3:1. Concomitance with other autoimmune diseases was observed in 36 % of patients, with thyroid disorders being the most prominent among them, and an 8 % indication for liver transplantation. In the evaluation of autoantibodies, ANA was detected in 52 %, followed by SMA at 20 %, and Anti-LKM-1 at 16 %. Liver biopsy findings were like the global literature, with interface hepatitis and lymphoplasmacytic infiltration observed. Immunohistochemical analysis showed immunostaining for NFκB, MAPK-p38, and TNF-α, corroborating the inflammatory and immunological characteristics of the disease. Immunoassay analysis in peripheral blood confirmed the activation of the MAPK-p38 signaling pathway, with a statistically significant difference between AIH patients and healthy controls. CONCLUSIONS: The epidemiological and histological findings of AIH in this study in Northeast Brazil were like global population data. Immunohistochemical analysis of liver tissue and immunoassay analysis in peripheral blood confirmed the activation of TNF-α and the NFκB and MAPK-p38 signaling pathways in AIH patients.

Histopathological Biocompatibility Evaluation of TheraCal PT, NeoMTA, and MTA Angelus in a Murine Model.

The aim of this study was to evaluate the biocompatibility of the regeneration of the dentin-pulp complex in a murine model with different treatments with MTA Angelus, NeoMTA, and TheraCal PT. An in vivo controlled experimental study of 15 male Wistar rats forming three study groups, the upper and lower central incisors were selected where pulpotomies were conducted, leaving a central incisor as control at 15, 30, and 45 days. For data analysis, these were expressed as mean ± standard deviation and were examined by Kruskal-Wallis test. Three factors were analyzed as follows: "inflammatory infiltrate; disorganization of pulp tissue, and the formation of reparative dentin". No statistical significance was found between the different groups (p > 0.05). Treatment with these three biomaterials (MTA, TheraCal PT, and Neo MTA) presented an inflammatory infiltrate and slight disorganization of the odontoblast layer in the pulp tissue of a murine model, with normal coronary pulp tissue and the formation of reparative dentin in the three experimental groups. Thus, we are able to conclude that all three are biocompatible materials.

DC-SIGN receptor is expressed by cells from cutaneous leishmaniasis lesions and differentially binds to Leishmania (Viannia) braziliensis and L. (Leishmania) amazonensis promastigotes

BACKGROUND Dendritic cells (DCs) specific intercellular adhesion molecule (ICAM)-3-grabbing non integrin receptor (DC-SIGN) binds to subgenera Leishmania promastigotes mediating its interaction with DC and neutrophils, potentially influencing the infection outcome. OBJECTIVES In this work, we investigated whether DC-SIGN receptor is expressed in cells from cutaneous leishmaniasis (CL) lesions as well as the in vitro binding pattern of Leishmania (Viannia) braziliensis (Lb) and L. (L.) amazonensis (La) promastigotes. METHODS DC-SIGN receptor was labeled by immunohistochemistry in cryopreserved CL tissue fragments. In vitro binding assay with CFSE-labeled Lb or La promastigotes and RAJI-transfecting cells expressing DC-SIGN (DC-SIGNPOS) or mock-transfected (DC-SIGNNEG) were monitored by flow cytometry at 2 h, 24 h and 48 h in co-culture. RESULTS In CL lesion infiltrate, DC-SIGNPOS cells were present in the dermis and near the epidermis. Both Lb and La bind to DC-SIGNPOS cells, while binding to DC-SIGNNEG was low. La showed precocious and higher affinity to DC-SIGNhi population than to DC-SIGNlow, while Lb binding was similar in these populations. CONCLUSION Our results demonstrate that DC-SIGN receptor is present in L. braziliensis CL lesions and interact with Lb promastigotes. Moreover, the differences in the binding pattern to Lb and La suggest DC-SIGN can influence in a difference way the intake of the parasites at the first hours after Leishmania infection. These results raise the hypothesis that DC-SIGN receptor could participate in the immunopathogenesis of American tegumentary leishmaniasis accounting for the differences in the outcome of the Leishmania spp. infection.

Comparison of Bio-C Pulpo and MTA Repair HP with White MTA: effect on liver parameters and evaluation of biocompatibility and bioactivity in rats.

AIM: To evaluate the tissue response promoted by Bio-C Pulpo (Bio), MTA Repair HP (MTA-HP) and White MTA (WMTA) and whether these materials cause liver changes in a rat experimental model. METHODOLOGY: Polyethylene tubes filled with Bio, MTA-HP and WMTA, and empty tubes (control group, CG) were implanted into the subcutaneous tissues of rats for 7, 15, 30 and 60 days. Inflammatory reaction score (IRS), capsule thickness, number of inflammatory cells (IC), von Kossa reaction, interleukin-6 (IL-6) and alkaline phosphatase (ALP) immunohistochemistry reactions were performed. Combined methods, von Kossa followed by immunohistochemistry for detection of ALP, were performed. At 60 days, the serum glutamic-oxaloacetic transaminase (GOT) and glutamic-pyruvic transaminase (GPT) levels were measured and liver fragments were collected for histological analysis; the data were assessed by one-way ANOVA analysis followed by Sidak's post-test. The biocompatibility and bioactivity data were subjected to the two-way ANOVA analysis followed by Tukey post hoc test, except the IRS. The IRS data were subjected to the Kruskal-Wallis ANOVA non-parametric test followed by Dunn's test (p ≤ .05). RESULTS: No significant difference was detected in serum GOT and GPT concentrations and in the number of hepatocytes among the experimental and CG samples. Although Bio-C Pulpo had the highest IC and IL-6-immunolabelled cells (p < 0.0001) at all periods, no significant difference was observed in the IRS among the materials, except at 60 days. In this period, the WMTA had lower IRS. All groups had a significant reduction in the capsule thickness and in the number of IC and IL-6-immunolabelled cells over time. Bio-C Pulpo, MTA-HP and WMTA specimens had greater immunoexpression of ALP than CG (p < .0001). At all periods, von Kossa-positive and birefringent structures were observed in the capsules around the materials. ALP-immunolabelled cells were also seen near von Kossa-positive structures. CONCLUSIONS: Bio-C Pulpo, MTA-HP and WMTA materials did not cause morphological changes in the liver and no significant alteration in the serum GOT and GPT levels. Moreover, these bioceramic materials were biocompatible and exhibited bioactive potential. However, Bio-C Pulpo induced greater inflammatory infiltrate than MTA-HP and WMTA at all periods.

Avaliação do microambiente tumoral em adenocarcinomas gástricos a partir de dados de expressão gênica / Evaluation of the tumor microenvironment in gastric adenocarcinomas using gene expression data

O câncer gástrico (CG) tem grande importância clínica devido a suas altas taxas de incidência e mortalidade estando entre os cinco tipos de câncer mais frequentes no mundo. O CG encontra-se associado ao diagnóstico tardio, que, somado a agressividade da doença, resulta em sobrevida mediana curta. A principal forma de tratamento é a cirurgia em conjunto com a quimioterapia. Mais recentemente, o tratamento com imunoterpia foi aprovado para casos de CG avançado metastático. O CG é usualmente classificado nos subtipos histológicos difuso e intestinal, porém, devido à sua alta heterogeneidade genética, pode ser também classificado em quatro subtipos moleculares (CIN, MSI, GS e EBV). Ambos aspectos de classificação estão associados a diferentes prognósticos e taxas de sobrevida, mas ainda não são suficientes para direcionar ou predizer quais pacientes respondem melhor a uma ou outra terapia. Neste contexto, outro fator relacionado à patogênese da doença, resposta a tratamento e prognóstico é o contexto imunológico do microambiente tumoral, como a composição e orientação funcional das células do infiltrado inflamatório associado ao tumor. Por meio de um método analítico de deconvolução aplicado a dados de expressão gênica é possível estimar a proporção de diferentes tipos celulares, além de seus aspectos funcionais. Neste trabalho, foi feito um levantamento das ferramentas e listas de genes marcadores de células imunes a fim de definir qual teria a melhor performance na estimação da proporção de células imunes em amostras de CG. Não observamos diferenças na composição do infiltrado inflamatório entre os subtipos histológios; já entre os subtipos moleculares de CG as proporções de linfóctios T CD8 se destacaram como maiores nos subtipos EBV e MSI, por exemplo. Foi então proposta uma classificação baseada em assinaturas mutacionais que conseguiu resgatar pacientes que apresentaram melhor prognóstico. O contexto imunológico nos pacientes com alta atividade mutacional (S4) apresentou elementos de que ser mais responsivo com maior expressão de genes associados à resposta imune citotóxica e do mecanismo de escape pelos checkpoints imunes. Além disso, há indícios de que há formação das estruturas terciárias de linfócitos pela correlação entre as quantidades estimadas de linfócitos T e B. Já o grupo com baixa atividade da assinatura (S4), associado a um pior prognóstico, apresentou maiores quantidades de linfócitos Treg. A avaliação global do microambiente tumoral, seus aspectos moleculares e imunológicos, proporciona dados mais informativos para estratificação dos pacientes quanto ao prognóstico e podem ser utilizadas em breve na prática clínica para direcionamento terapêutico.

Gastric cancer (GC) has great clinical importance due to its high incidence and mortality rates, being among the five most frequent types of cancer in the world. GC is associated with late diagnosis which, added to the aggressiveness of the disease, results in a short median survival. The main form of treatment is surgery in conjunction with chemotherapy. Recently, immunotherapy treatment has been approved for advanced metastatic GC. GC is usually classified into diffuse and intestinal histological subtypes, however, due to its high genetic heterogeneity, it can also be classified into four molecular subtypes (CIN, MSI, GS and EBV). Both classification are associated with different prognosis and survival rates, but they are still not sufficient to direct or predict which patients respond better to one or another therapy. In this context, another factor related to the pathogenesis of the disease, response to treatment and prognosis is the immunological context of the tumor microenvironment, such as the composition and functional orientation of the cells of the inflammatory infiltrate associated with the tumor. Through an analytical method of deconvolution applied to gene expression data, it is possible to estimate the proportion of different cell types, in addition to their functional aspects. In this work, the tools and lists of immune cell marker genes were surveyed in order to define which one would perform better in estimating the proportion of immune cells in GC samples. We did not observe differences in the composition of the inflammatory infiltrate between the histological subtypes; among the CG molecular subtypes, the proportions of T CD8 lymphocytes stood out as higher in the EBV and MSI subtypes, for example. A classification based on mutational signatures was then proposed, which managed to rescue patients with a better prognosis. The immunological context in these patients with high mutational activity (S4) showed elements of being more responsive with greater expression of genes associated with the cytotoxic immune response and the escape mechanism by immune checkpoints. In addition, there is evidence that there is formation of tertiary lymphocyte structures by the correlation between the estimated amounts of T and B lymphocytes. The group with low mutational signature activity (S4), on the other hand, associated with a worse prognosis, had higher amounts of Treg lymphocytes. The global assessment of the tumor microenvironment, its molecular and immunological aspects, provides more informative data for stratifying patients regarding prognosis and may soon be used in clinical practice for therapeutic guidance.

Dietary supplementation with multi-strain formula of probiotics modulates inflammatory and immunological markers in apical periodontitis

Abstract Objective The aim of this study was to evaluate whether probiotics multi-strain formula affects the development of apical periodontitis (AP) induced in rats. Methodology 16 Wistar rats were divided in two groups (n=8): rats with AP fed with regular diet (Control-C (CG)); rats with AP, fed with regular diet and supplemented with multi-strain formula (one billion colony-forming units (CFU)): GNC Probiotic Complex (PCG) ( Lactobacillus acidophilus, Lactobacillus salivaris, Lactobacillus plantarum, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium animalis subs. lactis and Streptococcus thermofilus ). AP was induced in the upper and lower first molars by dental pulp exposure to the oral environment. PCG was administered orally through gavage for 30 days during the AP development. After this period the animals were euthanized and the mandibles were removed and processed for histologic analysis, and immunochemical assays for interleukin (IL)-6, IL-10, IL-1β, RANKL, OPG, and TRAP. The Mann-Whitney U test and Student's t test were performed (P<.05). Results The CG showed more intense inflammatory infiltrate than the PCG group (P<.05). IL-1β, IL 6 and RANKL decreased in the PCG group compared with CG (P<.05). The IL-10 level increased in the PCG group (P<.05). The OPG level was similar in both groups (P>.05). The number of mature osteoclasts (TRAP-positive multinucleated cells) was lower in PCG group when compared to the CG (P<.05). Conclusion Probiotic Complex modulates inflammation and bone resorption in apical periodontitis.

Safety and bioactive potential of nanoparticles containing Cantaloupe melon (Cucumis melo L.) carotenoids in an experimental model of chronic inflammation.

The safety and bioactive potential of crude carotenoid extract from Cantaloupe melon nanoencapsulated in porcine gelatin (EPG) were evaluated in a chronic inflammatory experimental model. Animals were fed a high glycemic index and high glycemic load (HGLI) diet for 17 weeks and treated for ten days with 1) HGLI diet, 2) standard diet, 3) HGLI diet + crude carotenoid extract (CE) (12.5 mg/kg), and 4) HGLI diet + EPG (50 mg/kg). General toxicity signals were investigated, considering body weight, food intake, hematological, biochemical parameters, relative weight, morphology, and histopathology of organs. The biochemical parameters indicated the low toxicity of EPG. Acute hepatitis was observed in animals' livers, but CE and EPG groups presented improved tissue appearance. Chronic enteritis was observed in animals, with villi and intestinal glands preservation in the EPG group. The results suggest the safety and the bioactive effect of EPG, possibly related to its anti-inflammatory potential.

Effect of Saccharomyces cerevisiae UFMG A-905 in a murine model of food allergy.

Food allergy is triggered when there is an abnormal activation of the immune system by food allergens. Currently, there is no curative therapy for this pathological condition. Due to the immunomodulatory properties of probiotics they are potential candidates as therapeutic tools for food allergy. Therefore, the aim of this study was to evaluate the probiotic effect of Saccharomyces cerevisiae UFMG A-905 (905) in an in vivo model of food allergy. Probiotic effect was assessed by clinical, histological, immunological and microbiological parameters analysis. Furthermore, we also evaluated if 905 after inactivation has an effect, as well as if such an effect is dose dependent. Our results showed that oral administration of only viable 905 promotes a significant attenuation of tissue injury and myeloperoxidase (MPO) activity levels. Moreover, the treatment reduced interleukin 17 levels, and administration of the supernatant from the yeast culture also promoted a significant decrease in MPO levels. However, considering the systemic parameters, immunoglobulin (Ig)E and IgG anti-ovalbumin, which are essentials for triggering the allergic process, there was no effect, suggesting that the yeast promotes a local but not a systemic effect in the model evaluated. In addition, we found that only high doses of viable 905 were able to attenuate the signs of inflammation. In conclusion, oral administration of 905 led to a local effect that depends on the viability of the yeast.

Phenotypic characterization of immune cells in fetal tissues of cattle immunized and challenged with Neospora caninum.

The purpose of this work was to characterize the cellular phenotype in inflammatory infiltrates of fetal tissues from pregnant heifers immunized and experimentally challenged with Neospora caninum. Fetuses from 20 heifers separated into 5 groups were obtained. The experiment was designed as follow: Group A, heifers inoculated intravenously with live tachyzoites of Argentine strain NC-6 (n = 4); Group B heifers inoculated subcutaneously with soluble native antigen from the same strain formulated with immune stimulant complexes (ISCOMs) (n = 4); Group C heifers inoculated with recombinant proteins, rNcSAG1, rNcHSP20, rNcGRA7 formulated with ISCOMs (n = 4), Group D heifers inoculated subcutaneously with sterile phosphate buffered solution (n = 4) and Group E heifers inoculated subcutaneously with antigen-free ISCOMs (n = 4). Experimental challenge was performed at 70 days of gestation and all heifers were euthanized 34 days later. Fetal tissues were taken for histological studies. Inflammatory lesions were observed in brain and lung, and immunhistochemistry was used to identify CD3+, CD20+ and MHC II+ cells. The majority of the cells that infiltrate and circumscribe the lesions in the brain and lung tissue expressed MHC II antigen; varying between 70-90% of the total cellular infiltrate. CD3+ cells were also present within the lesions, contributing to up to 30% of the inflammatory cells. CD20+ cells appeared as a marginal group, in some cases, with a range between 10 and 25%. As expected, the immunolabeling of MHC II + and CD3 + cells in fetal tissues was associated with fetal infection with N. caninum. There were statistically significant differences in the distribution and population of the inflammatory infiltrate in relation to the immunogenic treatment and the type of tissue, with inflammatory cells being markedly less extensive fetuses from group A (dams previously exposed to N. caninum) and in brain tissue. This work showed that Neospora-infection induced MHC II+ and CD3+ cells in bovine fetuses from dams receiving experimental vaccines.

Chronic hepatitis B: The interplay between intrahepatic lymphocyte population and viral antigens in relation to liver damage.

In Chronic hepatitis B (CHB) infection, virus and immune response interplay is thought to be responsible for pathogenesis. Yet, the impact of each immune cell population and viral protein expression in liver damage is still unknown. Our aim was to study the interplay between intrahepatic immune response and viral activity in relation to CHB liver damage. Immunostaining was performed in 29 liver biopsies from untreated CHB patients to characterize liver infiltrate [Th (CD4+), CTL (CD8+), Treg (FoxP3+), Th17 (IL-17A+) and Th1 (T-bet+)] and viral antigen expression (HBsAg and HBcAg). Inflammatory activity and fibrosis were assessed using the HAI and METAVIR scoring system. All studied populations were identified in the portal-periportal (P-P) areas with a CD4+ lymphocyte predominance, while only CD8+ and FoxP3+ cells were observed in the intralobular area. Both P-P CD4+ and intralobular CD8+ cell frequencies were increased among severe hepatitis cases. Concerning HBsAg and HBcAg expression, a mutually exclusive pattern was observed. HBcAg was mainly detected among HBeAg-positive patients and was associated with hepatitis severity and higher frequency of P-P FoxP3+, intralobular CD8+ and FoxP3+ cells. HBsAg was identified among HBeAg-negative cases with less severe hepatitis grade and lower frequency of P-P CD4+ and intralobular FoxP3+ lymphocytes. In conclusion, the HBV antigen profile expression seen during CHB infection may be reflecting different stages of viral replication which impacts the host immune response and liver damage process. While HBcAg might be an inducer of a regulatory microenvironment, the intralobular CTL population seemed to have a key role in hepatitis severity.

Toxoplasma gondii causes lipofuscinosis, collagenopathy and spleen and white pulp atrophy during the acute phase of infection.

In this study, we evaluated homeostatic and functional disorders of the spleen in mice inoculated with Toxoplasma gondii. The kinetics of megakaryocyte and leukocyte production, body and spleen mass and certain histopathological aspects were analyzed. There was increased (P < 0.05) the accumulation of lipofuscin in the red pulp of the spleen, in the periods of 30 and 60 dpi of the infection, that is, in the chronification stage of the disease and decrease of the white pulp area. In addition, we observed (from 7dpi) a quantitative and qualitative increase (P < 0.05) in the deposition of collagen fibers in the spleen of all infected mice. Since resolution of the inflammatory process resulted in pathophysiological changes, we can suggest that the T. gondii invaded and multiplied in the cells of the white and red pulps of the spleen. Although we did not find the parasite in the spleen, this hypothesis is supported by the presence of diffuse inflammatory infiltrate, which extended through the spleen parenchyma of all inoculated mice. Taken together, our results suggest that T. gondii causes severe homeostatic disorders that have altered spleen physiology, including diffuse parenchymal inflammation, lipofuscinosis in histiocytes, early aging, collagenopathy, systemic sclerosis and spleen and white pulp atrophy.

El infiltrado tumoral del carcinoma ductal infiltrante en Holguín, Cuba / Ductal infiltrating carcinoma´s inflammatory tumor infiltrates in Holguín, Cuba

Introducción: el carcinoma ductal infiltrante es la neoplasia maligna mamaria más frecuente. Entre sus características anatomopatológicas se presenta el grado histológico de malignidad, como su factor pronóstico. Existen otras alteraciones histopatológicas, como la presencia e intensidad del infiltrado inflamatorio intratumoral y peritumoral, sobre lo cual es necesario profundizar. Objetivos: determinar la intensidad del infiltrado inflamatorio intraperitumoral y peritumoral, e identificar su relación con el grado histológico de malignidad. Métodos: el estudio de una serie de casos de 392 biopsias, diagnosticadas con carcinoma ductal infiltrante de mama, en el Departamento de Anatomía Patológica del Hospital Lenin, de Holguín, Cuba, desde el año 2011 al 2015. Se determinó la presencia e intensidad del infiltrado inflamatorio intratumoral y peritumoral, y su grado histológico de malignidad. Resultados: se logró diferenciar el 68,1% de los tumores de mama. Predominaron los infiltrados inflamatorios intratumoral y peritumoral ligeros, con 44,9% y 55,4% respectivamente. El 84,6% de los tumores bien diferenciados, mostraron un infiltrado inflamatorio intratumoral ligero. En el 47,2% de los tumores moderadamente diferenciados, y en el 41,7% de los tumores poco diferenciados, con un infiltrado inflamatorio severo; se realizaron infiltrados moderados. En los tumores bien diferenciados se observó un predominio del infiltrado inflamatorio peritumoral ligero, de hasta el 92,3%; mientras que el 48,3% de los tumores con poca diferenciación, mostraron una correlación positiva con el infiltrado inflamatorio severo. Conclusiones: en el infiltrado inflamatorio intratumoral y peritumoral, predominó la intensidad ligera, y apareció un factor pronóstico potencial: la correlación entre la intensidad del infiltrado inflamatorio y el grado histológico de malignidad.

Introduction: among most frequent malignant breast tumors, ductal carcinoma has prevailed. Its malignancy histological level has become the prognosis factor. However, histopathological abnormalities like intra and peritumoral inflammatory infiltrates intensity, still require serious studies. Objectives: to determine intra and peritumoral inflammatory infiltrates intensity, and to identify their relation to malignancy histological levels. Methods: a case series study with 392 infiltrating ductal carcinoma diagnosed biopsies, at Lenin Hospital Pathological Anatomy Department, from 2011 to 2015; in which presence and intensity of intratumoral, and peritumoral inflammatory infiltrates, and malignancy histological level, were found. Results: breast tumors were differentiated up to 68.1%. Intra and peritumoral lights inflammatory infiltrate prevailed, with 44.9% and 55.4% each. Well differentiated lights inflammatory infiltrated tumors reached 84.6%. Moderate differentiated tumors were moderate infiltrated, up to 47.2%; and poor diferentiated with severe inflamatory infíltrate, presented 41.7%. Among well differentiated tumors, peritumoral light inflamatory infiltrate revealed 92.3%. Those poor differentiated were only 48.3% positively related. Conclusions: intra and peritumoral light intensity inflammatory infiltrate was the most frequent, considered as the potential prognosis factor, because of its directly proportional relationship with malignancy histological level.

Avaliação clínica, endoscópica e histopatológica de cães com doença inflamatória intestinal / Clinical, endoscopic and histopathological evaluation of dogs with inflammatory bowel disease

O presente estudo teve como objetivo avaliar macro e microscopicamente a mucosa gastrintestinal de cães domiciliados, que apresentaram sinais gastrintestinais crônicos, por meio de exame endoscópico e da avaliação histopatológica. Os sinais clínicos, a macro e a microscopia das mucosas gástrica e duodenal foram correlacionados. Foram avaliados 20 cães de diferentes raças, sendo 7 machos e 13 fêmeas, com idade variando entre 1,7 a 15,8 anos, que apresentavam cronicidade dos sinais característicos da Doença Inflamatória Intestinal (DII). O estudo foi prospectivo e transversal, realizado em cães atendidos no Hospital Universitário de Medicina Veterinária da Universidade Federal Fluminense (HUVET - UFF) e em clínicas privadas do município do Rio de Janeiro. Amostras de mucosa gastroduodenal foram obtidas por meio de biópsia endoscópica e foram processadas por técnica histológica de rotina e coradas por Hematoxilina-Eosina e Giemsa. As amostras foram submetidas à análise histopatológica para pesquisa de bactérias espiraladas sugestivas de Helicobacter spp. O teste rápido da urease também foi realizado. Os sinais clínicos, e os achados macro e microscópicos foram confrontados e correlacionados. Neste estudo, foi possível estabelecer correlação positiva entre a perda de peso e a presença de enantema no antro, no corpo e no duodeno além de correlação entre alteração do apetite, a presença de infiltrado inflamatório gástrico e a positividade no teste da urease. Este estudo demonstrou a importância da observação de alguns sinais clínicos como a perda de peso e alterações do apetite no curso da Doença Inflamatória Intestinal. Desta forma, enfatiza-se que cães que apresentem tais sinais clínicos devem ter a Doença Inflamatória Intestinal incluída no grupo de diagnósticos diferenciais evitando, com isso, negligenciar uma enfermidade que, cada vez mais, acomete animais de pequeno porte.(AU)

The aim of this study was to evaluate macro and microscopic gastrointestinal mucosa of domiciled dogs, with chronic gastrointestinal symptoms by endoscopy and histopathological evaluation, correlating clinical signs, macro and microscopy characteristics/aspects/changes of duodenal and gastric mucosa. Twenty dogs of different breeds were evaluated with with chronic characteristic signs of Inflammatory Bowel Disease (IBD), being 7 males and 13 females, with ages ranging from 1.7 to 15.8 years old. The study was prospective and cross-sectional, conducted in dogs treated at the Teaching Veterinary Medicine Hospital of the Universidade Federal Fluminense (HUVET-UFF) and private clinics in the city of Rio de Janeiro. Gastroduodenal mucosal samples were obtained by endoscopic biopsy and were processed by routine histological technique and stained with hematoxylin- eosin and Giemsa. The samples were submitted to histopathological analysis for the detection of spiral bacteria suggestive of Helicobacter spp. The rapid urease test was also performed. The clinical signs, the macro and microscopic findings were compared and correlated. In this study, we established a positive correlation between weight loss and the presence of erythema in the antrum, body and duodenum as well as correlation between change in appetite and the presence of gastric inflammatory infiltrate and positivity in the urease test. This study demonstrated the importance of the observation of some clinical signs such as weight loss and appetite changes in the course of IBD. Thus, it is emphasized that dogs showing these clinical signs should have the IBD included in the differential diagnoses group avoiding thereby neglecting a disease that increasingly affects small animals.(AU)

Avaliação clínica, endoscópica e histopatológica de cães com doença inflamatória intestinal / Clinical, endoscopic and histopathological evaluation of dogs with inflammatory bowel disease

O presente estudo teve como objetivo avaliar macro e microscopicamente a mucosa gastrintestinal de cães domiciliados, que apresentaram sinais gastrintestinais crônicos, por meio de exame endoscópico e da avaliação histopatológica. Os sinais clínicos, a macro e a microscopia das mucosas gástrica e duodenal foram correlacionados. Foram avaliados 20 cães de diferentes raças, sendo 7 machos e 13 fêmeas, com idade variando entre 1,7 a 15,8 anos, que apresentavam cronicidade dos sinais característicos da Doença Inflamatória Intestinal (DII). O estudo foi prospectivo e transversal, realizado em cães atendidos no Hospital Universitário de Medicina Veterinária da Universidade Federal Fluminense (HUVET - UFF) e em clínicas privadas do município do Rio de Janeiro. Amostras de mucosa gastroduodenal foram obtidas por meio de biópsia endoscópica e foram processadas por técnica histológica de rotina e coradas por Hematoxilina-Eosina e Giemsa. As amostras foram submetidas à análise histopatológica para pesquisa de bactérias espiraladas sugestivas de Helicobacter spp. O teste rápido da urease também foi realizado. Os sinais clínicos, e os achados macro e microscópicos foram confrontados e correlacionados. Neste estudo, foi possível estabelecer correlação positiva entre a perda de peso e a presença de enantema no antro, no corpo e no duodeno além de correlação entre alteração do apetite, a presença de infiltrado inflamatório gástrico e a positividade no teste da urease. Este estudo demonstrou a importância da observação de alguns sinais clínicos como a perda de peso e alterações do apetite no curso da Doença Inflamatória Intestinal. Desta forma, enfatiza-se que cães que apresentem tais sinais clínicos devem ter a Doença Inflamatória Intestinal incluída no grupo de diagnósticos diferenciais evitando, com isso, negligenciar uma enfermidade que, cada vez mais, acomete animais de pequeno porte.(AU)

The aim of this study was to evaluate macro and microscopic gastrointestinal mucosa of domiciled dogs, with chronic gastrointestinal symptoms by endoscopy and histopathological evaluation, correlating clinical signs, macro and microscopy characteristics/aspects/changes of duodenal and gastric mucosa. Twenty dogs of different breeds were evaluated with with chronic characteristic signs of Inflammatory Bowel Disease (IBD), being 7 males and 13 females, with ages ranging from 1.7 to 15.8 years old. The study was prospective and cross-sectional, conducted in dogs treated at the Teaching Veterinary Medicine Hospital of the Universidade Federal Fluminense (HUVET-UFF) and private clinics in the city of Rio de Janeiro. Gastroduodenal mucosal samples were obtained by endoscopic biopsy and were processed by routine histological technique and stained with hematoxylin- eosin and Giemsa. The samples were submitted to histopathological analysis for the detection of spiral bacteria suggestive of Helicobacter spp. The rapid urease test was also performed. The clinical signs, the macro and microscopic findings were compared and correlated. In this study, we established a positive correlation between weight loss and the presence of erythema in the antrum, body and duodenum as well as correlation between change in appetite and the presence of gastric inflammatory infiltrate and positivity in the urease test. This study demonstrated the importance of the observation of some clinical signs such as weight loss and appetite changes in the course of IBD. Thus, it is emphasized that dogs showing these clinical signs should have the IBD included in the differential diagnoses group avoiding thereby neglecting a disease that increasingly affects small animals.(AU)

Involvement of mast cells, CD68+ and VEGF+ expressions in response to Himatanthus drasticus commercial latex in mice wound healing model / Envolvimento de mastócitos, expressão de CD68+ e VEGF+ em resposta ao látex comercial de Himatanthus drasticus em modelo de cicatrização em camundongos

This study aimed to evaluate Himatanthus drasticus latex in a mice wound healing experimental model. Animals were divided into four groups (n=7) according to the treatments: GI - saline 0.9% (control), GII - mineral oil (vehicle), GIII - H. drasticus commercial latex (HdCL) and GIV - H. drasticus mixed isolated fraction (MIF, 1 mg/mL). The treatments were applied topically once daily, 50 µL for 14 consecutive days. Macroscopic lesions were evaluated, considering parameters such as swelling, redness, granulation tissue and reepithelialization. VEGF+, CD68+ expressions and mast cells (Toluidin blue stain) were evaluated. HdCL induced higher contraction and exuberant granulation tissue (P > 0.05). HdCL showed a mild inflammatory process while MIF induced intense infiltrate inflammatory predominantly by lymphocytes, vascular congestion, bleeding and did not presented full reepithelialization. Reorganization of collagen fibers (red picrosirius stain) was observed. CD68+ expression and mast cells were presented as moderate, intense and mild in GI, GIII and GIV, respectively. Neovascularization occurred in all groups, while VEGF+ expression was intense in MIF in relation to HdCL. We concluded that HdCL presents wound healing potential, through modulation of mast cells, CD68+ and VEGF+ expressions that can be associated to triterpenes presence according MIF isolated from HdCL.(AU)

Objetivou-se avaliar o látex de Himatanthus drasticus em feridas induzidas experimentalmente em camundongos. Os animais foram divididos em quatro grupos (n=7): GI - salina 0,9% (controle), GII - óleo mineral (veículo), GIII - látex comercial de H. drasticus (HdCL) e GIV - fração isolada mista de H. drasticus (MIF, 1mg/mL). Os tratamentos foram aplicados topicamente uma vez ao dia (50µL), durante 14 dias consecutivos. Lesões macroscópicas, as expressões de VEGF+, CD68+ e a participação dos mastócitos (coloração azul de toluidina) foram avaliadas. HdCL induziu maior contração e tecido de granulação exuberante (P >0,05). HdCL induziu leve processo inflamatório enquanto MIF promoveu intenso infiltrado inflamatório predominantemente linfocítico, congestão vascular, hemorragia e reepitelização parcial. Observou-se reorganização das fibras colágenas (coloração picrosírius). A expressão de CD68+ e os mastócitos apresentaram-se moderados, intensos e leves em GI, GIII e GIV, respectivamente. A neovascularização foi observada em todos os grupos, enquanto a expressão de VEGF+ foi mais intensa em MIF em relação a HdCL. Conclui-se que HdCL apresenta potencial de cicatrização por meio da modulação dos mastócitos e das expressões de CD68+ e VEGF+, o que pode estar associado à presença de triterpenos de acordo com MIF isolada de HdCL.(AU)

Involvement of mast cells, CD68+ and VEGF+ expressions in response to Himatanthus drasticus commercial latex in mice wound healing model / Envolvimento de mastócitos, expressão de CD68+ e VEGF+ em resposta ao látex comercial de Himatanthus drasticus em modelo de cicatrização em camundongos

This study aimed to evaluate Himatanthus drasticus latex in a mice wound healing experimental model. Animals were divided into four groups (n=7) according to the treatments: GI - saline 0.9% (control), GII - mineral oil (vehicle), GIII - H. drasticus commercial latex (HdCL) and GIV - H. drasticus mixed isolated fraction (MIF, 1 mg/mL). The treatments were applied topically once daily, 50 µL for 14 consecutive days. Macroscopic lesions were evaluated, considering parameters such as swelling, redness, granulation tissue and reepithelialization. VEGF+, CD68+ expressions and mast cells (Toluidin blue stain) were evaluated. HdCL induced higher contraction and exuberant granulation tissue (P > 0.05). HdCL showed a mild inflammatory process while MIF induced intense infiltrate inflammatory predominantly by lymphocytes, vascular congestion, bleeding and did not presented full reepithelialization. Reorganization of collagen fibers (red picrosirius stain) was observed. CD68+ expression and mast cells were presented as moderate, intense and mild in GI, GIII and GIV, respectively. Neovascularization occurred in all groups, while VEGF+ expression was intense in MIF in relation to HdCL. We concluded that HdCL presents wound healing potential, through modulation of mast cells, CD68+ and VEGF+ expressions that can be associated to triterpenes presence according MIF isolated from HdCL.(AU)

Objetivou-se avaliar o látex de Himatanthus drasticus em feridas induzidas experimentalmente em camundongos. Os animais foram divididos em quatro grupos (n=7): GI - salina 0,9% (controle), GII - óleo mineral (veículo), GIII - látex comercial de H. drasticus (HdCL) e GIV - fração isolada mista de H. drasticus (MIF, 1mg/mL). Os tratamentos foram aplicados topicamente uma vez ao dia (50µL), durante 14 dias consecutivos. Lesões macroscópicas, as expressões de VEGF+, CD68+ e a participação dos mastócitos (coloração azul de toluidina) foram avaliadas. HdCL induziu maior contração e tecido de granulação exuberante (P >0,05). HdCL induziu leve processo inflamatório enquanto MIF promoveu intenso infiltrado inflamatório predominantemente linfocítico, congestão vascular, hemorragia e reepitelização parcial. Observou-se reorganização das fibras colágenas (coloração picrosírius). A expressão de CD68+ e os mastócitos apresentaram-se moderados, intensos e leves em GI, GIII e GIV, respectivamente. A neovascularização foi observada em todos os grupos, enquanto a expressão de VEGF+ foi mais intensa em MIF em relação a HdCL. Conclui-se que HdCL apresenta potencial de cicatrização por meio da modulação dos mastócitos e das expressões de CD68+ e VEGF+, o que pode estar associado à presença de triterpenos de acordo com MIF isolada de HdCL.(AU)

Local and hematological alterations induced by Philodryas olfersii snake venom in mice.

Envenomation by the South American opisthoglyphous snake Philodryas olfersii causes local pain, edema, erythema and ecchymosis; systemic envenomation is rare. In this work, we examined the inflammatory activity of P. olfersii venom (10, 30 and 60 µg) in mouse gastrocnemius muscle 6 h after venom injection. Intramuscular injection of venom did not affect hematological parameters such as red cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. The venom caused thrombocytopenia (at all three doses), leukopenia and lymphopenia (both at the two highest doses), as well as neutrophilia (30 µg), monocytosis (30 µg) and basophilia (10 µg). Of the cytokines that were screened [IL-1ß, IL-6, IL-10, IL-13, IL-17, TNF-α, IFN-γ, MIP-2 and KC] and IGF-1, only IGF-1 showed a significant increase in its circulating concentration, seen with 60 µg of venom; there were no significant changes in the cytokines compared to control mice. Histological analysis revealed the presence of edema, an inflammatory infiltrate and progressive myonecrosis. Edema and myonecrosis were greatest with 60 µg of venom, while the inflammatory infiltrate was greatest with 10 µg of venom. All venom doses caused the migration of polymorphonuclear and mononuclear leukocytes into muscle, but with no significant dose-dependence in the response. These findings show that, at the doses tested, P. olfersii venom does not cause hematological alterations and has limited effect on circulating cytokine concentrations. These data also confirm that the principal effects of the venom in mice are local edema, inflammatory cell infiltration and myonecrosis.

Parasitism and inflammation in ear skin and in genital tissues of symptomatic and asymptomatic male dogs with visceral leishmaniasis.

Canine visceral leishmaniasis (CVL) is transmitted through vector, although venereal transmission has been suggested. This study aimed to compare the parasitic loads and inflammatory processes in genital tissues with ear skin from seropositive male dogs. Forty-five seropositive dogs were separated into groups containing symptomatic (n = 23) and asymptomatic (n = 22) animals. The control group (n = 2) healthy animals with seronegative and negative results in direct parasitological test. Samples of ear tip skin, prepuce, glans penis, testis, epididymis, and prostate were collected for evaluation of parasitic load and inflammatory infiltrate. Although ear tip skin was the most intensely parasitized, prepuce and epididymis revealed no difference in parasitism when compared with ear tip skin (P > 0.05). Parasitic loads in testis and prostate were lower than other tissues (P < 0.05). Parasitism in glans penis was high, similar to prepuce and epididymis, but lower than ear tip skin. High parasitism was more frequent in symptomatic dogs than asymptomatic animals. Severe inflammatory processes were more frequent within the symptomatic animals compared with asymptomatic and more predominant in prepuce and epididymis. Ear tip skin and genital tissues presented signs of chronic inflammation. There were weak and moderate positive correlations between parasitic loads and inflammatory processes. Our results demonstrate that, likewise with the ear tip skin, the genital of seropositive dogs can carry a large number of Leishmania infantum amastigotes and this process are more intense in symptomatic animals. These data have important implications for understanding the possibility of venereal transmission of CVL.

Light-emitting diodes at 940nm attenuate colitis-induced inflammatory process in mice.

Inflammatory bowel disease (IBD) presents intense inflammatory infiltrate, crypt abscesses, ulceration and even loss of function. Despite the clinical relevance of IBD, its current therapy remains poorly effective. Infrared wavelength phototherapy shows therapeutic potential on inflammation. Our goal was to evaluate whether light-emitting diodes (LED) at 940nm are capable of mitigating the colitis-induced inflammatory process in mice. Forty male Swiss mice were assigned into five groups: control; control treated with LED therapy; colitis without treatment; colitis treated with LED therapy; colitis treated with Prednisolone. Experimental colitis was induced by acetic acid 7.5% (pH2.5) rectal administration. LED therapy was performed with light characterized by wavelength of 940nm, 45nm bandwidth, intensity of 4.05J/cm(2), total power of 270mW and total dose of 64.8J for 4min in a single application. Colitis-induced intestinal transit delay was inhibited by LED therapy. Colitis caused an increase of colon dimensions (length, diameter, total area) and colon weight (edema), which were inhibited by LED therapy. LED therapy also decreased colitis-induced tissue gross lesion, myeloperoxidase activity, microscopic tissue damage score and the presence of inflammatory infiltrate in all intestinal layers. Furthermore, LED therapy inhibited colitis-induced IL-1ß, TNF-α, and IL-6 production. We conclude LED therapy at 940nm inhibited experimental colitis-induced colon inflammation in mice, therefore, rendering it a promising therapeutic approach that deserves further investigation.