The endothelial layer, formed by endothelial cells, performs crucial functions in maintaining homeostasis. The endothelial integrity and function might be compromised due to various causes, including infection by Toxoplasma gondii, leading to an endothelial dysfunction. Toxoplasma gondii is an Apicomplexa parasite that infects a broad range of animals, including humans. This parasite can invade all nucleated cells, as well as endothelial cells. The interaction between this protozoan and endothelial cells can be mediated by different molecules, such as extracellular vesicles (EVs), which may either favor or hinder the infectious process. To investigate this interaction, we evaluated the infection of T. gondii on human brain microvascular endothelial cells (HBMEC) and human umbilical vein endothelial cells (HUVEC), in addition to assessing transcriptional changes. We also featured the EVs secreted by T. gondii and by infected and non-infected HBMEC and HUVEC. Finally, we evaluated the infection of cells stimulated with EVs of parasitic or cellular origin. Our results demonstrated that HUVEC not only exhibit a higher infection rate than HBMEC but also display a more pro-inflammatory transcriptional profile, with increased expression of interleukin-6 (IL6), interleukin-8 (IL8), and monocyte chemotactic protein-1 (MCP1) following infection. Additionally, we observed few differences in the concentration, distribution, and morphology of EVs secreted by both cell types, although their properties in modulating infection varied significantly. When cells were EVs stimulated, EVs from T. gondii promoted an increase in the HBMEC infection, EVs from infected or uninfected HBMEC reduced the infection, whereas EVs from HUVEC had no effect on the infectious process. In conclusion, our data indicate that T. gondii infection induces distinct changes in different endothelial cell types, and EVs from these cells can contribute to the resolution of the infection.

Extracellular vesicles (EVs) are produced by all kinds of cells, including endothelial cells. It has been observed that EVs present in fetal bovine serum (FBS), broadly used in cell culture, can be a confounding factor and lead to misinterpretation of results. To investigate this phenomenon, human brain microvascular endothelial cells (HBMECs) were cultured for 2 or 24 h in the presence of EV-depleted FBS (EVdS). Cell death, gene and protein expression, and the presence of EVs isolated from these cells were evaluated. The uptake of EVs, intercellular adhesion molecule 1 (ICAM-1) expression, and monocyte adhesion to endothelial cells exposed to EVs were also evaluated. Our results revealed higher apoptosis rates in cells cultured with EVdS for 2 and 24 h. There was an increase in interleukin 8 (IL8) expression after 2 h and a decrease in interleukin 6 (IL6) and IL8 expression after 24 h of culture. Among the proteins identified in EVs isolated from cells cultured for 2 h (EV2h), several were related to ribosomes and carbon metabolism. EVs from cells cultured for 24 h (EV24h) presented a protein profile associated with cell adhesion and platelet activation. Additionally, HBMECs exhibited increased uptake of EV2h. Treatment of endothelial cells with EV2h resulted in greater ICAM-1 expression and greater adherence to monocytes than did treatment with EV24h. According to our data, HBMEC cultivated with EVdS produce EVs with different physical characteristics and protein levels that vary over time.

Lytic vertebral lesions cause significant pain and variable neurological deficits can cause sensory, motor and proprioceptive loss, and are causes of bones lysis such as discospondylitis, vertebral tumors, bone cysts and osteomyelitis. This study aimed to investigate the prevalence of vertebral lytic lesions in the canine population, from computed tomographic (CT) image analyses. A retrospective study, based on the descriptive findings from simple and contrast enhanced CT scans of dogs, between the years 2012 to 2017, was performed. The study consisted of the sequential analysis of 371 reports to identify vertebral lytic lesions, in areas with osteopenia and hypoattenuation, based on Hounsfield Unit scores. The Fisher’s exact test and descriptive statistics were used for statistical analyses. Twenty-eight cases (7.54%) of vertebral lytic lesion were observed; 46.42% of these had characteristics of discospondylitis, with a prevalence in males, in the cranial and caudal ends of the vertebrae. Vertebral tumors were identified in 37.71% of the cases, with a higher incidence in defined medium sized breeds, with unifocal lesions in the vertebral body. Single lesion bone cysts were observed in 10.71% of cases and osteomyelitis in 7.14% of cases. A significant association between the number of vertebrae and degree of impairment (unifocal or multifocal) was observed. CT imaging has become a useful tool for evaluating the architecture and vertebral bone density. This study provides perspectives for the analysis of vertebral lytic lesions. In addition to showing the prevalence of lytic lesions, this study emphasizes the importance of using advanced diagnostic imaging techniques, such as CT imaging, for observing and identifying such lesions, thus directing further examinations such as bone biopsy, culture, and antibiogram or histopathology. It can be concluded that lytic lesions have a considerable incidence, impair the architecture of the vertebra and CT imaging is an effective tool for diagnosing them. Prevalence was observed in male dogs, neutered, with defined breed, of medium size, with unifocal lesion and lytic lesions associated with the discspondylitis.

Provides simple and intuitive functions for basic statistical analyses. Methods include the t-test (Student 1908 ), the Mann-Whitney U test (Mann and Whitney 1947 ), Pearson's correlation (Pearson 1895 ), and analysis of variance (Fisher 1925, ). Functions are compatible with 'ggplot2' and 'dplyr'.

Intestinal gas results in low quality imaging in abdominal ultrasound in domestic animals. On dogs, suggested preparation protocols are varied and low studied. The aim of this work was comparing the efficacy between two preparations in improving the complete abdominal ultrasound exam and pancreas preview. 40 dogs were enrolled in this study. They were separated in two groups (NS diet: fasting; WS diet: fasting added to simethicone). The pancreas images were taken from left lateral decubitus. A score of one to three (1: bad, 2: fair and 3: good) measured separately the quality of the complete exam, left and right limbs and pancreatic body. In both treatments, there were results above 50% of good images of the complete exam and right limb of the pancreas. Otherwise, there were results above 90% of bad images of the right limb and pancreatic body. According to the fact of there is no statistically significant difference between the diets, as well submitted animals to the NS diet as the WS diet, all the results obtained from this work (bad or good images) could be acquired from any of both diets. The good abdominal exam preview as well submitted animals to the simethicone as those without the medication refute this medicine requirement to the abdominal ultrasound exam, but not to the evaluation of the left limb and pancreatic body.