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Spontaneous lymphatic revascularization is a challenge and the establishment of new therapeutic strategies may improve life quality for patients suffering from lymphatic disorders. This study was designed to verify if VEGFC treatment improves lymphatic vascularization in a time‐dependent manner in mouse hindlimb (HL) after resection of the inguinal lymph node. Lymphatic vascular density (Vv) and length (Lv) were evaluated by stereology after immunohistochemistry. The control Group (CG) was not manipulated but received saline instead of VEGFC treatment. The surgery Group (SG) had the left inguinal lymph node resected but did not received VEGFC treatment. VEGFC Treated Group (TG) had the node resected and received VEGFC treatment. VEGFC and VEGFR3 local expression were assessed by qPCR. There was an effect of time over Vv and Lv in the SG and significant difference between CG and SG in the regions studied (proximal, medium and distal regions) of the left HL (LHL). The Lv showed significant difference between CG and SG only in the medium region. The Vv and the Lv for TG were higher than the other groups. VEGFC and VEGFR3 gene expression presented time effect in all regions of the LHL for SG and TG. Both VEGFC and VEGFR3 gene expression presented significant difference between CG and SG, between SG and TG and between CG and TG. This study showed significant decrease in lymphatic vascularization in the left hindlimb of mice after surgical removal of the inguinal lymph node and adjacent lymphatic vessels. Exogenous VEGFC could recover lymphatic vascularization through stimulating neolymphangiogenesis.

O estômago de ruminantes é um órgão complexo subdividido em quatro compartimentos: o rúmen, o retículo, o omaso e o abomaso, sendo os três primeiros os pré-estômagos ou pró-ventrículos. Cada um possui sua própria característica, responsável por mecanismos de ruminação essenciais. Junções gap são as únicas especializações na membrana celular que permitem a comunicação direta entre células adjacentes, enquanto que as junções de adesão são especializações que mantém a integridade tecidual. Sabe-se que elas contribuem para a homeostase tissular e são compostas por proteínas transmembrânicas chamadas conexinas (junções gap) e caderinas (junções de adesão). Além disso, essas junções estão também envolvidas no controle da proliferação celular, no controle de crescimento e diferenciação, apoptose, e a sincronização de funções eletrotônicas e metabólicas. Assim, o rúmen de bovinos pode ser um modelo para compreender a contribuição das conexinas e caderinas durante a diferenciação do epitélio ruminal a um estado funcional, que o transforma de epitélio de revestimento para um epitélio absortivo com funções de metabolização de substâncias, em especial o ácido lático e ácidos graxos. Dessa maneira, nossa hipótese é que as diferentes conexinas mudem sua dinâmica de expressão quanto ao tipo e localização, com repercussões na morfologia epitelial. Os objetivos do presente estudo foram a avaliação da expressão e localização das Conexinas 26, 32, 40 e 43 e da E-caderina no rúmen de fetos bovinos, bezerros recém-nascidos e bovinos adultos, além da avaliação morfológica do epitélio ruminal de através da microscopia óptica e eletrônica. Também possui como objetivo a avaliação da expressão gênica dessas proteínas no rúmen desses animais. Para isso, foi realizada a coleta de amostras do saco dorsal e do saco ventral do rúmen de fetos bovinos, bezerros recém-nascidos e bovinos adultos. Nos materiais coletados foram realizadas as técnicas de coloração com Hematoxilina-Eosina e morfometria, imunofluorescência, microscopia eletrônica de transmissão, extração de RNA total e sequenciamento genético. Na coleta do material as posições anatômicas observadas foram exatas às já descritas na literatura. O tecido ruminal apresentou diferenças morfométricas em relação às mensurações realizadas no tecido epitelial e na camada basal, nas idades estudadas. Além disso, foram observadas modificações teciduais ao longo do desenvolvimento ruminal na microscopia eletrônica de transmissão e na expressão das conexinas no tecido ruminal na imunofluorescência. A respeito da expressão das conexinas estudadas, ela difere em relação à idade dos animais e à localização no tecido ruminal, tornando evidente a necessidade de cada uma das conexinas em relação à idade, desenvolvimento e manutenção do tecido ruminal. A expressão tecidual da E-caderina apresentou-se semelhante nas idades estudadas. Em relação à expressão gênica e sequenciamento genético dessas proteínas, suas mudanças acompanharam as diferenças observadas na expressão tecidual através da imunofluorescência. Desse modo, pode-se concluir que ocorrem mudanças tanto na expressão tecidual quanto na expressão gênica dessas proteínas durante o desenvolvimento ruminal e sua manutenção enquanto adulto. The stomach of ruminants is a complex organ subdivided into four compartments: the rumen, the reticulum, the omasum and the abomasum, the first three being the prestomachs or pro-ventricles. Each one has its own characteristic, responsible for essential rumination mechanisms. Gap junctions are the only cell membrane specializations that allow direct communication between adjacent cells, whereas adhesion junctions are specializations that maintain tissue integrity. It is known that they contribute to tissue homeostasis and are composed of transmembrane proteins called connexins (gap junctions) and cadherins (adhesion junctions). In addition, these junctions are also involved in the control of cell proliferation, growth and differentiation, apoptosis, and the synchronization of electrotonomic and metabolic functions. Thus, the bovine rumen may be a model for understanding the contribution of connexins and cadherins during the differentiation of the ruminal epithelium into a functional state, which transforms it from coating epithelium to an absorptive epithelium with metabolizing functions of substances, especially the lactic acid and fatty acids. Therefore, our hypothesis is that the different connexins change their expression dynamics regarding type and location, with repercussions on epithelial morphology. The objective of the present study was to evaluate the expression and location of Connexins 26, 32, 40 and 43 and E-cadherin in the rumen of bovine fetuses, newborn calves and adult bovines, as well as the morphological evaluation of ruminal epithelium through optical and electronic microscopy. It also aims to evaluate the gene expression of these proteins in the rumen of these animals. For that, samples were collected from the dorsal and the ventral sac from the rumen of bovine fetuses, newborn calves and adult bovines. Hematoxylin-Eosin staining techniques and morphometry, immunofluorescence, transmission electron microscopy, total RNA extraction and genetic sequencing were performed in the collected materials. During the collection of the material it was observed that anatomical positions were exact to those already described in the literature. The ruminal tissue presented morphometric differences in relation to the measurements made in the epithelial tissue and the basal layer, in the studied ages. In addition, tissue changes were observed along ruminal development in transmission electron microscopy and in the expression of connexins in ruminal tissue in immunofluorescence. Regarding the expression of the connexins studied, it differs in relation to the age of the animals and the location in the ruminal tissue, making evident the necessity of each of the connexins in relation to the age, development and maintenance of ruminal tissue. The tissue expression of E-cadherin was similar in the studied ages. In relation to the gene expression and genetic sequencing of these proteins, their changes accompanied the observed differences in tissue expression through immunofluorescence. Thus, it can be concluded that changes occur in both the tissue expression and the gene expression of these proteins during ruminal development and its maintenance as an adult.

The bovine whey consists of more than 200 different types of proteins, of which β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulins and lactoferrin predominate. However, their concentrations are not stable due to the existence of protein dynamics during a transition from colostrum secretion to mature milk. To evaluate the dynamics of whey proteins of Jersey cows during a colostral phase and first month of lactation and an influence of the number of lactations, 268 milk samples from 135 Jersey cows were selected through a clinical evaluation. Whey was obtained by rennet coagulation of the mammary secretion. The concentration of total proteins was determined by the biuret method and their fractions were identified by 12% dodecyl sulfate-polyacrylamide gel electrophoresis (12% SDS-PAGE). Maximum concentrations of all protein fractions were observed in the first 12 h of lactation, reducing over the course of the study. Modification of the protein predominance was also observed. The transition from colostrum secretion to milk occurred between 24 and 72 h postpartum. There was an influence of the number of lactations on the dynamics of whey proteins, indicating that multiparous cows had better immunological and nutritional quality when compared to primiparous cows.

Pequi (Caryocar brasiliense Camb.), a fruit from Brazil's central region, was evaluated for its chemopreventive effects on preneoplastic liver lesions induced by the carcinogen diethylnitrosamine (DEN) in mice. BALB/c mice, 14 days of age, received an intraperitoneal injection at 10 µg/g of DEN. The mice received either of two doses of pequi oil (100 or 400 mg/kg) daily from the age of 30 days and were killed at the age of 189 days. Stereological parameters, including the volume density (Vv) and the total volume (Vtot) of the lesions (preneoplastic and adenomas), were measured and the expression of cytokeratins CK8/18 was evaluated. The total volume of lesions and adenomas was reduced by 51% in the group treated with the carcinogen and 400 mg/kg of pequi oil administered daily by an oral gavage for 25 consecutive weeks. In addition, some mice in this group did not develop lesions. Among the remaining preneoplastic lesions in this group, the number of remodelled profiles increased by 2.4-fold in the 400-mg pequi oil-treated mice relative to the 100-mg-treated mice. Our results show that pequi oil exerts a hepatoprotective effect against DEN-induced development of preneoplastic lesions and adenoma in mice and the potential for its use in the prevention of liver cancer.

Resumo: A indoleamina 2,3-dioxigenase (IDO) é uma enzima que cataboliza o aminoácido triptofano, levando à inibição da proliferação de linfócitos T, seja pela exaustão desse aminoácido no ambiente, ou pela indução via catabólitos induzindo-os a apoptose. Em mamíferos, esta enzima atua em diversas condições do organismo como a gestação, infecções, inflamações crônicas, transplantes e tumores, atuando na regulação imunológica. Estudos recentes identificaram a presença de moléculas homólogas a IDO em espécies filogeneticamente inferiores, cuja função parece estar restrita ao metabolismo do triptofano como fonte de energia. Este estudo teve por objetivo averiguar a expressão da IDO em células sanguíneas e órgãos hematopoiéticos de truta arco-íris pela imuno-histoquímica, buscando evidências de que a mesma poderia, nesta espécie, estar relacionada ao sistema imune. A expressão de IDO foi observada nos órgãos hematopoiéticos estudados incluindo o rim cefálico que apresentou marcação em células interrenais e leucócitos; baço, na qual a marcação restringiu à alguns leucócitos; no fígado a marcação ficou limitada à apenas algumas células dentro dos vasos sanguíneos e nas extensões sanguíneas pode-se visualizar a marcação de alguns leucócitos como os monócitos, linfócitos e neutrófilos. A predominância da marcação da IDO nesses tecidos pode constituir uma evidência de que a IDO identificada na O. mykiss esteja relacionada ao sistema imunológico nessa espécie. Abstract: Indoleamine 2,3-dioxygenase (IDO) is an enzyme that catabolizes the amino acid tryptophan, leading to inhibition of T lymphocyte proliferation, whether by depletion of this amino acid in the environment, or by induction via the catabolites inducing apoptosis. In mammals, this enzyme acts on various conditions of the body such as pregnancy, infections, chronic inflammation, transplantation and tumors, acting in immune regulation. Recent studies have identified the presence of homologous molecules IDO lower phylogenetically related species, whose function appears to be confined to the tryptophan metabolism as an energy source. This study aimed to investigate the expression of IDO in blood cells and hematopoietic organs of rainbow trout by immunohistochemistry, seeking evidence that it could, this species is related to the immune system. The expression of IDO was observed in hematopoietic organs studied including head kidney that show labeling in interrenal cells and leukocytes; spleen, in which the marking restricted to a few leukocytes in the liver;, labeling was restricted to only certain cells within the blood vessels and the blood extensions can view the marking of some leukocytes including monocytes, lymphocytes and neutrophils. The predominance of IDO marking these tissues may constitute evidence that IDO identified in O. mykiss is related to the immune system in this species.

This paper is a review of physiological and behavioral data on motor cortex stimulation (MCS) and its role in persistent neuropathic pain. MCS has been widely used in clinical medicine as a tool for the management of pain that does not respond satisfactorily to any kind of conventional analgesia. Some important mechanisms involved in nociceptive modulation still remains unclear. The aim of this study was to describe the mechanisms involved in neuropathic pain and introduce the effectiveness of electrical stimulation of the motor cortex used in the treatment of this disease. The ascending pain pathways are activated by peripheral receptors, in which there is the transduction of a chemical, physical or mechanical stimulus as a nerve impulse, where this impulse is transmitted to the dorsal horn of the spinal cord, which connects with second-order neurons and ascends to different locations in the central nervous system where the stimulus is perceived as pain. Because MCS has been proved to modulate this pathway in the motor cortex, it has been studied to mimic its effects in clinical practice and improve the treatments used for chronic pain. MCS has gained much attention in recent years due to its action in reversing chronic neuropathic pain, this being more effective than electrical stimulation at different locations and related pain nuclei. O presente trabalho é uma revisão de dados fisiológicos e comportamentais sobre a estimulação elétrica do córtex motor (ECM) e seu papel durante a dor neuropática persistente. A ECM tem sido amplamente utilizada na clínica médica como ferramenta para controle da dor que não respondem satisfatoriamente a nenhum tipo de analgesia convencional. Alguns importantes mecanismos envolvidos na modulação nociceptiva não foram, até o momento, esclarecidos. O objetivo deste estudo foi descrever os mecanismos envolvidos durante a dor neuropática e apresentar a eficiência da estimulação elétrica do córtex motor utilizada no tratamento desta doença. As vias ascendentes da dor são ativadas por receptores periféricos, onde há a transdução do estímulo químico, físico ou mecânico, em impulso nervoso, transmitindo este até a coluna posterior da medula espinal, onde ocorre conexão com neurônios de segunda ordem e ascendem à diferentes locais do sistema nervoso central, onde o estímulo periférico é percebido como dor. Por ser comprovada grande modulação deste sentido no córtex motor, vem sendo estudado o efeito da ECM para mimetizar os efeitos na pratica clinica e aperfeiçoar os tratamentos empregados durante a dor crônica. A ECM ganhou uma atenção especial nos últimos anos devido a sua ação em reverter quadros de dor neuropática de origem crônica, esta sendo mais eficiente do que a estimulação elétrica em diferentes locais e núcleos relacionados a dor.

The morphology and location of lymph nodes from seven species of Odontocetes, of both sexes and different age groups, were described. All animals were derived from stranding events along the North and Northeastern coasts of Brazil. After the identification of lymph nodes in situ, tissue samples were analyzed for light and electron microscopy. Vascular volume density (VVD) and vascular length density (VLD) were evaluated in the mesenteric lymph nodes. Lymph nodes occurred as solitary nodules or in groups, varying in shape and size. In addition to using the nomenclature recommended by Nomina Anatomica Veterinaria, new nomenclatures were suggested based on the lymph nodes topography. Lymph nodes were covered by a highly vascularized and innervated capsule of dense connective tissue, below which muscle fibers were observed, inconsistently, in all studied species. There was no difference in VLD among different age groups. However, VVD was higher in adults. Lymph nodes parenchyma was divided into an outer cortex, containing lymph nodules and germinal centers; a paracortical region, transition zone with dense lymphoid tissue; and an inner medulla, composed of small irregular cords of lymphatic tissue, blood vessels, and diffuse lymphoid tissue. Abundant collagen fibers were observed around arteries and arterioles. Germinal centers were more evident and developed in calves and young animals, being more discrete and sparse in adults. The morphology of lymph nodes in Odontocetes was typical of that observed in other terrestrial mammals. However, new groups of lymph nodes were described for seven species occurring in the Brazilian coast.

The study of stem cells has evolved rapidly in recent decades. The importance is given to the concept that these cells are potentially able to become any cell type and have the power of self-renewal throughout the life of the organism. Mesenchymal stem cells (MSCs) can be isolated from various organs of the body such as bone marrow, adipose tissue, synovium, muscle and dermis, deciduous teeth, umbilical cord, placenta, liver, spleen and thymus. After their isolation in vitro, mesenchymal stem cells have the capacity to differentiate into various mesenchymal lineages and various tissues after the use of appropriate cultures. Studies have reported that mesenchymal stem cells from adipose tissue have the potential to differentiate themselves, like the cells commonly studied bone marrow. Adipose tissue is attractive due to its easy access, rapid expansion in vitro and only one collects the large amount of tissue. This review intends to show the protocols for isolation, cell culture and means of commercial cellular differentiation most widely used with emphasis on adipose tissue.

Objective To check if ovariectomy produces alterations in the articular cartilage structure of the tibial proximal epiphysis and if aerobic physical exercise has any influence on these alterations. Methods Six-month-old female Wistar rats were distributed in three groups: Control group (CG), without ovariectomy and physical activity; Oophorectomy and sedentary group (OSG), with bilateral ovariectomy and without physical activity, and Oophorectomy and trained group (OTG), with bilateral ovariectomy and physical activity during 3 months. All rats were euthanised at 9 months of age. The results were obtained by using histological sections stained with Hematoxylin and Eosin, and Picrosirius stain. Results In relation to the thickness of the articular cartilage an increase was observed in the superficial layer in OSG in relation to OTG. The number of chondrocytes did not show any significant difference. The nuclear volume of chondrocytes in the superficial layer was higher in CG than in OSG . In the medium layer, the volume of chondrocytes was higher in CG than in OSG and the OTG. The same occurred when the values of the epiphysial cartilage was considered as a whole. For the collagen, the values of volume density of the collagen fibers were higher in the OSG and in the OTG than the CG. The same occurred with the OTG in relation to OSG. Conclusions The depression of estrogen levels produced important changes in the tibial cartilage of Wistar rats mainly on the superficial layer and physical activity seemed to relieve these effects. Microsc. Res. Tech. 76:381–387, 2013. © 2013 Wiley Periodicals, Inc.

A revascularização linfática é um desafio e o estabelecimento de novas estratégias terapêuticas podem melhorar a qualidade de vida de pessoas que sofrem de distúrbios linfáticos. O objetivo deste estudo foi verificar a capacidade de tratamento com VEGFC exógeno na melhoria da vascularização linfática de uma maneira dependente do tempo em membros pélvicos (MP) de camundongos após a remoção do linfonodo inguinal. O linfonodo inguinal esquerdo foi removido cirurgicamente para mimetizar patologias com diminuição da vascularização linfática. Densidade vascular linfática (Vv) e de comprimento (Lv) foram avaliadas por imunohistoquímica, seguidas de estereologia, após a cirurgia com ou sem o tratamento com VEGFC exógeno. O grupo controle não foi manipulado, mas recebeu soro fisiológico em vez de tratamento com VEGFC exógeno. As expressões do VEGFC e FLT4 local foram avaliadas por qPCR. Houve efeito do tempo sobre Vv e Lv no Grupo Cirurgia e diferença significativa entre os grupos Controle e Cirurgia nas três regiões estudadas (região proximal, média e distal) do MP esquerdo (MPE). A Lv mostrou diferença significativa entre os grupos Controle e Cirurgia somente na região média do MPE. A Vv e a Lv para o Grupo Tratamento foram maiores do que os outros grupos em todas as regiões do MPE. A expressão gênica do VEGFC e do FLT4 apresentou efeito do tempo em todas as regiões do MPE para os grupos Cirurgia e Tratamento. Ambas as expressões gênicas do VEGFC e do FLT4 apresentaram diferença significativa entre os grupos Controle e Cirurgia, entre os grupos Cirurgia e Tratamento e entre os grupos Controle e Tratamento. Os resultados mostraram que os camundongos são bons modelos experimentais para o uso de VEGFC exógeno como terapia de revascularização linfática, e o tratamento com VEGFC exógeno aumenta vascularização linfática já após 3 dias de dano linfático. Lymphatic revascularization is a challenge and the establishment of new therapeutic strategies may improve quality of life from those suffering from lymphatic disorders. The objective of this study was to verify the VEGFC treatment capacity in improving lymphatic vascularization in a time-dependent manner in mouse hind limb (HL) after removal of inguinal lymphnode. The left inguinal lymphnode was surgically removed to mimetize pathologies with decreased lymphatic vascularization. Lymphatic vascular density (Vv) and length (Lv) were evaluated by immunohistochemistry followed by stereology after surgery and/or VEGFC treatment. Control group was not manipulated but received saline instead of VEGFC treatment. VEGFC and FLT4 local expression were assessed by qPCR. There was effect of time over Vv and Lv in the SG and significant difference between CG and SG in the three studied regions (proximal, medium and distal region) of the left HL (LHL). The Lv showed significant difference between CG and SG only in the medium region. The Vv and the Lv for TG were higher than the other groups in all regions of LHL. VEGFC and FLT4 gene expression presented time effect in all regions of the LHL for SG and TG. Both VEGFC and FLT4 gene expression presented significant difference between CG and SG, between SG and TG, and between CG and TG. The results show that mice are good experimental models for VEGFC use as therapy for lymphatic revascularization, and VEGFC treatment increased the lymphatic vasculature already after 3 days of lymphatic damage.

The morphology and location of lymph nodes from seven species of Odontocetes, of both sexes and different age groups, were described. All animals were derived from stranding events along the North and Northeastern coasts of Brazil. After the identification of lymph nodes in situ, tissue samples were analyzed for light and electron microscopy. Vascular volume density (VVD) and vascular length density (VLD) were evaluated in the mesenteric lymph nodes. Lymph nodes occurred as solitary nodules or in groups, varying in shape and size. In addition to using the nomenclature recommended by Nomina Anatomica Veterinaria, new nomenclatures were suggested based on the lymph nodes topography. Lymph nodes were covered by a highly vascularized and innervated capsule of dense connective tissue, below which muscle fibers were observed, inconsistently, in all studied species. There was no difference in VLD among different age groups. However, VVD was higher in adults. Lymph nodes parenchyma was divided into an outer cortex, containing lymph nodules and germinal centers; a paracortical region, transition zone with dense lymphoid tissue; and an inner medulla, composed of small irregular cords of lymphatic tissue, blood vessels, and diffuse lymphoid tissue. Abundant collagen fibers were observed around arteries and arterioles. Germinal centers were more evident and developed in calves and young animals, being more discrete and sparse in adults. The morphology of lymph nodes in Odontocetes was typical of that observed in other terrestrial mammals. However, new groups of lymph nodes were described for seven species occurring in the Brazilian coast.

To check if ovariectomy produces alterations in the articular cartilage structure of the tibial proximal epiphysis and if aerobic physical exercise has any influence on these alterations.Six-month-old female Wistar rats were distributed in three groups: Control group (CG), without ovariectomy and physical activity; Oophorectomy and sedentary group (OSG), with bilateral ovariectomy and without physical activity, and Oophorectomy and trained group (OTG), with bilateral ovariectomy and physical activity during 3 months. All rats were euthanised at 9 months of age. The results were obtained by using histological sections stained with Hematoxylin and Eosin, and Picrosirius stain.In relation to the thickness of the articular cartilage an increase was observed in the superficial layer in OSG in relation to OTG. The number of chondrocytes did not show any significant difference. The nuclear volume of chondrocytes in the superficial layer was higher in CG than in OSG . In the medium layer, the volume of chondrocytes was higher in CG than in OSG and the OTG. The same occurred when the values of the epiphysial cartilage was considered as a whole. For the collagen, the values of volume density of the collagen fibers were higher in the OSG and in the OTG than the CG. The same occurred with the OTG in relation to OSG.The depression of estrogen levels produced important changes in the tibial cartilage of Wistar rats mainly on the superficial layer and physical activity seemed to relieve these effects.