Propolis anti-inflammatory effects on MAGE-1 and retinoic acid-treated dendritic cells and on Th1 and T regulatory cells

Background: Propolis exhibits huge potential in the pharmaceutical industry. In the present study, its effects were investigated on dendritic cells (DCs) stimulated with a tumor antigen (MAGE-1) and retinoic acid (RA) and on T lymphocytes to observe a possible differential activation of T lymphocytes, driving preferentially to Th1 or Treg cells. Methods: Cell viability, lymphocyte proliferation, gene expression (T-bet and FoxP3), and cytokine production by DCs (TNF-α, IL-10, IL-6 and IL-1ß) and lymphocytes (IFN-γ and TGF-ß) were analyzed. Results: MAGE-1 and RA alone or in combination with propolis inhibited TNF-α production and induced a higher lymphoproliferation compared to control, while MAGE1 + propolis induced IL-6 production. Propolis in combination with RA induced FoxP3 expression. MAGE-1 induced IFN-γ production while propolis inhibited it, returning to basal levels. RA inhibited TGF-ß production, what was counteracted by propolis. Conclusion: Propolis affected immunological parameters inhibiting pro-inflammatory cytokines and favoring the regulatory profile, opening perspectives for the control of inflammatory conditions.(AU)

Concomitant occurrence of multicentric hemangiosarcoma and histiocytic sarcoma in a dog

Histiocytic sarcoma (HS) is uncommon malignant neoplasia of round cells with marked predilection in Rottweiler and Bernese Mountain. The disseminated form, which mainly affects the spleen, lungs, lymph nodes, bone marrow, skin, and subcutis, presents a quick and aggressive clinical behavior. Hemangiosarcoma (HAS) is a malignant neoplasm of endothelial vessel cells commonly reported to affect the right atrium of dogs' hearts. A male Rottweiler, five years old, presented flaccid paraplegia and progressive muscular atrophy in the temporal, masseter, and limbs muscles; Due to the clinical stage of the animal, euthanasia was conducted. During the necroscopic examination, it was noticed that several masses presented different sizes; some were whitish, and others were reddish and spread in multiple organs (lungs, heart, spleen, stomach, kidneys, brain, medulla, skeletal muscle, and pre-scapular lymph node). Microscopically, in some organs such as the stomach, right ventricle, lungs, and medulla, it was noticed a proliferation of myeloid cells, highly cellular, with poor demarcation, no encapsulation, and with the infiltrative growth pattern of cells with high pleomorphism. Numerous tumoral emboli were observed in the spleen, brain, skeletal muscle, and lymph node. These cells were submitted for immunohistochemistry and were positive for CD18 (HS antibody). In the right atrium, liver, and kidney it was observed malignant and infiltrative endothelial proliferation (HSA) and emboli in the medulla. Therefore, we conclude that both neoplasms (HS and HSA) cause the animal's paraplegia due to their embolism and metastasis to the spinal cord and skeletal muscle.(AU)

Morphological aspects and germ cells of male reproductive tract of river stingray, Potamotrygon amandae

The family Potamotrygonidae are the only species of stingrays restricted to fresh water and located exclusively in South America. The objective of this research was to analyze the morphological aspects and germ cells of the male reproductive tract of Potamotrygon amandae. The samples were fixed in 10% formalin, and then dehydrated in an ascending ethanol series (70 to 100%). To carry out light microscopy analyses, they were embedded in paraffin, cut and stained; as for scanning electron microscopy analyses, the samples were dried, glued in metallic bases and metalized. The gross morphology consisted of the following paired organs: testis, epididymis, deferent duct, Leydig gland, seminal vesicle, clasper, and the clasper gland. Microscopically, several stages of spermatogenesis were observed in the testis, occurring in spherical follicles, similar to other stingrays. The epididymis was formed by one duct subdivided in various tubules. The deferent ducts were continuous with the epididymis, and the lumen was full of spermatozoa. The Leydig glands consisted of glandular units with eosinophilic content in the lumen of some, and the deferent ducts ran parallel to the ventral portion. The seminal vesicles possessed numerous compartments to store the sperm, with a wall similar to a hive, and the lumen was full of spermatozoa. Alcian Blue (AB) and Periodic Schiff-Acid (PAS) performed in the Leydig Gland, deferens ducts and seminal vesicle was positive only in the connective tissue, the cilia were PAS+ and the nuclei stained weakly for AB. The clasper gland was composed of unit glands and was covered with striated muscle externally. It stained very well with Periodic Schiff-Acid. The morphological aspects of the male reproductive tract of Potamotrygon amandae were similar to other stingrays.(AU)

Hematology and serum biochemistry of broilers fed diets supplemented with chondroitin and glucosamine sulfates

The objective of this study was to evaluate the hematology and serum biochemistry of broilers fed diets supplemented with chondroitin and glucosamine sulfates. An experiment was laid out in a completely randomized design with a 3 × 3 factorial arrangement (three levels of chondroitin sulfate: 0, 0.05 and 0.10%; and three levels of glucosamine sulfate: 0, 0.15, and 0.30%), with each treatment involving six replicates of 30 birds. Hematology (red blood cells, hemoglobin, hematocrit, total plasma protein [TPP], thrombocytes, white blood cells, eosinophils, monocytes, heterophils, and lymphocytes) and serum biochemistry (totalinteraction between sulfates influenced (p < 0.05) total calcium by 21 days, ionic calcium by 21 and 42 days, and phosphorus, chlorides, and sodium by 42 days. Supplementation with chondroitin and glucosamine sulfates in the broilers' diet favored their immune system and mineral metabolism, increasing serum concentrations of calcium, phosphorus, and sodium. serum protein [TSP], albumin, globulins, aspartate aminotransferase [AST], gamma-glutamyltransferase [GGT], alkaline phosphatase [AP], total calcium, ionic calcium, phosphorus, sodium, potassium, and chlorides) variables were evaluated at 21 and 42 days. Data were subjected to analysis of variance. When the means differed significantly by the F-test, orthogonal analysis was performed to test the linear and quadratic effects of chondroitin and glucosamine sulfate levels. Glucosamine sulfate reduced the number of monocytes linearly, by 42 days (p = 0.0399). There was an interaction effect between the sulfates on total white blood cells (p = 0.0099) and lymphocytes (p = 0.0004) by 21 days. Chickens supplemented with 0.10% chondroitin sulfate showed a linear increase in white blood cells (p = 0.0287) and lymphocytes (p = 0.0144) with the addition of glucosamine sulfate. Chondroitin sulfate supplementation increased serum albumin linearly (p = 0.0099) and TSP quadratically (p = 0.0140), by 21 days. Glucosamine sulfate induced a quadratic response (p < 0.05) in albumin by 42 days, with the lowest value found with the inclusion of 0.06%. Glucosamine sulfate reduced chlorides linearly (p = 0.0237) by 21 days and increased calcium linearly (p = 0.0012) by 42 days. The interaction between sulfates influenced (p < 0.05) total calcium by 21 days, ionic calcium by 21 and 42 days, and phosphorus, chlorides, and sodium by 42 days. Supplementation with chondroitin and glucosamine sulfates in the broilers' diet favored their immune system and mineral metabolism, increasing serum concentrations of calcium, phosphorus, and sodium.(AU)

Objetivou-se avaliar a hematologia e a bioquímica sérica de frangos de corte suplementados com sulfatos de condroitina e de glucosamina na ração. Foi conduzido um experimento em delineamento inteiramente casualizado, em esquema fatorial 3 x 3 (três níveis de sulfato de condroitina: 0; 0,05 e 0,10%; e três níveis de sulfato de glucosamina: 0; 0,15 e 0,30%), cada tratamento com seis repetições de 30 aves. Foram avaliadas as variáveis de hematologia (hemácias, hemoglobina, hematócrito, proteínas plasmáticas totais [PPT], trombócitos, leucócitos, eosinófilos, monócitos, heterofilos e linfócitos) e bioquímica sérica (proteínas séricas totais [PST], albumina, globulinas, aspartato aminotransferase [AST], gama glutamiltransferase [GGT], fosfatase alcalina [FA], cálcio total, cálcio iônico, fósforo, sódio, potássio e cloretos) aos 21 e 42 dias. Os dados foram submetidos à análise de variância. Quando as médias diferiram significativamente pelo teste F, a análise ortogonal foi realizada para testar os efeitos lineares e quadráticos dos níveis dos sulfatos de condroitina e de glucosamina. Observou-se efeito linear decrescente (p = 0,0399) do sulfato de glucosamina na quantidade de monócitos aos 42 dias. Houve interação dos sulfatos para leucócitos totais (p = 0,0099) e linfócitos (p = 0,0004) aos 21 dias. Frangos suplementados com 0,10% de sulfato de condroitina mostraram um aumento linear dos leucócitos (p = 0,0287) e dos linfócitos (p = 0,0144) com a inclusão de sulfato de glucosamina. A suplementação com sulfato de condroitina aumentou linearmente (p = 0,0099) a albumina sérica e afetou de forma quadrática (p = 0,0140) as PST aos 21 dias. O sulfato de glucosamina demonstrou um efeito quadrático (p < 0,05) sobre a albumina aos 42 dias, o menor valor foi encontrado para a inclusão de 0,06%, respectivamente. O sulfato de glucosamina reduziu linearmente (p = 0,0237) os cloretos aos 21 dias e aumentou linearmente (p = 0,0012) o cálcio total aos 42 dias. Verificouse interação (p < 0,05) dos sulfatos para cálcio total aos 21 dias, cálcio iônico aos 21 e 42 dias e para fósforo, cloretos e sódio aos 42 dias. A suplementação com os sulfatos de condroitina e de glucosamina na ração de frangos de corte favoreceram o sistema imune e o metabolismo de minerais, com aumento nas concentrações séricas de cálcio, fósforo e sódio.(AU)

Canine mixed tumors as one of the mammary neoplasia varieties

The most significant interest in veterinary oncology is occupied by tumors of a mixed nature, which are represented by the direct interaction of epithelial and mesenchymal components. According to some data, the formation of cell lineage of mesenchymal cells occurs due to the transformation of epidermal basket cells surrounding the glandular epithelium. In this case, the formation of a cartilage anlage with its further transformation into differentiated bone tissue occurs. The article contains information about the structure and features of the location and interaction of mixed tumors' glandular and stromal components in female dogs (n=29). It was revealed that in addition to simple mixed neoplasms, tumors with proplasia of the mesenchymal component into highly differentiated cartilage or bone tissue were also often registered. To confirm subcellular changes in the expression of certain types of proteins, such as a-SMA and vimentin, an IHC study was used. The expression of the studied biomarkers was established in mesenchymal fibroblastic differon cells and myoepithelial cells. The intensity of immunoreactive material expression ranged from moderate (2+) to strong (3+), indicating changes in myoepithelial cells' genotype during the formation of mixed mammary neoplasms.(AU)

Treatment of chronic immunomediated thrombocytopenia with mesenquimal stem cells in dog: case report / Tratamento de trombocitopenia imunomediada crônica com células-tronco mesenquimais em cão: relato de caso

Immunomediated thrombocytopenia is a systemic metabolic disorder in which the platelet count falls below reference values, as the patient's immune system destroys them. The main clinical signs in thrombocytopenia are petechial, hemorrhages, ecchymoses and suffusions. Hematomas can also occur in coagulation disorders. The diagnosis is based on clinical findings and hematological examinations. The treatment consists of the use of corticosteroids and immunosuppressants, delaying cell destruction, and may last for months, not always obtaining a cure for the disease. The present work reports the use of therapy with allogeneic mesenchymal stem cells, derived from the adipose tissue of dogs, for the treatment of chronic immunomediated thrombocytopenia, with an evolution of more than one year, in a Pinscher dog. The alternative treatment showed a good evolution, keeping platelets within the reference values during the treatment, giving the patient quality of life and removing the need for continuous medication for homeostasis after treatment.

A trombocitopenia imunomediada é uma desordem metabólica sistêmica, na qual a contagem plaquetária fica abaixo dos valores de referência, pois o sistema imunológico do paciente a destrói. O principal sinal clínico na trombocitopenia são hemorragias, petequiais, equimoses e sufusões. Hematomas podem ocorrer também em alterações da coagulação. O diagnóstico baseia-se nos achados clínicos e nos exames hematológicos. O tratamento consiste na utilização de corticosteroides e imunossupressores, o que retarda a destruição celular, mas pode se prolongar por meses, nem sempre obtendo cura da doença. O presente trabalho relata a utilização da terapia com células-tronco mesenquimais alogênicas, oriundas do tecido adiposo de cães, para tratamento de trombocitopenia imunomediada crônica, com evolução de mais de um ano, em um cão da raça Pinscher. O tratamento alternativo revelou boa evolução, pois manteve as plaquetas dentro dos valores de referência durante o tratamento, o que proporcionou qualidade de vida ao paciente e tornou desnecessárias medicações de uso contínuo para a homeostase após o tratamento.

Zootechnical performance, degree of steatosis and the genotoxic potential in yellowtail tetra Astyanax lacustris fed with different levels of L-carnitine / Desempenho zootécnico, grau de esteatose e potencial genotóxico de lambaris Astyanax lacustris alimentados com diferentes níveis de L-carnitina

L-carnitine perform a major role in transporting long-chain fatty acids into the mitochondria, where they are oxidized. It has been used in animal diets to decrease fat and increase muscle protein. The aim of this study was to evaluate the zootechnical performance, degree of steatosis in the liver, and genotoxic potential in Astyanax lacustris fed with different levels of L-carnitine (LC). Yellowtail tetra juveniles (n = 140) were distributed in 20 tanks of 70 L, with seven fish in each, in a water recirculation system with controlled temperature (27±0.1°C). The treatments with different levels of L-carnitine supplementation were: 0 (control), 250, 500, 750, and 1000 mg of LC per kg of food. The diets were provided twice a day for 60 days. The results showed that the different levels of LC did not affect (P>0.05) weight gain, survival, viscerosomatic index, and the liver hepatocytes showed a normal appearance. However, the use of LC supplementation showed genotoxic potential with a significant difference (P<0.05) for cell alterations when compared to the control at concentrations above 500mg kg-1.

A L-carnitina exerce um papel importante no transporte de ácidos graxos de cadeia longa até a mitocôndria para serem oxidados e tem sido incorporada em rações para animais com o objetivo de diminuir a deposição de gordura e aumentar a proteína muscular. O objetivo deste trabalho foi avaliar o desempenho zootécnico, o grau de esteatose no fígado e o potencial genotóxico em Astyanax lacustris alimentados com diferentes níveis de L-carnitina (LC). Juvenis de lambari-do-rabo-amarelo (n=140) foram distribuídos em 20 caixas de 70L, sete peixes em cada, em um sistema de recirculação de água com temperatura controlada (27±0,1°C). Os tratamentos com os níveis de suplementação foram: 0 (controle), 250, 500, 750 e 1000 mg de LC kg-1 de ração. As dietas foram fornecidas duas vezes ao dia, durante 60 dias. Os resultados mostraram que os diferentes níveis de LC não influenciaram (P>0,05) o ganho de peso; a sobrevivência, o índice viscerossomático e os hepatócitos do fígado apresentaram-se com aparência normal. No entanto, a suplementação com LC apresentou potencial genotóxico com diferença significativa (P<0,05) para alterações celulares quando comparada ao controle em concentrações superiores a 500mg kg-1.

Probiotics supplementation reduces cigarette smoke-induced damage in the respiratory micro-architecture of mice / Suplementação com probióticos reduz os danos causados pela fumaça de cigarro na microarquitetura respiratória de camundongos

This study evaluated the protective efficacy of probiotics supplementation against cigarette smoke-induced lung emphysema, inflammation, and loss of cilia in mice. Probiotics are known to promote mucosal tolerance and mitigate respiratory injuries. Twenty-four adult mice were randomly divided into three groups: control (Ctr), cigarette smoke (CS), and cigarette smoke + probiotics (CS+P). Probiotics were given for 7 days before exposure to smoke in the CS+P group. Tissue samples of the trachea (goblet cell count and index, loss of cilia), lungs (airspace distention), and bronchoalveolar lavage fluid (BALF) were collected and processed. The results showed a significant increase in acidic and neutral goblet cells in the CS group compared to the Ctr and CS+P groups (P < 0.05). Overall, goblet cell number and index were lower in the CS+P group (41.71 ± 5.76, 0.67 ± 0.073) than CS group (56.28 ± 5.34, 1.31 ± 0.28). Inflammatory cells and loss of cilia significantly decreased in mice fed probiotics before exposure to cigarette smoke (P < 0.05). Lung emphysema was also significantly reduced in the CS+P group compared to the CS group (P < 0.05). In conclusion, dietary supplementation of probiotics reduced lung emphysema, inflammatory cells, goblet cell index, and loss of cilia under conditions of cigarette smoke exposure in mice.(AU)

O presente trabalho avaliou a eficácia da suplementação com probióticos contra a indução de enfisema, inflamação e perda de cílios por fumaça de cigarro em camundongos. Sabe-se que os probióticos promovem a tolerância da mucosa e mitigam as injúrias respiratórias. Vinte e quatro camundongos foram divididos, ao acaso, em três grupos: controle (CTR), fumaça de cigarro (CS) e fumaça de cigarro + probióticos (CS+P). Os probióticos foram fornecidos por sete dias antes da exposição à fumaça no grupo CS+P. Foram colhidas e processadas amostras de tecidos da traqueia (contagem de células caliciformes e index, perda de cílios), pulmões (distensão do espaço aéreo) e fluido de lavagem broncoalveolar. Os resultados obtidos revelaram um aumento significativo em células caliciformes acidificadas e neutralizadas no grupo CS quando comparado aos grupos Ctr e CS+P (P<0,05). Os números global e o índice de células caliciformes foram menores no grupo CS+P (41,71+ 5,76; 0,67+ 0,073) que no grupo CS (56,28+ 5,34 e 1,131+ 0,28). As células inflamatórias e a perda de cílios decresceram em camundongos alimentados com probióticos antes da exposição à fumaça de cigarro (P<0,05). O enfisema pulmonar também foi significativamente reduzido no grupo CS+P quando comparado ao grupo CS (P<0,05). A conclusão obtida foi que o fornecimento de dieta suplementada com probióticos reduziu o enfisema pulmonar, as células inflamatórias, o índice de células caliciformes e a perda de cílios nos camundongos expostos à fumaça de cigarro.(AU)

Cell therapy with adipose tissue-derived human stem cells in the urinary bladder improves detrusor contractility and reduces voiding residue / A terapia celular com células-tronco humanas derivadas do tecido adiposo na bexiga urinária melhora a contratilidade do detrusor e reduz o resíduo miccional

Detrusor hypocontractility (DH) is a disease without a gold standard treatment in traditional medicine. Therefore, there is a need to develop innovative therapies. The present report presents the case of a patient with DH who was transplanted with 2 x 106 adipose tissue-derived mesenchymal stem cells twice and achieved significant improvements in their quality of life. The results showed that cell therapy reduced the voiding residue from 1,800 mL to 800 mL, the maximum cystometric capacity from 800 to 550 mL, and bladder compliance from 77 to 36.6 mL/cmH2O. Cell therapy also increased the maximum flow from 3 to 11 mL/s, the detrusor pressure from 08 to 35 cmH2O, the urine volume from 267 to 524 mL and the bladder contractility index (BCI) value from 23 to 90. The International Continence on Incontinence Questionnaire - Short Form score decreased from 17 to 8. Given the above, it is inferred that the transplantation of adipose tissue-derived mesenchymal stem cells is an innovative and efficient therapeutic strategy for DH treatment and improves the quality of life of patients affected by this disease.

A Hipocontratilidade Detrusora (HD) é uma doença sem um tratamento padrão-ouro na medicina tradicional. Logo, há a necessidade de desenvolvimento de terapias inovadoras. O presente relato apresenta um caso de paciente com HD transplantado duas vezes com 2 x 106 células-tronco mesenquimais derivadas do tecido adiposo que obteve melhoras significativas em sua qualidade de vida. Os resultados demonstraram que a terapia celular reduziu o resíduo miccional de 1.800mL para 800mL; a Capacidade Cistométrica Máxima de 800 para 550mL; a complacência de 77 para 36,6mL/cmH2O. A terapia celular também aumentou o fluxo máximo de 3 para 11mL/s; a pressão detrusora de 08 para 35cmH2O; o volume urinado de 267 para 524mL e o índice de contratilidade vesical (BCI) de 23 para 90. O score do International Continence on Incontinence Questionare - Short Form passou de 17 para 8. Diante do exposto, infere-se que o transplante de células-tronco mesenquimais derivadas do tecido adiposo é uma estratégia terapêutica inovadora e eficiente para o tratamento da HD e para melhoria da qualidade de vida de pacientes acometidos por essa doença.

Intertubular morphometric and ultrastructural testes analyses in mdx mice

Duchenne Muscular Dystrophy (DMD) reproductive alterations and the influence of antioxidant treatments may aid in understanding morphometry testicular quantification. In this context, the aim of the present study was to characterize the intertubular compartment (ITC) morphometry of animal testes in mdx mice supplemented with ascorbic acid (AA). Sixteen mice were used, namely the C57BL/10 (non-dystrophic) and C57BL/10Mdx (dystrophic) lineages, distributed into the following groups: Control (C60), Dystrophic (D60), Control supplemented with AA (CS60), Dystrophic supplemented with AA (DS60). A total of 200 mg/kg of AA were administered to mice for 30 days. Subsequently, the testicles were collected, weighed, and fragmented. The obtained fragments were fixed in Karnovsky's solution (pH 7.2) and embedded in historesin for morphometric and transmission electron microscopy assessments. Leydig cells were hypertrophic in the D60 group, but was reverted by AA supplementation in the DS60 group. The DS60 group also exhibited increased intertubular volume compared to the CS60 group. The ultrastructural images identified multilamellar bodies in dystrophic animals (lipid storage) and telocyte cells (transport substances) in both control and dystrophic animals. Morphometric alterations were, therefore, noted in the intertubular compartment due to Duchenne muscular dystrophy (DMD), with AA administration capable of altering Leydig cells in this condition.(AU)

Ultrastructural study of the esophagus and stomach of Arapaima gigas (Schinz 1822), juvenile paiche, created excavated tank

Paiche (Arapaima gigas) belongs to the Kingdom Animalia, Phylum Chordata, Class Actinopterygii, Order Osteoglossiformes, Family Arapaimidae, Genus Arapaima, and its origin may date to the Jurassic period. The species has natural habitat in the Amazonian rivers, found mainly in marginal lakes, being considered an important fishing resource, with high market value and high demand for meat and leather in both Brazilian and international trade. This study aims to report the morphology of the esophagus and stomach by light microscopy and scanning electronics microscopy. The esophagus was presented as muscular, short, tubular and fan-shaped in the cranial portion, also presenting deep longitudinal folds, and the entire mucosa is covered by mucus secretory cells with distinct morphological characteristics. Pirarurcu's stomach has a J-shape divided into three regions: cardiac with a lighter aspect, fundus portion with few folds in the mucosa, and pyloric with deeper folds, also presenting gastroliths in fundus and pyloric portions. Both microscopy studies highlighted three glandular regions, composed by mucoid columnar epithelial cells, gastric crypts with different shapes and sizes depending on each portion, in which the different shapes of the mucosal folds in each region of the stomach were evident, and digitiform microsaliences were found in the cardiac region, and micro-orifices and desmosome in the fundus region. Also, fundus and pyloric portions produce more mucus than the cardiac. Then morphology found was consistent with the eating habits and management of distinct characteristics of the digestive tract.(AU)

Prenatal origins of productivity and quality of beef

The productive traits of beef cattle are orchestrated by their genetics, postnatal environmental conditions, and also by the intrauterine background. Both under- or overnutrition, as specific dietary components, are able to promote persistent effects on the offspring. This occurs because dietary factors act not only affecting the availability of substrates for fetal anabolism and oxidative metabolism, but also as signals that regulate several events toward fetal development. Therefore, this study aimed to summarize the gestational nutrition effects on the offspring performance and meat quality in a long term. Overall, studies have shown that many of these alterations are under the control of epigenetic mechanisms, as DNA methylation, histones modification, and non-coding RNA. The current knowledge has indicated that the fetal programming responses are dependent on the window of fetal development in which the dietary treatment is applied, the intensity of maternal nutritional stimuli, and the treatment application length. Collectively, studies demonstrated that muscle cell hyperplasia is impaired when maternal requirements were not achieved in the second third of gestation, which limits the formation of a greater number of muscle fibers and the offspring growth potential in a long term. Changes in muscle fibers metabolism and in collagen content were also reported as consequence of a dietary perturbation during pregnancy. In contrast, a maternal overnutrition during the late pregnancy has been associated with beneficial responses on meat quality. In summary, ensuring an adequate maternal environment during the fetal development is crucial to enhance the productive responses in beef cattle operations.(AU)

Regulation of progesterone during follicular development by FSH and LH in sheep

Progesterone (P4) can participate in the development of female mammalian antral follicles through nuclear receptor (PGR). In this experiment, the differences of P4 synthesis and PGR expression in different developmental stages of sheep antral follicles (large > 5mm, medium 2-5mm, small < 2mm) were detected by enzyme-linked immunosorbent assay, immunohistochemistry, qRT-PCR and Western blotting. Secondly, sheep follicular granulosa cells were cultured in vitro. The effects of different concentrations of FSH and LH on P4 synthesis and PGR expression were studied. The results showed that acute steroid regulatory protein (StAR), cholesterol side chain lyase (P450scc) and 3β Hydroxysteroid dehydrogenase (3β-HSD) and PGR were expressed in antral follicles, and with the development of antral follicles in sheep, StAR, P450scc and the expression of 3β-HSD and PGR increased significantly. In vitro experiments showed that FSH and LH alone or together treatment could regulate P4 secretion and PGR expression in sheep follicular granulosa cells to varying degrees, hint P4 and PGR by FSH and LH, and LH was the main factor. Our results supplement the effects of FSH and LH on the regulation of P4 synthesis during follicular development, which provides new data for further study of steroid synthesis and function in follicular development.(AU)

Effect of In Ovo Ascorbic Acid Injection and Thermal Manipulation During Incubation on Intestine Morphology of Broilers Reared under Cold and Heat Stress

It was investigated if pre-incubation ascorbic acid (AA) injection in fertile eggs incubated at high temperature impacts the performance, the yield of carcass and parts, and the intestine morphology of broilers reared under heat stress. Three thousand Cobb® fertile broiler eggs were randomly distributed according to weight into three incubations treatments (eggs not injected with AA and incubated at 37.5°C; eggs not injected with AA and incubated at 39°C; and eggs injected with 6 µg AA/100 µL water prior to incubation and incubated at 39ºC). The hatched birds were reared at thermoneutral, cold, and hot house temperatures. Broilers reared under hot temperature presented lower feed intake and weight gain than the broilers of the different rearing temperatures. Egg incubation at 39.0 ºC and 39.0 ºC + AA reduced broiler viability. Carcass and cut yields were not influenced by incubation and rearing procedures. Duodenal goblet cell count was lower in broilers from eggs of the treatment 39ºC + AA than in broilers from the other incubation treatments and in broiler rearing in hot temperature. In the jejunum, the goblet cell counts were higher in broilers that were reared under hot than thermoneutral temperatures. The incubation treatment of 39 ºC+AA increased the goblet cell counts in the ileum of broilers reared under cold temperatures. Rearing temperature influenced the duodenal villi counts, which were lower under cold rearing conditions than in the two other rearing temperatures. The results showed that egg incubation at 39°C, independently of ascorbic acid injection, did not produce an effective epigenetic heat adaptation in broilers.(AU)

Comparison of three different strategies to treat sciatic nerve regeneration: an experimental study

Purpose: To compare the effect of vein conduit filled with adipose tissue stem cells (ASC) on peripheral nerve injury regeneration. Methods: We analyzed 30 male Wistar rats surgically submitted to a 5-mm gap on the sciatic nerve. Then, the animals were divided into three groups: nerve autografting (AG, n=10), autogenous inverted glycerol-conserved vein (VG, n=10), and autogenous inverted glycerol-conserved vein + ASC (VASCG, n=10). The study endpoints were neuromotor functional analysis, gastrocnemius muscle weight, and sciatic nerve graft histomorphometry analysis. In the histologic analysis, we added a control group (naïve nerve). Results: Regarding functional analysis (Walking tract- score), the findings at week 3 showed a difference between the AG and the VG (-96.6 vs. -59.6, p=0.01, respectively) and between the VG and the inverted vein + VASCG (-59.9 vs. -88.92, p=0.02). At week 12, this study showed a difference between the AG and the VG (-64.8 vs. -47.3, p=0.004, respectively), and also a difference between the VG and the VASCG (-47.3 vs. -57.4, p=0.02, respectively). There was no difference in the histomorphometry analysis (nerve diameter, Schwann cells counting). The gastrocnemius muscles on the intervention side were more atrophic when compared to the gastrocnemius muscles on the control side. Conclusions: Our results suggested better functional recovery in the inverted vein group when compared to control group, and inverted vein + ASC group.

Sheep corneal endothelium morphology - evaluation with trypan blue and alizarin red

Background: The endothelium is a layer fundamental to maintaining corneal transparency. In ophthalmology, sheep eyes have been used as a model in research related to corneal transplantation. Different techniques have been used to evaluate the corneal endothelium. Concerning vital dyes, corneal endothelial cell analyses have not yet been studied in ovines. The purpose of the present study was to evaluate the morphology of endothelial cells from different regions of the cornea of sheep after staining with alizarin red and trypan blue using an optical microscope. Materials, Methods & Results: Twenty healthy eyes of 10 male sheep obtained from a licensed commercial slaughterhouse were studied. The study was approved by the Research Committee of the Faculty of Veterinary at UFRGS and followed the ethical standards of the Association for Research in Vision and Ophthalmology (ARVO). Immediately after the slaughter, the eyes were enucleated and underwent eye examination. The corneal endothelium was stained with trypan blue and alizarin red and examined and photographed using an optical microscope. The central, superior, inferior, nasal and temporal areas of the cornea were evaluated for cell morphology. Data were compared by t-tests. Differences were considered statistically significant at P < 0.05. Immediately after staining the corneal endothelium, it was possible to examine with an optical microscope, obtain images and analyse the shape of endothelial cells from all regions of the sheep cornea. Polygonal, uniform and continuous cells were observed in all samples studied. Considering all the corneas analysed, cells with 6 sides (75.11%), 5 sides (12.76%) and 4 sides (12.12%) were found. In the central region of the cornea 75.91% of cells with 6 sides, 12.6% of cells with 5 sides and 11.48% with 7 sides were found. In the superior region of the cornea 76.07% of cells with 6 sides, 13.25% with 5 sides and 10.68% with 7 sides were found. In the lower region were found 74.72% of cells with 6 sides, 13% with 5 sides and 12.27% with 7 sides. In the temporal region, 74.14% were 6-sided cells, 11.42% had 5 sides, and 14.43% had 7 sides. Furthermore, in the nasal region, 74.72% of the cells had 6 sides, 13.54% had 5 sides, and 11.73% had 7 sides. No significant differences were found between cell morphology in all corneal regions evaluated. In addition, no significant difference was found when comparing the right eye with the left eye. Discussion: Different methods are used for the analysis of corneal endothelium. For ex vivo research optical microscopy after endothelial staining is an alternative low-cost technique that allows the analysis of all regions of the cornea. Quantitative analyses must characterise the endothelial parameters of the different species. The analysis of the morphology of corneal endothelium with an optic microscope after staining with alizarin red has been described as an effective, rapid and cost-efficient method, since this dye blends with the borated cells, allowing identification. In the present study, using optical microscopy and coloration with alizarin red it was possible to explore and obtain images of the ovine endothelium of all regions of the cornea. In the current study, the endothelium had a predominance of cells will 6 sides in all regions studied. This study allowed us to obtain images of the endothelium as well as quantitative data on the morphology of the different regions of the sheep cornea. This study demonstrated that morphology did not differ between the central and peripheral regions. The findings of this study represent a further source of reproducible data that should be considered when using sheep cornea as ex vivo model for experimental research.

Canine mammary neoplasms - evaluation of tumor microenvironment

Background: The tumor microenvironment is an important target of studies in different types of neoplasms. Understanding the role of general components such as immune, vascular and fibroblastic cells has the objective of contributing to prognosis and treatment. The aim of this study was to evaluate the relationship between mast cells and angiogenesis in benign and malignant mammary neoplasms by investigating the role of degranulation and microlocation of mast cells and neoformed vessels in canine mammary neoplasms. Materials, Methods & Results: Mammary glands (n = 122) from 50 female dogs submitted to mastectomy without chemotherapy were evaluated and categorized into 3 groups: control group (n = 46); malignant group (n = 57) and benign group (n = 19). Lymph nodes without changes (n = 59) and with metastases (n = 6) were also evaluated. To evaluate the MCD (mast cell density) and angiogenesis, Toluidine Blue (0.1%) and Gomori’s Trichrome techniques were performed and adapted from previous studies. Photomicrographs of 10 hotspot areas on a 40x objective lens of the mammary glands and lymph nodes were captured to assess MCD and angiogenesis. In the absence of these areas, random fields were captured. For the mammary glands of the malignant and benign groups, 20 fields were analyzed, as the analysis considered the microlocation (peritumoral and intratumoral). Counting was performed manually using ImageJ software version 1.42q by 2 observers. The statistical analysis were performed using SPSS software version 19.0. The most frequent histological type in the malignant group was carcinoma in mixed tumor (68.42%; 39/57) and in the benign group was benign mixed tumor (57.89%; 11/19). Female dogs without breed pattern were more frequently affected represented 70% of the animals and the mean age was 9 years and 8 months ± 3 years and 1 month...

Canine mammary neoplasms - evaluation of tumor microenvironment

Background: The tumor microenvironment is an important target of studies in different types of neoplasms. Understanding the role of general components such as immune, vascular and fibroblastic cells has the objective of contributing to prognosis and treatment. The aim of this study was to evaluate the relationship between mast cells and angiogenesis in benign and malignant mammary neoplasms by investigating the role of degranulation and microlocation of mast cells and neoformed vessels in canine mammary neoplasms. Materials, Methods & Results: Mammary glands (n = 122) from 50 female dogs submitted to mastectomy without chemotherapy were evaluated and categorized into 3 groups: control group (n = 46); malignant group (n = 57) and benign group (n = 19). Lymph nodes without changes (n = 59) and with metastases (n = 6) were also evaluated. To evaluate the MCD (mast cell density) and angiogenesis, Toluidine Blue (0.1%) and Gomoris Trichrome techniques were performed and adapted from previous studies. Photomicrographs of 10 hotspot areas on a 40x objective lens of the mammary glands and lymph nodes were captured to assess MCD and angiogenesis. In the absence of these areas, random fields were captured. For the mammary glands of the malignant and benign groups, 20 fields were analyzed, as the analysis considered the microlocation (peritumoral and intratumoral). Counting was performed manually using ImageJ software version 1.42q by 2 observers. The statistical analysis were performed using SPSS software version 19.0. The most frequent histological type in the malignant group was carcinoma in mixed tumor (68.42%; 39/57) and in the benign group was benign mixed tumor (57.89%; 11/19). Female dogs without breed pattern were more frequently affected represented 70% of the animals and the mean age was 9 years and 8 months ± 3 years and 1 month...(AU)

Anatomia e histologia do esôfago e vasos sanguíneos em Cães ­ Revisão de literatura / Anatomy and histology of the esophagus and blood vessels in dogs ­ Literature review

O sistema digestório é composto por órgãos de morfologia tubular, e glândulas associadas; tendo como principal função o processamento de alimentos e a absorção de nutrientes dos mesmos. Há um padrão estrutural comum entre órgãos tubulares deste sistema, o conhecimento desse padrão, facilita a compreensão das particularidades de cada órgão. Já os vasos sanguíneos são caracterizados e definidos por sua posição dentro do circuito vascular, pela estrutura individual, e mecanismos de controle da função vascular. Dessa forma, o objetivo do presente trabalho é aprofundar conhecimentos a respeito de anatomia e histologia do esôfago e vasos sanguíneos em cães.(AU)

The digestive system is composed of tubular morphology organs and associated glands; with the main function of food processing and nutrient absorption. There is a common structural pattern among tubular organs of this system, knowledge of this pattern facilitates the understanding of the particularities of each organ. Blood vessels are characterized and defined by their position within the vascular circuit, their individual structure, and the mechanisms that control vascular function. Thus, the objective of the present work is to deepen knowledge about the anatomy and histology of the esophagus and blood vessels.(AU)

Plasmocitoma extramedular em cão positivo para leishmaniose: Relato de caso / Extramedullary plasmacytoma in a dog positive for leishmaniasis: case report

Os plasmocitomas extramedulares são uma proliferação excessiva de células plasmocíticas, que pode acometer nos cães, o sistema cutâneo e mucocutâneo. Classificados como benignos, já que por sua vez possuem um baixo potencial metastático, porém o local onde se infiltra se comporta de forma agressiva. A formação dos nódulos é isolada, mas podem ser múltiplos, apresentam coloração avermelhada e possuem tamanhos médios de 1 a 2 cm. O histopatológico evidencia plasmócitos, em arranjo de cordão e entremeados a uma discreta quantidade de tecido conjuntivo. O objetivo deste relato é identificar e diagnosticar um caso de plasmocitoma extramedular em cão positivo para leishmaniose.(AU)

The extramedullary plasmacytomas are an excessive proliferation of plasmacytic cells, which can affect the cutaneous and mucocutaneous systems in dogs. Classified as benign, as they in turn have a low metastatic potential, but the site where they infiltrate behaves aggressively. The formation of nodules is isolated, but they can be multiple, present with a reddish color and have average sizes of 1 to 2 cm. Histopathology shows plasma cells, in a cord arrangement and intermingled with a small amount of connective tissue. The purpose of this report is to identify and diagnose a case of oral plasmacytoma in a dog.(AU)