The extracellular matrix protein pattern in the canine neoplastic mammary gland.

Extracellular matrix (ECM) proteins in the mammary gland provide structure and regulate its development and homeostasis. Alterations in its structure can regulate and support pathogenesis, like breast tumors. Aiming to identify the health and tumoral canine mammary ECM scaffold protein profile by immunohistochemistry, the decellularization process was carried out to remove the cellular content. Additionally, it was verified the influence of health and tumoral ECM on the attachment of health and tumoral cells. The types I, III, IV, and V structural collagens were scarce in the mammary tumor, and ECM fibers were disorganized. Vimentin and CD44 were more common in mammary tumor stroma, suggesting a role in cell migration that results in tumor progression. Elastin, fibronectin, laminin, vitronectin, and osteopontin were similarly detected under healthy and tumor conditions, providing the attachment of normal cells in healthy ECM, while tumoral cells were able to attach in tumoral ECM. The protein pattern demonstrates ECM alteration in canine mammary tumorigenesis, presenting new knowledge on mammary tumor ECM microenvironment.

Morphoquantitative evaluation of the heart of cutia (Dasyprocta agouti) and capybara (Hydrochoerus hydrochaeris) / [Avaliação morfoquantitativa do coração da cutia (Dasyprocta agouti) e da capivara (Hydrochoerus hydrochaeris)]

The cutia (Dasyprocta agouti) and capybara (Hydrochoerus hydrochaeris ) are hystricomorphic rodents of the Brazilian fauna. The wild animal anatomy is essential to the clinic and surgery practices and conservation programs. This work aimed to evaluate the heart quantitative morphology of the cutia and the capybara. For this, three hearts of cutia and three of capybaras were used from the collection of the Anatomy Sector of Domestic and Wild Animals of the Faculty of Veterinary Medicine and Animal Science of the University of São Paulo. The hearts were weighted and measured with a pachymeter. The left ventricle volume was estimated through the Cavalieri method. The hearts' mean weight was 16.76g (cutias) and 104.2g (capybaras). The mean values for the axis, width and base dimensions were of the cutia 4.3cm, 3.3cm and 3.3cm and for the capybara were 7.6cm, 6.0cm and 6.1cm, respectively. The mean volume of the left ventricle of the cutia and the capybara was 5.03cm3 and 54.55cm3, respectively. The quantitative results of the hearts were compatible with the average body weight of the rodents. Thus, these numerical data can be applied to veterinary cardiology.(AU)

A cutia (Dasyprocta agouti) e a capivara (Hydrochoerus hydrochaeris) são roedores histricomorfos da fauna brasileira. A anatomia de animais silvestres é essencial para a prática clínico-cirúrgica e programas conservacionistas. Este trabalho objetivou avaliar a morfologia quantitativa do coração da cutia e da capivara. Para isso, foram utilizados três corações de cutias e três de capivaras, provenientes do acervo do Setor de Anatomia dos Animais Domésticos e Silvestres da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo. Os corações foram pesados e as mensurações foram realizadas com paquímetro. O volume do ventrículo esquerdo foi calculado pelo método de Cavalieri. O peso médio dos corações foi de 16,76g (cutias) e de 104,2g (capivaras). Os valores médios para as dimensões eixo, largura e base foram, para a cutia, de 4,3cm, 3,3cm e 3,3cm e para a capivara, de 7,6cm, 6,0cm e 6,1cm, respectivamente. O volume médio do ventrículo esquerdo da cutia e da capivara foi respectivamente de 5,03cm3 e de 54,55cm3. Os resultados quantitativos dos corações foram compatíveis com o peso corporal médio dos roedores. Assim, esses dados numéricos podem ser aplicados à cardiologia veterinária.(AU)

Wistar rat dermis recellularization.

Skin lesions are normal to all species, regardless of gender or age. The skin, the largest organ of the body, has function as a primary barrier to the chemical, physical and biological aggressions of the environment. In animals, these lesions may be due to fights and/or predations, also as in humans, there is a very common cause of dermal lesions that are caused by burns and carcinomas. Looking for new techniques of tissue bioengineering, studies have been shown promising results for formulations of acellular biological scaffolds from tissue decellularization for the reconstitution of these lesions. The decellularization has its proof by a varied range of tests such as scanning electron microscopy and residual genomic DNA tests. Subsequently the tissue can go through the process of recellularization using cells of interest, even the animal that will receive this tissue, reducing the risks of rejection and improving the response to tissue transplantation. Thus, this manuscript aimed at the decellularization of the tissue with the use of chemical and physical means followed by sterilization and the establishment of a protocol for the recellularization of a decellularized scaffold from the Wistar rat dermis using murine fibroblasts and mesenchymal stem cells from canine adipose tissue for 7 days. After efficacy tests, the tissue recellularization were confirmed by immunofluorescence assays and scanning electron microscopy where the adherence of the cells in the biological scaffold was observed.

Efficacy and limitations of different approaches to anticipate the diagnosis of pregnancy in cattle / Eficácia e limitações de diferentes abordagens para antecipar o diagnóstico de gestação em bovinos

The study evaluated sonographic and serologic exams performed for early (20 to 30d) diagnosis of pregnancy. One hundred-twenty (n= 120) bovine recipients were synchronized (estrous=D0) and timed embryo transferred (TET, D7) with fresh in vitro produced embryos. In the first trial (n= 46), diagnosis of pregnancy was performed on day 20 (D20) by detecting CL blood flow (BF) and by Pregnancy-Associated Glycoproteins (PAGs) serology. In the second trial (n= 30), pregnancy diagnosis was performed on D25 by ultrasound visualization of uterine contents and by PAGs serology. In the last trial, PAG's serology was performed on D30. Ultrasonographic detection of the uterine contents and embryo viability performed on D30 (DG30) was considered the gold standard. The PROC FREQ procedure was used to test the agreement between diagnostic methods. On D20, the Doppler ultrasonography of the CL had showed high sensitivity (100%), but only moderate specificity (53.3%). On the same day, serologic diagnostic had no agreement (k= -0.08, P< 0.46) with the gold standard, with very low sensitivity (6.3%). However, the sensitivity of the serologic exam increased dramatically (from 6.3 to 100%) from D20 to D25, and it contributed to detect false negatives from the ultrasound diagnosis, improving the overall accuracy from 90% to 96.7%.(AU)

O estudo foi planejado para correlacionar exames ultrassonográficos e sorológicos realizados para o diagnóstico precoce (20 a 30d) de gestação. Cento e vinte (n= 120) receptoras bovinas foram sincronizadas (estro=D0), e embriões frescos produzidos in vitro foram transferidos em tempo fixo (TETF, D7). No experimento 1 (n= 46), o diagnóstico de gestação foi realizado no D20, pela detecção do fluxo sanguíneo do CL e pela sorologia de glicoproteínas associadas à gestação (PAGs). No experimento 2 (n= 30), a detecção da gestação foi realizada por meio da visualização do conteúdo do útero e também pela sorologia para PAGs. No experimento 3, a sorologia para PAGs foi realizada no D30. Em todos os experimentos, a visualização ultrassonográfica da vesícula e da viabilidade embrionária, realizada no D30, foi considerada padrão-ouro. O procedimento PROC FREQ testou o nível de concordância dos métodos diagnósticos. No D20, o diagnóstico baseado na vascularização do CL mostrou alta sensibilidade (100%) e apenas moderada especificidade (53,3%). Nesse mesmo dia, o diagnóstico sorológico não apresentou concordância (k=-0,08, P<0,46) com o padrão-ouro, além de baixa sensibilidade (6,3%). No entanto, a sensibilidade do exame sorológico aumentou drasticamente (6,3 para 100%) do D20 para o D25, contribuindo para detectar falsos negativos diagnosticados pela ultrassonografia, melhorando a acurácia (90 para 96,7%).(AU)

Concentration of collagen fibers in the musculature of broiler chickens fed with cupuacu seed by-product / Concentração de fibras colágenas na musculatura de frangos alimentados com farinha de cupuaçu

In this essay, we quantify the concentration of collagen fibers in broiler chickens exposed to increasing concentrations of cupuacu seed by-product. Collection of material was carried out in five chickens per treatment at 70 days old in the groups: control, 5% and 10% inclusion of cupuacu seed by-product. Fragments of Thoracic Pectoralis (PT) and Iliotibial lateralis (ITL) muscles were prepared for light and electronic microscopy. The amount of collagen fibers in the muscle groups was 1.08±0.61% in the PTC group; 6.24±2.58% in PT5% and 7.30±2.75% in PT10%. In the Iliotibial Lateralis groups, the results were 6.96±3.14% in the ITLC; 7.43±4.22% in the ITL5% and 8.66±2.35% in ITL10%. The amount of collagen fibers in the ITL5% and ITL10% groups showed no significant statistical difference. However, when compared to the ILTC group, there was a significant statistical difference. The PT muscle responds to standard nutritional changes, unlike the ILT muscle, which requires a high-nutrient formulation. The use of 5% cupuacu seed by-product has proven to be a viable alternative source of animal feed, as it promotes an increase in the concentrations of collagen fibers in the musculature of broiler chickens and is possibly the determining factor in meat texture.(AU)

Neste estudo, foram quantificadas as concentrações de fibras colágenas de frangos expostos a crescentes concentrações de farinha de cupuaçu. A coleta de material foi realizada em cinco animais por tratamento, aos 70 dias de idade, nos grupos: controle, inclusão de 5% e de 10% de farinha de cupuaçu. Fragmentos dos músculos peitoral torácico (PT) e iliotibial lateral (ITL) foram preparados para microscopia de luz e eletrônica. A quantidade de fibras colágenas nos grupos foi: 1,08±0,61% no grupo PTC; 6,24±2,58% em PT5% e 7,30±2,75% em PT10%. Nos grupos iliotibial lateral, os resultados foram: 6,96±3,14% no ITLC; 7,43±4,22% no ITL5% e 8,66±2,35% em ITL10%. A quantidade de fibras colágenas nos grupos ITL5% e ITL10% não apresentou diferença estatística significativa. No entanto, quando comparados ao grupo ILTC, houve diferença estatística significativa. O músculo PT responde a mudanças nutricionais padrão, ao contrário do músculo ILT, que requer alta formulação nutricional. O uso de 5% de farinha de cupuaçu provou ser uma fonte alternativa viável de alimentação animal, pois promove um aumento nas concentrações de fibras de colágeno na musculatura de frangos de corte e é possivelmente um fator determinante na textura da carne.(AU)

Bone marrow mononuclear cells versus mesenchymal stem cells from adipose tissue on bone healing in an Old World primate: can this be extrapolated to humans? / Células mononucleares de medula óssea versus células-tronco mesenquimais de tecido adiposo em cicatrização de lesão óssea em primata: pode ser extrapolado para humanos?

In veterinary medicine, the cell therapy is still unexplored and there are many unanswered questions that researchers tend to extrapolate to humans in an attempt to treat certain injuries. Investigating this subject in nonhuman primates turns out to be an unparalleled opportunity to better understand the dynamics of stem cells against some diseases. Thus, we aimed to compare the efficiency of bone marrow mononuclear cells (BMMCs) and mesenchymal stem cells (MSCs) from adipose tissue of Chlorocebus aethiops in induced bone injury. Ten animals were used, male adults subjected, to bone injury the iliac crests. The MSCs were isolated by and cultured. In an autologous manner, the BMMCs were infused in the right iliac crest, and MSCs from adipose tissue in the left iliac crest. After 4.8 months, the right iliac crests fully reconstructed, while left iliac crest continued to have obvious bone defects for up to 5.8 months after cell infusion. The best option for treatment of injuries with bone tissue loss in old world primates is to use autologous MSCs from adipose tissue, suggesting we can extrapolate the results to humans, since there is phylogenetic proximity between species.(AU)

Na medicina veterinária, a terapia celular ainda é inexplorada e há muitas perguntas não respondidas, o que leva os pesquisadores a uma tendência a estender a terapia para os seres humanos, na tentativa de tratar certas lesões. Investigar esse assunto em primatas não humanos revela-se uma oportunidade sem precedentes para compreender melhor a dinâmica das células-tronco contra algumas doenças. Assim, objetivou-se comparar a eficiência das células mononucleares de medula óssea (BMMCs) e das células-tronco mesenquimais (MSCs) do tecido adiposo de Chlorocebus aetiops na lesão óssea induzida. Foram utilizados 10 animais, adultos do sexo masculino, submetidos à lesão óssea nas cristas ilíacas. As MSCs foram isoladas e cultivadas; de forma autóloga, as BMMCs foram infundidas na crista ilíaca direita e as MSCs de tecido adiposo na crista ilíaca esquerda. Após 4,8 meses, a crista ilíaca direita foi totalmente reconstruída, enquanto a crista ilíaca esquerda continuou apresentando defeito ósseo evidente por até 5,8 meses após a infusão. A melhor opção para o tratamento de lesões com perda de tecido ósseo em primatas do Velho Mundo é a utilização de MSCs autólogas de tecido adiposo, sugerindo que se podem estender os resultados para seres humanos, uma vez que há proximidade filogenética entre as espécies.(AU)

Decellularization of placentas: establishing a protocol

Biological biomaterials for tissue engineering purposes can be produced through tissue and/or organ decellularization. The remaining extracellular matrix (ECM) must be acellular and preserve its proteins and physical features. Placentas are organs of great interest because they are discarded after birth and present large amounts of ECM. Protocols for decellularization are tissue-specific and have not been established for canine placentas yet. This study aimed at analyzing a favorable method for decellularization of maternal and fetal portions of canine placentas. Canine placentas were subjected to ten preliminary tests to analyze the efficacy of parameters such as the type of detergents, freezing temperatures and perfusion. Two protocols were chosen for further analyses using histology, scanning electron microscopy, immunofluorescence and DNA quantification. Sodium dodecyl sulfate (SDS) was the most effective detergent for cell removal. Freezing placentas before decellularization required longer periods of incubation in different detergents. Both perfusion and immersion methods were capable of removing cells. Placentas decellularized using Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, and 0.5% antibiotic) preserved the ECM structure better, but Protocol I was less efficient to remove cells and DNA content from the ECM than Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, and 0.5% antibiotic).

Histomorphometrics and quantitative unbiased stereology in canine uteri treated with medroxyprogesterone acetate.

This article describes the effects of MPA use on the canine uterus using stereological methods. Entire reproductive tracts were removed from normal healthy canine bitches (Canis lupus familiaris) and grouped as: nulliparous (n = 11), multiparous (n = 11) and MPA-treated (n = 11; nulliparous; two treatments; 5 mg/kg). 1 cm samples were cut from the corpus, horn and uterine tube and fixed in 10% formaldehyde. Sections of each were mounted on slides and stained with hematoxylin-eosin. We assessed the fraction area for components of endometrium and myometrium and VV (volume density) and SV (surface density) of the gland and stroma using the M36 test system provided by the STEPanizer Stereological Tool. No gross histological differences were observed between study groups in the uterine tube, uterine corpus and horn. The wall of the uterine corpus and horn in MPA-treated bitches was characterized as being thicker than in the other groups. A cross-section of the uterine corpus revealed no differences between components of uterine wall in the corpus and horn; however, differences were observed in the volume density [VV; %] in variables such as: VV[str.vasc/uterus] (nulliparous vs. multiparous; p = 0.0019) and VV[str.supravasc/uterus] (multiparous vs. nulliparous and MPA; p = 0.0035). In the endometrial gland, differences were detected in SV[gland/endom] (multiparous vs. MPA, p = 0.0442). In the uterine horn, differences were only observed in the variable VV[lumen.gland/endom] (multiparous vs. MPA; p = 0.0019). This study shows quantitative changes in the architecture of the endometrium and myometrium in all the uterine segments, mainly morphological endometrial gland changes of the uterine corpus, increasing the surface area per unit of volume; however, these changes usually do not differ quantitatively from those observed in the uterus of multiparous bitches.

Morphological study of the eye bulb of the hammerhead shark, Sphyrna lewini (Elasmobranch: Carcharhinidae) / Estudio morfológico del bulbo ocular del tiburón martillo, Sphyrna lewini (Elasmobranch: Carcharhinidae)

The hammerhead sharks shows a head laterally expanded with eyes and nostrils on the edges, which gives the species a hammer appearance. Another strand of studies indicates that the hypothesis of head shape alterations for better adaptions in the sensorial system with the development of structures associated to binocular sight and others sensorial organs. Given the variety of characteristics, the study aim was to describe the anatomical constitution of the hammerhead shark eye bulb. The bulb and its annexes exenteration was performed, and further dissection; the morphological description of the muscle insertions and eye bulb components were based on direct observation and were further photographed and catalogued. The eye bulb fragments and its annexes were histological technical. Between the sclera and the choroid it was possible to observe, by the electronical scanning microscopy, the thin layer called supra-choroid, in this region, spaces often filled by lymphatic vessels allied to a matrix formed by loose conjunctive tissue are found. In the choroid, a layer which is rich in blood vessels, loose conjunctive tissue and collagen fibers, was observed, besides pigmentary cells full of melanin in its interior, which result in its layer's dark color. Ciliary body is a choroid's dilatation; it has the aspect of a thick ring in finger-like shape, pigmented, covering the sclera surface and containing pigmentary cells. The crystalline capsule, which shows an acellular covering that, is hyaline and homogeneous. In the electronical scanning microscopy, it was observed that the capsule is extremely thick especially in the anterior face. The capsule is very elastic, constituted mainly by thin lamellae of collagenous fibers, as illustrated by the electronically scanning microscopy. Anatomic variations related mainly to the position of the eye bulb in the skull, fibrous tunica and lens call the attention and must be related to its habitat.

El tiburón martillo tiene la cabeza lateralmente expandida con los ojos y la nariz en sus márgenes. Estudios indican que alteraciones de la forma de la cabeza de estos animales son adaptaciones que mejoran el desarrollo del sistema sensorial y estructuras asociadas a la visión binocular y otros órganos sensoriales. Dada la variedad de características, el objetivo del estudio fue describir la constitución anatómica del bulbo ocular. Se disecó el bulbo ocular junto a sus anexos. La descripción morfológica de las inserciones musculares y componentes del bulbo ocular se basaron en la observación directa, siendo éstas fotografiadas y catalogadas. Entre la esclera y la coroides se observó através de microscopía electrónica de barrido, una delgada capa supra-coroides, y vasos linfáticos junto a la matriz formada por tejidos conectivos. La coroides, era abundante en vasos sanguíneos y fibras de tejido conectivo. Se observaron escasas células pigmentarias llenas de melanina, lo que se asemeja a un anillo en forma de dedo, de manera que cubren la superficie de la esclera y células que contienen pigmentos. En la cápsula del lente, fue posible observar un recubrimiento celular, hialino y homogéneo. En la microscopía electrónica de barrido, se observó una cápsula muy gruesa, principalmente en la región frontal. La cápsula elástica,estaba constituida por láminas delgadas, principalmente, por fibras de colágeno. Se concluyó que el bulbo ocular de estos animales posee variaciones anatómicas relacionadas, principalmente, con la posición del bulbo ocular en el cráneo, túnica fibrosa y lente, lo que puede estar directamente relacionado con su hábitat.

Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects / Glândulas adrenais de preás (Galea spixii, Wagler, 1831): aspectos morfológicos e morfométricos

Abstract Considering the physiological importance and need of greater morphophysiological knowledge of adrenal glands, the aims of present study were compare the morphometric data between left and right adrenal of male and female; perform a histological, scanning and transmission electron microscopy study showing tissue constitution of glands; finally, in order to define the presence and correct site of the cytochrome P450c17 expression in adrenal glands, immunohistochemical study of this enzyme was performed in 18 adrenal glands (right n=9 and left n=9) of nine adult Galea spixii (four males and five females). Right adrenal was more cranially positioned than left adrenal; dimensions (weight, length and width) of right adrenal was larger than left adrenal; no differences between male and female body and adrenal measurements were found; the morphology of cells and different amounts of lipid droplets may be related to the different demands of steroid hormones production, related to each zone of the adrenal cortex; and, the cytochrome P450c17 immunolocalization in fasciculate and reticular zone may be related with synthesis of 17-hydroxy-pregnenolone, 17-hydroxy-progesterone, dehydroepiandrosterone or androstenedione.

Resumo Considerando a importância fisiológica e a necessidade de melhor conhecimento morfofisiológico das glândulas adrenais, os objetivos do presente estudo foram comparar dados morfométricos entre adrenais direitas e esquerdas de machos e fêmeas; realizar estudos histológicos e microscopia eletrônica de varredura e transmissão revelando a constituição tecidual das glândulas; finalmente, foi realizado estudo imunohistoquímico com objetivo de definir a presença e correto sítio de expressão da citocromo P450c17 nas glândulas adrenais em 18 glândulas adrenais (direita n=9 e esquerda n=9) de nove Galea spixii adultos (quatro machos e cinco fêmeas). A glândula adrenal direita estava posicionada mais cranialmente que a adrenal esquerda; as dimensões (peso, comprimento e largura) da glândula adrenal direita foram maiores que as da esquerda; não houve diferença entre as medidas corpóreas e das adrenais entre machos e fêmeas; a morfologia das células e diferentes quantidades de gotículas lipídicas podem estar relacionadas a diferentes demandas na produção de hormônios esteroides relacionados a cada zona do córtex adrenal; e, a imunolocalização da citocromo P450 na zona fasciculada e reticular pode estar relacionado com a síntese de 17-hidroxi-pregnenolona, 17-hidroxi-progesterona, dehidroepiandrosterona ou androstenediona.

Morphological Aspects of Oral Denticles in the Sharpnose Shark Rhizoprionodon lalandii (Müller and Henle, 1839) (Elasmobranchii, Carcharhinidae).

The oral denticles of some elasmobranchs are found on the surface of the oral cavity and are homologous to those on the body surface, being well developed, independent and non-growing, with varying morphology and distribution depending on the species. The structural and three-dimensional characteristics of oral denticles from the rostro-ventral surface of the sharpnose shark Rhizoprionodon lalandii were described following imaging by both light and scanning electron microscopy. The light microscopy results showed that the triangular shape of the denticles consisted of a base and an apex. Picrosirius staining showed the arrangement of collagen fibres and oral denticles, and a predominance of type-I collagen was found in both structures under polarized light. There was a broad homogeneous distribution of denticles on the ventral surface, forming a leaf-like shape with the cusp facing the caudal region. Interlocking, hexagonal, geometric structures on its rostral side and ridges on the rostral side of the oral denticles were observed under increased magnification. We concluded that the denticle morphology found in R. lalandii differ of others analysed species, and the descriptions of these structures therefore provide important information for the classification of the species. In this species, the main functions can be assigned to help reduce hydrodynamic drag, particularly by this being a species that uses ram ventilation, and to protect the epithelium of the oropharynx of abrasion and parasites.

The Amniotic Membrane: Development and Potential Applications - A Review.

Foetal membranes are essential tissues for embryonic development, playing important roles related to protection, breathing, nutrition and excretion. The amnion is the innermost extraembryonic membrane, which surrounds the foetus, forming an amniotic sac that contains the amniotic fluid (AF). In recent years, the amniotic membrane has emerged as a potential tool for clinical applications and has been primarily used in medicine in order to stimulate the healing of skin and corneal diseases. It has also been used in vaginal reconstructive surgery, repair of abdominal hernia, prevention of surgical adhesions and pericardium closure. More recently, it has been used in regenerative medicine because the amniotic-derived stem cells as well as AF-derived cells exhibit cellular plasticity, angiogenic, cytoprotective, immunosuppressive properties, antitumoural potential and the ability to generate induced pluripotent stem cells. These features make them a promising source of stem cells for cell therapy and tissue engineering. In this review, we discussed the development of the amnion, AF and amniotic cavity in different species, as well as the applicability of stem cells from the amnion and AF in cellular therapy.

Isolation and characterization of mesenchymal stem cells from the yolk sacs of bovine embryos.

The yolk sac (YS) represents a promising source of stem cells for research because of the hematopoietic and mesenchymal cell niches that are present in this structure during the development of the embryo. In this study, we report on the isolation and characterization of YS tissue and mesenchymal stem cells (MSCs) derived from bovine YSs. Our results show that the YS is macroscopically located in the exocoelomic cavity in the ventral portion of the embryo and consists of a transparent membrane formed by a central sac-like portion and two ventrally elongated projections. Immunohistochemistry analyses were positive for OCT4, CD90, CD105, and CD44 markers in the YS of both gestational age groups. The MSCs of bovine YS were isolated using enzymatic digestion and were grown in vitro for at least 11 passages to verify their capacity to proliferate. These cells were also subjected to immunophenotypic characterization that revealed the presence of CD90, CD105, and CD79 and the absence of CD45, CD44, and CD79, which are positive and negative markers of MSCs, respectively. To prove their multipotency, the cells were induced to differentiate into three cell types, chondrocytes, osteoblasts, and adipocytes, which were stained with tissue-specific dyes (chondrogenic: Alcian Blue, osteogenic: Alizarin Red, and adipogenic: Oil Red O) to confirm differentiation. Gene expression analyses showed no differences in the patterns of gene expression between the groups or passages tested, with the exception of the expression of SOX2, which was slightly different in the G1P3 group compared to the other groups. Our results suggest that YS tissue from bovines can be used as a source of MSCs, which makes YS tissue-derived cells an interesting option for cell therapy and regenerative medicine.

Bovine yolk sac: from morphology to metabolomic and proteomic profiles.

In several species, placentation involves the presence of two different membranes responsible for maternal-fetal exchanges: the yolk sac and the chorioallantoic placenta. The yolk sac plays important roles in embryonic survival, mainly during the early stages of gestation. In bovine, it is a transitional membrane that is present until day 50-70 of pregnancy. Herein, we evaluated the morphological and molecular aspects of the yolk sac of bovine embryos during 24 to 52 days of gestation. A total of 69 embryos were allocated into three groups according to the crown-rump length and estimated ages. Yolk sac samples were then subjected to morphological and molecular analysis using mass spectrometry and nuclear magnetic resonance techniques. In contrast to alanine, which was observed only in Group I, during all gestational stages, we identified important metabolites such as aspartate, taurine, glycerophosphocholine, creatinine, creatine, hydrouracil, glutamate, glutamine, lactate, lysine, valine, myo-inositol, cadaverine, and choline. In addition, 314 random sequences of proteins were identified in the bovine yolk sac, and 47 of these were considered to be specific. Changes in alpha-fetoprotein and carcinoembryonic antigen concentrations during gestation were also evaluated. In conclusion, the majority of these proteins are related to the development of secondary metabolites that are involved in the activation of other proteins and metabolites, and in signaling pathways that are responsible for maternal-fetal exchanges, activation of programmed cell death mechanisms, and cellular differentiation, and also in proteins that are responsible for the yolk sac involution that is required to establish chorioallantoic placentation.

Morphological and Morphometric Description of Female Reproductive Tract of Sapajus apella (Capuchin monkey).

Forest destruction has progressively hampered the survival of many species, and this is why it is so important to study of the lives of primates in captivity. This study aimed to describe the morphological aspects of the female reproductive tract of Sapajus apella. We used five animals obtained from the National Primate Center, Ananindeua - PA. The ovaries were paired, compact and symmetrical and had a smooth surface. The uterine tubes were bilateral and convoluted in adult animals and straight in young individuals. The uterus was simple and located in the pelvic region. The vagina was a long structure due to the position of the uterus. The external genitalia were located in the urogenital perineum and consisted of dark pigmented labia majora and labia minora, a vaginal vestibule as long as the vagina and a well-developed clitoris. The results showed that the genitals of S. apella resemble those of other Neotropical primates.

Tongue papillae morphology of brown-throated sloth Bradypus variegatus (SCHINZ, 1825) / Morfologia das papilas linguais de preguiça comum Bradypus variegatus (SCHINZ, 1825)

The Bradypus variegatus inhabits the forests of South America and feeds from leaves, branches and sprouts from different plants. Due to its diet and the lack of literature on the morphological aspect of Xenarthras, five Bradypus variegatus tongues from animals which died from natural causes were evaluated, and they came from Pará State Museum Emílio Goeldi and were donated to the Laboratory of Animal Morphological Research (LaPMA) from UFRA, for revealing the different types of papillae and epithelial-connective tissue. Macroscopically, the tongues presented elongated shape, rounded apex, body, root, median sulcus in the root's apex, and two vallate papillae. The mucous membrane of the tongue revealed a keratinized stratified pavement epithelium, while the ventral surface of the tongue was thin and smooth, not provided with any type of papillae. However, the dorsal surface of the tongue was irregular with the presence of three types of papillae: filiform, fungiform and vallate papillae. The filiform papillae found were of a simple type, presenting a rounded base, irregularly distributed with a larger concentration and development on the tongue's apex and body. The fungiform papilla showed a practically smooth surface with irregular format, with the presence of gustatory pores; these were found all over the dorsal surface, with larger concentration at the rostral part of the apex. Only two vallate papillae were observed disposed in the root of the tongue, surrounded by a deep groove, and revealing several taste buds. The tongues from Bradypus variegatus presented gustatory papillae similar to the ones described for other Xenarthras species and wild mammals...

A Bradypus variegatus habita florestas da América do Sul e alimenta-se de folhas, ramos e brotos de várias plantas. Em decorrência de sua dieta, bem como da escassez de literatura sobre aspectos morfológicos dos Xenarthras, avaliaram-se cinco línguas de Bradypus variegatus, que foram a óbito por causas naturais, provenientes do Museu Paraense Emílio Goeldi e doados ao Laboratório de Pesquisa Morfológica Animal (LaPMA) da UFRA, a fim de se elucidarem os diferentes tipos de papilas e a interface epitélio-tecido conjuntivo. Macroscopicamente, as línguas apresentaram formato alongado, ápice arredondado, corpo, raiz, sulco mediano do ápice à raiz e duas papilas valadas. A mucosa da língua revelou epitélio de revestimento pavimentoso estratificado queratinizado, enquanto a face ventral era delgada e lisa, desprovida de qualquer tipo de papila. Já a face dorsal apresentou-se irregular, com a presença de três tipos papilares: papilas filiformes, fungiformes e valadas. As papilas filiformes encontradas eram do tipo simples, com base circular, distribuídas irregularmente, com maior concentração e desenvolvimento no ápice e no corpo da língua. A papila fungiforme mostrou-se com superfície praticamente lisa e de formato irregular, com a presença de poros gustativos. Estas foram encontradas por toda a superfície dorsal, com maior concentração na parte rostral do ápice. Foram observadas apenas duas papilas valadas, dispostas na região da raiz da língua, com corpo rodeado por sulcos profundos e vários botões gustativos. As línguas de Bradypus variegatus apresentaram papilas linguais similares às descritas para outras espécies de Xenarthras e mamíferos selvagens...

Histomorphometrical and proliferative aspects of placenta and uterus of the collared peccary (Tayassu tajacu).

The histomorphometric and proliferative characteristics of the collared peccary (Tayassu tajacu) placenta and uterus were analyzed. The material was examined by standard histological techniques and histochemistry (PAS, Perls and Alcian Blue pH 0.5 and 2.5%) and the cellular proliferation by AgNORs and flow cytometry. All the analyzed morphometric variables differed between pregnant and non-pregnant uteri in the luteal phase using the Dunnet test. Height and gland diameter of uterine glands increased linearly during pregnancy, with an intense positive PAS and Perls reaction in all stages. The cells with more than seven AgNORs per nuclei and the cells in the G2M cell cycle phase in the maternal tissue also increased after 70 days of pregnancy. The uteroplacental ridges had a linear increase in size with two distinct areas, base and top, with uterine epithelium and trophoblastic cells changing their morphology following the placental ridge development. Flow cytometry analysis showed the percentage of cells in each cell cycle phase with a quadratic behavior for stages G2/M in the maternal tissue, suggesting an increase in proliferative capacity of maternal tissue after 65 days of pregnancy. The same quadratic effect was observed in the G0/G1 phase in both maternal and fetal tissues. Cells in apoptosis showed cubic behavior in both tissues. The morphometric and cellular dynamic aspects observed in this study have not been previously described and they extend our knowledge of functions relating to maternal-fetal dynamics in this species.

Morphology and vascularization of the gastric compartments in three-toed sloth (Bradypus torquatus Illiger, 1811) / Morfología y vascularización de los compartimentos gástricos en osos perezosos de tres dedos (Bradypus torquatus Illiger, 1811)

The macroscopic anatomy and vascularization of the stomach compartments of Bradypus torquatus were examined in five animals from the University of São Paulo College of Veterinary Medicine and Zootechny collection. The method included aqueous perfusion of the arterial network, colored latex injection, fixation in formaldehyde (10 percent) and preservation in Laskovisk solution. Dissections were performed under mesoscopic light and photo documentations were performed for description and data analysis. In these animals, the largest abdominal organ was the stomach, which internally presented the cardiac, fundic and prepyloric regions, subdivided in six compartments (cardiac right, middle and left; fundic; pre-pyloric I and II). The stomach was irrigated by the left gastric and celiac arteries, which are ventral visceral branches of the abdominal aorta. These arteries emerged in the retroperitoneal region and reached the viscera through the mesogastric region, distributed in the large and small stomach curvatures, in the spleen and the pancreas. The primary collateral branches of the left gastric artery are directed to the large stomach curvature, and the celiac artery irrigated the spleen, the pancreas and the small stomach curvature. The vascular pattern differed in some aspects from that observed in the other multi-cavity stomachs of recent vertebrates.

Fue descrita la anatomía macroscópica y vascularización de los compartimientos del estómago en 5 Bradypus torquatus, animales pertenecientes al acervo de la Facultad de Medicina Veterinaria y Zootecnia de la Universidad de São Paulo - Brazil. El método incluyó: perfusión acuosa de la red arterial, inyección de látex coloreado, fijación en formaldehido (10 por ciento) y conservación en solución de Laskovisk. Para la descripción y análisis de los datos se realizaron disecciones bajo mesoscopía de luz y archivos fotográficos. Los animales presentaron el estómago como la víscera abdominal más abundante, que posee internamente las regiones: cardiaca, fúndica y pre-pilórica, subdivididas en seis compartimientos (cardiaco, derecho, medio e izquierdo; fúndico; pre-pilórico I y II). Estaba irrigado por las arterias gástricas izquierda y celiaca, que son ramas viscerales ventrales de la parte abdominal de la aorta. Estas arterias emergían en el retroperitoneo y a tráves del mesogastrio alcazaban las vísceras distribuyéndose en la gran y pequeña curvatura del estómago, bazo y páncreas. Las ramas colaterales primarias de la arteria gástrica izquierda estaban destinadas a la gran curvatura del estómago y los de la arteria celíaca irrigaban el bazo, páncreas y pequeña curvatura del estómago. El estándar vascular del Bradypus torquatus más conocido como oso perezoso de tres dedos demostró ser diferente, en algunos aspectos, al de otros estómagos pluricavitarios en vertebrados.

Ossificação endocondral em embriões e fetos de bovinos / Endocondral ossification in bovine embryos and fetus

Estudou-se a ossificação endocontral de 18 embriões e 12 fetos de até três meses de gestação, os quais foram coletados de úteros gestantes em frigoríficos e abatedouros. Os úteros foram dissecados e, em seguida, realizou-se uma incisão dorsal até o cérvix para avaliações macroscópicas dos embriões e fetos. Para o estudo microscópico foram realizadas técnicas de inclusão, seguidas de marcação dos depósitos de cálcio e fósforo, responsável pela ossificação dos moldes de cartilagem. Foram identificados hipertrofia da cartilagem e morte dos condrócitos e aumento da área de depósito de cálcio e fósforo, por volta da 10ª semana gestacional (74 dias). Durante a 11ª semana de gestação (81 dias), os grupamentos de carbonato de cálcio e fósforo espalharam-se por todo o osso, sendo mais intenso na diáfise.

The endocondral ossification process was analyzed in 18 embryos and 12 fetus until three months of pregnancy, which had been collected from pregnant uterus at slaughterhouse. Later, the uterus was separated followed by a dorsal incision from the cervix for better gross evaluations of the embryos and fetus. For the microscopical study, histological techniques had been performed followed by staining of the deposits of calcium and phosphorus match, responsible for the ossification of the cartilage casts, used as Von Kossa stain. Cartilage hypertrophy and death of chondrocytes, enlargement of the area of calcium and phosphorus deposit were foundon the 10th week of pregnancy (74 d). During the 11th week of pregnancy (81 d), the calcium carbonate and phosphorus had spread to all bones, being more intense in diaphysis.

Avaliação da técnica de arteriografia renal guiada por fluoroscopia em suínos / Description of the fluoroscopy-guided renal arteriography in swine

The fluoroscopy-guided renal arteriography was evaluated in order to be used as an auxiliary method in investigations and as a way to experimentally induce kidney diseases in swine. The technique was effective to obtain sharp images as well as to determine the area of renal irrigation. Despite its easiness, trained professionals are required to perform it.