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)

Development of the central nervous system in equine twin fetuses derived by somatic cell nuclear transfer.

Because of the growing importance of horses in leisure and several sports, somatic cell nuclear transfer (SCNT) is being used more frequently for cloning animals for performance and reproductive purposes. However, because of the need to perforate the zona pellucida during microsurgical reconstruction of the oocyte, it is possible that SCNT-derived embryos undergo premature hatching, resulting in embryo bisection and twinning. Therefore, because equine twin pregnancies often lead to abnormal embryo development and pregnancy failure, we performed a detailed comparative assessment of equine twin fetuses derived by SCNT with particular attention on the development of the central nervous system at 40 and 60 days gestation. The results of this study indicate that although cloned twin embryos show small differences in size, they do not exhibit apparent macro- or microscopic developmental discrepancies in the central nervous system, suggesting that the twining phenomenon resulting from SCNT does not affect fetal differentiation.

Reproductive system development in male and female horse embryos and fetuses: Gonadal hyperplasia revisited.

In horses, pregnancy is characterized by high levels of maternal estrogens that are produced largely by the interstitial tissue inside the gonads of the offspring, associated with a physiological gonadal hyperplasia, that is uncommon in other species. However, a detailed structural-functional understanding of the early stages of gonadal development and hyperplasia has remained elusive in horse pregnancy because of the lack of substantial data. The goal of this study was to describe the genital organs' development in 19 early horse embryos and fetuses (days 20-140 of gestation) of both sexes by means of anatomy, histology, stereology, and immunohistochemistry, with a specific focus on gonadal hyperplasia and interstitial tissue development. Gonadal hyperplasia with similar amounts of interstitial cells was observed in both sexes, but only during the early stage of development (days 40-90). Surprisingly, a higher degree of hyperplasia, characterized by larger amounts of interstitial cell-rich areas, was seen in fetal ovaries from 90 days of gestation onwards. Another novel aspect was that parallel to the hyperplasia of the interstitial cells, a much more precocious and pronounced differentiation of germinal cells was seen in the ovary, characterized by an earlier peak and decrease of DAZL and OCT protein immune markers. In conclusion, a reduced degree of hyperplasia and interstitial tissue in the fetal testis after 90 days of gestation suggests the existence of a more efficient mechanism regarding the synthesis of estrogen precursors as a structural or physiological difference between both fetal sexes, which warrants further investigation.

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).

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.

Morphometric developmental pattern of bovine handmade cloned concepti in late pregnancy.

Cloning procedures often interfere with conceptus growth and life ex utero, in a set of symptoms known as abnormal offspring syndrome (AOS). The aim of the present study was to compare the developmental pattern of in vivo-derived (IVD), IVF-derived and handmade cloning-derived (NT-HMC) Day 225 bovine concepti using established procedures. Pregnancy diagnosis was performed on Day 30 following blastocyst transfer on Day 7. Conceptus morphometry was assessed by ultrasonography on Day 51, and on Day 225 pregnant cows were killed for morphological examination of concepti. Pregnancy outcome was similar between groups, with greater pregnancy losses in the first trimester (70.6%) and smaller fetuses on Day 51 in the NT-HMC group than in the IVD (14.3%) and IVF (20.0%) groups. However, NT-HMC-derived concepti were twofold larger on Day 225 of gestation than controls. A higher frequency (63.5%) of placentomes larger than the largest in the IVD group was observed in the NT-HMC group, which may be relevant to placental function. Conceptus traits in the IVF group were similar to the IVD controls, with only slight changes in placentome types. Morphological changes in cloned concepti likely affected placental function and metabolism, disrupting the placental constraining mechanism on fetal growth in mid- to late pregnancy.

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.

Extracellular matrix in epitheliochorial, endotheliochorial and haemochorial placentation and its potential application for regenerative medicine.

A placenta is defined as structural approximation of maternal and foetal tissues to perform physiological exchange. Associated processes of differentiation and the establishment of its cells take place within the extracellular matrix (ECM) that provides a rich environment of collagens, fibronectins, cytokines and other components. Placental ECM is promising for tissue regeneration purposes, because it has immune tolerance capacities that may cause only minimal rejections of transplants with immunological differences between donor and recipient. However, specific characteristics of ECM during evolution of the structurally very diverse mammalian placenta are not yet revealed. We here address the major aspects of placental types, that is non-invasive (epitheliochorial), medium (endotheliochorial)-to-high (haemochorial) invasive nature of the interhemal barrier between the foetal and maternal blood system as well as their main components of ECM with special reference to species that are commonly used as animal models for human placentation and in the potential applications for regenerative medicine.

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.

Development of the cardiorespiratory system in dogs from days 16 to 46 of pregnancy.

Dogs have been studied for several reasons, such as the genetic improvement, their use as experimental models, in zoonotic research, cell therapy and as a model for human diseases. However, many features relating to the embryonic development of dogs remain unknown because of the absence of embryological studies. Considering the importance of the cardiorespiratory system in the development of embryos, the aim of this study was to investigate the development of the main cardiorespiratory organs of dog embryos and foetuses with estimated gestational ages from 16 to 46 days using macro- and microscopic descriptions. On day 16 of development, the neural tube and crest were formed, the anterior and posterior neuropore closure had begun and the somites had developed. Between days 22 and 27 of gestation, the lung buds and the initial formation of the primary bronchi and heart chambers were observed. The heart chambers exhibited the endo-, myo- and epicardial layers but did not have obvious differences in thickness among each other. Between days 41 and 46 of gestation, the nasal conchae and septa and trachea were formed, which exhibited characteristic epithelia. The lung formation and lobation were complete. The heart and major vessels exhibited mature histological architecture when their anatomical development was complete. The results of this study contribute to a more accurate definition of the embryonic and foetal developmental stages in dogs.

Buffalo (Bubalus bubali) Late Embryo and Foetus Development: A Morphological Analysis.

Many researches describe the embryonic developmental features in domestic animals; however, in farm animals, they are scarce. Most farm animal studies are related to assisted reproduction and embryos transfer techniques. But, morphological features and size measure to estimate the age gestation are rarely reported in literature. Thus, in this study, we described the developmental changes in the bubaline (Bubalus bubali) concepts from 21 to 60 days of gestation. Our results revealed that buffalo embryos similar morphological characteristics similar to other mammalian species. Also, similarities between bovine and bubaline persist; except on foetal stages when buffalos have a faster development than bovine. Therefore, buffalo's gestation period exhibits some varieties and accurate embryo age is more difficult. Yet, when we use a combination of the crown-rump, macroscopic analysis and alizarin red, it is possible to describe better the whole embryogenesis stages of the buffalo and which can contribute for future reproduction researches and applications in veterinary practice.

Adrenal glands of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831): morphological and morphometric aspects.

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.

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.