Full confluency, serum starvation, and roscovitine for inducing arrest in the G0/G1 phase of the cell cycle in puma skin-derived fibroblast lines

The puma population is constantly decreasing, and cloning by somatic cell nuclear transfer can be used to conserve the species. One of the factors determining the success of the development of cloned embryos is the cell cycle stage of the donor cells. We evaluated the effects of full confluency (~100%), serum starvation (0.5% serum), and roscovitine (15 µM) treatments on the cell cycle synchronization in G0/G1 of puma skin-derived fibroblasts by flow cytometric analysis. Also, we assessed the effects of these synchronization methods on morphology, viability, and apoptosis levels using microscopy tools. The results showed that culturing the cells to confluence for 24 h (84.0%), 48 h (84.6%), and 72 h (84.2%) and serum starvation for 96 h (85.4%) yielded a significantly higher percentage of cells arrested in the G0/G1 (P < 0.05) phase than cells not subjected to any cell cycle synchronization method (73.9%). Nevertheless, while serum starvation reduced the percentage of viable cells, no difference was observed for the full confluence and roscovitine treatments (P > 0.05). Moreover, roscovitine for 12 h (78.6%) and 24 h (82.1%) was unable to synchronize cells in G0/G1 (P > 0.05). In summary, full confluency induces puma fibroblast cell cycle synchronization at the G0/G1 stage without affecting cell viability. These outcomes may be valuable for planning donor cells for somatic cell nuclear transfer in pumas.(AU)

Métodos ultrassonográficos na avaliação das afecções prostáticas em cães ­ revisão de literatura / Ultrasonographic methods in the evaluation of prostatic disorders in dogs ­ paper review

O objetivo desta revisão foi descrever as principais técnicas de avaliação e achados ultrassonográficos da próstata canina. A ultrassonografia é o método de escolha para avaliação da glândula prostática, sendo imprescindível como auxílio no diagnóstico na detecção de anormalidades, principalmente quando há suspeita da presença de tumores. Com inovação dos equipamentos, surgiram técnicas complementares à ultrassonografia modo Bidimensional (B), tais como Doppler, ultrassonografia contrastada e elastografia, que aumentam a acurácia diagnóstica. O Doppler fornece informações sobre arquitetura vascular e aspectos hemodinâmicos dos vasos sanguíneos. A ultrassonografia contrastada permite determinar parâmetros relacionados à perfusão sanguínea das estruturas pelos agentes de contraste (ex.: microbolhas), definindo, portanto, padrões de alta e baixa intensidade da arquitetura vascular. A elastografia proporciona a avaliação da rigidez tecidual tanto de forma qualitativa, por meio de elastogramas, quanto quantitativa, por meio das velocidades de cisalhamento. Espera-se que esta revisão possa contribuir com informações relevantes aos leitores e veterinários da área de ultrassonografia e reprodução animal.(AU)

The purpose of this review was to describe the main assessment techniques and ultrasound findings of the canine prostate. Ultrasonography is the method of choice for assessing the prostate gland, being essential as an aid in the diagnosis in the detection of abnormalities, especially when the presence of tumors is suspected. With the innovation of the equipment, complementary techniques to B-mode ultrasonography have emerged, such as Doppler, contrasted ultrasound and elastography, which increase the diagnostic accuracy. Doppler provides information on vascular architecture and hemodynamic aspects of blood vessels. Contrast ultrasonography allows to determine parameters related to the blood perfusion of structures by contrast agents (eg, microbubbles), thus defining high and low intensity patterns of vascular architecture. ARFI elastography provides assessment of tissue stiffness both qualitatively, using elastograms, and quantitative, using shear speeds. It is hoped that this review can contribute with relevant information to readers and veterinarians in the area of asound and animal reproduction.(AU)

Transgênese e clonagem adotadas como biotécnicas reprodutivas em pequenos ruminantes / Transgenesis and cloning as reproductive biotechniques in small ruminants

Ovinos e caprinos são espécies que devido ao seu tamanho adequado, curto período gestacional e produção leiteira, tornaram-se importantes modelos na pesquisa pecuária, farmacêutica e biomédica. Assim, a engenharia genética tem sido amplamente aplicada à pesquisa com pequenos ruminantes. Caprinos e ovinos modificados geneticamente fornecem modelos valiosos para pesquisa sobre as funções dos genes, melhorando a exploração, obtendo produtos farmacêuticos no leite e aumentando a resistência às doenças. Além disso, o uso conjunto da clonagem e transgênese já foi responsável por marcos importantes na biotecnologia animal. Esta revisão destaca os avanços da clonagem e da engenharia genética nessas espécies obtidos nas últimas quatro décadas, com ênfase nas pesquisas realizadas no Brasil.

Sheep and goats are valuable species that due to their suitable size, short pregnancy period, and secretion of milk, have become important model in agricultural, pharmaceutical, and biomedical research. Thus, genome engineering has been widely applied to sheep and goat research. Gene-edited sheep and goats provide valuable models for investigations on gene functions, improving animal breeding, producing pharmaceuticals in milk and improving disease resistance. Additionally, the joint use of cloning and transgenesis has already been responsible for major milestones in animal biotechnology. This review highlights the advances of cloning and genome engineering in these species over the past four decades with particular emphasis on research in Brazil.

Meta-analysis of melatonin treatment and porcine somatic cell nuclear transfer embryo development

Abstract Porcine somatic cell nuclear transfer (SCNT) plays an important role in many areas of research. However, the low efficiency of SCNT in porcine embryos limits its applications. Porcine embryos contain high concentrations of lipid, which makes them vulnerable to oxidative stress. Some studies have used melatonin to reduce reactive oxygen species damage. At present there are many reports concerning the effect of exogenous melatonin on porcine SCNT. Some studies suggest that the addition of melatonin can increase the number of blastocyst cells, while others indicate that melatonin can reduce the number of blastocyst cells. Therefore, a meta-analysis was carried out to resolve the contradiction. In this study, a total of 63 articles from the past 30 years were analyzed, and six papers were finally selected. Through the analysis, it was found that the blastocyst rate was increased by adding exogenous melatonin. Melatonin had no effect on cleavage rate or the number of blastocyst cells, but did decrease the number of apoptotic cells. This result is crucial for future research on embryo implantation.

Meta-analysis of melatonin treatment and porcine somatic cell nuclear transfer embryo development

Porcine somatic cell nuclear transfer (SCNT) plays an important role in many areas of research. However, the low efficiency of SCNT in porcine embryos limits its applications. Porcine embryos contain high concentrations of lipid, which makes them vulnerable to oxidative stress. Some studies have used melatonin to reduce reactive oxygen species damage. At present there are many reports concerning the effect of exogenous melatonin on porcine SCNT. Some studies suggest that the addition of melatonin can increase the number of blastocyst cells, while others indicate that melatonin can reduce the number of blastocyst cells. Therefore, a meta-analysis was carried out to resolve the contradiction. In this study, a total of 63 articles from the past 30 years were analyzed, and six papers were finally selected. Through the analysis, it was found that the blastocyst rate was increased by adding exogenous melatonin. Melatonin had no effect on cleavage rate or the number of blastocyst cells, but did decrease the number of apoptotic cells. This result is crucial for future research on embryo implantation.(AU)

Evaluation of corpus luteum vascularization in recipient mares by using color Doppler ultrasound

Background: Embryo transfer is one of the most commonly used reproductive biotechnique. The success of embryotransfer is also affected by the synchrony of estrus and ovulation between donor and recipient animals. In horse reproduction, ultrasonography has been used, among other purposes, to diagnose early pregnancy. However, only the color Dopplerimaging mode makes it possible to evaluate the vascular architecture and the hemodynamic aspects of the vessels in severalorgans, especially the corpus luteum. The objective of this study was to evaluate, based on the color Doppler ultrasound,the corpus luteum vascularization and function from recipient mares at embryo transfer timing.Materials, Methods & Results: Mangalarga Machador mares from 5 to 10-year-old and a range of live weights of between350 to 450 kg were used for this experiment, kept in pasture-based on mombaça grass (Panicum maximum) and were givenad libitum access to water and mineral supplementation. The animals (n = 15) were gynecologically examined and uterineconsistency was evaluated by rectal palpation the same operator using an ultrasound system (SonoScape®) with a lineartransducer, and operating frequency ranging from 5 to 10 Mhz. The uterine tone was classified between grades 1 and 4 andsubjected to ovulation induction. The objective and subjective vascular perfusion of the corpus luteum was evaluated bycolor Doppler ultrasound on the day of embryo transfer and endometrium. The determination progesterone concentrationon the day of the embryo transfer was performed by direct chemiluminescence assay. The arcsine (√P/100) transformation was applied to the percentage data, and the results were expressed as mean (.) ± standard error of the mean (SEM).Further, the assumptions of normality and homoscedasticity were verified, respectively, based on the Shapiro-Wilk and...

Evaluation of corpus luteum vascularization in recipient mares by using color Doppler ultrasound

Background: Embryo transfer is one of the most commonly used reproductive biotechnique. The success of embryotransfer is also affected by the synchrony of estrus and ovulation between donor and recipient animals. In horse reproduction, ultrasonography has been used, among other purposes, to diagnose early pregnancy. However, only the color Dopplerimaging mode makes it possible to evaluate the vascular architecture and the hemodynamic aspects of the vessels in severalorgans, especially the corpus luteum. The objective of this study was to evaluate, based on the color Doppler ultrasound,the corpus luteum vascularization and function from recipient mares at embryo transfer timing.Materials, Methods & Results: Mangalarga Machador mares from 5 to 10-year-old and a range of live weights of between350 to 450 kg were used for this experiment, kept in pasture-based on mombaça grass (Panicum maximum) and were givenad libitum access to water and mineral supplementation. The animals (n = 15) were gynecologically examined and uterineconsistency was evaluated by rectal palpation the same operator using an ultrasound system (SonoScape®) with a lineartransducer, and operating frequency ranging from 5 to 10 Mhz. The uterine tone was classified between grades 1 and 4 andsubjected to ovulation induction. The objective and subjective vascular perfusion of the corpus luteum was evaluated bycolor Doppler ultrasound on the day of embryo transfer and endometrium. The determination progesterone concentrationon the day of the embryo transfer was performed by direct chemiluminescence assay. The arcsine (√P/100) transformation was applied to the percentage data, and the results were expressed as mean (.) ± standard error of the mean (SEM).Further, the assumptions of normality and homoscedasticity were verified, respectively, based on the Shapiro-Wilk and...(AU)

Nellore cloned calf viability during the postnatal period / Viabilidade de bezerros nelores clonados durante o período pós natal imediato

This study evaluated the viability of Nellore cloned calves derived from somatic cell nuclear transfer (SCNT) and compare their viability with animals of the same breed derived from in vitro fertilization (IVF). Thus, two groups were formed. Group I (GI) consisted of 10 calves derived from SCNT and group II (GII) consisted of 10 calves derived from IVF. The differences detected between the groups were in the physical examination of the respiratory tract in GI, which represented the most common clinical-pathological disturbances. The Apgar index score indicated that 80% of GI animals were depressed and all had pale mucous membranes. Thus, anemia was reported in GI. In GII, this started at 12 h of life and was probably caused by an iron deficiency. Moreover, total calcium and ionized calcium levels were higher in GI immediately after birth. These alterations probably resulted in a high incidence of mortality in GI, reaching 90% of the calves, whereas mortality was only 20% for the calves in GII. In conclusion, cloned calves, which were derived from SCNT, had physiological and metabolic alterations after delivery, leading to a higher mortality rate during the perinatal period.(AU)

O objetivo deste trabalho foi avaliar a viabilidade de bezerros da raça Nelore oriundos da técnica da transferência nuclear de células somáticas (TNCS), no período pós natal imediato, comparando-a com animais desta mesma raça, oriundos de fertilização in vitro (FIV). Para tanto, os animais foram alocados em dois grupos, a saber: Grupo I (GI) - 10 animais frutos de TNCS; e, Grupo II (GII) - 10 animais oriundos de FIV. Nos respectivos bezerros, todos obtidos por cesariana, foram realizadas as avaliações físicas, escore de APGAR, bem como coleta de amostras de sangue nos momentos 0 (ao nascimento), às 2, 4, 6 e 12 horas de vida, a fim de avaliar os resultados de eritrograma, análises bioquímicas e hormonais, comparando-os entre os grupos e momentos. Nos animais que vieram a óbito foi realizada a necropsia para investigar a causa mortis. As diferenças observadas foram em relação aos achados clínico-patológicos, envolvendo, principalmente, o sistema respiratório caracterizado por bradpneia associada à dispneia, e a presença de edema e atelectasia pulmonar observadas no GI. Ademais, após a colostragem notou-se que 80% dos animais avaliados não foram capazes de manter a glicemia sendo mais evidentes nos animais do GII, possivelmente devido à hiperinsulinemia que se manifestou neste grupo ao longo de todo o período experimental. ...(AU)

Bancos de recursos somáticos: Como eles contribuem para a conservação de grandes felídeos? / Somatic resource bank: How they contribute to the conservation of large felines?

Distintas estratégias de conservação têm sido adotadas visando a manutenção e recuperação da biodiversidade, especialmente de espécies que se encontram em diferentes níveis de ameaça à extinção. Neste grupo de espécies, encontram-se os grandes felídeos, os quais em virtude das ações antrópicas, como a destruição e a fragmentação de habitat, necessitam de esforços voltados para a conservação de seu material genético. Uma estratégia empregada para essa finalidade consiste nos bancos de recursos somáticos, os quais são definidos como a criopreservação de tecidos e células somáticas oriundas de diferentes populações. Essas amostras biológicas, quando adequadamente conservadas, são elementoschave para a multiplicação das espécies por meio de seu emprego na clonagem por transferência nuclear e indução de células à pluripotência. Assim, o objetivo desta revisão é abordar os aspectos relevantes envolvidos na formação de bancos de recursos somáticos em grandes felídeos, destacando os estudos desenvolvidos pelo Laboratório de Biotecnologia Animal (LBA), da Universidade Federal Rural do Semi-Árido (UFERSA) em onças-pintadas e onças-pardas.

Different conservation strategies have been adopted to maintain and recover biodiversity, especially for species that are at different levels of threat to extinction. In this group of species are the large felids, which, due to anthropic actions, such as habitat destruction and fragmentation, require efforts aimed at the conservation of their genetic material. A strategy employed for this purpose consists of somatic resource banks, which are defined as the cryopreservation of tissues and somatic cells from different populations. These biological samples, when properly preserved, are key elements for the multiplication of species through their use in cloning by nuclear transfer and induction of cells to pluripotency. Thus, the aim of this review is to address the relevant aspects involved in the formation of somatic resource banks in large felids, highlighting the studies carried out by the Laboratory of Animal Biotechnology (LBA) of the Federal Rural University of the Semi-Arid (UFERSA) in jaguars and pumas.

Transferência Nuclear de Células Somáticas interespecífica (TNCSi) na conservação de cervídeos em risco de extinção / Interspecific Somatic Cell Nuclear Transfer (TNCSi) in the conservation of endangered species of deer

A Transferência Nuclear de Células Somáticas interespecífica (TNCSi) envolve a transferência de um núcleo ou célula de uma espécie para o citoplasma de um oócito enucleado de outra espécie. Após ativação, os complexos carioplasto-citoplasto reconstruídos podem ser cultivados in vitro até o blastocisto, o estádio final de desenvolvimento pré-implantação. Esta pode ser uma estratégia interessante na tentativa de conservação de espécies em risco de extinção. Esta revisão tem por objetivo apresentar alguns detalhes da técnica e relatar a experiência de nosso grupo usando como modelo o veado-catingueiro.(AU)

Interspecies Somatic Cell Nuclear Transfer (iSCNT) involves the transfer of a nucleus or cell from one species into the cytoplasm of an enucleated oocyte from another. Once activated, reconstructed caryoplast-cytoplast complexes can be cultured in vitro to blastocyst, the final stage of preimplantation development. This may be an interesting strategy in attempting to conserve endangered species. The objective of this review is to present some details of the technique and to report the experience of our group using as model the brown brocket deer.(AU)

Transferência Nuclear de Células Somáticas interespecífica (TNCSi) na conservação de cervídeos em risco de extinção / Interspecific Somatic Cell Nuclear Transfer (TNCSi) in the conservation of endangered species of deer

A Transferência Nuclear de Células Somáticas interespecífica (TNCSi) envolve a transferência de um núcleo ou célula de uma espécie para o citoplasma de um oócito enucleado de outra espécie. Após ativação, os complexos carioplasto-citoplasto reconstruídos podem ser cultivados in vitro até o blastocisto, o estádio final de desenvolvimento pré-implantação. Esta pode ser uma estratégia interessante na tentativa de conservação de espécies em risco de extinção. Esta revisão tem por objetivo apresentar alguns detalhes da técnica e relatar a experiência de nosso grupo usando como modelo o veado-catingueiro.

Interspecies Somatic Cell Nuclear Transfer (iSCNT) involves the transfer of a nucleus or cell from one species into the cytoplasm of an enucleated oocyte from another. Once activated, reconstructed caryoplast-cytoplast complexes can be cultured in vitro to blastocyst, the final stage of preimplantation development. This may be an interesting strategy in attempting to conserve endangered species. The objective of this review is to present some details of the technique and to report the experience of our group using as model the brown brocket deer.

Radionuclide migration and organoleptic characteristics of beef in the adjacent areas to the former Semipalatinsk nuclear test site

This article presents the results of the migration of radionuclides along the soil-water-feed-animal livestock chain in the adjacent territories of the former Semipalatinsk nuclear test site (SNTS). To analyze the radio ecological situation in the territory adjacent to the SNTS, 6 stationary control points were created, from which samples of soil, water, vegetation, and animal products were taken. The radiometric method was used to determine the radioactive background of the studied areas. Gamma-, alpha-spectrometry methods were used to study the content of radionuclides. The radionuclide transfer along the soil-water-feed-animal livestock chain wasdetermined by a calculation method and also veterinary and sanitary examination of the beef selected from these items. The research results show that in all the test points under studythe EDR values, the beta-particles flux density is below the limit of permissible values. The samples detected radionuclides as Am-241, Cs-137 and Pu-239/240 not exceeding the standards established by the Hygienic standardsof the Republic of Kazakhstan. The chain of radionuclides was transferred to animal products. The organoleptic assessment of the quality of cattle meat showed that inconsistencies are observed: a slight increase in lymph nodes, muddy broth, and surface moisture of muscle.(AU)

Radionuclide migration and organoleptic characteristics of beef in the adjacent areas to the former Semipalatinsk nuclear test site

This article presents the results of the migration of radionuclides along the soil-water-feed-animal livestock chain in the adjacent territories of the former Semipalatinsk nuclear test site (SNTS). To analyze the radio ecological situation in the territory adjacent to the SNTS, 6 stationary control points were created, from which samples of soil, water, vegetation, and animal products were taken. The radiometric method was used to determine the radioactive background of the studied areas. Gamma-, alpha-spectrometry methods were used to study the content of radionuclides. The radionuclide transfer along the soil-water-feed-animal livestock chain wasdetermined by a calculation method and also veterinary and sanitary examination of the beef selected from these items. The research results show that in all the test points under studythe EDR values, the beta-particles flux density is below the limit of permissible values. The samples detected radionuclides as Am-241, Cs-137 and Pu-239/240 not exceeding the standards established by the Hygienic standardsof the Republic of Kazakhstan. The chain of radionuclides was transferred to animal products. The organoleptic assessment of the quality of cattle meat showed that inconsistencies are observed: a slight increase in lymph nodes, muddy broth, and surface moisture of muscle.

Estabelecimento de linhagens celulares em felídeos silvestres Uma revisão dos efeitos das condições de cultivo e criopreservação sobre a qualidade celular / Establishment of cell lines in wild felids A review of the effects of culture and cryopreservation conditions on cell quality

Linhagens celulares, principalmente fibroblastos derivados da pele, podem ser ferramentas interessantes para a conservação e multiplicação de felídeos silvestres ameaçados de extinção. Esses animais em virtude de seu quantitativo reduzido têm nos bancos de células somáticas uma alternativa para a conservação de material biológico derivado de pequenas populações, garantindo assim o armazenamento da diversidade genética de diferentes grupos de indivíduos. Isso porque tais linhagens, quando apropriadamente estabelecidas por meio das técnicas de cultivo in vitro e criopreservação, permitem seu emprego na obtenção de embriões por clonagem por transferência nuclear de célula somática e produção de células induzidas à pluripotência. Assim, uma das etapas essenciais para o uso dessas linhagens de maneira eficiente consiste na avaliação dos efeitos das condições de cultivo in vitro e criopreservação das células, visando reconhecer os danos gerados pelas manipulações e identificar os ajustes necessários aos protocolos empregados. Portanto, o objetivo desta revisão consiste em reunir e explorar informações úteis sobre as condições de cultivo in vitro e criopreservação de células derivadas de felídeos silvestres, visando o estabelecimento de linhagens celulares na conservação e multiplicação destas espécies.(AU)

Cell lines, mainly fibroblasts derived from the skin, can be interesting tools for the conservation and multiplication of endangered wild felids. These animals, due to their reduced quantity, have somatic cell banks as an alternative for the conservation of biological material derived from small populations, thus guaranteeing the storage of the genetic diversity of different groups of individuals. This is because such lines, when properly established through in vitro culture and cryopreservation techniques, allow their use in obtaining embryos by cloning by somatic cell nuclear transfer and production of cells induced to pluripotency. Thus, one of the essential steps for the use of these lines in an efficient way consists of the evaluation of the effects of the conditions of in vitro culture and cryopreservation of the cells, aiming to recognize the damages generated by the manipulations and to identify the necessary adjustments to the employed protocols. Therefore, the aim of this review is to gather and explore useful information about in vitro culture and cryopreservation conditions of cells derived from wild felids, aiming at the establishment of cell lines in the conservation and multiplication of these species.(AU)

Estabelecimento de linhagens celulares em felídeos silvestres – Uma revisão dos efeitos das condições de cultivo e criopreservação sobre a qualidade celular / Establishment of cell lines in wild felids – A review of the effects of culture and cryopreservation conditions on cell quality

Linhagens celulares, principalmente fibroblastos derivados da pele, podem ser ferramentas interessantes para a conservação e multiplicação de felídeos silvestres ameaçados de extinção. Esses animais em virtude de seu quantitativo reduzido têm nos bancos de células somáticas uma alternativa para a conservação de material biológico derivado de pequenas populações, garantindo assim o armazenamento da diversidade genética de diferentes grupos de indivíduos. Isso porque tais linhagens, quando apropriadamente estabelecidas por meio das técnicas de cultivo in vitro e criopreservação, permitem seu emprego na obtenção de embriões por clonagem por transferência nuclear de célula somática e produção de células induzidas à pluripotência. Assim, uma das etapas essenciais para o uso dessas linhagens de maneira eficiente consiste na avaliação dos efeitos das condições de cultivo in vitro e criopreservação das células, visando reconhecer os danos gerados pelas manipulações e identificar os ajustes necessários aos protocolos empregados. Portanto, o objetivo desta revisão consiste em reunir e explorar informações úteis sobre as condições de cultivo in vitro e criopreservação de células derivadas de felídeos silvestres, visando o estabelecimento de linhagens celulares na conservação e multiplicação destas espécies.

Cell lines, mainly fibroblasts derived from the skin, can be interesting tools for the conservation and multiplication of endangered wild felids. These animals, due to their reduced quantity, have somatic cell banks as an alternative for the conservation of biological material derived from small populations, thus guaranteeing the storage of the genetic diversity of different groups of individuals. This is because such lines, when properly established through in vitro culture and cryopreservation techniques, allow their use in obtaining embryos by cloning by somatic cell nuclear transfer and production of cells induced to pluripotency. Thus, one of the essential steps for the use of these lines in an efficient way consists of the evaluation of the effects of the conditions of in vitro culture and cryopreservation of the cells, aiming to recognize the damages generated by the manipulations and to identify the necessary adjustments to the employed protocols. Therefore, the aim of this review is to gather and explore useful information about in vitro culture and cryopreservation conditions of cells derived from wild felids, aiming at the establishment of cell lines in the conservation and multiplication of these species.

Spinal cord meningioma in a Boxer breed dog

Meningiomas are one of the most common primary tumors of the canine central nervous system. They usually grow within the cranial cavity but occasionally they are found in the spinal cord. A case of a Boxer breed with a spinal cord transitional meningioma is reported. Two years history presenting progressive neurological signs is described. The MRI examination revealed a focal, intradural extramedullary lesion within the right side of the spinal canal, compromising the C1and C2 vertebral segments. Cytoreductive surgery with a dorsolateral laminectomy approach was performed to decompressthe spinal cord and to obtain a definitive diagnosis. Histopathological studies diagnosed a grade I transitional meningioma. Immunohistochemistry and immunofluorescence analysis detected positive cells to vimentin (VIM), pan-cytokeratin (pCk),neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and doublecortin (DXC). Although apparently benign, positivity to pCK and DXC suggest a possible transition into a malignant tumor. Although variations in the behavior, histology and the immunoreactive profile of these tumors are reported, the latter constitutes a good indicator for the diagnosis of the patient.

Spinal cord meningioma in a Boxer breed dog

Meningiomas are one of the most common primary tumors of the canine central nervous system. They usually grow within the cranial cavity but occasionally they are found in the spinal cord. A case of a Boxer breed with a spinal cord transitional meningioma is reported. Two years history presenting progressive neurological signs is described. The MRI examination revealed a focal, intradural extramedullary lesion within the right side of the spinal canal, compromising the C1and C2 vertebral segments. Cytoreductive surgery with a dorsolateral laminectomy approach was performed to decompressthe spinal cord and to obtain a definitive diagnosis. Histopathological studies diagnosed a grade I transitional meningioma. Immunohistochemistry and immunofluorescence analysis detected positive cells to vimentin (VIM), pan-cytokeratin (pCk),neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and doublecortin (DXC). Although apparently benign, positivity to pCK and DXC suggest a possible transition into a malignant tumor. Although variations in the behavior, histology and the immunoreactive profile of these tumors are reported, the latter constitutes a good indicator for the diagnosis of the patient.(AU)

Epigenetic characterization of the H19/IGF2 locus in calf clones placenta / Caracterização epigenética do locus H19/IGF2 na placenta de bezerros clones

Somatic Cell Nuclear Transfer (SCNT-Cloning) is a promising technique in many areas and is based on genetically identical individuals. However, its efficiency is low. Studies suggest that the leading cause is inadequate epigenetic reprogramming. This study aimed to characterize the methylation pattern of the exon 10 regions of the IGF2 gene and the Imprinting Control Region (ICR) of the H19 gene in the placenta of cloned calves. For this study, female and male cloned calves presenting different phenotypes were used. Genomic DNA from these animals' placenta was isolated, then treated with sodium bisulfite and amplified to the ICR/H19 and IGF2 loci. PCR products were cloned into competent bacteria and finally sequenced. A significant difference was found between controls and clones with healthy phenotypes for the ICR/H19 region. In this region, controls showed a hemimethylated pattern, as predicted in the literature due to this region has an imprinted control, while clones were showed less methylated. For the IGF2, no significant differences were found between controls and clones. These results suggest that different genomic regions in the genome may be independently reprogrammed and that failures in reprogramming the DNA methylation patterns of imprinted genes may be one of the causes of the low efficiency of SCNT.(AU)

A Transferência Nuclear de Células Somáticas (TNCS-Clonagem) é uma técnica promissora em várias áreas, e se baseia na produção de indivíduos geneticamente idênticos. No entanto, sua eficiência é baixa. Estudos sugerem que a principal causa seja uma reprogramação epigenética inadequada. O objetivo desse trabalho é caracterizar o padrão de metilação da região éxon 10 do gene IGF2 e da Região Controladora de Imprinting (ICR) do gene H19 na placenta de bezerros clonados. Para a execução do trabalho foram selecionados clones bovinos fêmeas e machos, apresentando diferentes fenótipos. O DNA da placenta desses animais foi extraído, e em seguida foi tratado com bissulfito de sódio e amplificado para os loci ICR/H19 e IGF2. Os produtos da PCR foram clonados em bactérias competentes e, por fim, sequenciados. Foi encontrada uma diferença significativa entre os controles e os clones com fenótipos saudáveis para a região da ICR/H19. Nesta região, os controles tiveram um padrão hemimetilado, como previsto pela literatura, devido essa região ser imprinted. Enquanto os clones encontravam-se menos metilados. Para a região do éxon 10 do IGF2, não foi encontrada diferença significativa entre controles e clones. Estes resultados sugerem que as diferentes regiões do genoma podem se reprogramar independente umas das outras e que falhas na reprogramação do padrão de metilação do DNA de genes imprinted podem ser uma das causas da baixa eficiência da TNCS.(AU)

Pathological features of cloned calves that died in the neonatal period / Aspectos patológicos de bezerros clonados com óbito no período neonatal

Somatic-cell nuclear transfer is a cloning technique that enables the creation of a viable embryo from a donor adult to produce a genetically identical individual. This technique opens numerous potential possibilities for medicine and animal reproduction. However, several reports have documented cloning-related issues. Embryo and fetal losses remain significantly higher than in other techniques, and there is a high incidence of dystocia and hydrops, which decreases efficiency and increases costs. Animals delivered at term often exhibit a syndrome known as macrosomia and experience difficulties in adapting to life outside the uterus, and death is a common outcome. In the present study, 41 cloned calves that died in the neonatal period were subjected to gross and histopathological examination. Most important gross lesions were found in the liver (enlargement, congestion, yellowish color), kidneys (brownish color at surface and cut, and cysts), lungs (atelectasis, parenchymal consolidation, and secretions in bronchi and bronchioles), and heart (concentric and eccentric hypertrophy, hematic cysts, persistence of ductus arteriosus). Primary microscopic findings were seen in the liver, kidneys, and lungs from neonatal calves. In the liver, 85% of the animals exhibited hepatic degeneration. The presence of a brownish pigment within the cortical tubules of the kidneys was found in approximately 90% of the samples; the presence of this pigment has not been previously reported in cloned calves. In the lungs, a large number of animals exhibiting lesions characteristic of pneumonia (55%). These changes were the pivotal causes of death, mainly due to problems in adapting to life outside the uterus and opportunistic infections in the neonatal period. Further investigation focusing on pathological anatomical changes is necessary to map these abnormalities in cloned animals.(AU)

A transferência nuclear de células somáticas ou clonagem é uma técnica que permite produzir um indivíduo geneticamente igual a um outro indivíduo adulto. Esta técnica abre inúmeras possibilidades para a medicina e para a reprodução animal. Porém, existem inúmeros relatos de problemas associados à clonagem. A taxa de perda nos períodos embrionário e fetal ainda é muito alta quando comparada a outras biotécnicas; além disso, há uma maior incidência de hidropsias e distocias, diminuindo a eficiência e aumentando o custo da técnica. Os animais que vem a termo frequentemente apresentam uma síndrome chamada de macrossomia, e apresentam dificuldades de adaptação à vida extrauterina e, por isso, o óbito é um desfecho comum. No presente trabalho realizou-se necropsia e coleta de fragmentos de órgãos para avaliação histopatológica de 41 bezerros com óbito neonatal. As lesões macroscópicas mais importantes foram encontradas no fígado (hepatomegalia, congestão e coloração amarelada), rins (coloração amarronzada na superfície e ao corte, e cistos), pulmões (atelectasia, parênquima consolidado, e secreções nos brônquios e bronquíolos), e coração (hipertrofia concêntrica e excêntrica, cistos hemáticos e persistência de ducto arterioso). As principais alterações microscópicas observadas foram presença de pigmento acastanhado no interior dos túbulos corticais renais (aproximadamente 90% dos animais), degeneração hepática (85% das amostras avaliadas) e lesões características de pneumonia (55% dos animais). A pigmentação acastanhada no interior dos túbulos corticais é uma alteração que ainda não havia sido relatada anteriormente em animais clonados. As alterações observadas nestes órgãos foram determinantes para o óbito, e devem ter ocorrido sobretudo devido a problemas na adaptação ao ambiente extrauterino e em decorrência de infecções adquiridas no período neonatal. Os achados encontrados no presente trabalho denotam a necessidade de investigação anatomopatológica detalhada de animais clonados inviáveis, na tentativa de mapear as anormalidades apresentadas por eles.(AU)

Pathological features of cloned calves that died in the neonatal period / Aspectos patológicos de bezerros clonados com óbito no período neonatal

Somatic-cell nuclear transfer is a cloning technique that enables the creation of a viable embryo from a donor adult to produce a genetically identical individual. This technique opens numerous potential possibilities for medicine and animal reproduction. However, several reports have documented cloning-related issues. Embryo and fetal losses remain significantly higher than in other techniques, and there is a high incidence of dystocia and hydrops, which decreases efficiency and increases costs. Animals delivered at term often exhibit a syndrome known as macrosomia and experience difficulties in adapting to life outside the uterus, and death is a common outcome. In the present study, 41 cloned calves that died in the neonatal period were subjected to gross and histopathological examination. Most important gross lesions were found in the liver (enlargement, congestion, yellowish color), kidneys (brownish color at surface and cut, and cysts), lungs (atelectasis, parenchymal consolidation, and secretions in bronchi and bronchioles), and heart (concentric and eccentric hypertrophy, hematic cysts, persistence of ductus arteriosus). Primary microscopic findings were seen in the liver, kidneys, and lungs from neonatal calves. In the liver, 85% of the animals exhibited hepatic degeneration. The presence of a brownish pigment within the cortical tubules of the kidneys was found in approximately 90% of the samples; the presence of this pigment has not been previously reported in cloned calves. In the lungs, a large number of animals exhibiting lesions characteristic of pneumonia (55%). These changes were the pivotal causes of death, mainly due to problems in adapting to life outside the uterus and opportunistic infections in the neonatal period. Further investigation focusing on pathological anatomical changes is necessary to map these abnormalities in cloned animals.(AU)

A transferência nuclear de células somáticas ou clonagem é uma técnica que permite produzir um indivíduo geneticamente igual a um outro indivíduo adulto. Esta técnica abre inúmeras possibilidades para a medicina e para a reprodução animal. Porém, existem inúmeros relatos de problemas associados à clonagem. A taxa de perda nos períodos embrionário e fetal ainda é muito alta quando comparada a outras biotécnicas; além disso, há uma maior incidência de hidropsias e distocias, diminuindo a eficiência e aumentando o custo da técnica. Os animais que vem a termo frequentemente apresentam uma síndrome chamada de macrossomia, e apresentam dificuldades de adaptação à vida extrauterina e, por isso, o óbito é um desfecho comum. No presente trabalho realizou-se necropsia e coleta de fragmentos de órgãos para avaliação histopatológica de 41 bezerros com óbito neonatal. As lesões macroscópicas mais importantes foram encontradas no fígado (hepatomegalia, congestão e coloração amarelada), rins (coloração amarronzada na superfície e ao corte, e cistos), pulmões (atelectasia, parênquima consolidado, e secreções nos brônquios e bronquíolos), e coração (hipertrofia concêntrica e excêntrica, cistos hemáticos e persistência de ducto arterioso). As principais alterações microscópicas observadas foram presença de pigmento acastanhado no interior dos túbulos corticais renais (aproximadamente 90% dos animais), degeneração hepática (85% das amostras avaliadas) e lesões características de pneumonia (55% dos animais). A pigmentação acastanhada no interior dos túbulos corticais é uma alteração que ainda não havia sido relatada anteriormente em animais clonados. As alterações observadas nestes órgãos foram determinantes para o óbito, e devem ter ocorrido sobretudo devido a problemas na adaptação ao ambiente extrauterino e em decorrência de infecções adquiridas no período neonatal. Os achados encontrados no presente trabalho denotam a necessidade de investigação anatomopatológica detalhada de animais clonados inviáveis, na tentativa de mapear as anormalidades apresentadas por eles.(AU)