Tratamento cirúrgico de fibrossarcoma cutâneo em uma égua Paint Horse: relato de caso

Os fibrossarcomas são neoplasias mesenquimais malignas primárias de baixa incidência em equinos e que esporadicamente produzem metástases. O uso de tecnologias de imagem e de citologia aspirativa é bastante relevante na identificação da lesão, porém o exame histopatológico ainda é o método de eleição para o diagnóstico definitivo. Nesse contexto, o presente estudo teve por objetivo relatar o caso de um fibrossarcoma em uma égua. Foi solicitado atendimento, devido ao desenvolvimento de uma massa crânio-lateral à glândula mamária direita, para uma égua Paint Horse de nove anos de idade. À palpação, verificou-se que a massa era circunscrita, possuía consistência firme e aderência à musculatura abdominal. Como tratamento, foi realizada exérese cirúrgica com margens amplas da neoformação. Durante o procedimento observou-se que a massa não tinha continuidade com o tecido mamário e pouca área de aderência ao tecido muscular abdominal. A massa foi encaminhada para análise histopatológica em que foi constatada proliferação neoplásica mesenquimal compatível com fibrossarcoma. A paciente foi acompanhada periodicamente por dois anos e, durante esse período, não foi verificada nenhuma recidiva tanto no local do procedimento cirúrgico como em áreas distantes. Assim sendo, concluímos que o atendimento precoce, a exérese cirúrgica com margens amplas, a identificação histopatológica e o acompanhamento periódico foram ferramentas essenciais para o sucesso no tratamento da paciente.The present study aims to report the case of a mare whose care was requested, due to the development of a craniolateral mass to the right mammary gland. At palpation, it was verified that the mass had firm consistency and apparently did not adhere. During the surgical excision of the neoformation, it was observed that the mass had no continuity with the mammary tissue and had little area of adhesion to the abdominal muscle tissue. Macroscopically, it was observed that the material was of a rigid consistency to the cut, of yellowish color and presented cystic area in the central region. Microscopically, fibrosarcoma compatible tissue was observed. The neoplasm was removed with a good margin of safety and did not recur for a period of two years after the surgical procedure

Efeitos de reguladores do metabolismo lipídico no desenvolvimento in vitro de embriões bovinos e na sobrevivência à vitrificação

Embryos produced in vitro (IVP) have low survival to conventional cryopreservation methods. In attempt to improve the production and the rate of embryo survival to the cryopreservation process, was used the metabolic regulators, L-carnitine (CA), forskolina (FO) and the conjugated linoleic acid isomers trans-10, cis-12 (CLA), separately or in combination in different doses, starting the fourth (D4) or sixth (D6) days of the embryo culture development in vitro. In experiment 1, the best results of production (62,0 ± 12,5%, P = 0,02), expansion (60,7%, P = 0,009), and hatching (41,1 % P = 0,04) was obtained using 2,5 mM of CA. In experiment 2, the production of embryos was not influenced by supplementation with FO (P=0,2), however, the dose of 15,0μM was the best result of expansion (53,2%) and hatching (46,8%) (P=0,001). In experiment 3, the production of embryos was not influenced by supplementation with CLA (P=0,4), however, the dose of 150,0 μM resulted in higher rates of expansion (85,5%, P=0,001) and hatching (70,9%, P=0,001). In the experiment 4, the embryo production was not influenced by the supplementations (P=0,16), however the best result of the production was using 15,0μM of FO individually. The expansion and hatching rates was influenced by supplementation with the metabolism regulators added individually or in a combination (P=0,001). The best expansion rate was observed with the association CA+FO+CLA (90,05%), that was not different (P>0,05) of the CLA (86,4%) or CA+CLA (87,9%) groups. However, the best hatching rate was obtained with CLA (71,8%), which was higher to the other treatments (P=0.01). In conclusion, the use of the metabolic regulators CA, FO and CLA from the fourth day of the embryo culture in vitro increase ...Embriões produzidos in vitro (PIV) apresentam baixa sobrevivência aos métodos convencionais de criopreservação. Na tentativa de melhorar a produção e taxa de sobrevivência embrionária após a vitrificação foram utilizados reguladores metabólicos L-carnitina (CA), Forskolina (FO) e Ácido linoleico conjugado trans-10; cis-12 (CLA), separadamente ou em associação em diferentes doses, à partir do quarto (D4) ou sexto (D6) dia do cultivo de desenvolvimento embrionário. No experimento 1 o uso de 2,5 mM de CA influenciou positivamente na produção (62,0±12,5%, P=0,02), expansão (60,7%, P=0,009) e eclosão (41,1%, P=0,04). No experimento 2, a produção de embriões não foi influenciada pela suplementação com FO (P=0,2), entretanto, a dose de 15,0μM afetou positivamente a expansão (53,2%, P=0,001) e eclosão (46,8%, P0,001). No experimento 3, a produção de embriões não foi influenciada pela suplementação do meio de cultivo com CLA (P=0,4), no entanto a dose de 150,0 μM afetou positivamente a expansão (85,5%, P=0,001) e eclosão (70,9%, P=0,001) dos embriões reaquecidos e cultivados por 48 horas. No experimento 4, a associação dos reguladores de metabolismo lipídico utilizados não influenciou a produção de embriões (P=0,16), entretanto o melhor resultado de produção observado foi no tratamento com 15,0 μM de FO individualmente. As taxas de expansão e eclosão foram influenciadas pela suplementação com reguladores de metabolismo adicionados individualmente ou em associação (P=0,001). A melhor taxa de expansão foi observada na suplementação associando CA+FO+CLA, com 90,05%, que não diferiu (P>0,05) da suplementação com CLA (86,4) e da associação CA+CLA (87,9%). Entretanto, a melhor taxa de eclosão foi obtida com CLA (71,8%, P=0,01), superior aos demais tratamentos. Foi concluído que o uso dos..

Estudo comparativo entre fontes de macromoléculas na produção in vitro de embriões bovinos e seus reflexos na criopreservação

Dentre as biotecnologias aplicadas à reprodução animal, atualmente um dos maiores desafios é a criopreservação de embriões PIV. Esses embriões apresentam alta criosensibilidade, devido principalmente a excessiva deposição lipídica no citoplasma, e que por sua vez está relacionada com a utilização de SFB nos meios. Neste trabalho foram avaliados os efeitos da suplementação protéica com BSA, SFB e FE e suas associações na PIV de embriões bovinos durante as etapas de MIV e CIV, visando melhorar sua criotolerância. Na MIV foram utilizadas sete diferentes combinações (SFB, BSA, FE, BSA+SFB, BSA+FE, SFB+FE e BSA+SFB+FE), e no CIV quatro tratamentos (BSA, BSA+SFB, BSA+FE e BSA+SFB+FE). Os embriões em estádios de Bl e Bx foram vitrificados e após seu reaquecimento foram avaliadas as taxas de eclosão embrionária em 24 e 48 horas. Dos oócitos colocados em maturação, obteve-se um total de 85,4% de clivagem e 35,52% de produção embrionária. Ao analisar os efeitos da fonte protéica na MIV notou-se que o grupo tratado com BSA apresentou as piores taxas de clivagem (80,96%) (p<0,05) em relação aos demais grupos, o melhor resultado de produção embrionária foi obtido com BSA+SFB (46,69%) e os melhores índices de eclosão embrionária pós reaquecimento foram nos tratamentos BSA+FE (44,35%), SFB+FE (44,90%) e BSA+SFB+FE (42,48%). Quando se avaliou os efeitos de fonte protéica na CIV, não houve diferença significativa entre os tratamentos quanto a clivagem, o BSA apresentou o pior desempenho na produção de embriões (31,88%) (p<0,05) em relação aos demais grupos que não diferiram entre si, e os melhores índices de eclosão embrionária 24 e 48h pós reaquecimento foram obtidos com o tratamento BSA+FE (37,64% e 51,34%, respectivamente)Among the biotechnology applied to animal reproduction, nowadays one of the biggest challenges is the cryopreservation of embryos produced in vitro. These embryos have a high cryosensitivity, mainly due to excessive fat deposition in the cytoplasm, which in turn is related with the FCS used in the mediums for embryo production in vitro. The aim of this study was to evaluate the effects of protein supplementation with BSA, FBS and FE, and their associations in IVP bovine embryos during the stages of IVM and IVC, to improve their cryotolerance. For IVM were used seven different combinations (FCS, BSA, FE, BSA + FCS, BSA + FE, FE and FBS + BSA + FCS + FS), and four treatments during IVC (BSA, BSA + FCS, BSA + FE, BSA+ FE+ FCS). Embryos in stage of Bl and Bx were vitrified and after their reheating the rates of hatched blastocysts at 24 and 48 hours were evaluated. Of the oocytes placed into maturation, we obtained 85.4% of cleavage and 35.52% of embryo produced. When were evaluated the effects of protein source in IVM was noted that the BSA-treated group showed the worst rates of cleavage (80.96%) (p <0.05) compared to other groups, the best result of embryo production was obtained with FCS + BSA (46.69%) and the highest rates of hatched blastocysts after rewarming were obtained with FE + BSA (44.90%), FE + FCS (44.35%) and FS + FCS + BSA ( 42.48%). When we assessed the effects of protein source in the IVC, there was no significant difference between treatments for the cleavage, the BSA group had the worst performance in the production of embryos (31.88%) (p <0.05) compared to other groups that not differ, and the highest hatched blastocysts in 24 and 48 hours after rewarming were obtained by treating BSA + FE (37.64% and 51.34%, respectively)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

USO DE ANTIOXIDANTES PARA MELHORAR A EFICIÊNCIA REPRODUTIVA DE REBANHO BOVINO SUBMETIDO A PROTOCOLO DE SINCRONIZAÇÃO COM PROGESTERONA (P4)

Estrus synchronization with progesterone is commonly used in farms with better reproductive control. Free radicals production, i.e. oxidative stress, is associated with progesterone levels. The oxidative stress is responsible for aggression to the cellular membrane, leading to a lise and lipoperoxide formation. In this work, the antioxidant compensatory effect (Vitamins C and E) associated with the exogenous progesterone implant (P4), used in estrus synchronization protocols in cattle, was evaluated. Twenty-five cows were randomly selected in 5 different groups: 1) control without P4, 2) control with P4, 3) P4 + vitamin C and E, 4) P4 + vitamin E, 5) P4 + vitamin C. The lipid lipoperoxidation was measured trough Thiobarbituric acid reactive substances (TBA-RS) and Glutatione Peroxidase enzyme (GSHpx) at days 0 and 7 of the estrus synchronization protocol. The use of vitamin E, in this experiment, showed a better pregnancy rate, however, the results must be validated before orienting the use of the vitamin in cows synchronized with P4

Effects of fatty acid suplementation in Holstein cows at pre and post partum period, on estrous cycle return and in vitro production of embryos

The supplementation of dairy cattle with sources of polyunsaturated fatty acids (PUFA) can be use to increase the energy level of the diet in addition to having positive effects on reproductive functions of important tissues including the hypothalamus, pituitary, ovaries and uterus. The aims of this study were to evaluate the reproductive conditions of the postpartum, number of follicles, corpus luteum (CL) presence, concentration of progesterone (P4), aspirated oocytes, amount of viable oocytes and in vitro production of embryos (IVPE) of the Holstein multiparous donors supplemented with rich diet in protected PUFA (especially linoleic acid - n- 6) and non-protected (especially linolenic acid - n-3) during pre and post partum. The diets had been given for pre-partum during 30 d and post partum 60 d. The donors were divided into three groups: Control (n=6), Megalac-E® (n=5; supplemented with protected fat source 100 g/donor/ day in pre-partum and 300 g/donor/day in postpartum) and linseed (n=5; supplemented with fat source unprotected containing 1 kg/donor/day pre-partum and 1.5 kg/donor/day in postpartum). The animals were submitted to ovum pick-up (OPU) on days 30, 45 and 60 d postpartum. The recovered oocytes were selected and the viable ones were submitted to IVPE procedures. The data were analyzed by the method of least squares variance using the GLM protocol. The differences between averages were compared by Tukey test with 5% significance. There was no detectable effect of treatment, aspirations of postpartum days and interactions on variables: CL presence, concentration of P4, amount of viable oocytes, viable oocytes rate, IVPE and embryos production rate. However, was observed in the group supplemented with linseed more follicles and total oocytes than Megalac-E® and Control group. Supplementation with PUFA didn't increase the number of viable oocytes and IVPE.</p

Supplementation with the histone deacetylase inhibitor trichostatin A during in vitro culture of bovine embryos

Summary Trichostatin A (TSA) is a histone deacetylase inhibitor that induces histone hyperacetylation and increases gene expression levels. The aim of the present study was to establish a suitable condition for the use of TSA in in vitro cultures of bovine embryos, and to determine whether TSA would increase blastocyst rates by improvement of chromatin remodelling during embryonic genome activation and by increasing the expression of crucial genes during early development. To test this hypothesis, 8-cell embryos were exposed to four concentrations of TSA for different periods of time to establish adequate protocols. In a second experiment, three experimental groups were selected for the evaluation of embryo quality based on the following parameters: apoptosis, total cell number and blastocyst hatching. TSA promoted embryonic arrest and degeneration at concentrations of 15, 25 and 50 nM. All treated groups presented lower blastocyst rates. Exposure of embryos to 5 nM for 144 h and to 15 nM for 48 h decreased blastocyst hatching. However, the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay revealed similar apoptosis rates and total cell numbers in all groups studied. Although, in the present study, TSA treatment did not improve the parameters studied, the results provided background information on TSA supplementation during in vitro culture of bovine embryos and showed that embryo quality was apparently not affected, despite a decrease in blastocyst rate after exposure to TSA. © Cambridge University Press 2011

Effect of pre- anc postcalving supplementation of primiparous Nellore donor cows with sources of polyunsarurated fatty acids on the number of oocytes obtained in vivo and in vitro embryo production

The objective of this study was to evaluate the effects of feeding primiparous Nellore cows supplements rich in protected or unprotected linoleic (n-6) and linolenic (n-3) acids before and after calving on the number of follicles, total number of cumulus-oocyte complexes (COCs) collected by aspiration (OPU) and oocytes suitable for culture (grades I, II and III), and in vitro embryo production (IVEP). The donor cows were randomly divided into three groups: control (n=7), Megalac-E® (n=8; 100 g/donor/day), and flaxseed (n=7; 1.0 kg/donor/day). The diets were offered at least 30 days precalving and 75 days postcalving. The animals were submitted to OPU on postcalving days 30, 45, 60 and 75. Recovered oocytes were selected and those considered suitable for culture were submitted to the IVEP procedures. The data were analyzed using a completely randomized design with repeated measures over time. There was no effect of supplementation on the number of follicles, the number of recovered COCs and those suitable for culture, or IVEP. However, the rate of oocytes suitable for culture recovered by OPU was higher on postcalving days 60 and 75 when compared to days 30 and 45. Additionally, the rate of embryos produced in vitro increased after postcalving day 45. In conclusion, pre- and postcalving supplementation with 100 g/day Megalac-E® or 1.0 kg/day flaxseed does not alter the number of oocytes obtained in vivo or IVEP rates of lactating primiparous Nellore donor cows. However, IVEP improves when the programs are carried out 45 days after calving.</span

Chemically Assisted Enucleation Results in Higher G6PD Expression in Early Bovine Female Embryos Obtained by Somatic Cell Nuclear Transfer

Despite extensive efforts, low efficiency is still an issue in bovine somatic cell nuclear transfer (SCNT). The hypothesis of our study was that the use of cytoplasts produced by chemically assisted enucleation (EN) would improve nuclear reprogramming in nuclear transfer (NT)-derived embryos because it results in lower damage and higher cytoplasm content than conventional EN. For that purpose, we investigated the expression of two X-linked genes: X inactive-specific transcript (XIST) and glucose 6-phosphate dehydrogenase (G6PD). In the first experiment, gene expression was assessed in day-7 female blastocysts from embryonic cell NT (ECNT) groups [conventional, ECNT conv; chemically assisted, ECNT deme (demecolcine)]. Whereas in the ECNT conv group, only one embryo (25%; n = 4) expressed XIST transcripts, most embryos showed XIST expression (75%; n = 4) in the ECNT deme group. However, no significant differences in transcript abundance of XIST and G6PD were found when comparing the embryos from all groups. In a second experiment using somatic cells as nuclear donors, we evaluated gene expression profiles in female SCNT-derived embryos. No significant differences in relative abundance (RA) of XIST transcripts were observed among the groups. Nonetheless, higher (p < 0.05) levels of G6PD were observed in SCNT deme and in vitro-derived groups in comparison to SCNT conv. To know whether higher G6PD expression in embryos derived from SCNT chemically assisted EN indicates higher metabolism in embryos considered of superior quality or if the presence of higher reactive oxygen species (ROS) levels generated by the increased oxygen consumption triggers G6PD activation, the expression of genes related to stress response should be investigated in embryos produced by that technique.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP