Bovine somatic cell nuclear transfer using mitomycin C-mediated chemical oocyte enucleation.

SummaryChemical oocyte enucleation holds the potential to ease somatic cell nuclear transfer (SCNT), although high enucleation rates remain limited to micromanipulation-based approaches. Therefore, this study aimed to test mitomycin C (MMC) for use in bovine functional chemical oocyte enucleation. Incubation of denuded eggs in 10 µg ml-1 MMC for different periods did not affect most maturation rates (0.5 h: 85.78%A, 1.0 h: 72.77%B, 1.5 h: 83.87%A, and 2.0 h: 82.05%A) in comparison with non-treated controls (CTL; 85.77%A). Parthenogenetic development arrest by MMC was efficient at cleavage (CTL: 72.93%A, 0.5 h: 64.92%A,B, 1.0 h: 60.39%B,C, 1.5 h: 66.35%A,B, and 2.0 h: 53.84%C) and blastocyst stages (CTL: 33.94%A, 0.5 h: 7.58%B, 1.0 h: 2.47%C, 1.5 h: 0.46%C, and 2.0 h: 0.51%C). Blastocysts were obtained after nuclear transfer (NT) using MMC enucleation [NT(MMC): 4.54%B] but at lower rates than for the SCNT control [NT(CTL): 26.31%A]. The removal of the meiotic spindle after MMC incubation fully restored SCNT blastocyst development [NT(MMC+SR): 24.74%A]. Early pregnancies were obtained by the transfer of NT(MMC) and NT(MMC+SR) blastocysts to synchronized recipients. In conclusion, MMC leads to functional chemical oocyte enucleation during SCNT and further suggests its potential for application towards technical improvements.

Effects of beta-glucans ingestion (Saccharomyces cerevisiae) on metabolism of rats receiving high-fat diet.

We investigated the effects of beta-glucans (Saccharomyces cerevisiae) ingestion on metabolic parameters of Wistar rats receiving high-fat diet. The experimental period was divided into two stages: in the first one, the animals were divided into two groups containing 12 animals each. The first group received commercial feed and the second received high-fat diet containing 20% of pork fat during 60 days. At the end of this period, body weight, blood glucose and Lee index were assessed. In the second stage, those 24 animals were redivided into four groups: (C) - control diet; (CB) - control diet and treated with Beta-glucan (BG); (O) - obese animals and (OB) - obese animals treated with BG. Animals from groups CB and OB received 30 mg/kg of BG dissolved in saline solution by gavage. Animals from groups C and O received only saline solution for 28 days. The design used was totally randomized in 2 × 2 factorial scheme. Data were submitted to analysis of variance (anova). Animals from OB group showed inferior levels (p < 0.05) of total cholesterol (13.33%), triacylglycerols (16.77%) and blood glucose (23.97%) when compared to the animals from group O. The use of BG has provided smaller increase in Lee index (p < 0.05), without promoting alteration in feed and water consumption, organs weight, HDL-C, LDL+VLDL-C, carcass composition, villus/crypt ratio, and pancreas, kidney and stomach histology. BG from S. cerevisiae promoted beneficial metabolic effects in rats receiving high-fat diet.

Effects of in ovo feeding with glycerol for broilers.

The objective was to evaluate the effect of in ovo feeding with glycerol on post-hatch development in broiler chicks. A total of 408 fertile eggs were divided into six experimental groups consisting of five 0.9% saline solutions containing various concentrations of glycerol (12.5, 25.0, 37.5 and 50.0 nmol/ml), and a placebo group (inoculation with saline only) and a control group (without inoculation). Inoculations were performed at 17 days of incubation for the evaluation of hatchability, embryo mortality, body and viscera weights, intestinal epithelium morphometry, blood glucose and liver glycerol kinase activity of chicks at hatching. Inoculation of solutions containing glycerol did not influence body weight at hatching and relative weights of liver, pancreas, intestine and breast. There was a quadratic effect of glycerol levels on the weights of yolk residue and gizzard and on blood glucose, and an increasing linear effect on spleen and heart weights. Higher duodenum and ileum villous height and deeper jejunum and ileum crypts were obtained with 50.0 nmol/ml of glycerol. A linear increasing effect was also observed in liver glycerol kinase activity; however, lower blood glucose was observed with 37.5 and 50 nmol/ml of glycerol. It is therefore concluded that glycerol may be used at doses of 25 nmol/ml as a substrate in in ovo feeding of broiler chickens. However, further studies must be conducted not only to establish an optimal dose but also to evaluate the combination of this substrate with other nutrients used in the in ovo feeding.

Meiotic arrest as an alternative to increase the production of bovine embryos by somatic cell nuclear transfer.

This study aimed to evaluate the effect of meiotic arrest using phosphodiesterase type 3A (PDE 3A) inhibitors, cilostamide and C-type natriuretic peptide (NPPC), on pre-maturation (PM) of oocytes to be used in the production of cloned embryos. Nuclear maturation, in vitro embryo production (IVP), somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA), and total cells number of cloned embryos were evaluated. The results were analysed by chi-squared and Kruskal-Wallis test with a P-value 0.05) between control and PM, both for cleavage (78.2% and 76.9%) and blastocyst (35.5% and 29.3%) rates. After SCNT, cleavage rate was also similar (P > 0.05) between control and PM (66% and 51.9%) however, blastocyst rate was lower (P < 0.05) in the PM group than in the control group (7.4% and 30.2%). After 6 h of PM with 100 nM of NPPC, approximately 84.9% of the oocytes remained at GV. No difference was found between control and PM in cleavage (69.2% and 76.1%) and blastocyst rates (37,4% and 35%) after IVP. Similarly, no differences between PM and control groups were observed for cleavage (69.2% and 68.4%) and blastocyst (24.4% and 21.5%) rates. SCNT and PA embryos from control or PM oocytes had similar total cell number. It can be concluded that PM for 6 h with 100 nM NPPC is feasible for cloned embryo production without affecting embryo outcome.

Equilíbrio acidobásico, parâmetros urinários e sanguíneos de gatos induzidos ao estresse e suplementados com composto antioxidante / Acid-base balance, urinary and blood parameters of cats stress-induced cats and supplemented with antioxidant compound

Este estudo teve como objetivo avaliar a suplementação do composto antioxidante comercial EconomasE (Alltech, Brasil ( AOX) sobre o equilíbrio acidobásico, os parâmetros urinários, o hemograma completo e a fragilidade osmótica de eritrócitos (FOE) de gatos estressados. Foram utilizados 24 gatos adultos (3,49±0,87kg), distribuídos em delineamento inteiramente ao acaso, com quatro níveis (0, 250, 500, 750mg de AOX/kg de alimento na matéria seca) e seis repetições, durante o período de 80 dias. Os gatos foram induzidos ao estresse por meio da presença de cães próximo ao recinto experimental (agente estressor; AE), do 61º dia até o final do experimento. A pressão parcial de dióxido de carbono (pCO2) e o bicarbonato (HCO3 -) aumentaram linearmente (P<0,05) conforme o aumento da ingestão de AOX. Os maiores valores de dióxido de carbono total (tCO2) (P<0,05) foram observados nos gatos alimentados com 500 e 750mg de AOX/kg de dieta. As concentrações de hemoglobina foram maiores nos animais alimentados com 250 e 500mg de AOX/kg de dieta. Os parâmetros urinários e da FOE foram semelhantes nos gatos em todas as dietas. Esses dados indicam que a suplementação com AOX apresenta efeitos benéficos no equilíbrio acidobásico e na concentração de hemoglobina de gatos induzidos ao estresse.(AU)

This study aimed to evaluate the supplementation of a commercial antioxidant EconomasE (Alltech, Brazil - AOX) on the acid-base balance, urinary parameters, red blood count (RBC) and erythrocyte osmotic fragility (EOF) of stressed cats. Twenty-four adult cats (3.49±0.87kg) were distributed in a completely randomized design with four levels (0, 250, 500, 750mg AOX/kg feed dry matter) and six replicates for a period of 80 days. Cats were induced to stress through the presence of dogs in the experimental environment (stressor agent; AE) from the 61th day to the end. Partial pressure of carbon dioxide (pCO2) and bicarbonate (HCO3) increased linearly (P<0.05) with increased intake of AOX. The highest values of total carbon dioxide (tCO2) (P<0.05) were observed in cats fed the 500 and 750mg AOX/kg diet. Hemoglobin concentration was higher (P<0.05) in animals fed the 250 and 500mg AOX/kg diet. The urinary parameters and EOF were similar among all diets. These data indicate that AOX supplementation has beneficial effects in acid-base balance and hemoglobin concentration of stress-induced cats.(AU)

Tratamento alternativo em gatos acometidos por DITUIF / Alternative treatment in cats affected by FLUTD

This report describes two cases of male cats affected by FLUTD (Feline lower urinary tract disease). The first patient had been affected by numerous relapses after passing through various therapeutic treatments and surgical interventions, and was subjected to the alternative protocol after being recommended for euthanasia, the second patient had no history of urinary tract disorders in the past, and was not subjected to any kind of previous treatment. The two patients had a significant improvement in their clinical condition, were not affected by relapses after participation in the alternative design, and were accompanied for about a year after their treatment.(AU)

Characterization of allele-specific expression of the X-linked gene MAO-A in trophectoderm cells of bovine embryos produced by somatic cell nuclear transfer.

Somatic cell nuclear transfer (SCNT) may affect epigenetic mechanisms and alter the expression of genes related to embryo development and X chromosome inactivation (XCI). We characterized allele-specific expression of the X-linked gene monoamine oxidase type A (MAO-A) in the trophectoderm (TF) of embryos produced by SCNT. Total RNA was isolated from individual biopsies (N = 25), and the allele-specific expression assessed by reverse transcription-polymerase chain reaction-restriction fragment length polymorphism. Both paternal and maternal alleles were expressed in the trophectoderm. However, a higher frequency of the mono-allelic expression of a specific allele was observed (N = 17; 68%), with the remaining samples showing the presence of mRNA from both alleles (N = 8; 32%). Considering that MAO-A is subject to XCI in bovine, our results suggest that SCNT may influence XCI because neither an imprinted (mono-allelic expression in all samples) nor a random (presence of mRNA from both alleles in all samples) pattern of XCI was observed in TF. Due to the importance of XCI in mammalian embryo development and its sensitivity to in vitro conditions, X-linked genes subject to XCI are candidates for use in the development of embryo quality molecular markers for assisted reproduction.

Metabolic parameters in rats receiving different levels of oral glycerol supplementation.

The use of glycerol in the diets for animals is of interest because it is a residue of biodiesel production and rich in energy. Thus, this study aimed to evaluate metabolic and physiological parameters of rats receiving supplemental pure glycerol by gavage. We used 30 Wistar rats (initial weight 202.7 ± 29.98 g) receiving 0 (control/saline), 200, 400, 800 and 1600 mg glycerol/kg of body weight (bidistilled glycerine, 99.85% glycerol) beside food and water ad libitum for 28 days. We used a completely randomised design with five treatments and six replicates. At the end of the experiment, the animals were killed, and the results showed that there was no change (p > 0.05) in the intake and excretion of water, the average daily weight gain, dry matter, ash and crude protein in the carcass or plasma triacylglycerols. There was a beneficial effect (p < 0.05) up to a dose of 800 mg/kg glycerol on feed intake, percentage of carcass fat, plasma levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), high-density lipoprotein (HDLc) and low-/very low-density lipoprotein (LDLc + VLDLc). The levels of total cholesterol and glucose were increased with up to a dose of 800 mg/kg glycerol (but remained within the normal range); they were reduced with the dose of 1600 mg/kg. The total leucocyte count tended to be reduced, although it was within the reference values for rats. There were no renal or pancreatic lesions. In conclusion, glycerol presented as a safe supplement at the studied doses, even having some beneficial effects in a dose-dependent manner in rats.

Fatty acid profile, oxidative stability of pork lipids and meat quality indicators are not affected by birth weight.

The aim of this study was to investigate whether fatty acid (FA) profile, oxidative stability of lipids and other meat quality traits differed between high (HW: 1.8 to 2.2 kg) and low (LW: 0.8 to 1.2 kg) birth weight piglets. Forty new-born male pigs (n=20 HW, n=20 LW) were reared in separate pens until the finishing period, when they were slaughtered at 150 days of age, and pH and temperature were measured in the carcass. Afterwards, the Longissimus dorsi muscle was excised from the carcass, and samples were collected for subsequent meat quality analyses (thaw loss, cooking loss, shear force, chemical analysis and sensory analysis for tenderness). Birth weight had minor impacts on meat quality traits, which were limited to higher shear force in the LW group (P<0.01). Chemical components (moisture, protein, fat, ash), cholesterol levels and lipid oxidation (thiobarbituric acid-reactive substances) were not affected by birth weight (P>0.05). FA profile and the amount of saturated, monounsaturated and polyunsaturated fatty acids were similar, but HW pigs had higher atherogenic index than their LW counterparts (P<0.01). Notwithstanding the higher shear force presented by the lower birth weight pigs, in the sensory test, the panelists did not detect any differences in the tenderness of pork from HW and LW animals. Therefore, our results suggest that low birth weight has minimal impact on meat quality.

Effect of ractopamine on the chemical and physical characteristics of pacu(Piaractus mesopotamicus) steaks / Efeito da ractopamina sobre as características físicas e químicas de filés de pacu (Piaractus mesopotamicus)

The objective was to evaluate the use of ractopamine (RAC) in the diet for pacu (Piaractus mesopotaminus) in the finishing phase on some quality parameters of the fillets. Thirty-five animals weighing 0.868±0.168kg were distributed in a completely randomised design with five treatments (0.0 - control; 11.25, 22.50, 33.75 and 45 ppm of RAC) and seven replicates with two fillets obtained from the same animal. The diets were isocaloric and isoprotein and experimental time was 90 days. RAC did not affect (P>0.05) the initial pH or ph after 24 hours of the fillets. Compared to the control, RAC increased (P<0.05) the moisture content of the fillets in natura and lipid oxidation of samples stored for 12 days in the refrigerator or freezer for 60 days. The RAC in 11.25 ppm reduced (P<0.05) the lipid content, while 45 ppm reduced (P<0.05) the crude protein in the fillets. Considering only RAC, there was a linear increase (P<0.05) in the lipid content (P<0.05) and a linear reduction in crude protein and weight loss after cooking the fillets. There was a quadratic effect (P<0.05) on the ash content, weight loss and lipid oxidation in fillets stored in the refrigerator or freezer. A RAC dose of 33.75 ppm resulted in a lower lipid oxidation index. In conclusion, ractopamine at 11.25 ppm is effective for reducing the fat content in fillets of pacu, although it increases the formation of peroxides in samples kept in the freezer for longer than 60 days. At 33.75 ppm, ractopamine is effective in reducing the effect of oxidation during storage in the refrigerator or freezer.

O objetivo foi avaliar a influência do uso de ractopamina na alimentação de pacus (Piaractus mesopotaminus) na fase de terminação sobre alguns parâmetros de qualidade dos filés. Trinta e cinco animais com peso de 0,868±0,168kg foram distribuídos em delineamento inteiramente ao acaso com cinco tratamentos (0,0 - controle; 11,25; 22,50; 33,75 e 45ppm de ractopamina) e sete repetições, sendo a parcela representada por dois filés provenientes de um animal. As rações foram isoenergéticas e isoproteicas, e o tempo experimental foi de 90 dias. A ractopamina não influenciou (P>0,05) o pH inicial dos filés e nem após 24 horas. Comparado ao controle, a RAC aumentou (P<0,05) a umidade dos filés in natura e a oxidação lipídica dos mesmos quando armazenados por 12 dias em geladeira ou 60 dias em freezer. Na dose de 11,25 reduziu (P<0,05) o teor de extrato etéreo e, na dose de 45 ppm, reduziu (P<0,05) o de proteína bruta dos filés. Considerando somente a RAC, houve aumento linear (P<0,05) do teor de gordura e redução linear (P<0,05) da proteína bruta e perda de peso por cocção. Houve efeito quadrático (P<0,05) sobre o teor de cinzas, perda de peso por descongelamento e oxidação lipídica para os filés armazenados em geladeira ou em freezer, sendo 33,75 ppm o nível de RAC que resultou em menor índice de oxidação lipídica. Conclui-se que a ractopamina a 11,5 ppm é efetiva em reduzir o teor de lipídeos em filés de pacu, embora aumente a formação de peróxidos em amostras mantidas em freezer após 60 dias. A 33,75 ppm a ractopamina é efetiva em reduzir os efeitos da oxidação durante o armazenamento em geladeira ou freezer.

Effect of the addition of IGF-I and vitamin E to stored boar semen.

The objective of this study was to evaluate the addition of IGF-I to pig insemination doses stored at 15°C, in conjunction with the addition of different amounts of vitamin E (α-tocopherol). Semen samples (n = 12) from four boars were treated by the addition of different concentrations of vitamin E, ranging up to 400 µg/ml. Immediately after processing and after the doses had been stored at 15°C for 24 or 72 h, samples were warmed at 37°C and 30 ng/ml of IGF-I was added. The assessments were made after 10 and 120 min of IGF-I addition. There was a minor effect of the vitamin E added before cooling and IGF-I added after storage on sperm quality. The addition of 400 µg/ml of vitamin E to diluted semen reduced (P < 0.01) the malondialdehyde (MDA) production in boar semen stored at 15°C for 72 h, regardless of the addition of IGF-I as additive during a 120 min incubation period at 37°C. In these conditions, IGF-I also reduced (P < 0.05) the MDA production in semen samples without addition of vitamin E. IGF-I in the presence of vitamin E reduced (P = 0.03) the glucose intake in freshly diluted boar semen samples before cooling. It was concluded that the addition of 400 µg/ml of vitamin E reduces the MDA production in boar semen stored at 15°C for 72 h, regardless of the presence of IGF-I additive. The addition of IGF-I in doses stored for 72 h with vitamin E ensures higher sperm motility after 120 min of incubation at 37°C.

Dental trauma among Brazilian schoolchildren: prevalence, treatment and associated factors.

AIM: To assess the prevalence of traumatic dental injury (TDI), as well as associated factors, behaviour of affected schoolchildren and normative treatment needs. METHODS: The present cross-sectional study involved 590 children aged 7-14 years at state schools in Campina Grande, Brazil. The O'Brien classification [1994] was used for the diagnosis of TDI and the body mass index (BMI) was used as an indicator of overweight/obesity. Clinical tests were carried out by two duly calibrated examiners (intra-observer and inter-observer agreement: 0.87 and 0.90, respectively). The Chi-square test was used (5% level of significance) to determine whether TDI was associated with age, gender, ethnicity, overweight/obesity, lip seal and overjet. Backward stepwise multivariate regression analysis was performed. Normative treatment needs were determined based on the criteria of the International Association of Dental Traumatology (IADT). When the absence of treatment was detected, each child/adolescent was asked about the reason for non-treatment. RESULTS: The prevalence of TDI was 12.7%. The most common type of trauma was enamel fracture (67.0%), followed by enamel-dentine fracture (25.3%). TDI was 4.9-fold greater (95% CI: 1.6-14.4) among children aged 13 and 14 years, 1.9- fold greater (95% CI: 1.1-3.2) among males and 2.6-fold greater (95% CI: 1.2-5.4) among those with inadequate lip seal. The majority of schoolchildren did not undergo treatment (82.6%) due to a belief that it was unnecessary (53.2%). The normative clinical evaluation revealed that adhesive restoration was the most common form of treatment (84.0%). CONCLUSION: The prevalence of TDI was low. Age, gender and inadequate lip seal were associated with dental trauma. A significant number of schoolchildren did not receive treatment for dental trauma.

Avaliação preliminar in vivo do efeito hipocolesterolêmico do polvilho da fruta-de-lobo (Solanum lycocarpum A. St.-Hil. ) / Preliminary evaluation of in vivo hypocholesterolemic effect of the flour of fruit-of-wolf (Solanum lycocarpum A. St.-Hil. )

O polvilho da fruta-de-lobo é um produto extraído da polpa da fruta-de-lobo verde (Solanum lycocarpum A. St.-Hil.), popularmente utilizado. Pouco se conhece a respeito desse polvilho, mas são atribuídos a ele vários efeitos terapêuticos, dentre eles a redução do colesterol. Este trabalho teve o objetivo de verificar a ação da administração do polvilho da fruta-de-lobo sobre animais hipercolesterolêmico. Utilizou-se 24 ratos Wistar fêmeas distribuídas em delineamento ao acaso, em três grupos contendo 8 animais em cada grupo. Estes grupos foram definidos como: controle normal (CN), controle hipocolesterolêmico (CH) e hipocolesterolêmico tratado (HT). O grupo CN recebeu dieta comercial, os grupo CH e HT receberam a dieta comercial enriquecida de colesterol e ácido cólico e o grupo HT recebeu também, 100 mg de polvilho da fruta-lobo, diariamente, por sonda orogástrica. O experimento teve uma duração de 6 semanas onde se avaliou o colesterol total sérico semanalmente, peso dos animais semanalmente e o consumo diário de ração. Ao término do experimento, foram avaliados os seguintes parâmetros: frações séricas de colesterol HDL, LDL+VLDL, peso do fígado, colesterol total hepático, lipídeos totais hepático e lâminas de microscopia foram feitas para avaliação dos hepatócitos. Não houve diferença significativa em relação ao peso corporal dos animais, ao consumo da dieta e nas análises de colesterol total sérico entre os grupos estudados. Avaliando-se a relação VLDL +LDL/HDL, os níveis encontrados no grupo HT foram significativamente menores que o grupo CH. Já em relação ao colesterol hepático, o grupo HT mostrou níveis menores de colesterol que o CH. Observou-se nos lipídeos hepáticos que não houve diferença significativa entre os grupo CH e HT, e no peso do fígado houve diferença significativa entre os grupos avaliados. Em relação à microscopia, os grupos hipercolesterolêmicos apresentaram discreta vacuolização no citoplasma dos hepatócitos. Concluiu-se, que o polvilho da fruta-de-lobo não influenciou o colesterol sérico dos animais, entretanto, reduziu os níveis de colesterol hepático.

The fruit-of-wolf flour is a product extracted from the pulp of green fruit-of-wolf (Solanum lycocarpum A. St .- Hil.) and is commonly used. Little is known about this flour, but several therapeutic effects, including cholesterol reduction, are attributed to it. This study aimed to verify the action of the administration of fruit-of-wolf flour to hypercholesterolemic animals. We used 24 female Wistar rats allocated in randomized design to three groups containing 8 animals each. These groups were defined as normal control (CN), hypocholesterolemic control (CH) and hypocholesterolemic treated rats (HT). The CN group received a commercial diet, while the CH and the HT group received the commercial diet enriched with cholesterol and cholic acid; the HT group also received 100 mg of fruit-of-wolf flour, daily, by orogastric tube. The experiment lasted for six weeks and the following characteristics were evaluated: weekly total serum cholesterol, weekly weight of animals and daily food intake. At the end of the experiment, we assessed the following parameters: serum cholesterol fractions HDL, LDL + VLDL, liver weight, liver total cholesterol, liver total lipids and microscopic slides were prepared for the evaluation of hepatocytes. There was no significant difference in body weight of animals, diet consumption and analysis of serum total cholesterol among the studied groups. Assessing the relationship VLDL + LDL / HDL, the levels found for the HT group were significantly lower than those for the CH group. As regards liver cholesterol, the HT group showed lower cholesterol levels than the CH group. For liver lipids there was no significant difference between the CH and the HT group, and for liver weight there was no significant difference among the studied groups. As to microscopy, the hypercholesterolemic groups showed slight vacuolization in the cytoplasm of hepatocytes. It was concluded that fruit-of-wolf flour did not influence the serum cholesterol of animals but reduced the levels of liver cholesterol.

Avaliação da adequação legal de receitas e notificações de receita de medicamentos sujeitos a controle especial dos setores públicos e privados / Evaluation of the legal prescription and notifications adequacy for special control medicinesin the public and private sectors

Os objetivos desta pesquisa foram avaliar a adequação legal de receitas de controle especial e notificações de receitas dispensadas em uma Drogaria de Varginha ? MG, estabelecer uma comparação entre as prescrições provenientes da rede pública e privada e detectar as classes de medicamentos mais prescritas. Analisaram-se 960 prescrições através de um formulário com itens obrigatórios de acordo com a legislação vigente. Os dados obtidos foram analisados pelo Teste Qui-quadrado, utilizando a origem das notificações e receitas como desfecho. Considerando as receitas; a única variável que apresentou diferença entre os setores público e privado foi a descrição por extenso da quantidade de medicamento, que foi mais adequadamente cumprida pelo setor privado. Em relação às notificações, a legibilidade foi significativamente mais freqüente em notificações provenientes de centros públicos, enquanto a designação da Unidade Federativa e a descrição por extenso da quantidade de medicamento foram mais presentes no setor privado. Por outro lado, a descrição completa da posologia, da data da notificação, a presença de carimbo e descrição da forma farmacêutica foram mais adequadas no setor público. Nenhuma das prescrições foi considerada totalmente adequada aos dados exigidos pela Portaria 344/98. As classes medicamentosas mais prescritas foram os benzodiazepínicos em notificações de receita e os antidepressivos em receituário de controle especial. Concluiu-se que a prescrição de medicamentos sujeitos a controle especial apresenta falhas de acordo com a legislação vigente.

The objectives of the present study were to evaluate the legal prescription adequacy in one drugstore located in the south of Minas Gerais state, in order to establish comparisons between the prescriptions of public and private services and to detect the most commonly classes of medicines prescribed. It was analyzed 960 prescriptions in accordance with the current law. The data were analyzed by descriptive statistics and compared using frequency distribution test(Chi-square test) considering private and public sources of prescription as the dependent variable. Considering special control prescriptions, the extensive description of the amount of medicine was fulfilled more frequently by the private sector. However, for the notifications; legibility, dosage, pharmaceutical form, date of issue and presence of stamp were more frequently present in the public service, while assignment of the Federative Union, the extensive description of the amount of medicine and the dosage was more present in the private sector. None of the evaluated prescriptions was considered totally adjusted with law 344/98. The prescribed medicines more commonly found were benzodiazepines for prescription notifications and antidepressants in special control prescriptions. It was concluded that the medicine prescriptions subjected to special control are frequently not adequate to the current law.

Recovery, cryopreservation and fertilization potential of bovine spermatozoa obtained from epididymides stored at 5 degrees C by different periods of time.

The objetive of the present study was to evaluate the effect of the interval between animal's death and sperm recovery on the freezability and fertilizing ability of spermatozoa from bull epididymides stored for different periods of time. Testis from 25 bulls were collected at the abattoir 2h after the slaughter. In the laboratory spermatozoa from one epididymis were recovered and analysed for motility. The remaining epididymis was stored for 24h (G24), 48h (G48) and 72h (G72) at 5 degrees C. At the end of each time period, spermatozoa were recuperated and cryopreserved in Tris-egg yolk and glycerol. Pre-freeze and post-thaw sperm samples were taken to assess total and progressive motility, concentration, membrane integrity and acrosome integrity. For evaluation of fertilizing ability, in each time period five straws of each bull were thawed, pooled and used for in vitro embryo production. The results showed that after 48h of storage there was a decline in total motility, which did not change until 72h. Progressive motility, plasma membrane and acrosome integrity were not affected by any of the storage periods. Conversely, all sperm parameters, except progressive motility, were reduced after cryopreservation. Embryo production was less (P<0.05) in the treatments than in the reference group. However, there was no differences (P>0.05) in blastoycst rate among experimental groups. Considering all the embryos produced by epididymal spermatozoa a greater proportion of female embryos was observed, which was similar to the reference embryos. The shift observed on sex ratio toward female for those two groups was also observed when they were compared with the expected 1:1 ratio (P<0.05). The results showed the possibility to produced in vitro embryos using cryopreseved spermatozoa from epididymides and stored for long period of time at 5 degrees C. These procedures became an important tool for animal preservation when the sperm cells cannot be cryopreserved immediately after the animal's death.

Hypoglycemic activity of polysaccharide fractions containing beta-glucans from extracts of Rhynchelytrum repens (Willd.) C.E. Hubb., Poaceae.

Beta-glucans are soluble fibers with physiological functions, such as interference with absorption of sugars and reduction of serum lipid levels. The objective of the present study was to analyze the distribution of beta-glucans in different tissues of the African grass species Rhynchelytrum repens and also to evaluate their hypoglycemic activity. Leaf blades, sheaths, stems, and young leaves of R. repens were submitted to extraction with 4 M KOH. Analysis of the fractions revealed the presence of arabinose, glucose, xylose, and traces of rhamnose and galactose. The presence of beta-glucan in these fractions was confirmed by hydrolyzing the polymers with endo-beta-glucanase from Bacillus subtilis, followed by HPLC analysis of the characteristic oligosaccharides produced. The 4 M KOH fractions from different tissues were subjected to gel permeation chromatography on Sepharose 4B, with separation of polysaccharides with different degrees of polymerization, the highest molecular mass (above 2000 kDa) being found in young leaves. The molecular mass of the leaf blade polymers was similar (250 kDa) to that of maize coleoptile beta-glucan used for comparison. The 4 M KOH fraction injected into rats with streptozotocin-induced diabetes showed hypoglycemic activity, reducing blood sugar to normal levels for approximately 24 h. This performance was better than that obtained with pure beta-glucan from barley, which decreased blood sugar levels for about 4 h. These results suggest that the activity of beta-glucans from R. repens is responsible for the use of this plant extract as a hypoglycemic drug in folk medicine.

Hypoglycemic activity of polysaccharide fractions containing ß-glucans from extracts of Rhynchelytrum repens (Willd.) C.E. Hubb., Poaceae

ß-Glucans are soluble fibers with physiological functions, such as interference with absorption of sugars and reduction of serum lipid levels. The objective of the present study was to analyze the distribution of ß-glucans in different tissues of the African grass species Rhynchelytrum repens and also to evaluate their hypoglycemic activity. Leaf blades, sheaths, stems, and young leaves of R. repens were submitted to extraction with 4 M KOH. Analysis of the fractions revealed the presence of arabinose, glucose, xylose, and traces of rhamnose and galactose. The presence of ß-glucan in these fractions was confirmed by hydrolyzing the polymers with endo-ß-glucanase from Bacillus subtilis, followed by HPLC analysis of the characteristic oligosaccharides produced. The 4 M KOH fractions from different tissues were subjected to gel permeation chromatography on Sepharose 4B, with separation of polysaccharides with different degrees of polymerization, the highest molecular mass (above 2000 kDa) being found in young leaves. The molecular mass of the leaf blade polymers was similar (250 kDa) to that of maize coleoptile ß-glucan used for comparison. The 4 M KOH fraction injected into rats with streptozotocin-induced diabetes showed hypoglycemic activity, reducing blood sugar to normal levels for approximately 24 h. This performance was better than that obtained with pure ß-glucan from barley, which decreased blood sugar levels for about 4 h. These results suggest that the activity of ß-glucans from R. repens is responsible for the use of this plant extract as a hypoglycemic drug in folk medicine.

Development of bovine embryos reconstructed by nuclear transfer of transfected and non-transfected adult fibroblast cells

An association of two techniques, nuclear transfer (NT), and transfection of somatic animal cells, has numerous potential applications and considerable impact, mainly in agriculture, medicine, pharmacy, and fundamental biology. In addition, somatic cell nuclear transfer is the most efficient alternative to produce large transgenic animals. We compared in vitro and in vivo developmental capacities of NT using fibroblast cells isolated from a 14-month-old cloned Simmental heifer (FCE) vs the same line transfected with a plasmid containing neomycin-resistant genes (TFCE). There were no significant differences (P > 0.5) in either fusion (116/149 = 78% vs 216/301 = 72%), cleavage (78/116 = 67% vs 141/216 = 65%) and blastocyst (35/116 = 30% vs 52/216 = 24%) rates or in pregnancy rate at 30 to 35 days after embryo transfer (2/17 vs 3/17) between NT using FCE and TFCE, respectively. Transfection and long-term in vitro culture of transfected cells did not affect developmental capacity of NT embryos up to 40 days of gestation

Isolation of transfected fibroblast clones for use in nuclear transfer and transgene detection in cattle embryos

Transgenesis in cattle has provided numerous opportunities for livestock production. The development of nuclear transfer (NT) technology has improved the production of transgenic livestock. However, the isolation of pure colonies from a single transfection event remains laborious and can be a constraint in the production of transgenic livestock. We used 96-well cell culture plates to isolate cell lineages obtained from a single fibroblast transfected with the pCi-Neo plasmid. Since single mammalian cells do not grow well in fresh medium, we evaluated the use of conditioned medium. The neomycin phosphotransferase gene was detected in isolated colonies and NT embryos were produced from these cells. Multiplex-PCR assays were performed to detect the transfected fragment as well as autosomal satellite DNA in single NT embryos. This approach provided a reliable method for isolating transfected mammalian cells and for diagnosing the incorporation of desirable vectors in NT embryos. This method can reduce the time and cost of transgenic livestock production.

Composiçäo de ácidos graxos da gordura na carcaça de frangos de corte sob dietas com diferentes fontes de energia / Carcass fatty acid composition of broiler fed different sources of energy

Avaliaram-se os níveis de energia metabolizável normalmente utilizados nas raçöes de frangos de corte, obtidos pela inclusäo de óleos vegetais (soja, canola e palma) e seus efeitos sobre o perfil de ácidos graxos da pele e dos músculos da coxa e do peito, levando-se em consideraçäo a linhagem e o sexo. Foram utilizados 2400 pintos de um dia, machos e fêmeas, das linhagens comerciais Hubbard e Avian Farms. As aves receberam raçäo inicial e final com níveis de energia metabolízavel de 3050 e 3150kcal/kg, respectivamente, obtidos pela inclusäo dos óleos vegetais, fornecedores de ácidos graxos, inclusive "w-3". Utilizou-se um delineamento inteiramente ao acaso em um arranjo fatorial com 16 tratamentos (4 raçöes x 2 sexos x 2 linhagens) com duas repetiçöes cada. Diferenças entre médias foram testadas pela dms. Näo houve efeito significativo da inclusäo de óleos na raçäo sobre o perfil de ácidos graxos na carcaça. O mesmo ocorreu quanto ao efeito de sexo e de linhagem