A biotechnological tool for glycoprotein desialylation based on immobilized neuraminidase from Clostridium perfringens.

BACKGROUND: Sialic acids are widely distributed in nature and have biological relevance owing to their varied structural and functional roles. Immobilized neuraminidase can selectively remove terminal N-acetyl neuraminic acid from glycoproteins without altering the protein backbone while it can be easily removed from the reaction mixture avoiding sample contamination. This enables the evaluation of changes in glycoprotein performance upon desialylation. METHODS: Neuraminidase was immobilized onto agarose activated with cyanate ester groups and further used for desialylation of model glycoproteins, a lysate from tumour cells and tumour cells. Desialylation process was analysed by lectin binding assay, determination of sialyl-Tn or flow cytometry. RESULTS: Clostridium perfringens neuraminidase was immobilized with 91 % yield and expressed activity yield was of 41%. It was effective in the desialylation of bovine fetal serum fetuin, bovine lactoferrin and ovine submaxilar mucin. A decrease in sialic-specific SNA lectin recognition of 83% and 53 % was observed for fetuin and lactoferrin with a concomitant increase in galactose specific ECA and PNA lectin recognition. Likewise, a decrease in the recognition of a specific antibody (82%) upon mucin desialylation was observed. Moreover, desialylation of a protein lysate from the sialic acid-rich cell line TA3/Ha was also possible leading to a decrease in 47 % in SNA recognition. Immobilized neuraminidase kept 100% of its initial activity upon five desialylation cycles. CONCLUSIONS: Immobilized neuraminidase is an interesting as well as a robust biotechnological tool for enzymatic desialylation purposes. GENERAL SIGNIFICANCE: Immobilized neuraminidase would contribute to understand the role of sialic acid in biological processes.

Xylanase and B-glucanase in maize- and soybean meal-based diets for broilers / Xilanase e B-glucanase em dietas a base de milho e de farelo de soja para frangos de corte

The objective of this study was to evaluate the effect of adding different xylanase and β-glucanase enzyme blends to maize- and soybean meal-based diets on performance and energy metabolizability in broilers. Two experiments were carried out with broilers of the COBB 500 strain. In the first experiment, 1960 chicks were assigned to a completely randomized design with a 2 × 3 + 1 factorial arrangement, totaling seven treatments, namely, T1- Positive control (PC); T2 - Negative control 1 (NC1; PC minus 200 kcal kg-¹ ME); T3 - NC1 + Blend A; T4 - NC1 + Blend B; T5 - Negative control 2 (NC2; PC minus 167 kcal kg-¹ ME and 5% amino acids); T6 - NC2 + Blend A; and T7 - NC2 + Blend B. Fourteen replicates were used per treatment and 20 birds per experimental unit. The parameters evaluated at 21 and 42 days of age were weight gain (WG), feed intake (FI) and feed conversion (FC). At 42 days, production efficiency index (PEI), viability and the yields of cuts were also calculated. Birds that received diets with a reduced nutritional value showed a reduction in WG and PEI and worsened FC as compared those of PC treatment (p 0.05) from 1 to 21 days of life. For the yield of thigh + drumstick, the factors were statistically similar (p > 0.05) to those observed in the PC birds. In the second experiment, 432 fourteen-day-old chicks were distributed in a completely randomized design with seven treatments, with eight replicates per treatment and six birds per experimental unit. The apparent metabolizable energy (AME) and nitrogen-corrected AME (AMEn) values were determined. Overall, the NC2 diet with Blend B provided the highest AME and AMEn values; however, NC1 with the same enzyme blend was the treatment which provided the lowest values. The addition of xylanase [...].

Objetivou-se avaliar o efeito da adição de diferentes blends enzimáticos de xilanase e β-glucanase em dietas a base de milho e de farelo de soja sobre o desempenho e metabolizabilidade da energia para frangos de corte. Foram realizados dois experimentos com frangos de corte da linhagem COBB500. No primeiro experimento, foram utilizados 1960 pintos distribuídos em delineamento inteiramente casualizado em arranjo fatorial 2 x 3 + 1, totalizando sete tratamentos: T1- Controle Positivo (CP); T2- Controle negativo 1 (CN1; CP menos 200 kcal Kg-¹ de EM); T3 - CN1 + Blend A; T4 - CN1 + Blend B; T5 - Controle negativo 2 (CN2; CP menos 167 kcal Kg-¹ de EM e 5% de aminoácidos); T6 - CN2 + Blend A e T7 - CN2 + Blend B. Foram utilizadas 14 repetições por tratamento e 20 aves por unidade experimental. Os parâmetros avaliados aos 21 e aos 42 dias de idade dos frangos foram o ganho de peso (GP), o consumo de ração (CR) e a conversão alimentar (CA). Aos 42 dias calculou-se o índice de eficiência produtiva (IEP), viabilidade (VIAB) e rendimento de cortes. Foi observado que as aves que receberam dietas com reduzido valor nutricionais apresentaram redução no GP e IEP e piora na CA comparadas ao CP (p 0,05) no período de um a 21 dias de vida. Para o rendimento de coxa mais sobrecoxa (RCS), foi possível observar que os fatores apresentaram estatisticamente (p >0,05) semelhantes ao CP. No segundo experimento foram utilizados 432 pintos com 14 dias de idade, distribuídas em um delineamento inteiramente casualizado com sete tratamentos com 8 repetições por tratamento e 6 aves por unidade experimental. Foram determinados os valores de energia metabolizável aparente (EMA) e aparente corrigida para balanço de nitrogênio (EMAn). Em geral, a dieta CN2 como Blend B apresentou o maior valor de EMA e EMAn, porém, [...].

Xylanase and B-glucanase in maize- and soybean meal-based diets for broilers / Xilanase e B-glucanase em dietas a base de milho e de farelo de soja para frangos de corte

The objective of this study was to evaluate the effect of adding different xylanase and β-glucanase enzyme blends to maize- and soybean meal-based diets on performance and energy metabolizability in broilers. Two experiments were carried out with broilers of the COBB 500 strain. In the first experiment, 1960 chicks were assigned to a completely randomized design with a 2 × 3 + 1 factorial arrangement, totaling seven treatments, namely, T1- Positive control (PC); T2 - Negative control 1 (NC1; PC minus 200 kcal kg-¹ ME); T3 - NC1 + Blend A; T4 - NC1 + Blend B; T5 - Negative control 2 (NC2; PC minus 167 kcal kg-¹ ME and 5% amino acids); T6 - NC2 + Blend A; and T7 - NC2 + Blend B. Fourteen replicates were used per treatment and 20 birds per experimental unit. The parameters evaluated at 21 and 42 days of age were weight gain (WG), feed intake (FI) and feed conversion (FC). At 42 days, production efficiency index (PEI), viability and the yields of cuts were also calculated. Birds that received diets with a reduced nutritional value showed a reduction in WG and PEI and worsened FC as compared those of PC treatment (p < 0.05). However, the birds that consumed the NC2 diet with Blend B exhibited a similar WG to those in PC group (p > 0.05) from 1 to 21 days of life. For the yield of thigh + drumstick, the factors were statistically similar (p > 0.05) to those observed in the PC birds. In the second experiment, 432 fourteen-day-old chicks were distributed in a completely randomized design with seven treatments, with eight replicates per treatment and six birds per experimental unit. The apparent metabolizable energy (AME) and nitrogen-corrected AME (AMEn) values were determined. Overall, the NC2 diet with Blend B provided the highest AME and AMEn values; however, NC1 with the same enzyme blend was the treatment which provided the lowest values. The addition of xylanase [...].(AU)

Objetivou-se avaliar o efeito da adição de diferentes blends enzimáticos de xilanase e β-glucanase em dietas a base de milho e de farelo de soja sobre o desempenho e metabolizabilidade da energia para frangos de corte. Foram realizados dois experimentos com frangos de corte da linhagem COBB500. No primeiro experimento, foram utilizados 1960 pintos distribuídos em delineamento inteiramente casualizado em arranjo fatorial 2 x 3 + 1, totalizando sete tratamentos: T1- Controle Positivo (CP); T2- Controle negativo 1 (CN1; CP menos 200 kcal Kg-¹ de EM); T3 - CN1 + Blend A; T4 - CN1 + Blend B; T5 - Controle negativo 2 (CN2; CP menos 167 kcal Kg-¹ de EM e 5% de aminoácidos); T6 - CN2 + Blend A e T7 - CN2 + Blend B. Foram utilizadas 14 repetições por tratamento e 20 aves por unidade experimental. Os parâmetros avaliados aos 21 e aos 42 dias de idade dos frangos foram o ganho de peso (GP), o consumo de ração (CR) e a conversão alimentar (CA). Aos 42 dias calculou-se o índice de eficiência produtiva (IEP), viabilidade (VIAB) e rendimento de cortes. Foi observado que as aves que receberam dietas com reduzido valor nutricionais apresentaram redução no GP e IEP e piora na CA comparadas ao CP (p < 0,05), porém as aves que consumiram a dieta CN2 com o Blend B, que apresentaram GP semelhante ao CP (p > 0,05) no período de um a 21 dias de vida. Para o rendimento de coxa mais sobrecoxa (RCS), foi possível observar que os fatores apresentaram estatisticamente (p >0,05) semelhantes ao CP. No segundo experimento foram utilizados 432 pintos com 14 dias de idade, distribuídas em um delineamento inteiramente casualizado com sete tratamentos com 8 repetições por tratamento e 6 aves por unidade experimental. Foram determinados os valores de energia metabolizável aparente (EMA) e aparente corrigida para balanço de nitrogênio (EMAn). Em geral, a dieta CN2 como Blend B apresentou o maior valor de EMA e EMAn, porém, [...].(AU)

Blood cells and endothelial barrier function.

The barrier properties of endothelial cells are critical for the maintenance of water and protein balance between the intravascular and extravascular compartments. An impairment of endothelial barrier function has been implicated in the genesis and/or progression of a variety of pathological conditions, including pulmonary edema, ischemic stroke, neurodegenerative disorders, angioedema, sepsis and cancer. The altered barrier function in these conditions is often linked to the release of soluble mediators from resident cells (e.g., mast cells, macrophages) and/or recruited blood cells. The interaction of the mediators with receptors expressed on the surface of endothelial cells diminishes barrier function either by altering the expression of adhesive proteins in the inter-endothelial junctions, by altering the organization of the cytoskeleton, or both. Reactive oxygen species (ROS), proteolytic enzymes (e.g., matrix metalloproteinase, elastase), oncostatin M, and VEGF are part of a long list of mediators that have been implicated in endothelial barrier failure. In this review, we address the role of blood borne cells, including, neutrophils, lymphocytes, monocytes, and platelets, in the regulation of endothelial barrier function in health and disease. Attention is also devoted to new targets for therapeutic intervention in disease states with morbidity and mortality related to endothelial barrier dysfunction.

Recommendations for abortion surveys using the ballot-box technique / Recomendações para inquéritos sobre aborto usando a técnica de urna

The article lists recommendations for dealing with methodological aspects of an abortion survey and makes suggestions for testing and validating the survey questionnaire. The recommendations are based on the experience of the Brazilian Abortion Survey (PNA), a random sample household survey that used the ballot-box technique and covered adult women in all urban areas of the country.

O artigo lista recomendações para lidar com aspectos metodológicos de um inquérito sobre aborto e faz sugestões para testar e validar o questionário do levantamento. As recomendações baseiam-se na experiência da Pesquisa Nacional de Aborto (PNA), uma pesquisa domiciliar baseada em amostra aleatória da população urbana do Brasil que utilizou a técnica de urna.

Cirurgia por orifícios naturais transcolônica: acesso NOTES peri-retal (PNA) para excisão mesoretal total / Transcolonic natural orifice surgery: peri-rectal NOTES access (PNA) for total mesorectal excision

OBJETIVOS: Cirurgia por orifícios naturais tem sido recentemente aplicada em series clínicas para cirurgia abdominal. Apesar de potenciais vantagens do acesso NOTES transcolônico para doenças colorretais, este ainda não havia sido utilizado clinicamente. O presente trabalho descreve a primeira aplicação bem-sucedida de NOTES transcolônico da literatura, em uma nova abordagem de excisão mesoretal total (TME) para cancer de reto. MÉTODOS: Foi obtida aprovação de Comitê de Ética em Pesquisa para cirurgias por orifícios naturais, e o paciente assinou termo de consentimento informado. Em um paciente de 54 anos portador de adenocarcinoma de reto, o procedimento de retossigmoidectomia e linfadenectomia, com excisão mesoretal total foi realizada utilizando um acesso posterior transcolônico pouco acima da borda anal. A dissecção mesorretal foi conseguida utilizando um colonoscópio flexível e instrumentos endoscópicos, com assistência laparoscópica. O espécime foi retirado via transanal, e anastomose foi transorificial, com estoma proximal de proteção. RESULTADOS: O tempo operatório foi de 350 min, não ocorrendo complicações operatórias. A evolução pós-operatória foi favorável, e o paciente recebeu alta no sexto dia de pós-operatório com dieta plena. CONCLUSÃO: Este primeiro relato bem sucedido de cirurgia NOTES transcolônica traz potencialmente novas fronteiras de aplicações clínicas na cirurgia minimamente invasiva. O tratamento de doenças colorretais utilizando o novo acesso flexível PNA (Perirectal NOTES Access) é uma promissora nova abordagem, paralelamente à laparoscopia e cirurgia aberta, para melhoria do tratamento dos pacientes.

OBJECTIVES: Clinical natural orifice surgery has been applied for abdominal surgery in recent years. Despite potential advantages of transcolonic NOTES for colorectal diseases, it was since now not yet clinically applied. The study describes the first successful human application of transcolonic NOTES in the literature, in a new transrectal TME procedure for rectal cancer. METHODS: IRB approval was obtained at the institution for the study, and the patient signed informed consent. In a 54yrs-old male patient with a rectal adenocarcinoma, total mesorectal resection and rectosigmoidectomy with lymphadenectomy was performed using a posterior transcolonic access 3cm from the anal verge. Mesorectal dissection was achieved using a flexible colonoscope and endoscopic instrumentation and laparoscopic assistance. The specimen was extracted transanally, and transorificial anastomosis was performed, with proximal stoma. RESULTS: Operative time was 350 min, no intraoperative complications occurred. The postoperative course was uneventful, patient was discharged after 6 days. CONCLUSION: Successful first human report on Transcolonic NOTES potentially brings new fronteers and applications for minimally invasive surgery. The treatment of colorectal diseases through a flexible Perirectal NOTES Access (PNA) is a promising new approach besides existing laparoscopic and open surgery to improve patient care.

Estimativa de ingestão protéica de pacientes em hemodiálise comparação entre registro alimentar e equivalente protéico deaparecimento de nitrogênio (pna) / Estimation of protein intake of hemodialysis patients comparison between food record and protein equivalent ofnitrogen appearance (pna)

Introdução: A avaliação do consumo alimentar, particularmente da ingestão de proteínas, constitui uma ferramenta importante na prevenção, tratamento emonitoramento da desnutrição nos pacientes em hemodiálise. Para tanto, os métodos mais comumente utilizados na prática clínica são o registro alimentar eo equivalente protéico do aparecimento de nitrogênio (PNA). Objetivo: Avaliar a concordância entre o registro alimentar e o PNA para a estimativa da ingestãoprotéica nos pacientes em hemodiálise. Casuística e Métodos: Foram estudados 39 pacientes (67% homens, 46,8±14,7anos, IMC 23,5±4,6kg/m2) emtratamento hemodialítico. Os critérios de exclusão foram presença de doenças catabólicas, função renal residual e uso de glucocorticóides. O registro alimentarfoi preenchido durante três dias e o PNA foi calculado conforme recomendado pelo NKF-DOQI. Resultados: A ingestão protéica obtida pelo registro alimentarfoi menor que aquela obtida pelo PNA (56,3±23,4 vs 67,9±23,9g/dia, respectivamente; P≤0,001). Os métodos apresentaram uma boa correlação (r=0,54;P<0,001), porém uma grande variabilidade individual foi observada (-51,7 a +28,5g/dia). A ingestão protéica obtida por ambos os métodos correlacionou-sepositivamente com o IMC (registro alimentar: r=0,41 e PNA r=0,80; P<0,05). A diferença entre os métodos (registro alimentar - PNA) correlacionou-senegativamente com o IMC (r=-0,49; P<0,01). A análise por subgrupos de IMC mostrou que o registro alimentar subestimava a ingestão protéica em relação aoPNA apenas nos pacientes com IMC≥25kg/m2. A regressão linear múltipla apontou o IMC≥25kg/m2 como único fator independentemente associado àsvariações entre os métodos. [β =-16,9 (-31,1 a -2,8); P=0,02] Conclusão: O método do registro alimentar exibiu valores menores de proteínas em relação aométodo do PNA nos pacientes em hemodiálise, sendo a condição de sobrepeso/obesidade um importante determinante destas variações.

Introduction: The assessment of food consumption, particularly of protein intake, constitutes an important tool in the prevention, treatment, and monitoringof malnutrition in hemodialysis patients. Food record and protein equivalent of nitrogen appearance (PNA) are the most commonly used methods for suchpurpose in the clinical practice. Aim: To evaluate the agreement between food record and PNA for the assessment of protein intake in hemodialysis patients.Subjects and Methods: Thirty-nine hemodialysis patients (67% male, 46.8 ± 14.7 years old, BMI 23.5 ± 4.6 kg/m2) were studied. The exclusion criteriawere catabolic illnesses, residual renal function and use of glucocorticoids. Food intake was recorded 3 days and PNA was calculated as proposed by theNKF-DOQI. Results: The protein intake assessed by food record was lower than that obtained by PNA (56.3 ± 23.4 vs 67.9 ± 23.9 g/day, respectively;P≤0.001). The methods were well correlated (r=0.54; P<0.001), however, a large individual variability was observed (-51.7 to +28.5 g/day). The protein intakeobtained by both methods correlated positively with BMI (food record: r=0.41 and PNA: r=0.80; P<0.05). The difference between the methods (food record,PNA) correlated negatively with BMI (r=-0.49; P<0.001). The analysis by subgroups of BMI showed that the food record method underestimated proteinintake in relation to PNA only in patients with BMI ≥25kg/m2. The multiple linear regression analysis pointed to BMI ≥25kg/m2 as a factor independentlyassociated with variations between the methods. Conclusion: The food record method showed lower values of protein in comparison with the PNA methodin hemodialysis patients, and overweight/obesity status was an important determinant of these variations.

Evaluation of acrosomal status and sperm viability in fresh and cryopreserved specimens by the use of fluorescent peanut agglutinin lectin in conjunction with hypo-osmotic swelling test

OBJECTIVE: In this study, we evaluated whether the hypo-osmotic swelling test (HOST) can be used as a vital marker in combination with peanut agglutinin (PNA) - labeling in fresh and cryopreserved spermatozoa. MATERIALS AND METHODS: Human sperm populations were exposed to a hypo-osmotic medium for 60 minutes, and then incubated in a 1 µg/mL solution of the fluorescent dye Hoescht 33258 (H33258) for 10 minutes. Excess stain was removed by washing in phosphate-buffered saline (PBS) solution, and the pellet was resuspended in 100 µL of culture medium. Twenty microliters of this solution were subsequently smeared on a microscope slide, and fixed in ice-cold methanol to permeabilize the sperm membranes. The fixed smears were finally incubated in a 40-µg/mL FITC-PNA solution for 20 minutes. Simultaneous assessment of acrosome and viability scores was done in a fluorescent microscope equipped with appropriate filters and phase contrast illumination. The same slide was examined for FITC-PNA labeling, tail swelling, and for Hoechst-33258 staining by interchanging the filters and phase contrast optics. RESULTS: In fresh specimens, HOST was found to provide viability assessments comparable to those obtained using the H33258 method (r = 0.95). However, the results of HOST and H33258 were not correlated in cryopreserved specimens (r = 0.22). There was no alteration of PNA-labeling due to the HOST or H33258. CONCLUSIONS: FITC-PNA labeling in conjunction with the visualization of the morphological change induced by exposure to hypo-osmotic solution provides a simple but effective method for establishing the state of acrosomal membrane and viability in fresh human spermatozoa, but this technique is not reliable for cryopreserved ones.

Comportamiento de parámetros dialíticos y nutricionales en peritoneo diálisis pediátrica / Behaviour of dialysis and nutritional parameters in paediatric peritoneal dialysis (PD)

El manejo de pacientes pediátricos en diálisis peritoneal requiere la vigilancia continua de la dosis de diálisis y el apoyo nutricional, para lograr el mejor crecimiento y desarrollo posibles en el niño urémico. Para esto se requiere medir una serie de variables dialíticas y nutricionales que permitirán un ajuste continuo de nuestra terapia. Objetivos: 1) conocer el valor de diferentes parámetros dialíticos y nutricionales en niños en diálisis peritoneal crónica, 2) calcular la dosis de diálisis (Kt/V), la velocidad de catabolismo proteico (VCP), el Equivalente de la Aparición de Nitrógeno Ureico (PNA), el Balance nitrogenado, el test de Equilibrio Peritoneal (PET) y 3) calcular la correlación entre ellas. Pacientes y Método: Se evaluó en forma prospectiva 15 niños en diálisis peritoneal crónica, realizando 59 mediciones en 6 meses de seguimiento. Se controló mensualmente las variables clínicas: peso, talla, superficie corporal, volumen urinario y volumen de dializado de 24 hrs; se midió en plasma, orina y dializado en forma simultánea: creatinina, proteína, albúmina y nitrógeno ureico; y estudio ácido base y electrolitos en plasma. Se mantuvo un aporte proteico calórico según recomendaciones RDA (Recommended Dietary Allowances), y se calculó el Kt/Vurea peritoneal y residual mensual, el PET cada 6 meses, y la VCP, Balance Nitrogenado y PNA mensuales. Resultados: El Kt/V urea promedio residual y total fue de 1,49 ± 1 y 3,41 ± 0,86 respectivamente. Se demostró una correlación positiva entre el Kt/V y la depuración de creatinina residual (p < 0,05). El PET al inicio y fin del estudio para D/ Pcreatinina hora 4 fue 0,78 ± 0,01 y 0,74 ± 0,13 respectivamente (p = n.s.), y para el D4/D0 glucosa los valores fueron 0,35 ± 0,11 y 0,34 ± 0,08 (p = n.s.). La ingesta diaria proteica fue de 3,32 ± 1,05 y el catabolismo proteico 1,32 ± 0,47 gr/kg/día, con un Balance Nitrogenado de + 2,0 gr/kg/día, y un PNAn de 0,94 ± 0,33. El Kt/ V residual y total se correlacionaron significativamente con el PNAn, y se encontró una correlación significativa pero negativa entre ingesta proteica y bicarbonato plasmático (p < 0,05). Conclusión: Las variables estudiadas muestran un balance proteico positivo y un Kt/V superior a las recomendaciones para adultos, sugiriendo que mayor dosis de diálisis se asocia a mejores indicadores nutricionales. La correlación negativa entre bicarbonato en plasma e ingesta proteica sugiere una relación entre el aporte proteico y acidosis, lo cual requiere una cuidadosa supervisión.

The management of children on PD requires constant monitoring of the dose of dialysis and nutritional support in order to maximise growth and development in the uraemic child. The measurement of a series of dialytic and nutritional variables allow the adjustment of treatment to the childs needs. Objectives: 1) to measure the different variables in child undergoing PD, 2) calculate the dialysis dose (Kt/V), the protein catabolic rate (PCR), the urea equivalent of nitrogen appearence (PNA), the nitrogen balance, the peritoneal equilibrium test (PET) and, 3) to determine the correlation between them. Patients and Methods: A prospective study of 15 children undergoing chronic PD, with 59 serial measurements in a 6 month period. Monthly measurements of weight/height, body surface area, urine volume, dialysis volume in 24 hrs, and the values of creatine, protein, albumin and urea in plasma, urine and dialysate, and in plasma the acid-base and electrolytes. A diet containing proteins according to RDA was given. Kt/V urea peritoneal and residual, PCR, PNA and nitrogen balance were measured monthly, PET every 6 months. Results: The mean Kt/V residual and total was 1.49 ± 1 and 3.41 ± 0.86 respectively. A positive correlation between Kt/v and residual creatine clearence was found (p < 0.05). There were no significant differences between the PET for creatinin (4th hour) or glucose D4/D0 at the beginning and end of the study, 0.78 ± 0.01 and 0.74 ± 0.13 for creatinine and 0.35 ± 0.11 and 0.34 ± 0.08 respectively. Dietary protein intake was 3.32 ± 1.05, protein catabolism 1.32 ± 0.47 gm/kg/day, with a nitrogen balance of 2 gm/kg/day and a PNAn of 0.94 ± 0.33. Total and residual Kt/V was correlated with the PNAn, and a negative correlation between protein intake and plama bicarbonate was found. Conclusions: The variables studied show a positive protein balance and a Kt/V greater than the recommended for adults, suggesting that bigger dialysis doses results in a better nutritional status. The negative correlation between plasma bicarbonate and protein intake suggests a relation between protein intake and acidosis, of which needs to be carefully monitored.

Efeito de três diferentes diluidores sobre o sêmen canino submetido a dois protocolos de descongelamento / Effect of three different diluent on canine semen submitted to two thawing protocols

Com objetivo de avaliar o efeito de dois protocolos de descongelação do sêmen canino congelado em três diluidores foram coletados ejaculados de nove cães. Cada ejaculado foi dividido em 3 amostras, centrifugado e os pellets tratados sob diferentes protocolos: 1°- meio à base de glicina e gema de ovo; 2° - meio à base de TRIS e 3° - meio M9. Foram envasadas 28 palhetas de O,5ml segundo cada protocolo. Amostras tratadas sob cada protocolo foram avaliadas (M1) quanto a motilidade, vigor e integridade das membranas. As palhetas foram refrigeradas a 5°C por 60 min. e congeladas em N2. Uma amostra de cada protocolo foi avaliada quanto: motilidade, vigor e integridade das membranas após o equilíbrio (M2). Eram sempre descongeladas três pares de palhetas, sendo que cada par havia sido congelado sob um mesmo protocolo. Uma das palhetas de cada par era descongelada a 37°C por 30 segundos e outra a 72°C por 8 segundos. Cada grupo foi avaliado após o descongelamento (M3) quanto a motilidade e vigor espermático, avaliação computadorizada do movimento, integridade das membranas e avaliação acrossomal por meio de FitcPNA e PI. As amostras foram mantidas em banho Maria a 37°C por 1h para o teste de longevidade espermática e reavaliadas (M4) quanto à motilidade e vigor espermáticos, integridade das membranas. Após análise estatística concluímos que as células espermáticas caninas descongeladas a 72°C por 8 segundos apresentaram um maior somatório de bons resultados quanto aos testes de viabilidade espermática in vitro empregados.(AU)

To verify the effect of three different extenders and two thawingtemperatures on frozen-thawed canine sperm characteristics, oneejaculate from nine dogs were separately collected and processed (n=9).Each ejaculate was divided into 3 equal samples and centrifuged. Thepellets were re-suspended using: 1st pellet Glicyne-egg yolk extender(GEY), 2nd pellet - TRIS extender and 3rd pellet - M9 extender and allsamples were filled in 0.5ml straws. A total of 84 straws (28 for eachprotocol) were done. The sperm motility, vigour and plasmamembrane integrity from each protocol were immediately evaluated(M1) and the straws were brought to a refrigerator at 5ºC for 60minutes. After that, a sample from each protocol was warmed up ina water bath 37ºC for 5 min. and sperm motility, vigour and plasmamembrane integrity were evaluated (M2). The straws were frozen inliquid nitrogen. One straw from each protocol was thawed at 37ºCfor 30 sec and another at 72ºC for 8 sec and the sperm motility;vigour, CASA and plasmatic membrane integrity and acrossomalstatus using FITC-PNA stain were evaluated (M3). Plasma membrane,sperm motility and vigour were evaluated after a 1-hour incubation at37ºC (M4). Statistical analysis showed that the higher temperaturehad positive effect on froze-thawed canine sperm characteristics. Thebest results were taken when canine semen was frozen in GEYextender and thawed 72ºC for 8 sec. (AU)