Effectiveness of tes-tris or tris association with low density lipoprotein on in vitro longevity of refrigerated buffalo semen / Eficácia da associação TES-TRIS ou TRIS com lipoproteína de baixa densidade sobre a longevidade in vitro de sêmen refrigerado de búfalos

This study investigated in vitro the efficacy of four different extenders (TES-TRIS and TRIS with LDL low-density lipoprotein at concentrations of 10 or 5%) on the longevity of buffalo sperm in the refrigeration process at 5ºC. Sperm motility was assessed every 24 hours up to 72 hours of incubation using computer assisted sperm analysis and sperm membrane integrity was examined by the hypoosmotic test (HOST) at T1, T24, T48 and T72 hours. Eleven buffaloes (1 ejaculate per buffalo) of the Murrah breed were used, ranging in age from 4 to 5 years. Immediately after collection, each ejaculate was fractionated into 4 aliquots, and each aliquot was diluted in one of four diluents to obtain 50x106SPTZ/mL. The samples were packed in 0.5mL straws and refrigerated (-0.25°C/min) to 5°C and maintained at this temperature until evaluation. Prior to evaluation the samples were heated at 37°C for 30 seconds. The statistical package used for analysis was STATA 12.0 "Statistical Analysis Software" and means were compared by the Friedman test (P<0.05). The results of sperm kinetics and HOST indicate that the TRIS diluent with 10% LDL could be a promising alternative for semen refrigeration at 5ºC, to be used in conventional and fixed time artificial insemination.(AU)

Este estudo investigou in vitro a eficácia de quatro diferentes extensores (TES-TRIS e TRIS com lipoproteína de baixa densidade - LDL, nas concentrações de 10 ou 5%) sobre a longevidade espermática de búfalos no processo de refrigeração a 5ºC. A motilidade espermática foi avaliada a cada 24 horas até 72 horas de incubação, por sistema computadorizado "CASA", e a integridade de membrana espermática foi examinada pelo teste hiposmótico (HOST) em T1, T24, T48 e T72 horas. Foram utilizados 11 búfalos (um ejaculado por búfalo) da raça Murrah, com idade variando de quatro a cinco anos. Imediatamente após a coleta, cada ejaculado foi fracionado em quatro alíquotas, e cada alíquota foi diluída em um dos quatro diluidores para a obtenção de 50x106 SPTZ/mL. As amostras foram envasadas em palhetas de 0,5 mL, refrigeradas (-0,25oC/minuto) até 5oC e mantidas nessa temperatura até a avaliação. Previamente à avaliação, as amostras foram aquecidas a 37oC por 30 segundos. O pacote estatístico utilizado para as análises foi o STATA 12.0 "Statistical Analysis Software", e as médias foram comparadas pelo teste de Friedman (P<0,05). Os resultados de cinética e HOST até o tempo de 48 horas indicam que o diluidor TRIS com 10% LDL seria uma alternativa promissora para a refrigeração do sêmen a 5ºC, a ser utilizado na inseminação artificial e na inseminação artificial em tempo fixo.(AU)

Dilución isotópica con deuterio para determinar ingesta de leche humana y composición corporal materna / Deuterium-oxide turnover technique to assess breast milk intake and maternal body composition / Diluição isotópica com deutério para determinar a ingestão de leite humano e a composição corporal materna

El objetivo del trabajo fue describir la aplicación de la técnica de dilución isotópica con deuterio de dosis a la madre para determinar la ingesta de leche materna y la composición corporal de las madres, en distintos tipos de lactancia. El método analítico se aplicó en cuatro casos modelo de pares madre-lactante en los cuales las madres recibieron una dosis oral de agua deuterada, recolectándose 6 muestras de saliva de ambos durante 15 días. El enriquecimiento de deuterio se determinó en un espectrómetro FTIR-Shimadzu-Affinity obteniéndose la ingesta de leche materna (ILM) y de agua de otras fuentes (Fd). Se observó una variación del enriquecimiento de deuterio en la saliva del lactante, asociada al tipo de lactancia recibida, siendo mayor en el caso de lactancia materna exclusiva (LME). Asimismo, a medida que aumentó Fd, disminuyó ILM. Además, fueron determinadas el agua corporal, la masa libre de grasa y la masa grasa materna. La transferencia de las habilidades técnicas y del conocimiento a través de metodologías innovadoras para determinar la ingesta de leche materna es de utilidad como herramienta de evaluación de la alimentación del lactante y para investigar en qué medida la lactancia natural es reemplazada por la ingesta de otros alimentos. Mejorar la estimación de la LME contribuye al conocimiento de la recomendación de OMS y UNICEF de mantener la misma hasta el sexto mes de vida.

The aim of this study was to describe the application of the dose-to-the-mother deuterium-oxide turnover technique to determine the breast milk intake and body composition of mothers in different types of breastfeeding. This analytical method was performed in four mother-infant pairs at 4 months from birth. Mothers received an oral dose of deuterated water, collecting 6 samples of saliva from mother and baby during a period of 15 days. Deuterium enrichment was determined in a Shimadzu FTIR-spectrometer-Affinity to obtain the intake of breast milk and water from non-breast milk sources. In this study, a variation of the enrichment of deuterium in the saliva of the infant was observed, being higher when the infant was exclusively breastfed. As non-breast milk water increased, the intake of human milk decreased. Furthermore, maternal total body water, fat free mass and fat mass were determined. To improve technical skills and knowledge through innovative methods of breast milk measurement can be useful as an assessment tool for evaluating infant feeding and investigating the extent to which breast milk is being replaced by the consumption of other foods in order to estimate exclusive breastfeeding in the future. This would contribute to the knowledge of maintaining breastfeeding until the sixth month of life, as it is recommended by WHO and UNICEF.

O objetivo deste estudo foi descrever a aplicação da técnica de diluição isotópica com deutério de dose à mãe para determinar a ingestão de leite materno e a composição corporal das mães, em diferentes tipos de aleitamento. O método analítico foi aplicado em quatro casos modelo de pares mães-lactante nos quais as mães receberam uma dose oral de água deuterada, coletando-se 6 amostras de saliva de ambos (mães e lactantes) durante 15 días. O enriquecimento de deutério foi determinado em um espectrômetro FTIR-Shimadzu-Affinity, sendo obtida a ingestão de leite materno (ILM) e de água proveniente de outras fontes (Fd). Observou-se uma variação do enriquecimento de deutério na saliva do lactante, associada ao tipo de aleitamento recebido, sendo maior no caso de aleitamento materno exclusivo (AME). Também, na medida que aumentou Fd, diminuiu ILM. Além disso, a água corporal, a massa livre de gordura y a massa gorda materna foram determinadas. A transferência das habilidades técnicas e do conhecimento através de metodologias inovadoras para determinar a ingestão de leite materno é de utilidade como ferramenta de avaliação da alimentação do lactante e para investigar em que medida o aleitamento natural é substituído pela ingestão de outros alimentos. Melhorar a avaliação do AME contribui ao conhecimento da recomendação da OMS e UNICEF de manter a mesma até o sexto mês de vida.

Deuterium oxide dilution and body composition in overweight and obese schoolchildren aged 6-9 years / Diluição do óxido de deutério e composição corporal em escolares de seis a nove anos com sobrepeso e obesidade

ABSTRACT OBJECTIVE: To correlate different methods of body composition assessment in overweight or obese schoolchildren, using deuterium oxide (D2O) dilution as a reference. METHODS: Percentage of total body water (%TBW), fat free mass (%FFM), and body fat (%BF) were assessed by D2O and tetrapolar electrical bioimpedance analysis (BIA) in 54 obese and overweight students aged 6-9 years. Skinfold thickness (ST), body mass index (BMI), conicity index (CI), waist circumference (WC), waist-to-height ratio (WHtR), and waist-to-hip (WHR) ratio were also used. RESULTS: Mean values for body composition were 38.4% ± 8.4% BF, 44.9% ± 6.1% TBW and 61.6% ± 8.4% FFM. There was no significant difference in body weight, body fat mass (FM), TBW, and FFM between genders. Regarding D2O, ST underestimated %BF, and overestimated %FFM in both genders (p < 0.05). BIA overestimated %TBW in the group as a whole and in males (p < 0.05). The only positive and strong correlations occurred in females regarding the WC (s= 0.679), CI (r = 0.634), and WHtR (r = 0.666). CONCLUSIONS: In this sample of obese and overweight children, there were strong correlations between body composition measured by D2O and some indices and anthropometric indicators in females, but there was no positive and strong correlation of fat tissue with the indices/indicators at all ages and in both genders.

RESUMO OBJETIVO: Correlacionar diferentes métodos para avaliação de composição corporal em escolares diagnosticados com sobrepeso e obesos com o uso como referência da diluição de óxido de deutério (D2O). MÉTODOS: O percentual de água corporal total (%ACT), massa livre de gordura (%MLG) e gordura corporal (%GC) foi obtido pelo D2O e pela bioimpedância elétrica tetrapolar (BIA), em 54 estudantes com sobrepeso e obesos, entre seis-nove anos. O método das dobras cutâneas (DC) com o uso de triciptal e panturrilha, índice de massa corporal (IMC), índice de conicidade (IC), circunferência de cintura (CC), relação cintura/estatura (RCE) e relação cintura/quadril (RCQ) também foi usado. RESULTADOS: Os valores médios para composição corporal aferidos pelo D2O foram 38,4 ± 8,4%GC, 44,9 ± 6,1%ACT e 61,6 ± 8,4%MLG. Não houve diferença significativa entre peso corporal, massa corporal de gordura (MG), ACT e MLG entre os sexos. Considerando o D2O, DC subestimou o %GC e superestimou o %MLG em ambos os sexos (p < 0,05); BIA superestimou %ACT no grupo como um todo e no masculino (p < 0,05). As únicas correlações fortes e positivas ocorreram no grupo feminino nas variáveis CC (s = 0,679), IC (r = 0,634) e RCE (r = 0,666). CONCLUSÕES: Nessa amostra de crianças obesas e com sobrepeso, houve fortes correlações entre a composição corporal mensurada pelo D2O e alguns índices e indicadores antropométricos nas meninas, mas nenhuma correlação forte e positiva do tecido adiposo foi encontrada com os índices/indicadores em todas as idades e ambos os sexos.

Validation of the Korean coefficient for the modification of diet in renal disease study equation

BACKGROUND/AIMS: Race and ethnicity are important determinants when estimatingglomerular filtration rate (GFR). The Korean coefficients for the isotope dilution mass spectrometry (IDMS) Modification of Diet in Renal Disease (MDRD) Study equations were developed in 2010. However, the coefficients have not been validated. The aim of this study was to validate the performance of the Korean coefficients for the IDMS MDRD Study equations. METHODS: Equation development and validation were performed in separate groups (development group, n = 147 from 2008 to 2009; validation group, n = 125 from 2010 to 2012). We compared the performance of the original IDMS MDRD equations and modified equations with Korean coefficients. Performance was assessed by comparing correlation coefficients, bias, and accuracy between estimated GFR and measured GFR, with systemic inulin clearance using a single injection method. RESULTS: The Korean coefficients for the IDMS MDRD equations developed previously showed good performance in the validation group. The new Korean coefficients for the four- and six-variable IDMS MDRD equations using both the development and validation cohorts were 1.02046 and 0.97300, respectively. No significant difference was detected for the new Korean coefficients, in terms of estimating GFR, between the original and modified IDMS MDRD Study equations. CONCLUSIONS: The modified equations with Korean coefficients for the IDMS MDRD Study equations were not superior to the original equations for estimating GFR. Therefore, we recommend using the original IDMS MDRD Study equation without ethnic adjustment in the Korean population.

Acupressure versus dilution of fentanyl to reduce incidence of fentanyl-induced cough in female cancer patients: a prospective randomized controlled study / 대한마취과학회지

BACKGROUND: Fentanyl-induced cough (FIC) is a transient condition with a reported incidence of 18% to 65% depending on the dose and route of administration of fentanyl. Nonpharmacological methods to prevent FIC are more cost-effective than medications. Dilution of fentanyl has a proven role in the prevention of FIC. Acupressure can also prevent FIC because it has a proven role in the treatment of cough. METHODS: This study included 225 female patients with an American Society of Anesthesiologists physical status of I or II who were randomly divided into 3 groups of 75 patients each. Patients in the control group received undiluted fentanyl at 3 µg/kg, patients in the acupressure group received undiluted fentanyl at 3 µg/kg with acupressure, and patients in the dilution group received diluted fentanyl at 3 µg/kg. Coughing was noted within 2 min of fentanyl administration. The severity of FIC was graded as mild (1-2 coughs), moderate (3-4 coughs), or severe (≥5 coughs). The timing of coughs was also noted. RESULTS: The incidence of FIC was 12.7% in the control group, 6.8% in the dilution group, and 1.3% in the acupressure group. The difference in the incidence of cough was statistically significant (P = 0.008) between the control and acupressure groups. The difference in the severity of cough among the groups was not statistically significant. The median onset time of cough among all groups was 9 to 12 seconds. CONCLUSIONS: The application of acupressure prior to administration of fentanyl significantly reduces the incidence of FIC. Dilution of fentanyl also reduces the incidence of FIC, but the difference is not statistically significant.

Autoantibodies with Mimicking Specificity Detected by the Dilution Technique in Patients with Warm Autoantibodies

BACKGROUND: The aim of this study was to investigate the frequency of autoantibodies with mimicking specificity by using the dilution technique, to assess the usefulness of the combination of the dilution technique and red blood cell (RBC) phenotyping, and to establish a pre-transfusion testing algorithm in patients with warm autoantibodies. METHODS: Serum samples from 71 patients with warm autoantibodies were tested using the dilution technique. Among them, 25 samples were adsorbed with allogeneic ZZAP (a combination of dithiothreitol and enzyme) or polyethylene glycol (PEG) and their RBC phenotypes were determined. Thirty-nine patients were transfused with our pre-transfusion testing algorithm using a combination of dilution technique and RBC phenotyping. RESULTS: Autoantibodies with mimicking specificity were detected by the dilution technique in 26.8% (19/71) of the patients and most of them were directed against Rh system antigens. The agreement of the results obtained with the dilution technique in combination with RBC phenotyping and those from ZZAP or PEG adsorption was 100% (18/18) in patients who have autoantibodies with mimicking specificity and/or alloantibodies. No clinical symptoms indicating severe acute or delayed hemolytic transfusion reactions were reported in the 39 patients transfused with our pre-transfusion testing algorithm. CONCLUSIONS: Autoantibodies with mimicking specificity detected by the dilution technique in patients with warm autoantibodies are relatively frequent, can be discriminated from alloantibodies by employing a combination of dilution technique and RBC phenotyping, and might not appear to cause severe acute or delayed hemolytic transfusion reactions.

Development and Evaluation of a Laboratory Information System-Based Auto-Dilution and Manual Dilution Algorithm for Alpha-Fetoprotein Assay

No abstract available.

Body composition measures of obese adolescents by the deuterium oxide dilution method and by bioelectrical impedance

The objectives of the present study were to describe and compare the body composition variables determined by bioelectrical impedance (BIA) and the deuterium dilution method (DDM), to identify possible correlations and agreement between the two methods, and to construct a linear regression model including anthropometric measures. Obese adolescents were evaluated by anthropometric measures, and body composition was assessed by BIA and DDM. Forty obese adolescents were included in the study. Comparison of the mean values for the following variables: fat body mass (FM; kg), fat-free mass (FFM; kg), and total body water (TBW; percent) determined by DDM and by BIA revealed significant differences. BIA overestimated FFM and TBW and underestimated FM. When compared with data provided by DDM, the BIA data presented a significant correlation with FFM (r = 0.89; P < 0.001), FM (r = 0.93; P < 0.001) and TBW (r = 0.62; P < 0.001). The Bland-Altman plot showed no agreement for FFM, FM or TBW between data provided by BIA and DDM. The linear regression models proposed in our study with respect to FFM, FM, and TBW were well adjusted. FFM obtained by DDM = 0.842 x FFM obtained by BIA. FM obtained by DDM = 0.855 x FM obtained by BIA + 0.152 x weight (kg). TBW obtained by DDM = 0.813 x TBW obtained by BIA. The body composition results of obese adolescents determined by DDM can be predicted by using the measures provided by BIA through a regression equation.

Prevalence of mutans streptococci isolated from complete dentures and their susceptibility to mouthrinses

The aims of this study were to evaluate the incidence of mutans streptococci (MS - sessile form) on complete maxillary dentures after use of a specific denture paste, and to determine the minimum inhibitory concentration (MIC) and maximum inhibitory dilution (MID) of 3 oral mouthrinses: Cepacol, Plax and Periogard. Seventy-seven complete denture wearers were randomly assigned into 2 groups, according to the product used for denture cleaning: Control group - conventional dentifrice (Kolynos-Super White); and Test group: experimental denture cleaning paste. Denture biofilm was collected at baseline and after 90 and 180 days after treatment by brushing the dentures with saline solution. After decimal serial dilution, samples were seeded onto agar sucrose bacitracin to count colonies with morphological characteristics of MS. MS identification was performed by the sugar fermentation tests. After this procedure, brain heart infusion broth (BHI) was added to oral mouthrinses (Plax, Cepacol e Periogard) and seeded on Petri dishes. The colonies were seeded using the Steers multiplier and, after the incubation, the MIC and MID of the mouthrinses were calculated. The results showed an incidence of 74.0 percent (n=57) of MS in the 77 complete dentures examined in the study, being 76.3 percent (n=29) of the Control group (conventional dentifrice) and 71.8 percent (28) of the Test group (experimental denture cleaning paste). In both groups, the number of positive cases for MS decreased from day 0 to day 180. In the Test group there was a slight decrease in the incidence of Streptococcus mutans 90 days after use of the experimental denture cleaning paste, which was not observed in the Control group. As regards to mouthrinses, for both groups, Periogard showed antimicrobial action with the highest dilution, followed by Cepacol and Plax. In conclusion, the incidence of MS in complete dentures was high and Periogard was the mouthrinse with the strongest antimicrobial action against MS. The experimental denture cleaning paste showed a slight action against S. mutans after 90 days of treatment.

O objetivo deste estudo foi avaliar a incidência de estreptococos do grupo mutans (forma séssil) de próteses totais superiores após o uso de uma pasta específica para higienização de próteses totais, bem como determinar a concentração inibitória mínima (CIM) e diluição inibitória máxima (DIM) de 3 enxaguatórios bucais: Cepacol, Plax e Periogard. Setenta e sete usuários de próteses totais foram distribuídos em 2 grupos: (A) grupo Controle: dentifrício convencional (Kolynos Super Branco) e (B) grupo Teste: pasta experimental para higiene de próteses (1). O biofilme das próteses foi coletado no início e após 90 e 180 dias dos tratamentos, por meio de escovação das próteses com solução salina. Após a diluição decimal seriada, as amostras foram semeadas em agar sacarose bacitracina para a contagem de colônias características de estreptococos do grupo mutans. A identificação dos estreptococos do grupo mutans foi realizada por meio de testes de fermentação de açúcares. Após este procedimento, o caldo de infusão de cérebro e coração (BHI) foi adicionado aos antissépticos (Plax, Cepacol e Periogard) e semeado em placas de Petri. As colônias foram semeadas usando o mutiplicador de Steers e após a incubação, a concentração inibitória mínima e a concentração inibitória máxima dos enxaguatórios foram calculadas. Os resultados mostraram que a incidência de estreptococos do grupo mutans nas 77 próteses totais examinadas foi de 74,0 por cento (n=57), sendo 76,3 por cento (n=29) do grupo Controle (pasta convencional) e 71,8 por cento (n=28) do grupo Teste (pasta experimental). Em ambos os grupos, o número de casos positivos para estreptococos do grupo mutans diminuiu, do início (0 dias) para 180 dias. Houve uma pequena diminuição da incidência de Streptococcus mutans após 90 dias do uso da pasta experimental, o que não foi observado no grupo controle. Com relação aos enxaguatórios, para ambos os grupos, o Periogard apresentou atividade antimicrobiana com diluições maiores, seguido do Cepacol e Plax. Pôde-se concluir que a incidência de estreptococos do grupo mutans em próteses totais foi elevada e que o Periogard foi o enxaguatório com melhor ação antimicrobiana contra os estreptococos do grupo mutans. A pasta experimental mostrou uma pequena ação contra S. mutans após 90 dias de tratamento.

Hemodilución normovolémica inducida: mejoría de los valores hemodinámicos y gasométricos en pacientes con claudicación a la marcha / Induced normovolemic hemodilution: improvement of the hemodynamic and gasometric values in patients with walk claudication

Se realizó un estudio de casos y controles de 49 pacientes con claudicación intermitente a la marcha, atendidos en la consulta de hemodilución del Hospital Provincial Docente Dr Joaquín Castillo Duany de Santiago de Cuba en un quinquenio, a fin de identificar los cambios observados en la hiperemia y gasometría de la población investigada. Los integrantes de la casuística fueron asignados a uno de 2 grupos: los tratados con hemodilución normovolémica inducida (grupo de estudio) y los que recibieron tratamiento convencional con vasodilatadores antiagregantes y ejercicio físico (grupo control), teniendo en cuenta la presencia de factores de riesgo en ambos. Se concluyó que los primeros mejoraron las condiciones hemodinámicas y de oxigenación, dadas por el aumento del índice de amplitud y de la relación consumo de oxígeno e índice de flujo eritrocitario.

A case-control study of 49 patients with intermittent walk claudication, assisted at the hemodilution service of Dr Joaquín Castillo Duany Teaching Provincial Hospital in Santiago de Cuba was carried out in a five year period, in order to identify the changes observed in the hyperemia and gasometry of the investigated population. The case material was assigned to one of 2 groups: the patients treated with induced normovolemic hemodilution (study group), and those who received conventional treatment with antiplatelet vasodilators and physical exercise (control group), keeping in mind the presence of risk factors for both. It was concluded that the first ones improved hemodynamics and oxygenation conditions, given by the increase of the width index and of the relationship between oxygen consumption and erythocytes flow index.

Development and validation of a liquid chromatography-isotope dilution tandem mass spectrometry for determination of olanzapine in human plasma and its application to bioavailability study / 药学学报

A simple, reliable and sensitive liquid chromatography-isotope dilution mass spectrometry (LC-ID/MS) was developed and validated for quantification of olanzapine in human plasma. Plasma samples (50 microL) were extracted with tert-butyl methyl ether and isotope-labeled internal standard (olanzapine-D3) was used. The chromatographic separation was performed on XBridge Shield RP 18 (100 mm x 2.1 mm, 3.5 microm, Waters). An isocratic program was used at a flow rate of 0.4 m x min(-1) with mobile phase consisting of acetonitrile and ammonium buffer (pH 8). The protonated ions of analytes were detected in positive ionization by multiple reactions monitoring (MRM) mode. The plasma method, with a lower limit of quantification (LLOQ) of 0.1 ng x mL(-1), demonstrated good linearity over a range of 0.1 - 30 ng x mL(-1) of olanzapine. Specificity, linearity, accuracy, precision, recovery, matrix effect and stability were evaluated during method validation. The validated method was successfully applied to analyzing human plasma samples in bioavailability study.

Early Vascular Access Blood Flow as a Predictor of Long-term Vascular Access Patency in Incident Hemodialysis Patients

The long-term clinical benefits of vascular access blood flow (VABF) measurements in hemodialysis (HD) patients have been controversial. We evaluated whether early VABF may predict long-term vascular access (VA) patency in incident HD patients. We enrolled 57 patients, of whom 27 were starting HD with arteriovenous fistulas (AVFs) and 30 with arteriovenous grafts (AVGs). The patients' VABF was measured monthly with the ultrasound dilution technique over the course of the first six months after the VA operation. During the 20.4-month observational period, a total of 40 VA events in 23 patients were documented. The new VA events included 13 cases of stenosis and 10 thrombotic events. The lowest quartile of average early VABF was related to the new VA events. After adjusting for covariates such as gender, age, hypertension, diabetes, VA type, hemoglobin levels, body mass index, parathyroid hormone, and calcium-phosphorus product levels, the hazard ratio of VABF (defined as <853 mL/min in AVF or <830 mL/min in AVG) to incident VA was 3.077 (95% confidence interval, 1.127-8.395; P=0.028). There were no significant relationships between early VABF parameters and VA thrombosis. It is concluded that early VABF may predict long-term VA patency, particularly VA stenosis.

Auto-antibody Showing Anti-Fy(b) Specificity as Proven by the Dilution Method in the Presence of Warm Autoantibodies: A Case Report / 대한수혈학회지

Several approaches have been introduced to detect allo-antibodies in the presence of warm auto-antibodies, and these methods include warm autoadsorption, cysteine-activated papain and dithiothreitol (ZZAP), and polyethylene glycol (PEG) and dilution of the patient's serum. Among them, the dilution technique is a simple and rapid method. During pretransfusion testing of a 33 year-old systemic lupus erythematosus (SLE) patient with warm auto-antibodies, antibody identification was done by the dilution technique with using serum diluted 1-in-8. The patient demonstrated an anti-Fy(b) pattern of reactivity in his sera. Contrary to our expectations, the phenotype of the erythrocytes was Fy(a+/b+) and the genotype, as assessed by performing PCR-restriction fragment length polymorphism (RFLP), was FY*A/FY*B. These results suggest that the antibody is an autoantibody showing anti-Fy(b) specificities. An antibody identification test using undiluted serum showed the same result when 40 days had passed. We report here on a case with auto-anti-Fy(b) proven by the dilution method in the presence of warm autoantibodies.

Análise TG-ROC de testes de imunofluorescência no diagnóstico de leishmaniose visceral canina / TG-ROC analysis of immunofluorescence assays in canine visceral leishmaniasis diagnosis

OBJETIVO: Analisar a acurácia do diagnóstico de dois protocolos de imunofluorescência indireta para leishmaniose visceral canina. MÉTODOS: Cães provenientes de inquérito soroepidemiológico realizado em área endêmica nos municípios de Araçatuba e de Andradina, na região noroeste do estado de São Paulo, em 2003, e área não endêmica da região metropolitana de São Paulo, foram utilizados para avaliar comparativamente dois protocolos da reação de imunofluorescência indireta (RIFI) para leishmaniose: um utilizando antígeno heterólogo Leishmania major (RIFI-BM) e outro utilizando antígeno homólogo Leishmania chagasi (RIFI-CH). Para estimar acurácia utilizou-se a análise two-graph receiver operating characteristic (TG-ROC). A análise TG-ROC comparou as leituras da diluição 1:20 do antígeno homólogo (RIFI-CH), consideradas como teste referência, com as diluições da RIFI-BM (antígeno heterólogo). RESULTADOS: A diluição 1:20 do teste RIFI-CH apresentou o melhor coeficiente de contingência (0,755) e a maior força de associação entre as duas variáveis estudadas (qui-quadrado=124,3), sendo considerada a diluição-referência do teste nas comparações com as diferentes diluições do teste RIFI-BM. Os melhores resultados do RIFI-BM foram obtidos na diluição 1:40, com melhor coeficiente de contingência (0,680) e maior força de associação (qui-quadrado=80,8). Com a mudança do ponto de corte sugerido nesta análise para a diluição 1:40 da RIFI-BM, o valor do parâmetro especificidade aumentou de 57,5 por cento para 97,7 por cento, embora a diluição 1:80 tivesse apresentado a melhor estimativa para sensibilidade (80,2 por cento) com o novo ponto de corte. CONCLUSÕES: A análise TG-ROC pode fornecer importantes informações sobre os testes de diagnósticos, além de apresentar sugestões sobre pontos de cortes que podem melhorar as estimativas de sensibilidade e especificidade do teste, e avaliá-los a luz do melhor custo-benefício.

OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis. METHODS: Dogs from the seroepidemiological survey conducted in an endemic area of the cities of Araçatuba and Andradina, in Northwestern São Paulo state, in 2003, and in a non-endemic area of the metropolitan region of São Paulo, were used to assess two protocols of indirect immunofluorescence assay (IFA) for leishmaniasis: one using a Leishmania major heterologous antigen (IFA-BM) and another using a Leishmania chagasi homologous antigen (IFA-CH). Two-graph receiver operating characteristic (TG-ROC) analysis was used to estimate accuracy. TG-ROC analysis compared 1:20 dilution readings of the homologous antigen (IFA-CH), considered as reference test, with IFA-BM dilutions (heterologous antigen). RESULTS: The 1:20 dilution used in the IFA-CH test showed the best contingency coefficient (0.755) and the highest strength of association between the two variables studied (chi-square=124.3). Thus, it was considered the test reference dilution in comparisons with different IFA-BM test dilutions. The best IFA-BM results were obtained from 1:40 dilutions with the best contingency coefficient (0.680) and highest strength of association (chi-square=80.8). With the change in the cut-off point, recommended for the IFA-BM 1:40 dilution in this analysis, the specificity parameter value rose from 57.5 percent to 97.7 percent, even though the 1:80 dilution showed the best sensitivity estimate (80.2 percent), with the new cut-off point. CONCLUSIONS: TG-ROC analysis can provide important information about diagnostic tests, in addition to offering suggestions on cut-off points that can improve test sensitivity and specificity estimates and assessing these tests in terms of the best cost-benefit ratio.

In vitro interference of Hyptis martiusii Benth. & chlorpromazine against an aminoglycoside - resistant Escherichia coli.

The antibacterial and synergistic activity of the ethanol extract from Hyptis martiusii Benth. was assayed by microdillution. The growth of two isolates of Escherichia coli tested was inhibited by the extract. The minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) values ranged from 512 and >1024 μg/ml for the E. coli 27 and 1024 and > 1024 μg/ml for the E. coli ATCC8539, respectively. A synergism between this extract and all aminoglycosides assayed was demonstrated. In the same form synergism between chlorpromazine and kanamycin, amikacin and tobramycin was observed, indicating the involvement of an effl ux system. Extracts from H. martiusii could be used as a source of plant derived natural products with modifying antibiotic activity and these products may interact and affect multidrug resistance systems (MDR) as efflux pumps.

Efeito da centrifugação sobre a qualidade do sêmen canino / Effect of centrifugation on quality of canine semen

Avaliou-se o efeito da centrifugação sobre a viabilidade do sêmen canino e compararam-se três meios de diluição pré-centrifugação. Utilizaram-se 10 ejaculados completos de 10 cães que, após a avaliação inicial, foram divididos em quatro porções (grupos). Uma das amostras, não centrifugada, formou o grupo-controle; as outras foram diluídas em três diferentes meios e centrifugadas a 800 x g por 15 minutos, formando os grupos: CPSA - constituído por sêmen centrifugado em plasma seminal autólogo; CLG - sêmen centrifugado em meio à base de leite desnatado e glicose (LG); e CPer- sêmen centrifugado em gradientes de Percoll (45 por cento e 90 por cento). Após a centrifugação e a eliminação do sobrenadante, procedeu-se à ressuspensão de todas as porções do ejaculado em LG e à imediata avaliação quanto à motilidade, vigor, aglutinação espermática e integridade das membranas espermáticas. Todas as suspensões foram, então, incubadas a 37ºC por 30 minutos e reavaliadas. O processo de centrifugação não causou danos aos espermatozoides e a centrifugação em meio LG melhorou a viabilidade espermática.

The effect of the centrifugation process on canine sperm viability was evaluated using three different precentrifugation extenders in the process. After an initial evaluation, ten complete ejaculates from ten dogs were used and subdivided in four groups. One sample of semen was not centrifuged and was used as control and the remaining samples of semen were diluted in three different extenders and centrifuged at 800 x g per 15 minutes, performing groups: CPSA- centrifuged in autologous seminal fluid, CLGcentrifuged in skim milk plus glucose extender (LG), and CPer- centrifuged under Percoll gradient (45 percent and 90 percent). After centrifugation, the resulting pellets were diluted in LG and evaluated for motility, viability, sperm agglutination, and spermatic membrane integrity. All the samples were incubated at 37ºC for 30 minutes and the evaluations were performed again. Centrifugation procedures did not induce damage to canine spermatozoa and samples centrifuged and diluted in skim milk plus glucose extender remained with better viability.

Ultradiluição homeopática de triiodotironina altera a apoptose celular da cauda de girinos de Rana catesbeiana: in vitro / Ultra high dilution of triiodothyronine modifies the cellular apoptosis of Rana catesbeiana tadpole tail: in vitro

Ultradiluição (UHD) é o efeito de uma solução, diluída acima do número de Avogrado, que na dependência da sua dinamização (diluição com sucussão) induz um efeito celular supressivo ou estimulante, com conseqüente obtenção de uma curva dose-efeito oscilatória. Por outro lado, a 3,3,5 Triiodo-L-Tironina (T3) é o hormônio mais importante na indução e manutenção das mudanças metamórficas dos girinos, nelas incluídas a absorção da cauda. O presente estudo, cego e randomizado, tem como objetivo comprovar que o T3 5.10-24M (10ª cH) altera a apoptose induzida pelo T3 100 nM na cauda de girinos de Rana catesbeiana, in vitro. Foram distribuídos 60 explantes em três grupos: Grupo A: sem o estímulo do T3 em dose farmacológica e em UHD; Grupo B (teste): sob a ação de T3 100 nM e T3 10ª cH (5.10-24 M); Grupo C (controle): sob a ação do T3 100 nM e etanol 70% sem sucussão. A análise estatística da área dos explantes, no primeiro e ultimo dia do experimento, e do índice apoptótico foi realizado através do teste t Student e foi considerado estatisticamente significante quando p<0,05. Embora sem diferenças significativas na área dos explantes do grupo teste e no grupo controle, um maior e significante índice apoptótico foi identificado nos explantes do grupo teste. Este resultado confirma que o T3 na 10ª cH altera a ação do T3 em dose farmacológica. Futuros experimentos serão realizados, com diferentes dinamizações, com o objetivo da parametrização da curva dose-efeito.

Ultra High Dilution (UHD) is the effect of a solution, beyond the Avogadro limits, that in the dependence of the applied dinamization (dilution with succussion) elicits a suppressive or a stimulant effect on a living cell, with a consequent generation of an oscillatory dose-effect curve. The entire process of anuran amphibian metamorphosis is under thyroid hormones control, included the complete resorption of the tadpole tail. A random and blind study was performed, with the intent to prove that T3 5.10-24 M (10ª cH) modifies the apoptosis induction of T3 100 nM in Rana catesbeiana tadpoles tail tips, in vitro. 60 Explants were distributed in three ways: Group A: without T3 action, at pharmacological and UHD dose; Group B (test): under the action of T3 100 nM and treated with T3 10ª cH (UHD); Group C (control): under the action of T3 100 nM and treated with ethanol 70% unsuccussed. In order to identify significant differences in the area of the remainder explants, at the first and final day of the experiment, and in the apoptotic index we used a student t-test. Although we didnt find statistical difference in macroscopic tadpoles tail tips area from test and control groups, a high and significant (p<0,05) index of apoptosis in histology was found in explants of test group. This data confirms that T3 10 cH modifies the effect of T3 at pharmacological dose. More studies will be necessary, using different dinamizations, to the parameterization of the dose-effect curve proceeding from these experiments.

Efeito do glicerol, etilenoglicol, acetamida e leite desnatado na criopreservação de espermatozóides eqüinos / Effects of glycerol, ethylene glycol, acetamide, and dried skim milk in cryopreservation of equine sperm

Avaliou-se a eficácia de diferentes associações de crioprotetores no congelamento de sêmen eqüino. Foram utilizados três ejaculados de oito garanhões para testar o diluidor lactose-EDTA-gema de ovo com as seguintes associações de macromoléculas e crioprotetores: T1 - glicerol 5 por cento (controle); T2 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 2,5 por cento; T3 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 3,5 por cento; T4 - metilcelulose 0,5 por cento, rafinose 0,15g e acetamida 5 por cento; T5 - glicerol 5 por cento e 2,4g de leite desnatado; T6 - 1 por cento de glicerol, 4 por cento de etilenoglicol e 2,4g de leite desnatado; T7 - 5 por cento de etilenoglicol e 2,4g de leite desnatado. O sêmen foi diluído em meio Kenney (1:1), centrifugado a 400 x g por 12 minutos, ressuspendido nos diluidores para atingir a concentração de 100X10(6)/ml, envasado em palhetas de 0,5ml e congelado 3cm acima do nível de nitrogênio líquido por 10 minutos. O descongelamento foi realizado em banho-maria a 75ºC por sete segundos. Após o descongelamento foram avaliados: motilidade total e progressiva, vigor, morfologia espermática e integridade estrutural e funcional da membrana plasmática. O T1 apresentou os maiores valores de motilidade total e progressiva (38,4 por cento e 33,8 por cento, respectivamente). O vigor e os resultados do teste HO não diferiram entre os tratamentos. Os diluidores contendo leite em sua composição (T5, T6 e T7) apresentaram maiores valores de integridade funcional e estrutural da membrana plasmática. Pode-se concluir que as modificações incorporadas aos meios diluidores testados não resultaram em melhor efeito crioprotetor que o meio à base de glicerol 5 por cento no congelamento do sêmen eqüino.

The efficacy of different combinations of cryoprotectants for equine semen was evaluated. Three ejaculates of eight stallions were used to test the lactose-EDTA-egg yolk extender with the following association of cryoprotectants: T1 - glycerol 5 percent (control group); T2 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 2.5 percent; T3 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 3.5 percent; T4 - methyl cellulose 0.5 percent, raffinose 0.15g, and acetamide 5 percent; T5 - glycerol 5 percent and 2.4g of dried skim milk; T6 - glycerol 1 percent, ethylene glycol 4 percent, and 2.4g of dried skim milk; T7 - ethylene glycol 5 percent and 2.4g of dried skim milk. After collection, Kenney extender was added to the semen 1:1, and centrifuged at 400 x g for 12 minutes. Sperm pellets were diluted to reach 100x10(6) cells/ml. Spermatozoa were frozen in 0.5ml straws 3cm above the nitrogen level, during 10 minutes. Thawing of samples was done at 75ºC for seven seconds followed by immersion of the straw in a water bath at 37ºC for 30 seconds. Post-thaw total and progressive motilities and sperm vigor were evaluated. Sperm membrane integrities of the tail and caput, respectively, were evaluated by the hypoosmotic swelling test and fluorescent dyes. T1 showed the highest post-thaw total and progressive motilities (38.4 percent and 33.8 percent, respectively). No significant difference was found among treatments for vigor and hypoosmotic swelling test. T5, T6, and T7 showed higher post-thaw values for sperm membrane integrity. It may be concluded that the association of cryoprotectants used in this experiment did not result in better cryoprotectant effect than 5 percent glycerol for equine semen cryopreservation.

Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates / Comparação entre E-test e o método da microdiluição do CLSI para teste de susceptibilidade a antifúngicos de isolados orais de Candida albicans

Thirty Candida albicans isolated from oral candidosis patients and 30 C. albicans isolated from control individuals were studied. In vitro susceptibility tests were performed for amphotericin B, fluconazole, 5-flucytosine and itraconazole through the Clinical and Laboratorial Standards Institute (CLSI) reference method and E test system. The results obtained were analyzed and compared. MIC values were similar for the strains isolated from oral candidosis patients and control individuals. The agreement rate for the two methods was 66.67 percent for amphotericin B, 53.33 percent for fluconazole, 65 percent for flucytosine and 45 percent for itraconazole. According to our data, E test method could be an alternative to trial routine susceptibility testing due to its simplicity. However, it can not be considered a substitute for the CLSI reference method.

Trinta Candida albicans isoladas de pacientes portadores de candidose oral e 30 Candida albicans isoladas de indivíduos controle foram estudadas. Testes de susceptibilidade in vitro foram realizados com anfotericina B, fluconazol, 5-flucitosina e itraconazol pelo método do Clinical and Laboratorial Standars Institute (CLSI) e por E-test. Os resultados obtidos foram analisados e comparados. Os valores de CIM foram semelhantes para amostras isoladas de pacientes portadores de candidose oral e indivíduos controle. A concordância entre os dois métodos foi de 66,7 por cento para a anfotericina B, 53,33 por cento para o fluconazol, 65 por cento para a flucitosina e 45 por cento para o itraconazol. De acordo com estes resultados, o método do E-test poderia ser uma alternativa para a triagem de casos de rotina pela sua simplicidade. Entretanto, este método não pode ser considerado como um substituto para o método de referência do CLSI.

Evaluación del cumplimiento de las indicaciones de dilución y administración de antibióticos intravenosos, según las recomendaciones emitidas por los laboratorios farmacéuticos, en pacientes del Servicio de Clínica 1 del Hospital de niños “Baca Ortiz” de Quito de julio a diciembre 2007

Partiendo de que una de las principales actividades del profesional de enfermería en su función asistencial es la administración de medicamentos, según indicaciones médicas, el mismo tendrá conocimientos sobre farmacología, el que adquiere de manera general durante su formación y de forma explícita durante el ejercicio de su profesión. Esto requiere de una guía protocolar donde se explique de forma detallada aquellos procedimientos esenciales referentes a la preparación, dilución y administración de los antibióticos. Esta investigación tuvo la finalidad de realizar una evaluación acerca del cumplimiento de las recomendaciones emitidas por los laboratorios farmacéuticos para la dilución y administración de los antibióticos intravenosos en el servicio de Clínica 1 del Hospital de Niños “Baca Ortiz”. Se utilizó un estudio de tipo epidemiológico, analítico, transversal y de punto, para lo cual se trabajó con todo el universo: 13 enfermeras de cuidado directo y todos los niños ingresados en este servicio durante las 24 horas del día durante el mes de noviembre 2007. Los datos obtenidos fueron procesados con ayuda del paquete estadístico: EPI INFO OV. 604 CD ATLANTA. Se confirman que en relación a la dilución se cumplen en 84.4% de las recomendaciones emitidas por los laboratorios farmacéuticos, pero en cuanto a la administración se cumple solamente el 34.1%, se obtuvieron además datos de un adecuado manejo en el almacenamiento de los antibióticos, no así en la conservación de los medicamentos sobrantes.