Objective: To evaluate the clinical usefulness of urinary N-acetyl-beta-D-glucosaminidase (NAG) excretion for the detection of early tubular damage in type 2 diabetes mellitus (T2DM). Subjects and methods: Thirty six patients with T2DM were divided into two groups based on urinary albumin to creatinine ratio (ACR): normoalbuminuria (ACR < 30 mg/g; n = 19) and microalbuminuria (ACR = 30-300 mg/g; n = 17). The following parameters were determined in both groups: urinary NAG and albumin, serum and urine creatinine, fasting plasma glucose and glycated hemoglobin (HbA 1c ). Results: Urinary NAG levels [Units/g creatinine; median (range)] were significantly increased in microalbuminuria group [17.0 (5.9 -23.3)] compared to normoalbuminuria group [4.4 (1.5 -9.2)] (P < 0.001). No differences between groups were observed in fasting glucose, HbA 1c , serum creatinine levels and estimated glomerular filtration rates (eGFR). Urinary NAG positively correlated with ACR (r = 0.628; p < 0.0001), while no significant association was observed between NAG and glycemia, HbA 1c , serum creatinine and eGFR. RESUMOObjetivo: Avaliar a utilidade clínica da excreção urinária da N-acetil-beta-D-glucosaminidase (NAG) para a detecção de dano tubular precoce no diabetes melito tipo 2 (DM2). Sujeitos e méto-dos: Foram estudados trinta e seis pacientes com DM2 que se dividiram em dois grupos com base na excreção urinária de albumina (EUA): normoalbuminúrico (EUA < 30 mg/g de creatinina; n = 19) e microalbuminúrico (EUA = 30-300 mg/g de creatinina; n = 17). Em ambos os grupos foram determinados os seguintes parâmetros: NAG e albumina urinária, creatinina sérica e urinária, glicemia de jejum e hemoglobina glicada (HbA 1c ). Resultados: Os níveis de NAG urinária [unidades/g de creatinina; mediana (intervalo interquartílico)] foram significativamente maiores no grupo microalbuminúrico [17,0 (5,9 -23,3)] em comparação com o grupo normoalbuminúrico [4,4 (1,5 -9,2)] (p < 0,001). Não se observaram diferenças significativas entre os dois grupos nos níveis de glicemia de jejum, HbA 1c , creatinina sérica e taxa de filtração glomerular estimada (TFGe). A NAG urinária se correlacionou positivamente com o EUA (r = 0,628, p < 0,0001), não sendo observada associação significativa da NAG com glicemia, HbA 1c , creatinina sérica e TFGe. Conclusões: O aumento da NAG urinária na fase de microalbuminúria da nefropatia diabética (ND) sugere que a disfunção tubular já está presente nesse período. A associação positiva significativa entre a excreção urinária da NAG e EUA indica a possível aplicação clínica da NAG urinária como marcador complementar para a detecção precoce da ND no DM2. Arq Bras Endocrinol Metab. 2014;58(8):798-801 Descritores Nefropatia diabética; diabetes melito tipo

RESUMEN "POSIBLE ROL DE LOS MACRÓFAGOS EN FLUIDO SEMINAL"Este trabajo se realizó para estudiar el rol de los macrófagos en semen y su asociación con: el porcentaje de espermatozoides con DNA nativo, morfología espermática y presencia de anticuerpos antiespermá-ticos (AAE).Se utilizó muestras de semen de 31 pacientes y se evaluaron según los criterios de la Organización Mundial de la Salud. La cromatina nuclear se estudió con Naranja de Acridina, fluorocromo que discrimina el DNA nativo, color verde, del DNA desnaturalizado, color anaranjado. La concentración de macrófagos se determinó utilizando Rojo Neutro para teñir las células fagocíticas y cámara de Neubauer para contar. La presencia de AAE se investigó mediante reacción de aglutinación mixta.Se encontró asociación entre la presencia de AAE y concentración de macrófagos (p<0,05). Hubo correlación significativa entre presencia de macrófagos y espermatozoides con DNA desnaturalizado (p<0,05). Pero no hubo asociación significativa entre concentración de macrófagos y morfología espermática (p:0,414). Concluimos que existe un fuerte indicio de que los macrófagos presentes en plasma seminal participan en el mecanismo fagocítico contribuyendo a la calidad seminal, facilitando la fertilización.PALABRAS CLAVE: Concentración de macrófagos. Anticuerpos antiespermáticos. Inmunovigilancia. DNA espermático.Morfología espermática. Infertilidad masculina. ABSTRACT"THE POSSIBLE MACROPHAGE ROLE IN SEMINAL FLUID" The present report was designed to study the macrophage role and their association with percentage of spermatozoa showing DNA native, morphologically normal sperm and occurrence of antibody-coated spermatozoa.Human sperm samples from 31 patients were used in this study. All samples were evaluated for standard semen parameters according to World Health Organization criteria. The sperm heads with abnormal versus normal chromatin structure were specified as orange-red versus green by Acridine Orange method. Macrophage concentration was assessed with Neutral Red in Neubauer hemocytometer. The technique for the determination of sperm bound antisperm antibodies was sperm mixed antiglobulin reaction. In our study the presence of antisperm antibodies in associated to macrophages concentration (p<0.05). The log of phagocytic cells were highly correlated with an increase of spermatozoa showing DNA denaturation (p<0.05). The log of phagocytic cells were highly correlated with an increase of spermatozoa showing DNA denaturation (p<0.05). There isn't significative association between the log macrophage concentration and morphologically normal sperm (p:0.414).Thus, the present study further emphasizes the importance of immunosurveillance of the macrophages, and the contribution at the semen quality for the success of the fertilization. Numerous studies have postulated the key factors responsible for the prediction of man potential fertility including spermatic count, acrosoma state, integrity of the cell membrane, morphology and morphometry of the whole spermatozoa and/or of the he...

RESUMEN "POSIBLE ROL DE LOS MACRÓFAGOS EN FLUIDO SEMINAL"Este trabajo se realizó para estudiar el rol de los macrófagos en semen y su asociación con: el porcentaje de espermatozoides con DNA nativo, morfología espermática y presencia de anticuerpos antiespermá-ticos (AAE).Se utilizó muestras de semen de 31 pacientes y se evaluaron según los criterios de la Organización Mundial de la Salud. La cromatina nuclear se estudió con Naranja de Acridina, fluorocromo que discrimina el DNA nativo, color verde, del DNA desnaturalizado, color anaranjado. La concentración de macrófagos se determinó utilizando Rojo Neutro para teñir las células fagocíticas y cámara de Neubauer para contar. La presencia de AAE se investigó mediante reacción de aglutinación mixta.Se encontró asociación entre la presencia de AAE y concentración de macrófagos (p<0,05). Hubo correlación significativa entre presencia de macrófagos y espermatozoides con DNA desnaturalizado (p<0,05). Pero no hubo asociación significativa entre concentración de macrófagos y morfología espermática (p:0,414). Concluimos que existe un fuerte indicio de que los macrófagos presentes en plasma seminal participan en el mecanismo fagocítico contribuyendo a la calidad seminal, facilitando la fertilización.PALABRAS CLAVE: Concentración de macrófagos. Anticuerpos antiespermáticos. Inmunovigilancia. DNA espermático.Morfología espermática. Infertilidad masculina. ABSTRACT"THE POSSIBLE MACROPHAGE ROLE IN SEMINAL FLUID" The present report was designed to study the macrophage role and their association with percentage of spermatozoa showing DNA native, morphologically normal sperm and occurrence of antibody-coated spermatozoa.Human sperm samples from 31 patients were used in this study. All samples were evaluated for standard semen parameters according to World Health Organization criteria. The sperm heads with abnormal versus normal chromatin structure were specified as orange-red versus green by Acridine Orange method. Macrophage concentration was assessed with Neutral Red in Neubauer hemocytometer. The technique for the determination of sperm bound antisperm antibodies was sperm mixed antiglobulin reaction. In our study the presence of antisperm antibodies in associated to macrophages concentration (p<0.05). The log of phagocytic cells were highly correlated with an increase of spermatozoa showing DNA denaturation (p<0.05). The log of phagocytic cells were highly correlated with an increase of spermatozoa showing DNA denaturation (p<0.05). There isn't significative association between the log macrophage concentration and morphologically normal sperm (p:0.414).Thus, the present study further emphasizes the importance of immunosurveillance of the macrophages, and the contribution at the semen quality for the success of the fertilization. Numerous studies have postulated the key factors responsible for the prediction of man potential fertility including spermatic count, acrosoma state, integrity of the cell membrane, morphology and morphometry of the whole spermatozoa and/or of the he...

The results show that tobacco alters sperm concentration and morphology with an increase of immature forms, demonstrating an altered spermatogenesis process. The consumption of tobacco should be evaluated to carry out the integral study of infertile man.

Foi desenvolvido e validado um ELISA tipo sanduíche "in house" para a quantificação de anticorpos (Ac) anti-anexina V em plasma. Os parâmetros de validação estudados foram: (i) precisão, expressado como coeficiente de variação (CV) inter-e intra-ensaio, (ii) exatidão, expressado como porcentagem de desvio entre o valor obtido e o valor real, (iii) limite de detecção (LOD), avaliado a partir do branco de reagentes e (iv) robusteza, obtida através da introdução propositada de pequenas variações em diferentes parâmetros. Além disso, a técnica "in house" foi comparada com um método comercial. Encontrou-se que ambos CV foram < 20%, a exatidão foi de 100 ± 20%, o limite de detecção foi menor que 1 U mL -1 e as pequenas variações na técnica não produziram variações significativas nos resultados. A comparação com o método comercial mostrou uma correlação aceitável. Concluiu-se que o método desenvolvido cumpre satisfatoriamente com os parâmetros de padronização e validação para imuno-análise.An "in house" sandwich ELISA for the quantification of plasma anti-annexin V antibodies was developed and validated. The validation parameters studied were: (i) precision, expressed either as the intra-or the inter-assay coefficient of variation (CV), (ii) exactitude, expressed as the percentage deviation between the obtained value and the real value, (iii) limit of detection (LOD), evaluated from the reagents blank and (iv) robustness, obtained by deliberately introducing slight variations in different parameters. Also, a comparison between the "in house" technique and a commercial method was performed. The research revealed that both CV were < 20%, exactitude was within the 100 ± 20% range, limit of detection was below 1 U mL -1 and that slight variations in the technique did not produce any significant variations in the results. Comparison with the commercial method showed an acceptable correlation. It was concluded that the method developed here satisfactorily accomplishes the parameters of standardization and validation for an immunoassay.Keywords: ELISA, annexin V, antibodies, anti-phospholipid syndrome, foetal losses IntroductionAnnexins belong to a family of proteins that are able to bind to negatively charged phospholipids and membrane bilayers through calcium dependent interactions. Though their fine structure has been well described, their functions have not been clearly identified yet.1 Like others, annexins constitute a group of ubiquitous cytoplasmic proteins involved in signal transduction. 2Annexin V is a 320-amino acid-residue, 36-kDa-protein that is folded into a planar cyclic arrangement of four repeats with each repeat composed of five alpha-helical segments. 3,4 It is expressed in various cell types, including placental trophoblasts and vascular endothelial cells. This protein is highly expressed in an apparently constitutive manner by placental trophoblasts and is displayed Quantification of Autoantibodies to Annexin V in Plasma by an "In House" Sandwich ELISA J. Braz. Chem. Soc. 454 on the apical membrane o...