RESUMO
Los antígenos especifícos de neutrófilos NA1 (HNA-1a), NA2 (HNA-1b) y SH (HNA-1c) son formas alotípicas del Fc gamma RIIIb y los blancos más frecuentes de los aloanticuerpos antigranulocitarios. El objetivo de este estudio fue determinar las frecuencias alélicas de los antígenos específicos de neutrófilos pertenecientes al sistema HNA-1 en donantes de sangre y amerindios de la etnia Toba de la ciudad de Rosario, Argentina. Se genotipificaron doscientos dieciocho individuos no relacionados para HNA-1a, HNA-1b y HNA-1c mediante reacción en cadena de polimerasa con cebadores secuencia específica (PCR-SSP). Las frecuencias alélicas en los donantes de sangre para HNA-1a y HNA-1b fueron 0,44 y 0,56 respectivamente y en la población amerindia Toba fueron 0,77 y 0,23 respectivamente. El alelo HNA-1c presentó una frecuencia de 0,023 en los donantes de sangre, pero no se detectó en ninguno de los individuos amerindios estudiados. Los presentes datos mostraron que las frecuencias de los alelos que codifican al sistema HNA-1 en la población mayoritaria de Rosario y en la minoritaria amerindia Toba son similares a las descriptas en europeos y otras poblaciones amerindias distantes, respectivamente. (AU)
The neutrophil-specific antigens NA1 (HNA-1a), NA2 (HNA-1b) and SH (HNA-1c) are allotypic forms of Fc gamma RIIIb and the most frequent targets of neutrophil alloantibodies. The aim of this study was to determine to gene frequencies of the neutrophil-specific antigens bolonging to the HNA-1 system in blood donors and Toba amerindians fron Rosario, Argentina. Two hundred and eighteen unrelated individual from Rosario were typed for HNA-1a, HNA-1b and HNA-1c, using polymerase chain reaction with sequence-specific primers (PCR-SSP). For the argentinean blood donors, the HNA-1a and HNA-1b gene frequencies were 0.44 and 0.56 and for the amerindians Toba were 0.77 and 0.23 respectively. The HNA-1c gene frequency in blood donors was 0.023 but the allele was absent within the amerindian individuals. The present data showed that the HNA-1 allele frequencies in the major population and the Toba amerindian from Rosario are similar to those described in European and others distant amerindians populations, respectively. (AU)
Assuntos
Humanos , Indígenas Sul-Americanos/genética , Frequência do Gene , Isoantígenos/genética , Neutrófilos/imunologia , Etnicidade/genética , Alelos , População , ArgentinaRESUMO
Los antígenos especifícos de neutrófilos NA1 (HNA-1a), NA2 (HNA-1b) y SH (HNA-1c) son formas alotípicas del Fc gamma RIIIb y los blancos más frecuentes de los aloanticuerpos antigranulocitarios. El objetivo de este estudio fue determinar las frecuencias alélicas de los antígenos específicos de neutrófilos pertenecientes al sistema HNA-1 en donantes de sangre y amerindios de la etnia Toba de la ciudad de Rosario, Argentina. Se genotipificaron doscientos dieciocho individuos no relacionados para HNA-1a, HNA-1b y HNA-1c mediante reacción en cadena de polimerasa con cebadores secuencia específica (PCR-SSP). Las frecuencias alélicas en los donantes de sangre para HNA-1a y HNA-1b fueron 0,44 y 0,56 respectivamente y en la población amerindia Toba fueron 0,77 y 0,23 respectivamente. El alelo HNA-1c presentó una frecuencia de 0,023 en los donantes de sangre, pero no se detectó en ninguno de los individuos amerindios estudiados. Los presentes datos mostraron que las frecuencias de los alelos que codifican al sistema HNA-1 en la población mayoritaria de Rosario y en la minoritaria amerindia Toba son similares a las descriptas en europeos y otras poblaciones amerindias distantes, respectivamente.
The neutrophil-specific antigens NA1 (HNA-1a), NA2 (HNA-1b) and SH (HNA-1c) are allotypic forms of Fc gamma RIIIb and the most frequent targets of neutrophil alloantibodies. The aim of this study was to determine to gene frequencies of the neutrophil-specific antigens bolonging to the HNA-1 system in blood donors and Toba amerindians fron Rosario, Argentina. Two hundred and eighteen unrelated individual from Rosario were typed for HNA-1a, HNA-1b and HNA-1c, using polymerase chain reaction with sequence-specific primers (PCR-SSP). For the argentinean blood donors, the HNA-1a and HNA-1b gene frequencies were 0.44 and 0.56 and for the amerindians Toba were 0.77 and 0.23 respectively. The HNA-1c gene frequency in blood donors was 0.023 but the allele was absent within the amerindian individuals. The present data showed that the HNA-1 allele frequencies in the major population and the Toba amerindian from Rosario are similar to those described in European and others distant amerindians populations, respectively.
Assuntos
Humanos , Frequência do Gene , Indígenas Sul-Americanos/genética , Isoantígenos/genética , Neutrófilos/imunologia , Alelos , Argentina , Etnicidade/genética , PopulaçãoRESUMO
La implementación de técnicas de amplificación de ácidos nucleicos (NAT) para la detección de Virus de Hepatitis C (VHC) y Virus de Inmunodeficiencia Humana (VIH) en el tamizaje de donantes de sangre permite acortar los períodos ventana para los ensayos utilizados en la detección de antígenos y/o anticuerpos, identificando donaciones infectivas y evitando su transfusión. Todos los usuarios de NAT deben proceder a su validación para investigar si los procedimientos involucrados cumplen y garantizan confiabilidad en la reducción del período ventana en unidades de hemocomponentes factibles de ser transfundidas. El objetivo de este trabajo fue mostrar los resultados obtenidos de la implementación de técnica NAT in house validadas frente a estándares internacionales para la detección de VHC y VIH en donantes de sangre. Los resultados fueron analizados con el software Probit analysis programs. Para un 95% de positividad, el límite de detección fue de 98,50 Ul/ml para VHC y 294, 84 Ul/ml para VIH. La especificidad fue del 100% y los procedimientos presentaron robustez. La validación de técnicas de PCR in house permite demostrar que, por sensibilidad y especificidad, son una alternativa válida y aplicable en el tamizaje de donantes de sangre.
The technical implementation of nucleic acid amplification (NAT) for detecting hepatitis C virus (HCV) and Human Immunodeficiency Virus (HIV) in blood donors screening can shorten the window periods of tests used in the detection antigen and/or antibodies, identifying and preventing infectious donations transfusion. All users of NAT should proceed to validation to investigate whether the procedures involved meet and ensure reliability in reducing the window period in units of transfused blood products to be feasible. The aim of this study was to report the results of the technical implementation of NAT in-house validated against international standards for the detection of HCV and HIV genomes in blood donors. The results were analyzed using Probit analysis software programs. For a 95% positivity, the detection limit was 98.50 IU/ml for HCV and 294. 84 IU/ml for HIV. The specificity was 100% and showed robust procedures. Validation of in-house PCR techniques can demonstrate that for sensitivity and specificity, are a valid and applicable in screening blood donors.
Assuntos
HIV , Doadores de Sangue , Hepacivirus/imunologia , Programas de Rastreamento , Bancos de Sangue , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Transfusão de Sangue , Técnicas de Amplificação de Ácido Nucleico/métodos , Viroses/diagnóstico , Viroses/prevenção & controleAssuntos
Humanos , Antígenos de Grupos Sanguíneos/genética , Antígenos de Grupos Sanguíneos/imunologia , Doadores de Sangue/estatística & dados numéricos , Sistema do Grupo Sanguíneo Rh-Hr , Argentina/etnologia , Frequência do Gene , Grupos Populacionais/genética , Isoantígenos/imunologia , PrevalênciaRESUMO
Un individuo con un fenotipo eritrocitario raro carece de uno o varios antígenos presentes en la mayor parte de la población de pertenencia. Cuando presenta el anticuerpo correspondiente, se pueden producir complicaciones perinatales, transfusionales y/o transplantológicas. Se presenta el caso de una embarazada aloinmunizada derivada a nuestro servicio en la semana 12 de su tercera gesta para su evaluación y seguimiento. El diagnóstico inmunohematológico le asignó el excepcional fenotipo "p" (aproximadamente 1/200 000 individuos), asociado con una mayor tasa de abortos espontáneos y a reacciones transfusionales graves cuando se transfunden unidades incompatibles. El estudio del gen A4GALT demostró la presencia de la mutación c.752C > T en doble dosis. Esta mutación lleva a un cambio de una prolina por una leucina en el residuo 251 de la 4-α-galactosiltransferasa. Por parto inducido por sufrimiento fetal, nace a las 36 semanas una bebé con prueba de antiglobulina (Coombs) directa negativa, eluido reactivo, con ictericia que requirió luminoterapia. Una semana después el neonato fue externado sin secuelas aparentes. Posteriormente, a raíz de una cirugía inminente y la improbabilidad de encontrar sangre compatible, se elaboró un plan para cubrir las posibles demandas. Este caso pone en evidencia la necesidad de contar a nivel nacional con un laboratorio de referencia de inmunohematología y un banco de sangre de grupos raros, que permita resolver con celeridad situaciones que requieran transfundir a estos individuos.
A rare blood group is usually defined as the absence of a high prevalence antigen or the absence of several antigens within a single blood group system. These individuals may develop clinically significant red cell antibodies to the high incidence red cell antigens they lack. A 33-year-old alloimmunized woman was referred to our center at the 12th week of her third pregnancy for evaluation and follow up. The laboratory work-up grouped her as belonging to "p" phenotype, associated with difficulties to find compatible blood for transfusion and a high incidence of recurrent miscarriage. At 36 weeks, a baby girl was born by induced labor due to fetal suffering. With a negative direct antiglobulin test but a positive elution test, she was in the neonatology ward for one week receiving luminotherapy. Homozygosity for a missense mutation at position 752 (c.752C > T) in the A4GALT gene was found to be responsible for the p phenotype. This mutation changes a proline to a leucine at codon 251 of the 4-α-galactosyltransferase. Recently, due to an imminent chirurgical intervention and the impossibility to have compatible blood available for transfusion, an autologous donation plan was designed to satisfy probable demand. This case showed the need for blood bank facilities capable to respond satisfactorily to these situations in Argentina. This would facilitate the storage of cryopreserved blood from individuals with rare blood groups for homologous use or to develop rare blood donors programs.
Assuntos
Adulto , Feminino , Humanos , Gravidez , Eritroblastose Fetal/sangue , Galactosiltransferases/genética , Mutação de Sentido Incorreto , Sistema do Grupo Sanguíneo P/genética , Fenótipo , Sequência de Bases , Transfusão de Sangue , Glicosiltransferases/análiseRESUMO
A rare blood group is usually defined as the absence of a high prevalence antigen or the absence of several antigens within a single blood group system. These individuals may develop clinically significant red cell antibodies to the high incidence red cell antigens they lack. A 33-year-old alloimmunized woman was referred to our center at the 12th week of her third pregnancy for evaluation and follow up. The laboratory work-up grouped her as belonging to "p" phenotype, associated with difficulties to find compatible blood for transfusion and a high incidence of recurrent miscarriage. At 36 weeks, a baby girl was born by induced labor due to fetal suffering. With a negative direct antiglobulin test but a positive elution test, she was in the neonatology ward for one week receiving luminotherapy. Homozygosity for a missense mutation at position 752 (c.752C > T) in the A4GALT gene was found to be responsible for the p phenotype. This mutation changes a proline to a leucine at codon 251 of the 4-?-galactosyltransferase. Recently, due to an imminent chirurgical intervention and the impossibility to have compatible blood available for transfusion, an autologous donation plan was designed to satisfy probable demand. This case showed the need for blood bank facilities capable to respond satisfactorily to these situations in Argentina. This would facilitate the storage of cryopreserved blood from individuals with rare blood groups for homologous use or to develop rare blood donors programs.
Assuntos
Eritroblastose Fetal/sangue , Galactosiltransferases/genética , Mutação de Sentido Incorreto , Sistema do Grupo Sanguíneo P/genética , Fenótipo , Adulto , Sequência de Bases , Transfusão de Sangue , Feminino , Glicosiltransferases/análise , Humanos , GravidezRESUMO
BACKGROUND: Hereditary non-polyposis colon cancer (HNPCC) is an autosomal dominant syndrome predisposing to the early development of various cancers including those of colon, rectum, endometrium, ovarium, small bowel, stomach and urinary tract. HNPCC is caused by germline mutations in the DNA mismatch repair genes, mostly hMSH2 or hMLH1. In this study, we report the analysis for genetic counseling of three first-degree relatives (the mother and two sisters) of a male who died of colorectal adenocarcinoma at the age of 23. The family fulfilled strict Amsterdam-I criteria (AC-I) with the presence of extracolonic tumors in the extended pedigree. We overcame the difficulty of having a proband post-mortem non-tumor tissue sample for MSI testing by studying the alleles carried by his progenitors. METHODS: Tumor MSI testing is described as initial screening in both primary and metastasis tumor tissue blocks, using the reference panel of 5 microsatellite markers standardized by the National Cancer Institute (NCI) for the screening of HNPCC (BAT-25, BAT-26, D2S123, D5S346 and D17S250). Subsequent mutation analysis of the hMLH1 and hMSH2 genes was performed. RESULTS: Three of five microsatellite markers (BAT-25, BAT-26 and D5S346) presented different alleles in the proband's tumor as compared to those inherited from his parents. The tumor was classified as high frequency microsatellite instability (MSI-H). We identified in the HNPCC family a novel germline missense (c.1864C>A) mutation in exon 12 of hMSH2 gene, leading to a proline 622 to threonine (p.Pro622Thr) amino acid substitution. CONCLUSION: This approach allowed us to establish the tumor MSI status using the NCI recommended panel in the absence of proband's non-tumor tissue and before sequencing the obligate carrier. According to the Human Gene Mutation Database (HGMD) and the International Society for Gastrointestinal Hereditary Tumors (InSiGHT) Database this is the first report of this mutation.
Assuntos
Neoplasias Colorretais Hereditárias sem Polipose/diagnóstico , Análise Mutacional de DNA/métodos , Mutação em Linhagem Germinativa , Repetições de Microssatélites , Proteína 2 Homóloga a MutS/genética , Adulto , Sequência de Aminoácidos , Análise Mutacional de DNA/normas , Feminino , Testes Genéticos , Instabilidade Genômica , Humanos , Masculino , Dados de Sequência Molecular , National Institutes of Health (U.S.) , Linhagem , Proteínas/genética , Padrões de Referência , Alinhamento de Sequência , Estados UnidosRESUMO
OBJECTIVE: Our purpose was to study, in semen samples of infertile patients, the relationship between the Modified Sperm Stress Test (MOST) and the presence of antisperm antibodies (ASA), macrophages concentration and the Hypoosmotic Swelling Test (HOST). METHODS: Semen samples from 42 men undergoing evaluation for infertility were examined according to WHO criteria. Twenty-five of them, whithout clumping non hyperviscosity, were selected. The MOST test was applied according to the author's original technique. ASA were determined with a direct mixed agglutination test, TAC II, developed and validate by our group. Macrophage concentration was evaluated with Neutral Red stain, and functional integrity of the sperm membrane with the Hypoosmotic Swelling Test using an hypoosmotic solution of 150 mOm/ml composed of equal parts of fructose and sodium citrate. RESULTS: The Chi square test was applied to the observational data obtaining the following results: There was a statistically significant association between the presence of ASA and altered MOST (p<0.001). In all samples with ASA, abnormal MOST values were obtained (MOST<0.39). Besides, there is a statistically significant association exists between the increased concentration of macrophages and abnormal MOST (p<0.01); and altered HOST was positively correlated with abnormal MOST (p<0.02). CONCLUSION: Results clearly demonstrate the high predictive power of MOST like a test of sperm resistance to the forced lipoperoxidation, offering conditions to become a good predictor of sperm performance. Understanding the sperm resistance to the ROS and their harmfull effects on the sperm functions, a proportion of infertile men can be succesfully treated.
Assuntos
Estresse Oxidativo , Espermatozoides/imunologia , Autoanticorpos/imunologia , Humanos , Testes Imunológicos , Macrófagos/imunologia , MasculinoRESUMO
OBJECTIVES: To investigate the presence of macrophages in human semen samples and the function they carry out in the seminal fluid. Their presence was studied in relation to spermatic morphology, percentage of spermatozoids with native DNA, and presence of antispermatic antibodies. METHODS AND RESULTS: The work was performed with semen samples from 31 unfertile males from 63 couples in which the "female factor" was ruled out as the cause of infertility. Sperm study according to WHO (1992) was carried out in all samples, in addition to: DNA study with acridine orange as fluorocrom, macrophage concentration by neutral red in a Neubauer camera, and detection of antispermatic antibodies with a mixed agglutination test (TAC II) (validated with Mar Screen-Fertility technologies). Sperm morphology was evaluated by Papanicolaou test. 19/31 selected sperm samples (61.3%) showed increased concentration of macrophages, 13 of them (41.9%) with denaturalized DNA, and 8 (25.8%) abnormal morphology. Six samples showed increased macrophage concentration and predominance of native DNA, whereas 11 samples showed increased macrophages and abnormal morphology. Among 18 (58.1%) samples showing antispermatic antibodies 14 (77.7%) had an increased concentration of macrophages. Statistical analysis resulted in a high correlation between macrophage concentration and increased percentage of spermatozoids with denaturalized DNA (p < 0.05). An increased concentration of macrophages is associated with the presence of antispermatic antibodies (p < 0.05). There was not evidence of significant association between concentration of macrophages and percentage of morphologically normal spermatozoids (p < 0.05). CONCLUSIONS: We can conclude that macrophages are present in human semen and participate in immunovigilance contributing to improve the seminal quality.
Assuntos
Macrófagos , Sêmen/citologia , Contagem de Células , Distribuição de Qui-Quadrado , Humanos , Masculino , Contagem de EspermatozoidesRESUMO
Las células normales y neoplásicas tienen similares necesidades de hierro, pero las últimas pueden exhibir mecanismos alterados de adquisición y secuestro del metal que les permiten continuar multiplicándose en tejidos del huésped, aun en condiciones de restricción del nutriente. El presente trabajo tuvo como objetivo investigar los parámetros séricos del metabolismo del hierro en pacientes con Mieloma Múltiple (MM), y compararlos con los correspondientes a una población normal. En 48 pacientes con el cáncer hematolègico se determinó Hierro (µ/dl) e Indice de Saturación de la Transferrina (por ciento) por métodos colorimétricos, Ferritina (ng/ml) por IRMA y Transferrina (mg/dl) por inmunodifusión radial. El grupo control estuvo constituido por 40 individuos sanos, correspondientes en edad y sexo. En el grupo pacientes se encontraron valores significativamente elevados (P<0,00001) de Ferritina sérica (x=504,6 ng/ml ± 386,2) y de Indice de Saturación de la Transferrina (x=45 por ciento ± 14,9) comparados con los controles (x=86,7 ng/ml ± 30 y x=29,9 ng/ml ± 8,36, respectivamente). Niveles significativamente más bajos de Transferrina sérica (P<0,00001) se observaron en el grupo pacientes (x=270 mg/dl ± 102,1) respecto a los controles (x=360 mg/dl ± 57,3).No hubo diferencias significativas (P=0,8189) entre la Ferremia del grupo pacientes (x=100,85 µg/dl ± 56,2) y los controles (x=98,6 µg/dl ± 32,4). Los resultados obtenidos mostraron un metabolismo del hierro aberrante en MM, que podría favorecer el crecimiento celular maligno y la susceptibilidad a infecciones, e inhibir funciones inmunes
Assuntos
Humanos , Masculino , Adulto , Feminino , Pessoa de Meia-Idade , Distúrbios do Metabolismo do Ferro , Mieloma Múltiplo/fisiopatologia , Estudos de Casos e Controles , Ferritinas , Ferro , Mieloma Múltiplo/complicações , Biomarcadores Tumorais , TransferrinaRESUMO
Las células normales y neoplásicas tienen similares necesidades de hierro, pero las últimas pueden exhibir mecanismos alterados de adquisición y secuestro del metal que les permiten continuar multiplicándose en tejidos del huésped, aun en condiciones de restricción del nutriente. El presente trabajo tuvo como objetivo investigar los parámetros séricos del metabolismo del hierro en pacientes con Mieloma Múltiple (MM), y compararlos con los correspondientes a una población normal. En 48 pacientes con el cáncer hematolÞgico se determinó Hierro (A/dl) e Indice de Saturación de la Transferrina (por ciento) por métodos colorimétricos, Ferritina (ng/ml) por IRMA y Transferrina (mg/dl) por inmunodifusión radial. El grupo control estuvo constituido por 40 individuos sanos, correspondientes en edad y sexo. En el grupo pacientes se encontraron valores significativamente elevados (P<0,00001) de Ferritina sérica (x=504,6 ng/ml ± 386,2) y de Indice de Saturación de la Transferrina (x=45 por ciento ± 14,9) comparados con los controles (x=86,7 ng/ml ± 30 y x=29,9 ng/ml ± 8,36, respectivamente). Niveles significativamente más bajos de Transferrina sérica (P<0,00001) se observaron en el grupo pacientes (x=270 mg/dl ± 102,1) respecto a los controles (x=360 mg/dl ± 57,3).No hubo diferencias significativas (P=0,8189) entre la Ferremia del grupo pacientes (x=100,85 Ag/dl ± 56,2) y los controles (x=98,6 Ag/dl ± 32,4). Los resultados obtenidos mostraron un metabolismo del hierro aberrante en MM, que podría favorecer el crecimiento celular maligno y la susceptibilidad a infecciones, e inhibir funciones inmunes (AU)
Assuntos
Humanos , Masculino , Adulto , Feminino , Pessoa de Meia-Idade , Idoso , Mieloma Múltiplo/fisiopatologia , Distúrbios do Metabolismo do Ferro/etiologia , Mieloma Múltiplo/complicações , Ferro/sangue , Transferrina/sangue , Ferritinas/sangue , Estudos de Casos e Controles , Biomarcadores TumoraisRESUMO
Se estudian anticuerpos anti-A y anti-B en moco cervical pre y postratamiento con solución de 2-mercaptoetanol, utilizado para romper puentes disulfuro intracatenario de los anticuerpos IgM (AU)
Assuntos
Humanos , Feminino , Muco do Colo Uterino/imunologia , Anticorpos/imunologia , Incompatibilidade de Grupos Sanguíneos/imunologia , Infertilidade/imunologia , Sistema ABO de Grupos Sanguíneos/imunologiaRESUMO
Se estudian anticuerpos anti-A y anti-B en moco cervical pre y postratamiento con solución de 2-mercaptoetanol, utilizado para romper puentes disulfuro intracatenario de los anticuerpos IgM