TLR2-TLR4/CD14 polymorphisms and predisposition to severe invasive infections by Neisseria meningitidis and Streptococcus pneumoniae.

PURPOSE: Streptococcus pneumoniae and Neisseria meningitidis are major causes of severe invasive bacterial infections in some individuals. Apparently the genetic is a major susceptibility determinant to these infectious diseases. We study if the functional polymorphisms within genes of the innate immune system (TLR2-TLR4 and CD14) are related to the predisposition to severe invasive infections caused by S. pneumoniae and N. meningitidis. MATERIAL AND METHODS: Prospective descriptive study. Sixty-six Caucasian healthy children and 173 consecutive Caucasian children with invasive bacterial infections by N. meningitidis (n=59) and S. pneumoniae (n=114) were enrolled between January 1, 2008 and December 31, 2010. All blood samples were genotyped with description of the coding polymorphisms in p.R753Q of TLR2 gene and p.D299G of TLR4 gene as well as the promotor polymorphism c.-159C>T of the CD14 gene. RESULTS: Compared to the controls the p.753Q allele of TLR2 and the allele c.-159T of CD14 were more frequent in patients with S. pneumoniae (p<0.0001 and p=0.0167) and meningococcal infections (p=0.0003 and p=0.0276 respectively). CONCLUSIONS: Genetical variations in the innate immune system by polymorphisms in the TLR2 and CD14, could be related with an increases susceptibility to severe invasive infections by S. pneumoniae and N. meningitidis.

Farmacogenética en el tratamiento del asma / Pharmacogenetics in the treatment of asthma

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Hiperbilirrubinemia neonatal. Peculiar debut del déficit de alfa-1-antitripsina / Neonatal hyperbilirrubinemia: a peculiar presentation of alpha-1-antitrypsin deficiency

La deficiencia de alfa-1-antitripsina (AlTT) es una enfermedad hereditaria que predispone a afección hepática en la infancia y a enfisema pulmonar en la edad adulta. Constituye la causa genética más común en el niño y de enfisema en el adulto; también es la que motiva más trasplantes hepáticos en niños y adolescentes. En nuestro medio presenta una prevalencia de 1 / 1.600-4.000 y una incidencia de 1 / 200 nacidos vivos. La A1TT es una glucoproteína inhibidora de proteasas séricas, codificada en el cromosoma 14 y que se hereda de forma autosómica recesiva. El fenotipo normal es el MM, y las mutaciones más frecuentes son las que asocian S y Z. Las manifestaciones clinicas más frecuentes en neonatos son la hiperbilirrubinemia conjugada, la colestasis y la hepatomegalia. Presentamos el caso de un recién nacido del sexo masculino ingresado por prematuridad e hiperbilirrubinemia directa, lo que motiva diferentes supuestos etiopatogénicos. Realizadas las pruebas sistemáticas y el estudio hepático, se determinó descenso de AlTT, y se decidió efectuar un estudio genético, en el que presentó un genotipo SS para el alelo de AlTT. La madre era SS y el padre MS, y ambos eran asintomáticos. En controles posteriores, se confirmó descenso en la cifra de AlTT, asociado a elevaciones puntuales de ésta como reactante de fase aguda, en relación con diversos procesos agudos. El interés de este caso radica en que, compartiendo la alteración genética con familiares de primer grado y, al ser éstos aparentemente asintomáticos, nuestro paciente debutó en el periodo neonatal inmediato; este hecho es peculiar en la bibliografía médica (AU)

[Neonatal screening for cystic fibrosis]. / Cribado neonatal de fibrosis quística.

OBJECTIVE: To analyze the efficiency of the method of neonatal screening for cystic fibrosis (CF) used in Castille and Leon (Spain), which is carried out with blood from Guthrie spots. MATERIAL AND METHODS: A total of 36,086 newborns were studied from January 1999 to June 2001. Immunoreactive trypsinogen (IRT) was quantified in all samples and genetic study covering 87.5 % of mutations in the CFTR gene was carried out when IRT levels were > 60 ng/mL. The sweat test was performed in all children in whom at least one mutation was detected. RESULTS: IRT values of > 60 ng/mL were found in 285 children (0.79 %). Of these, eight children (2.8 %) were diagnosed with CF and a further 11 children (3.9 %) with a negative sweat test were found to have one mutation and were thus classified as healthy carriers. To date, no false negatives have been detected. CONCLUSIONS: The two-stage screening method fulfills the required criteria. Its sensitivity is 98.5 % and the basic model can be used in other regions although genetic screening should be optimized by pilot programs to identify the local spectrum of CFTR mutations.

Cribado neonatal de fibrosis quística / Neonatal screening for cystic fibrosis

Objetivo: Analizar la eficacia del método utilizado para el cribado neonatal de fibrosis quística en Castilla y León, realizado con muestras de sangre impregnadas en papel absorbente. Material y métodos: Se estudiaron 36.086 recién nacidos desde enero de 1999 a junio de 2001 mediante cuantificación de tripsinógeno inmunorreactivo (TIR). Cuando los valores fueron superiores a 60 ng/ml se buscaron mutaciones en el gen CFTR con una cobertura del 87,5% de los alelos mutantes en nuestra población. Se solicitó estudio mediante test del sudor en todos los niños en los que se encontró una mutación. Resultados: Se detectaron valores de TIR > 60 ng/ml en 285 niños (0,79%), se diagnosticó fibrosis quística en 8/285 (2,8%) y en otros 11/285 (3,9%) se encontró una mutación, con test de sudor negativo, por lo que se consideraron portadores sanos. Hasta la actualidad no se tiene noticia de la aparición de ningún falso negativo. Conclusiones: El método de cribado en dos fases utilizado cumple los criterios exigibles con una sensibilidad del 98,5%. El modelo general del estudio puede utilizarse en otras comunidades, aunque el rastreo de mutaciones deberá ser optimizado realizando programas piloto que identifiquen el espectro local de mutaciones de fibrosis quística. (AU)

Eficacia de un test clínico como preselección de niños con sospecha de síndrome X frágil / Effectiveness of a clinical checklist in the preselection of children with suspicion of fragile x syndrome

Antecedentes: El síndrome X frágil (SXF) es la causa hereditaria más frecuente de retraso mental. Puede diagnosticarse con técnicas de genética molecular, pero su variada expresión dificulta la sospecha clínica. Objetivo: Se valora la utilidad de un test de seis criterios clínicos como método de preselección a los niños candidatos para estudio genético del síndrome. Pacientes y métodos: Se estudiaron 70 pacientes varones entre 2 y 10años, con retraso mental de causa desconocida, aplicándoseles un test con seis criterios clínicos (retraso mental, historia familiar de retraso mental, facies alargada, orejas grandes, conducta autística y déficit de atención) que se valoraron de 0 a 2 puntos. En todos se realizó estudio molecular del gen SXF usando reacción en cadena de la polimerasa y Southern-blot. Resultados: El estudio molecular confirmó la mutación completa (>200 repeticiones CGG) en 14/70 (20%) niños. La suma de 6puntos en el test fue el límite más discriminativo y fue alcanzado por los 14 enfermos con mutación (100%), pero sólo por 2 de 56 casos (3,5%) sin mutación. El mejor modelo diagnóstico fue la asociación del retraso mental, deficiencia de atención e hiperactividad, historia familiar de retraso mental y orejas grandes seguido de la facies alargada y la conducta autista. Conclusión: Un test clínico de 6 parámetros facilita la preselección de niños con sospecha de SXF para ser confirmados luego con técnicas de genética molecular (AU)

[Effectiveness of a clinical test in the preselection of children with suspected fragile X syndrome]. / Eficacia de un test clínico como preselección de niños con sospecha de síndrome X frágil.

BACKGROUND: Fragile X syndrome (FXS) is the most frequent hereditary cause of mental retardation. It can be diagnosed by molecular genetic techniques, but clinical suspicion is made less likely by it variable expression. OBJECTIVE: To assess the effectiveness of a six-item checklist in the preselection of children who are candidates for FXS genetic study. MATERIAL AND METHODS: We studied 70 male patients aged between 2 and 10 years with mental retardation of unknown cause. In all patients a checklist with six clinical criteria (mental retardation, history of familial mental retardation, long face, large ears, autistic-like behaviour, and attention deficit disorder with hyperactivity) measured from 0-2 points was applied and molecular genetic studies using polymerase chain reaction and Southern-blot were performed. RESULTS: In 14 of the 70 children (20%) molecular study confirmed full mutation (200 CGG repeats). A score of six points in the test had the greatest discriminatory power and was reached by 14 patients (100%) with mutation, but only by 2of 56patients (3.5%) without mutation. The most accurate diagnostic model was the association of mental retardation, attention deficit disorder with hyperactivity, large ears and a history of familial mental retardation followed by long face and autistic-like behaviour. CONCLUSIONS: The six-item checklist improved the preselection of children with suspicion of FXS, which was later confirmed by molecular genetic techniques.

[Agenesis of vas deferens and cystic fibrosis]. / Agenesia de conductos deferentes y fibrosis quística.

Congenital bilateral absence of vasa deferens appears in 6% of obstructive azoospermia, and 60-70% of these patients also have cystic fibrosis mutations. Unilateral aplasia or agenesia of vasa deferens occurs in less than 1% male individuals and some studies have found that up to 43% cases show mutations in the cystic fibrosis gen. We contribute four case reports of bilateral agenesia who were seen for infertility, all of which showed presence of mutation. In none of the two cases of unilateral agenesia, who consulted for vasectomy, a mutation in the cystic fibrosis gen was found. Patients with bilateral agenesia and their partners should be screened for cystic fibrosis, prior to spermatic microaspiration and assisted fecundation.

[Value of salivary antibodies for determining seropositivity to measles, rubella, and mumps in children and adults]. / Valor de los anticuerpos salivares para la determinación de la seropositividad frente a sarampión, rubéola y parotiditis en niños y adultos.

BACKGROUND: IgA and IgG antibodies can be detected in saliva in order to assess the immune status against measles, rubella and mumps. PATIENTS AND METHODS: Serum and saliva were simultaneously obtained from 50 adults between 19 and 52 years of age that were non-vaccinated and from 50 children from 15 months to 13 years of age that had been vaccinated against measles, rubella and mumps at 15 months of age. Specific IgG and IgA antibodies were determined by ELISA. Values higher than the 95% confidence interval obtained in 39 non-vaccinated and non-infected infants were considered as positive. RESULTS: In adults 96-100% and in children 90-98% were seropositive for the viral antigens studied. A positive result in saliva was always higher than 50%, with the percentage being higher in children than in adults and mainly for IgA antibodies. According to the present study, the combined determination of IgG and IgA antibodies in saliva would detect 86% of the children seropositive for measles, 87% for rubella and 82% to mumps, with these results being slightly lower in adults. Children without salivary antibodies were frequently younger than 3 years of age and were negative for more than one viral antigen. CONCLUSIONS: The study of salivary antibodies is a non-invasive method to assess seropositivity against measles, rubella and mumps, but it is advisable that both IgG and IgA antibodies be determined.

[Serum antibodies against Haemophilus influenzae in children from the province of Valladolid (Spain)]. / Anticuerpos séricos contra Haemophilus influenzae en niños de la provincia de Valladolid.

OBJECTIVE: Invasive infections by Haemophilus influenzae type b (Hib) in children are increasing in Spain. Moreover, they are becoming more resistant to antibiotics. Currently, effective vaccines against Hib are available and their inclusion in the routine schedule of immunizations is discussed. Knowledge of the childhood serology appears to be useful in decision making. PATIENTS AND METHODS: Two hundred and thirty-five normal children, between 1 month and 15 years of age, were studied. Thirty-six adults sera were also included as controls. Polysaccharide of Hib was conjugated with tyramine and used to coat polystyrene microtitration plates. The determination of antibodies was performed by ELISA and the results are expressed in optical density units. The mean level of antibodies present in 6 sera from a patient with Bruton syndrome (17 uu), just before degammaglobulinotherapy, was considered as the limit of protection. RESULTS: The antibody titer correlated with age (p < 0.001). The levels in children younger than 3 years (25.7 +/- 32.5) were lower than in older children (p < 0.001). Of the children younger than 3 years of age, 53% did not reach the protective level of antibodies, compared to 15.2% of those between 3-6 years and 22.7% of children between 6 and 14 years. Whereas the mean antibody level increased with age in positive sera, the percentage of seropositive children decreased after 3 years of age. CONCLUSIONS: The number of children spontaneously protected against Hib is low (47%) until 3 years of age. Nevertheless, this number increases rapidly after this age. Immunization must be restricted to this age group.

[Serum eosinophil cationic protein in children with allergic and nonallergic inflammation]. / Proteína catiónica eosinófila sérica en niños con inflamación alérgica y no alérgica.

Serum eosinophil cationic protein (ECP) levels increase in inflammation processes with activation of eosinophils. We studied serum ECP in (I) 32 pollinic children without symptoms, in June and October and (II) 10 children with acute asthma crisis. As control groups we included (III) 25 children sent to the hospital with suspected allergic diseases in which an IgE mediated process could be ruled out; (IV) 34 coeliac patients; (V) 15 children with cystic fibrosis and (VI) 48 normal children. The pollinic children had increased figures of ECP in June (21.2 +/- 9.2 micrograms/L) compared to normal controls (p < 0.001) and they continued to have high levels in October (13.5 +/- 9.2 micrograms/L, p < 0.05). The patients with very high ECP (> 20 micrograms/K), in spite of being asymptomatic, showed a negative correlation between ECP/peak-flow (p: 0.038). In addition, in these patients the ECP also had a negative correlation with the recovery of bronchospasm from June to October (p: 0.024). Some asthmatic children also had high ECP, but the results were too heterogeneous to draw any conclusions, possibly due to the drugs received. The ECP was independent of age and sex. It not correlated with serum IgE, nevertheless, in non-atopic patients it did correlate with blood eosinophilia (p < 0.005). In coeliac and cystic fibrosis patients, we did not find ECP to be increased. In conclusion, serum ECP increases in some allergic patients and suspected allergy, but not in all cases. It does not increase in other chronic mucosal inflammations, such as coeliac or cystic fibrosis. It correlates with bronchospasms and would have some value in predicting short-term evolution.

[Measurement and comparison of the rheumatoid factors class IgG, IgM and IgA in chronic juvenile rheumatoid arthritis]. / Medida y comparación del factor reumatoide de clase IgG, IgM e IgA en artritis crónica juvenil.

The rheumatoid factor (RF) was studied in 35 sera from 23 children with juvenile rheumatoid arthritis (JRA). The immunoglobulin class of RF was investigated and also its reactivity to both human and rabbit IgG. The RF of IgG class (IgG-RF) was more frequently positive than the IgM-RF and the IgA-RF. Nevertheless, against human IgG we found IgM-RF in the 51% of sera and IgA-RF in 48%, and against rabbit IgG in 65 and 37% respectively. All classes of RF were more frequent in rheumatoid patients than in normal controls (p less than 0.0005) although the IgA-RF increase was not significant in some groups. The specificity of the 5 RF types was always very high (92-100%). The sensibility ranged between 71% (IgG-RF against rabbit IgG) and the 37% (IgA-RF against rabbit IgG). Most sera simultaneously contained more than one class of RF. Against human IgG, the 37% had 2 classes. When we used rabbit IgG, the 31% had 3 classes and the 62% had 2 classes. The correlation of every RF class each other generally was very high (p less than 0.001). A correlation was also present in the seronegative and the systemic group, when we separately studied every clinic form. The ELISA allows detect positive IgG-RF and IgA-RF in seronegative cases by agglutination tests, therefore the seronegative concept must be reconsidered. The correlation among different RF classes are, frequently, very closed.