Neurofibromatosis type I: mutation spectrum of NF1 in spanish patients.

Neurofibromatosis type I (NF1) is one of the most common genetic disorders in humans. NF1, a tumor predisposition syndrome, is caused by heterozygous pathogenic variants in the NF1 gene. Molecular genetic testing of NF1 is complex, especially because of the presence of a high number of partial pseudogenes, some of them with a high percentage of sequence identity. In this study, we have analyzed the largest cohort of NF1 Spanish patients (150 unrelated individuals suspected of having NF1 and 53 relatives, making a total of 203 individuals). Mutation analysis of the entire coding region was performed in all unrelated index patients. Additionally, the Multiplex Ligation-dependent Probe Amplification (MLPA) test of the NF1 gene and SPRED1 gene analysis (sequencing and MLPA test) was performed in some of the negative patients for NF1 point mutations. When fulfilling the National Institutes of Health (NIH) criterion for the clinical diagnosis of NF1, the detection rate was 79%. Among the 80 genetically confirmed NF1 probands, we detected 69 different pathogenic variants. Two mutations (3%) were gross deletions of the whole gene, the remaining 78 mutations (97%) were small changes spread among all NF1 exons. Among these 69 different mutations detected, 42 mutations were described elsewhere, and 27 mutations were novel mutations. When segregation was studied, 67% of mutations resulted de novo variants. No genetic mosaicism was detected on patients' parents.

Highest Plasma Phenylalanine Levels in (Very) Premature Infants on Intravenous Feeding; A Need for Concern.

OBJECTIVE: To analyse the association in newborns between blood levels of phenylalanine and feeding method and gestational age. STUDY DESIGN: This observational, cross-sectional study included a sample of 11,829 infants between 2008 and 2013 in a Spanish region. Data were recorded on phenylalanine values, feeding method [breast, formula, mixed (breast plus formula), or partial or fully intravenous feeding], gestational age in weeks (<32, 32-37, ≥37), gender and days since birth at the moment of blood collection. Outcomes were [phenylalanine] and [phenylalanine] ≥95th percentile. Associations were analysed using multivariate models [linear (means difference) and logistic regression (adjusted odds ratios)]. RESULTS: Higher phenylalanine values were associated with lower gestational age (p<0.001) and with intravenous feeding (p<0.001). CONCLUSION: The degree of prematurity and intravenous feeding influenced the plasma concentration of phenylalanine in the newborn. Caution should be taken in [phenylalanine] for newborns with intravenous feeding, monitoring them carefully. Very preterm infants given the recommended amount of amino acids should also be strictly monitored. These findings should be taken into consideration and call for adapting the amounts to the needs of the infant.

Significance of calcium binding, tyrosine phosphorylation, and lysine trimethylation for the essential function of calmodulin in vertebrate cells analyzed in a novel gene replacement system.

Calmodulin (CaM) was shown to be essential for survival of lower eukaryotes by gene deletion experiments. So far, no CaM gene deletion was reported in higher eukaryotes. In vertebrates, CaM is expressed from several genes, which encode an identical protein, making it difficult to generate a model system to study the effect of CaM gene deletion. Here, we present a novel genetic system based on the chicken DT40 cell line, in which the two functional CaM genes were deleted and one allele replaced with a CaM transgene that can be artificially regulated. We show that CaM is essential for survival of vertebrate cells as they die in the absence of CaM expression. Reversal of CaM repression or ectopic expression of HA-tagged CaM rescued the cells. Cells exclusively expressing HA-CaM with impaired individual calcium binding domains as well as HA-CaM lacking the ability to be phosphorylated at residues Tyr(99)/Tyr(138) or trimethylated at Lys(115) survived and grew well. CaM mutated at both Ca(2+) binding sites 3 and 4 as well as at both sites 1 and 2, but to a lesser degree, showed decreased ability to support cell growth. Cells expressing CaM with all calcium binding sites impaired died with kinetics similar to that of cells expressing no CaM. This system offers a unique opportunity to analyze CaM structure-function relationships in vivo without the use of pharmacological inhibitors and to analyze the function of wild type and mutated CaM in modulating the activity of different target systems without interference of endogenous CaM.

Regulation of the ligand-dependent activation of the epidermal growth factor receptor by calmodulin.

Calmodulin (CaM) is the major component of calcium signaling pathways mediating the action of various effectors. Transient increases in the intracellular calcium level triggered by a variety of stimuli lead to the formation of Ca(2+)/CaM complexes, which interact with and activate target proteins. In the present study the role of Ca(2+)/CaM in the regulation of the ligand-dependent activation of the epidermal growth factor receptor (EGFR) has been examined in living cells. We show that addition of different cell permeable CaM antagonists to cultured cells or loading cells with a Ca(2+) chelator inhibited ligand-dependent EGFR auto(trans)phosphorylation. This occurred also in the presence of inhibitors of protein kinase C, CaM-dependent protein kinase II and calcineurin, which are known Ca(2+)- and/or Ca(2+)/CaM-dependent EGFR regulators, pointing to a direct effect of Ca(2+)/CaM on the receptor. Furthermore, we demonstrate that down-regulation of CaM in conditional CaM knock out cells stably transfected with the human EGFR decreased its ligand-dependent phosphorylation. Substitution of six basic amino acid residues within the CaM-binding domain (CaM-BD) of the EGFR by alanine resulted in a decreased phosphorylation of the receptor and of its downstream substrate phospholipase Cγ1. These results support the hypothesis that Ca(2+)/CaM regulates the EGFR activity by directly interacting with the CaM-BD of the receptor located at its cytosolic juxtamembrane region.

Incidencia de factores de riesgo cardiovasculares en niños y adolescentes con hipertensión arterial esencial en Artemisa / Incidence of cardiovascular risk factors in children and adolescent presenting with essential high blood pressure in Artemisa province

La Hipertensión Arterial del adulto está relacionada con la presencia de factores de riesgo de enfermedad cardiovascular y comienza en la niñez, muchas veces de manera asintomática, por lo que es importante diagnosticar los factores de riesgo que más se relacionan con la hipertensión en el niño, motivo por el cual realizamos esta investigación. Determinar la incidencia de factores de riesgo cardiovasculares en niños y adolescentes con Hipertensión Arterial esencial. Se estudiaron 100 hipertensos esenciales, de ambos sexos, en edades entre los 5 y 18 años. Los pacientes fueron atendidos en la Consulta de Cardiología Pediátrica del municipio Artemisa, Provincia La Habana de enero del 2006 a enero del 2010. Se utilizaron variables como: edad, sexo, peso, talla, peso al nacer, ejercicio físico, tabaquismo activo y pasivo, y además los antecedentes personales y familiares de hipertensión arterial, diabetes mellitus, hipercolesterolemia y obesidad. Encontramos una mayor frecuencia en el sexo masculino, así como en el grupo etáreo de 10 a 18 años. Con relación a los antecedentes familiares, aparece la Hipertensión Arterial como el factor de riesgo cardiovascular más frecuente. El sobrepeso corporal y el sedentarismo fueron los factores de riesgo personales que predominaron.(AU)

The high blood pressure in the adult is related to presence of risk factors of cardiovascular disease and it start in childhood often in a symptomatic way, thus authors carried out the present research because it is important to diagnose the risk factors more related to hypertension in the child. To determine the cardiovascular risk factors in children and adolescents of both sexes aged between 5 and 18. Patients were seen in the Pediatric Cardiology Consultation of the Artemisa province from January, 2006 to January, 2010. Variables included: age, sex, weight, height, birth weight, physical exercise, active and passive smoking and also the personal and family backgrounds of high blood pressure, diabetes mellitus, hypercholesterolemia and obesity. There was a great frequency in male sex, as well as in the age group from 10 to 18. With regard to the family backgrounds the high blood pressure appears as the more frequent cardiovascular risk factor. There was predominance of excess body weight and a sedentary life as risk factors.(AU)

Incidencia de factores de riesgo cardiovasculares en niños y adolescentes con hipertensión arterial esencial en Artemisa / Incidence of cardiovascular risk factors in children and adolescent presenting with essential high blood pressure in Artemisa province

La Hipertensión Arterial del adulto está relacionada con la presencia de factores de riesgo de enfermedad cardiovascular y comienza en la niñez, muchas veces de manera asintomática, por lo que es importante diagnosticar los factores de riesgo que más se relacionan con la hipertensión en el niño, motivo por el cual realizamos esta investigación. Determinar la incidencia de factores de riesgo cardiovasculares en niños y adolescentes con Hipertensión Arterial esencial. Se estudiaron 100 hipertensos esenciales, de ambos sexos, en edades entre los 5 y 18 años. Los pacientes fueron atendidos en la Consulta de Cardiología Pediátrica del municipio Artemisa, Provincia La Habana de enero del 2006 a enero del 2010. Se utilizaron variables como: edad, sexo, peso, talla, peso al nacer, ejercicio físico, tabaquismo activo y pasivo, y además los antecedentes personales y familiares de hipertensión arterial, diabetes mellitus, hipercolesterolemia y obesidad. Encontramos una mayor frecuencia en el sexo masculino, así como en el grupo etáreo de 10 a 18 años. Con relación a los antecedentes familiares, aparece la Hipertensión Arterial como el factor de riesgo cardiovascular más frecuente. El sobrepeso corporal y el sedentarismo fueron los factores de riesgo personales que predominaron(AU)

The high blood pressure in the adult is related to presence of risk factors of cardiovascular disease and it start in childhood often in a symptomatic way, thus authors carried out the present research because it is important to diagnose the risk factors more related to hypertension in the child. To determine the cardiovascular risk factors in children and adolescents of both sexes aged between 5 and 18. Patients were seen in the Pediatric Cardiology Consultation of the Artemisa province from January, 2006 to January, 2010. Variables included: age, sex, weight, height, birth weight, physical exercise, active and passive smoking and also the personal and family backgrounds of high blood pressure, diabetes mellitus, hypercholesterolemia and obesity. There was a great frequency in male sex, as well as in the age group from 10 to 18. With regard to the family backgrounds the high blood pressure appears as the more frequent cardiovascular risk factor. There was predominance of excess body weight and a sedentary life as risk factors(AU)

Prehipertensión arterial en adolescentes, epidemiología y otras variables de riesgo para la adultez temprana / Arterial pre-hypertension in adolescents, epidemiology and other risk variables for early adulthood

Se realizó una investigación de casos control doble ciego controlado para determinar factores de riesgos asociados a la prehipertensión en adolescentes, determinando edad según sexo, antecedentes patológicos familiares de HTA, circunferencia abdominal por Índice de masa corporal más presencia de hipertrofia ventricular izquierda. Se realizó un muestreo aleatorio definiéndose dos grupos de 70 pacientes: A prehipertensos, B negativos. Fueron discretamente mayoría los varones, aunque las hembras prehipertensas eran más jóvenes. El 87,1 por ciento de los prehipertensos tenían padres hipertensos con elevada fracción atribuible 79,4 por ciento. En general, los obesos y sobrepesos fueron la mayoría con gran cintura existiendo asociación entre variables según índice de masa corporal. El 41,4 por ciento de los prehipertensos tuvo hipertrofia ventricular izquierda. La prehipertensión, obesidad e hipertrofia ventricular izquierda son factores modificables que unidos con la genética familiar, ensombrecen el pronóstico en la población adulta joven de San Antonio de los Baños (AU)

It was conducted a case-control, double-blind controlled Investigation to determine risk factors associated with pre-hypertension in adolescents, determining age by gender, family pathological antecedents of AHT, abdominal circumference by body mass index plus presence of left ventricular hypertrophy. It was conducted a random sampling defining two groups of 70 patients: A (pre-hypertensive), B (negative). Males were discreetly the majority, although pre-hypertensive females were younger. 87.1 per cent of pre-hypertensive cases had hypertensive parents with high attributable fraction (79.4 per cent). In general, those obese and overweight were majority with a large waist, existing association between variables according to body mass index. The 41.4 per cent of the pre-hypertensive cases had left ventricular hypertrophy. Pre-hypertension, obesity and left ventricular hypertrophy are modifiable factors that together with the family genetics darken the prognosis in the young adult population of San Antonio de los Baños (AU)

Calmodulin-mediated regulation of the epidermal growth factor receptor.

In this review, we first describe the mechanisms by which the epidermal growth factor receptor generates a Ca(2+) signal and, subsequently, we compile the available experimental evidence regarding the role that the Ca(2+)/calmodulin complex, formed after the rise in cytosolic free Ca(2+) concentration, exerts on the receptor. We focus not only on the indirect action that Ca(2+)/calmodulin exerts on the epidermal growth factor receptor, as a result of the activation of distinct calmodulin-dependent kinases, but also, and more extensively, on the direct interaction of Ca(2+)/calmodulin with the receptor. We also describe several mechanistic models that could account for the Ca(2+)/calmodulin-mediated regulation of epidermal growth factor receptor activity. The control exerted by calmodulin on distinct epidermal growth factor receptor-mediated cellular functions is also discussed. Finally, the phosphorylation of this Ca(2+) sensor by the epidermal growth factor receptor is highlighted.