CALCA and TRPV1 genes polymorphisms are related to a good outcome in female chronic migraine patients treated with OnabotulinumtoxinA.

BACKGROUND: Some variables have been proposed as predictors of efficacy of OnabotulinumtoxinA in chronic migraine patients, but data available are inconclusive. We aimed to analyse the influence of single nucleotide polymorphisms in the response to OnabotulinumtoxinA. METHODS: We included 156 female patients treated with OnabotulinumtoxinA accordingly to PREEMPT paradigm in three headache units. OnabotulinumtoxinA was offered to patients that had not responded to topiramate and at least one other preventative. Age at first procedure was 43.7 ± 11.8 years (16-74). Patients with a reduction of at least 50% in the number of migraine days after two OnabotulinumtoxinA procedures were considered as responders. We analysed 25 polymorphisms selected for their relevance regarding migraine pathophysiology and their association with migraine according to previously published genome-wide association studies. Genotyping was performed using KASP probes and a LightCycler-480 (Roche-Diagnostics). Allelic, genotypic frequencies and dominance/recesivity hypothesis of the allelic variants were compared between responders and non-responders by Fisher's exact test. RESULTS: Response to treatment with OnabotulinumtoxinA was achieved in 120 patients (76,9%). Two polymorphisms showed differences: CALCA rs3781719, where allele C represents 26.9% in responders and 40.9% in non-responders (p = 0.007, OR = 3.11 (1.33-7.26)); and TRPV1 rs222749, where allele A represents 4.17% in responders and 12.5% in non-responders (p = 0.013, OR = 3.29 (1.28-8.43)). No significant differences in rest of polymorphisms or clinical or demographic variables were found. CONCLUSIONS: Polymorphic variations of CALCA and TRPV1 genes might play a role as prognostic markers of efficacy of OnabotulinumtoxinA in chronic migraine female patients in our population.

Familial Hemiplegic Migraine with Severe Attacks: A New Report with ATP1A2 Mutation.

Introduction. Familial hemiplegic migraine (FHM) is a rare disorder characterized by migraine attacks with motor weakness during the aura phase. Mutations in CACNA1A, ATP1A2, SCN1A, and PRRT2 genes have been described. Methods. To describe a mutation in ATP1A2 gene in a FHM case with especially severe and prolonged symptomatology. Results. 22-year-old woman was admitted due to migraine-type headache and sudden onset of right-sided weakness and aphasia; she had similar episodes in her childhood. Her mother was diagnosed with hemiplegic migraine without genetic confirmation. She presented with fever, decreased consciousness, left gaze preference, mixed aphasia, right facial palsy, right hemiplegia, and left crural paresis. Computed tomography (CT) showed no lesion and CT perfusion study evidenced oligohemia in left hemisphere. A normal brain magnetic resonance (MR) was obtained. Impaired consciousness and dysphasia began to improve three days after admission and mild dysphasia and right hemiparesis lasted for 10 days. No recurrences were reported during a follow-up of two years. We identified a variant in heterozygous state in ATP1A2 gene (p.Thr364Met), pathogenic according to different prediction algorithms (SIFT, PolyPhen2, MutationTaster, and Condel). Conclusion. Prolonged and severe attacks with diffuse hypoperfusion in a FHM seemed to be specially related to ATP1A2 mutations, and p.T364M should be considered.

Spectrum of mutations in the CFTR gene in cystic fibrosis patients of Spanish ancestry.

We analyzed 1,954 Spanish cystic fibrosis (CF) alleles in order to define the molecular spectrum of mutations in the CFTR gene in Spanish CF patients. Commercial panels showed a limited detection power, leading to the identification of only 76% of alleles. Two scanning techniques, denaturing gradient gel electrophoresis (DGGE) and single strand conformation polymorphism/hetroduplex (SSCP/HD), were carried out to detect CFTR sequence changes. In addition, intragenic markers IVS8CA, IVS8-6(T)n and IVS17bTA were also analyzed. Twelve mutations showed frequencies above 1%, p.F508del being the most frequent mutation (51%). We found that eighteen mutations need to be studied to achieve a detection level of 80%. Fifty-one mutations (42%) were observed once. In total, 121 disease-causing mutations were identified, accounting for 96% (1,877 out of 1,954) of CF alleles. Specific geographic distributions for the most common mutations, p.F508del, p.G542X, c.1811 + 1.6kbA > G and c.1609delCA, were confirmed. Furthermore, two other relatively common mutations (p.V232D and c.2789 + 5G > A) showed uneven geographic distributions. This updated information on the spectrum of CF mutations in Spain will be useful for improving genetic testing, as well as to facilitate counselling in people of Spanish ancestry. In addition, this study contributes to defining the molecular spectrum of CF in Europe, and corroborates the high molecular mutation heterogeneity of Mediterranean populations.

[Autosomal recessive diseases with mental retardation]. / Enfermedades autosómicas recesivas con retraso mental.

INTRODUCTION: Autosomal recessive diseases with mental retardation are disorders that affect autosomes, and their genetic expression occurs in individuals who are homozygotic for a mutation, while heterozygotic subjects are unaffected carriers. If both parents are carriers, the theoretical possibility of their children also being carriers is 50%, the risk of the children being affected by the disease is 25%, and there is a 25% chance of their being healthy. They are an important source of mental deficiencies and inborn errors of metabolism (IEM) are some of their characteristic syndromes. DEVELOPMENT: The genetic disorders known as IEM can be classified according to the metabolism they affect, that is, purines, pyrimidines, amino acids, and so on. One of the lysosomal disorders is Tay-Sachs disease, which is rare among the general population but is very frequent in populations with a high rate of consanguinity, such as the Ashkenazi Jews. One of the most notable disorders affecting the metabolism of amino acids is the case of phenylketonuria due to mutations in the phenylalanine hydroxylase gene (PAH). It accounts for 0.5-1% of mental diseases and appears with a frequency rate of between 1/11,500 and 1/14,000 in newborn infants. Its early diagnosis through neonatal screening programmes makes it possible to start administering a phenylalanine-free diet and thus prevent mental retardation. CONCLUSIONS: Knowledge of this kind of autosomal diseases with neurological involvement, together with their correct and early diagnosis, makes it possible to establish suitable treatment regimens in some cases and to carry out genetic counselling in all of them.

Enfermedades autosómicas recesivas con retraso mental / Autosomal recessive diseases with mental retardation

Introducción. Las enfermedades autosómicas recesivascon retraso mental son alteraciones que afectan a los autosomas ysu expresión genética se da en individuos que son homocigotos parauna mutación, y los heterocigotos son portadores no afectos.Con ambos padres portadores, la posibilidad teórica de que sushijos sean portadores es del 50%, un riesgo de 25% de hijos afectadospor la enfermedad, y otro 25% sanos. Son origen importantede deficiencias mentales, y algunos síndromes característicos sonlos errores congénitos del metabolismo (ECM). Desarrollo. Los trastornosgenéticos de los ECM se pueden clasificar de acuerdo con elmetabolismo alterado: purinas, pirimidinas, aminoácidos, etc. Dentrode los trastornos lisosomales se encuentra la enfermedad de Tay-Sachs, que es rara en la población general, pero con una alta frecuenciaen poblaciones de gran consanguinidad, como los judíosasquenazí. Entre las alteraciones que afectan al metabolismo delos aminoácidos, es especialmente relevante el caso de la fenilcetonuriapor mutaciones en el gen de fenilalanina hidroxilasa (PAH).Supone un 0,5-1% de las enfermedades mentales, y aparece conuna frecuencia de l/11.500-1/14.000 en recién nacidos. Su diagnósticoprecoz con los programas de cribado neonatal permite instaurarla administración de una dieta alimenticia carente de fenilalaninay evitar el retraso mental. Conclusiones. El conocimiento ycorrecto y precoz diagnóstico de este tipo de enfermedades autosómicascon afectación neurológica permite establecer unas pautasadecuadas de tratamiento en unos casos y de asesoramiento genéticoen todos

Introduction. Autosomal recessive diseases with mental retardation are disorders that affect autosomes, and theirgenetic expression occurs in individuals who are homozygotic for a mutation, while heterozygotic subjects are unaffectedcarriers. If both parents are carriers, the theoretical possibility of their children also being carriers is 50%, the risk of thechildren being affected by the disease is 25%, and there is a 25% chance of their being healthy. They are an important sourceof mental deficiencies and inborn errors of metabolism (IEM) are some of their characteristic syndromes. Development. Thegenetic disorders known as IEM can be classified according to the metabolism they affect, that is, purines, pyrimidines, aminoacids, and so on. One of the lysosomal disorders is Tay-Sachs disease, which is rare among the general population but is veryfrequent in populations with a high rate of consanguinity, such as the Ashkenazi Jews. One of the most notable disordersaffecting the metabolism of amino acids is the case of phenylketonuria due to mutations in the phenylalanine hydroxylase gene(PAH). It accounts for 0.5-1% of mental diseases and appears with a frequency rate of between 1/11,500 and 1/14,000 innewborn infants. Its early diagnosis through neonatal screening programmes makes it possible to start administering aphenylalanine-free diet and thus prevent mental retardation. Conclusions. Knowledge of this kind of autosomal diseases withneurological involvement, together with their correct and early diagnosis, makes it possible to establish suitable treatmentregimens in some cases and to carry out genetic counselling in all of them

Susceptibility genes for asthma and allergy: hits and questions.

Allergy and asthma are closely related conditions which result from a complex interaction between several genetic and environmental factors. On the basis of familial linkage analysis data from different populations, some chromosomal regions containing a series of susceptibility loci have been identified. To date, genome-wide search studies have reported a group of chromosomal regions which include most likely several candidate genes for asthma and allergy. Some of these genes are excellent candidates, as they are known to modulate the immune response and airways inflammation processes. So, by focusing efforts on the analysis of those chromosomal regions, the characterization of the potentially important genes can be achieved. This will undoubtedly provide a clue for a better understanding of those pathologic components involved in the onset and development of asthma and allergic disorders.

Increased TNFA*2, but not TNFB*1, allele frequency in Spanish atopic patients.

Tumor necrosis factor (TNF) is a potent proinflammatory cytokine involved in asthma and atopy. Increased TNF-alpha levels have been found in airway biopsies and bronchoalveolar lavage fluids from asthmatic patients. Constitutional variations in the TNF-alpha secretion levels in vitro are associated with molecular polymorphisms located within and around the TNF loci. Our study objective was to investigate the association between atopy and two described di-allelic polymorphisms in the TNF locus: a G to A transition at position -308 in the 5'-promoter region of the TNFA gene (TNFA*1 and TNFA*2 alleles) and an Ncol restriction fragment length polymorphism (RFLP) in the first intron of the TNFB gene (TNFB*1 and TNFB*2 alleles). The genetic study was performed in 65 unrelated atopic patients and 60 healthy controls. The regions of interest were amplified from genomic DNA using specific primers and polymerase chain reaction. SSP-PCR analysis for TNFA -308 polymorphism genotyping and endonuclease digestion analysis for the TNFB Ncol RFLP were used. The frequency of the TNFA*2 allele was significantly higher in atopic subjects compared to the control group (38.5% vs. 10.5%; chi2 = 32.06; p <0.0001). The TNFA*2 allele is associated with a higher risk for the development of atopy (risk ratio = 9.44; EF = 0.65; chi2 = 30.06 p <0.0005). On the other hand, no significant association between the TNFB alleles and atopy was found. In conclusion, the TNFA*2 allele could be also a genetic risk marker for the predisposition to atopy in our population, as has been reported in other studies. Either the TNFA gene itself or a linked gene on chromosome region 6p21, which has yet to be identified, is a candidate gene for susceptibility to atopy.

Spectrum of CFTR mutations in the Middle North of Spain and identification of a novel mutation (1341G-->A). Mutation in brief no. 252. Online.

We have analyzed 39 unrelated cystic fibrosis (CF) families by denaturing gradient gel electrophoresis (DGGE) and direct sequencing in order to determine the spectrum of CF mutations in our population. This approach has allowed us to detect 72 out of the 78 CF chromosomes (92.3%). The DF508 mutation was found to be present in 51/78 (65.4%) CF chromosomes, in accordance with the predicted Northwest-Southeast gradient within the European population. Another 14 known mutations, and the novel 1341G-->A mutation were identified. Nine out of fifteen non DF508 mutations were present in a single chromosome. The 1341G-->A mutation, found in 2 unrelated patients, is a new mutation associated to severe phenotype, causing pancreatic insufficiency and chronic lung infections. Our data suggest a different distribution of non-DF508 mutations in our population when compared with previous studies carried out in Spanish CF families. Six out of the 14 non-F508 in our study were not present in a recent study carried out in 640 Spanish families with CE These six mutations account for 29.6% non DF508 chromosomes in our sample.

Lack of association between atopy and RsaI polymorphism within intron 2 of the Fc(epsilon)RI-beta gene in a Spanish population sample.

BACKGROUND: The B genotype of RasI polymorphism located within intron 2 of the Fc-IgE receptor I (Fc(epsilon)RI) gene was previously found to be increased in atopic patients from a Japanese population sample. METHODS: We studied these A/B genotypes in 70 Spanish atopic patients, and the results were compared to those of 51 nonatopic controls. RasI polymorphisms were studied by specific digestion of polymerase chain reaction fragments followed by polyacrylamide gel electrophoresis. RESULTS: The polymorphism frequency (A/A: 25/70, A/B: 28/70, B/B: 17/70) found in patients did not differ from the frequency in nonatopic control subjects. CONCLUSIONS: We did not find RasI polymorphisms associated with atopic disease. The genetic findings in atopy and asthma may be very different according to ethnic and local characteristics, and they must be carefully verified in different population samples.

[Biological and genetic basis of atopy and asthma]. / Fundamentos biológicos y genéticos de la atopia y el asma.

Atopy is triggered by allergens and enhanced by environmental factors, but it has a clear genetic basis, as it is confirmed by the high incidence in siblings and twins. In the last few years, many authors have published genetic studies on asthmatic and atopic patients, generally with very controversial results. In the present article, IgE regulation and other immunological mechanisms which are assumed to be involved in the atopic reaction are reviewed. In the second part, the coding genes of factors, cytokines and receptors that take part in the atopy are commented, as well as review of the recent articles published about genetic markers or polymorphisms associated to these genes. The unveilling of the genetic background of atopy and asthma is a very difficult task, and it will need the definition of a specific atopic phenotype and a clear knowledge of the immunologic basis of atopy. Finally, due to racial and geographical variations a wide collaborative study of many research groups distributed all over the world will be needed.

Alpha-1 antichymotrypsin levels are actively increased in normal colostrum.

BACKGROUND: It is well known that human colostrum has important antiinflammatory functions. The purpose of the current study was to determine antiprotease levels in colostrum and serum and to assess the importance of local synthesis and the electrophoretic differences in both locations. METHODS: Five protease inhibitors were determined by radical immunodiffusion in colostrum and serum samples taken simultaneously from 50 healthy women, 36 to 72 hours after delivery. RESULTS: Antithrombin II, inter-alpha trypsin inhibitor, and alpha-2 macroglobulin levels were undetectable in colostrum. Mean antitrypsin levels in colostrum were 6% of serum levels, but colostrum alpha-1 antichymotrypsin was higher than expected (0.39+/-0.34 g/l) in relation according to the albumin passive transport, and their mean value was 41% of serum levels. Colostrum antichymotrypsin levels did not correlate with serum antichymotrypsin levels or with colostrum albumin levels. The antichymotrypsin molecule in colostrum had a slower electrophoretic mobility when compared with that of serum antichymotrypsin, and it showed a different pattern in Western blot analysis, with a predominating 80-kDa molecule. CONCLUSIONS: Although the origin of colostral antichymotrypsin is unclear, local production in breast epithelial cells is likely. Antichymotrypsin is increased in colostrum, and its molecule has some characteristic differences, suggesting that it has an important and specific role in infant nutrition during breast milk feeding.

HLA-DQA1*0501 and DQB1*02 homozygosity and disease susceptibility in Spanish coeliac patients.

Coeliac disease (CD) susceptibility is strongly associated with HLA-DQA1*0501 and DQB1*02 alleles. There are discordant reports on whether homozygosity increases the risk. We genotyped HLA-DQA1*0501 and DQB1*02 in 50 CD patients, 100 parents, and 50 controls. Most patients (96%) were positive for DQA1*0501 (RR = 18.07, p < 0.001), 94% for DQB1*02 (RR = 17.55, p < 0.001), and 92% for both alleles together (RR = 31.82, p < 0.001). DQA1*0501 was found in 52% of controls, DQB1*02 in 44%, and only 24% had both alleles. Patients homozygosity or heterozygosity was estimated by assessing-in each case-whether one or both parents were carriers of the allele of risk. The frequencies of parents both positive for DQA1*0501 (58%) and for DQB1*02 (53.1%) were higher than expected by the fact that the proband is a carrier. These findings suggest that the frequency of homozygosity is increased among CD patients, and therefore, homozygosity for either DQA1*0501 or DQB1*02 represents a risk factor added to the fact of being a carrier.

Antineutrophil cytoplasmic antibodies (ANCA) in idiopathic pulmonary hemosiderosis.

Four children were diagnosed with idiopathic pulmonary hemosiderosis (IPH), over a period of 4 years. Retrospectively, antineurtrophil cytoplasmic antibodies (ANCA) were studied by indirect immunofluorescence (IIF) and ELISA in 18 sera from these patients, stored at -20 degrees C. ANCA-positive sera, from 1/20 to 1/1, 200 dilution, were found in 3/4 of the patients, by IIF. The patient with the highest titre of ANCA died 3 months later during an acute crisis, the other two patients need a minimal dose of steroids. In one case only, a patient who is still without treatment, had no ANCA. The antibodies anti-myeloperoxidase and anti-proteinase-3 were negative or at border line levels. Rheumatoid factor, antinuclear (Hep-2), anti-endomysial, anti-reticulin and antibasement membrane antibodies were negative in all sera. The surviving patients were followed-up for more than 10 years with no systemic or renal disease appearances. The presence of serum ANCA may help to classify children with pulmonary haemorrhage and may have a prognostic value.

[Absence of fibronectin in a 40-day-old child who died as a result of septicemia with disseminated intravascular coagulation]. / Ausencia de fibronectina en un niño de 40 días fallecido durante una septicemia con coagulación intravascular diseminada.

A new born infant who died when he was 40 days old, after urinary infection, septicemia and disseminated intravascular coagulation (DIC) is reported. The concentration of fibronectin (FN) was undetectable (less than 1.1 mg/dl). His mother and one sister had also decreased levels (19 and 19.5 mg/dl), although inside normal limits, when they were compared to simultaneously studied normal controls (27 +/- 8). There were not infections, coagulation disturbs neither keloid scars in the family. A patient cousin also died at the first days of life. The deficiency of our case was much more important that FN levels found in 23 children with sepsis and DIC (range 6-38 mg/dl), therefore it is possible he had a primary deficiency. The investigation of FN levels in all newborns with severe infections or other disturbs is recommended, since these patients could be benefit from purified FN or cryoprecipitate therapy.