Implication of non-coding PAX6 mutations in aniridia.

There is an increasing implication of non-coding regions in pathological processes of genetic origin. This is partly due to the emergence of sophisticated techniques that have transformed research into gene expression by allowing a more global understanding of the genome, both at the genomic, epigenomic and chromatin levels. Here, we implemented the analysis of PAX6, whose coding loss-of-function variants are mainly implied in aniridia, by studying its non-coding regions (untranslated regions, introns and cis-regulatory sequences). In particular, we have taken advantage of the development of high-throughput approaches to screen the upstream and downstream regulatory regions of PAX6 in 47 aniridia patients without identified mutation in the coding sequence. This was made possible through the use of custom targeted resequencing and/or CGH array to analyze the entire PAX6 locus on 11p13. We found candidate variants in 30 of the 47 patients. 9/30 correspond to the well-known described 3' deletions encompassing SIMO and other enhancer elements. In addition, we identified numerous different variants in various non-coding regions, in particular untranslated regions. Among these latter, most of them demonstrated an in vitro functional effect using a minigene strategy, and 12/21 are thus considered as causative mutations or very likely to explain the phenotypes. This new analysis strategy brings molecular diagnosis to more than 90% of our aniridia patients. This study revealed an outstanding mutation pattern in non-coding PAX6 regions confirming that PAX6 remains the major gene for aniridia.

Genetics and epidemiology of aniridia: Updated guidelines for genetic study. / Genética y epidemiología de la aniridia congénita: actualización de buenas prácticas para el diagnóstico genético.

Aniridia is a panocular disease characterized by iris hypoplasia, accompanied by other ocular manifestations, with a high clinical variability and overlapping with different abnormalities of the anterior and posterior segment. This review focuses on the genetic features of this autosomal dominant pathology, which is caused by the haploinsufficiency of the PAX6 gene. Mutations causing premature stop codons are the most frequent among the wider mutational spectrum of PAX6, with more than 600 different mutations identified so far. Recent advances in next-generation sequencing (NGS) have increased the diagnostic yield in aniridia and contributed to elucidate new etiopathogenic mechanisms leading to PAX6 haploinsufficiency. Here, we also update good practices and recommendations to improve genetic testing and clinical management of aniridia using more cost-effective NGS analysis. Those new approaches also allow studying simultaneously both structural variants and point-mutations in PAX6 as well as other genes for differential diagnosis, simultaneously. Some patients with atypical phenotypes might present mutations in FOXC1 and PITX2, both genes causing a wide spectrum of anterior segment dysgenesis, or in ITPR1, which is responsible for a distinctive form of circumpupillary iris aplasia present in Gillespie syndrome, or other mutations in minor genes. Since aniridia can also associate extraocular anomalies, as it occurs in carriers of PAX6 and WT1 microdeletions leading to WAGR syndrome, genetic studies are crucial to assure a correct diagnosis and clinical management, besides allowing prenatal and preimplantational genetic testing in families.

Genetics and epidemiology of aniridia: Updated guidelines for genetic study.

Aniridia is a panocular disease characterized by iris hypoplasia, accompanied by other ocular manifestations, with a high clinical variability and overlapping with different abnormalities of the anterior and posterior segment. This review focuses on the genetic features of this autosomal dominant pathology, which is caused by the haploinsufficiency of the PAX6 gene. Mutations causing premature stop codons are the most frequent among the wider mutational spectrum of PAX6, with more than 600 different mutations identified so far. Recent advances in next-generation sequencing (NGS) have increased the diagnostic yield in aniridia and contributed to elucidate new etiopathogenic mechanisms leading to PAX6 haploinsufficiency. Here, we also update good practices and recommendations to improve genetic testing and clinical management of aniridia using more cost-effective NGS analysis. Those new approaches also allow studying simultaneously both structural variants and point-mutations in PAX6 as well as other genes for differential diagnosis, simultaneously. Some patients with atypical phenotypes might present mutations in FOXC1 and PITX2, both genes causing a wide spectrum of anterior segment dysgenesis, or in ITPR1, which is responsible for a distinctive form of circumpupillary iris aplasia present in Gillespie syndrome, or other mutations in minor genes. Since aniridia can also associate extraocular anomalies, as it occurs in carriers of PAX6 and WT1 microdeletions leading to WAGR syndrome, genetic studies are crucial to assure a correct diagnosis and clinical management, besides allowing prenatal and preimplantational genetic testing in families.

[Vaccination against hepatitis B at the Palma-Palmilla Health Center of Málaga]. / Vacunación contra la hepatitis B en el Centro de Salud Palma-Palmilla de Málaga.

AIM: To describe the results of a programme of anti-hepatitis B vaccination of high-risk groups. DESIGN: Observational descriptive study, of a retrospective character. SITE. At a community level within the confines of Primary Care in the Palma-Palmilla (Málaga) Health Centre, between June 1989 and March 1990. PATIENTS AND OTHER PARTICIPANTS: Individuals with a high risk of Hepatitis B infection, according to a modified CDC (Centre for Diseases Control) scale. The subjects were found during their attendance as patients and from among the Health Centre staff. INTERVENTIONS: The second generation vaccine developed by genetic engineering (Engerix B) was used. It was administered by intramuscular injection in a dosage of 20 mcg to those weighing more than 25 kilos; and of 10 mcg to those weighing less than 25 kilos. The vaccination pattern was of three doses in months 0, 1 and 6, followed by a monthly sero-conversion check. MEASUREMENTS AND MAIN FINDINGS: 169 individuals began the vaccination programme: 17.7% were health workers and 81.6% lived with carriers of the virus. 87.6% completed the vaccination programme. Sero-conversion in the individuals controlled was 95.5%. Only five patients were sero-negative after the third vaccination. Of these four cases achieved sero-conversion after a fourth or fifth dosage. CONCLUSIONS: We found there was a high rate of sero-conversion; and also high acceptance of the programme by those living with a carrier.

[Knowledge and attitudes of ambulatory patients regarding various aspects of pharmacologic treatment]. / Conocimientos y actitudes de pacientes ambulatorios frente a algunos aspectos del tratamiento farmacológico.

Recognizing the patients believes and attitudes towards their disease and treatment as well as facilitating basic information are essential for a satisfactory compliance. In this respect, and in order to know the state of things, personal interviews were carried out following a pre-established protocol and evaluating the degree of acceptance of information oriented to the outpatient. Two hundred and seven questionnaires were collected. Populations mean age was 50 years, 60% were women and there was a high percentage of illiterate patients or with only primary school level (69.8%), with a medium-low social status (98%). In general, they were aware of the indication of treatment (80%) although 56% of patients did not know the consequences of a poor control of their disease. Seventy eight percent wished to receive more information and 147 (96%) patients preferred the physician as the provider of it. The opinion regarding the information leaflets oriented towards the outpatient was unanimously favorable being the most interesting aspects those regarding the instructions on the use of drugs and adverse effects. The educational level was the factor influencing most significantly the patient's attitude. We conclude saying that in order to carry out a pharmacological treatment educational program it is necessary to evaluate the knowledge, needs and attitudes of the population towards whom it is directed.