A collection of three integration-free iPSCs derived from old male and female healthy subjects.

Here, we present the characterization of three iPSC lines derived from dermal fibroblasts of old healthy subjects. Fibroblasts were reprogrammed using Sendai viral vectors encoding OCT4, SOX2, KLF4 and c-MYC. The iPSCs expressed endogenous pluripotency markers, could generate the three germ layers (ectoderm, mesoderm and endoderm), maintained a stable karyotype, and were free from Sendai vectors and reprogramming factors. These integration-free iPSCs can serve for establishing control cell cultures in studies searching for phenotypes and mechanisms that could potentially be dysregulated in degenerative diseases.

A collection of integration-free iPSCs derived from Parkinson's disease patients carrying mutations in the GBA1 gene.

Mutations in the GBA1 gene, which encodes the lysosomal enzyme Glucocerebrosidase1 are major risk factors for Parkinson's disease (PD) and dementia with Lewy bodies (DLB). We have generated induced pluripotent stem cells (iPSCs) from fibroblasts of four PD patients carrying the N370S/wt or the L444P/wt heterozygous mutations in GBA1. The iPSCs presented a normal morphology, expressed endogenous pluripotency markers and could be differentiated into endodermal, mesodermal and ectodermal cells. These iPSCs were free from Sendai viral vectors and reprogramming factors, had a normal karyotype and maintained the original GBA1 genotype.

A collection of four integration-free iPSC lines derived from diagnosed sporadic Alzheimer's disease patients with different APOE alleles.

Genetic polymorphism of apolipoprotein E (APOE) confers differential susceptibility to late-onset Alzheimer's disease (LOAD). The ε3 allele of APOE, the most common isoform, does not represent a risk factor for LOAD. In contrast, the ε4 allele is the strongest genetic risk factor for this disease. Here, we present the characterization of four iPSC lines generated from dermal fibroblasts of diagnosed sporadic AD patients using Sendai viral vectors encoding OCT4, SOX2, KLF4 and c-MYC. The iPSCs expressed endogenous pluripotency markers, could be differentiated into the three germ layers, maintained the original genotypes, and were free from Sendai vectors and reprogramming factors.

An integration-free iPSC line, ICCSICi007-A, derived from a female Alzheimer's disease patient with the APOE-ε4/ε4 alleles.

The epsilon4 (ε4) allele of the APOE gene, which encodes the apolipoprotein E4 (ApoE4), is the strongest genetic risk factor known for late-onset Alzheimer´s disease (LOAD). Here, we present the characterization of an iPSC line generated from dermal fibroblasts of a female AD patient using Sendai viral vectors encoding the transcription factors OCT4, SOX2, KLF4 and c-MYC. The iPSCs maintained the original genotype, a normal karyotype, were free from Sendai viral vectors and reprogramming factors, presented a normal morphology, expressed endogenous pluripotency markers, and could be differentiated into ectodermal, mesodermal and endodermal cells, confirming its pluripotency.

Generation of an integration-free iPSC line, ICCSICi006-A, derived from a male Alzheimer's disease patient carrying the PSEN1-G206D mutation.

The familial form of Alzheimer's disease (FAD), which is caused by mutations in PRESENILIN 1 (PSEN1) and amyloid precursor protein (APP) genes, represents less than 5% of all AD cases and has an early-onset. We report the generation and characterization of an iPSC line derived from a FAD patient carrying the PSEN1-G206D mutation. The iPSC line maintained the original genotype, a normal karyotype, was free from Sendai viral vectors and reprogramming factors (OCT4, SOX2, KLF4 and c-MYC), presented a typical morphology, expressed endogenous pluripotency markers, and could be differentiated into ectodermal, mesodermal and endodermal cells, confirming its pluripotency.

Generation of an integration-free iPSC line, ICCSICi005-A, derived from a Parkinson's disease patient carrying the L444P mutation in the GBA1 gene.

The L444P mutation in the GBA1 gene which encodes ß-glucocerebrosidase-1, is a major risk factor for developing Parkinson's disease (PD) and dementia with Lewy bodies (DLB). We report the generation and characterization of an induced pluripotent stem cell (iPSC) line derived from a female PD patient carrying the L444P/wt mutation. The iPSC line presented a normal morphology, expressed endogenous pluripotency markers, could be differentiated into endodermal, mesodermal and ectodermal cells, was free from Sendai vectors and reprogramming factors, had a normal karyotype and maintained the original GBA1 genotype. Thus, this iPSC line can serve to establish cellular models of PD.

Characterization of patients' requests and pharmacists' professional practice in oropharyngeal condition in Spain.

BACKGROUND: A sore throat is the reason behind 4 million visits to health care services per year in Spain. The management of these ailments is usually associated with an inappropriate use of medicines. Community pharmacists are often the first point of contact for patients under the healthcare system and play a major role in the management of minor ailments. OBJECTIVE: To characterize the pharmacists' professional practice in oropharyngeal condition in terms of patients' requests and pharmacists' interventions performed. SETTING: Community pharmacies throughout Spanish territory. METHOD: Cross-sectional multicenter observational study, undertaken between November 2012 and March 2013. Patients were recruited consecutively in the participant pharmacies. Eligible patients were those making a consultation related to an oropharyngeal condition or requesting treatment for an oropharyngeal condition. A univariate descriptive analysis showing the frequency of occurrence of the different variables was performed. This was completed with a multivariate statistical analysis through a multiple correspondence method, in order to analyze the potential association between the pharmacist profile and the intervention provided. MAIN OUTCOME MEASURE: Professional Pharmacy Service requested (dispensing/counselling/medication review with follow-up), reason for consultation, source of the recommendation, Pharmacist's intervention and reason for referral. RESULTS: 710 pharmacies and 3,547 patients participated in the study. The most frequently requested service was dispensing (44.7 %), followed by counselling (31.8 %). Regarding dispensing, the majority of patients requested a throat preparation, mostly antiseptics. Symptoms associated with the pharmacy consultation were mainly throat symptoms (70.8 %), voice symptoms (24.9 %), and mouth/tongue/lip symptoms (10.1 %). The most common pharmacist intervention was "selection of pharmacological treatment", followed by "dispensing the requested medicine"; 6.51 % of the patients were referred to a general practitioner, mainly due to fever and other symptoms associated with infection. No relationship between the pharmacist profile and the intervention performed was found. CONCLUSION: The most frequently requested professional service was the dispensing service, mainly for treating throat symptoms. Community pharmacists play a major role in managing patients with an oropharyngeal condition. They can keep them out of general practice or act as referral agents when a more severe disease is identified.

PIAS4 is associated with macro/microcephaly in the novel interstitial 19p13.3 microdeletion/microduplication syndrome.

Array comparative genomic hybridization (aCGH) is a powerful genetic tool that has enabled the identification of novel imbalances in individuals with intellectual disability (ID), autistic disorders and congenital malformations. Here we report a 'genotype first' approach using aCGH on 13 unrelated patients with 19p13.3 submicroscopic rearrangement (11 deletions and 2 duplications) and review cases in the literature and in public databases. Shared phenotypic features suggest that these patients represent an interstitial microdeletion/microduplication syndrome at 19p13.3. Common features consist of abnormal head circumference in most patients (macrocephaly with the deletions and microcephaly with the duplications), ID with developmental delay (DD), hypotonia, speech delay and common dysmorphic features. The phenotype is associated with at least a ~0.113 Mb critical region harboring three strong candidate genes probably associated with DD, ID, speech delay and other dysmorphic features: MAP2K2, ZBTB7A and PIAS4, an E3 ubiquitin ligase involved in the ubiquitin signaling pathways, which we hypothesize for the first time to be associated with head size in humans.

An excess of chromosome 1 breakpoints in male infertility.

In a search for potential infertility loci, which might be revealed by clustering of chromosomal breakpoints, we compiled 464 infertile males with a balanced rearrangement from Mendelian Cytogenetics Network database (MCNdb) and compared their karyotypes with those of a Danish nation-wide cohort. We excluded Robertsonian translocations, rearrangements involving sex chromosomes and common variants. We identified 10 autosomal bands, five of which were on chromosome 1, with a large excess of breakpoints in the infertility group. Some of these could potentially harbour a male-specific infertility locus. However, a general excess of breakpoints almost everywhere on chromosome 1 was observed among the infertile males: 26.5 versus 14.5% in the cohort. This excess was observed both for translocation and inversion carriers, especially pericentric inversions, both for published and unpublished cases, and was significantly associated with azoospermia. The largest number of breakpoints was reported in 1q21; FISH mapping of four of these breakpoints revealed that they did not involve the same region at the molecular level. We suggest that chromosome 1 harbours a critical domain whose integrity is essential for male fertility.

[Antiphospholipid syndrome updating]. / Actualización del síndrome antifosfolipídico.

The antiphospholipid syndrome (APS) was defined 20 years ago by Hughes as a thrombosis (arterial and/or venous) state and/or recurrent pregnancy loss in the presence of antiphospholipid antibodies like lupic anticoagulant and/or anticardiolipin, which sometimes coexist with thrombocytopenia. Although the prevalence of APS is estimated to be 3 to 200 cases per 100,000 inhabitants, many cases remain without a proper diagnosis. Recurrent thrombosis events can happen at any site in the vascular tree, yet most common affected sites are deep veins and cerebral arteries. As a result of its vast and protean manifestations, APS is currently being dealt with by most medical specialties. But despite research advances and improvements in its knowledge, many features of this syndrome such as its etiology and pathophysiology are not well understood yet. Standard therapy for vascular and obstetric problems is difficult because of the absence of evidence-based guidance and continues to be based on antiplatelet and anticoagulation drugs. Yet some authors claim individual treatments according to the risk of thrombosis or fetal loss. In this review we offer, through an extensive bibliographical search of the subject over the last 2 years, a view of the state-of-the-art of APS including current criteria for diagnosis, molecular basis, main associated clinical manifestations, management and treatment.

Quantitative abnormalities of peripheral blood distinct T, B, and natural killer cell subsets and clinical findings in obstetric antiphospholipid syndrome.

OBJECTIVE: Few studies have assessed immunophenotypic abnormalities on lymphocyte subsets in patients with antiphospholipid syndrome (APS). We performed an extended immunological study to define alterations of distinct T, B, and natural killer (NK) cell subsets in obstetric patients with APS and their relationship with APS-associated complications. PATIENTS AND CONTROLS: 36 women with APS [Sydney criteria, Group A1 without thrombosis (n=26), Group A2 with thrombosis (n=10)]; and 36 age matched women with recurrent abortion without antiphospholipid antibodies (disease controls; Group B), 36 healthy parous women (healthy controls; Group C), and 36 healthy nonparous women (healthy controls; Group D). Thrombotic events occurred after history of abortions in all A2 women. Three-color whole-blood flow cytometry was used to characterize the distinct immunophenotypes. RESULTS: A1 patients had significantly higher percentages of CD4+CD45RA-CCR7+ central memory cells (A1 vs D), higher percentages of activated CD4+CD25+ T cells (A1 vs D), and lower percentages and absolute counts of CD4+CD45RA-CCR7- effector memory cells (A1 vs D). Group A2 patients had higher percentages and absolute numbers of CD19+CD27-IgD+ naive B cells (A2 vs A1 vs all controls), lower percentages and absolute numbers of CD3-CD56+CD16+ NK cells (A2 vs all controls), and higher percentages of activated CD4+DR+ (A2 vs all controls), CD8+DR+ (A2 vs A1 vs C vs D), CD4+CD38+DR+ (A2 vs D), and CD4+CD25+DR+ T cells (A2 vs all controls). Increased percentages of CD8+DR+ T cells [relative risk (RR) 2.43, 95% CI 1.09-5.44, p=0.02] and of naive B cells (RR 3.05, 95% CI 1.30-7.11, p=0.009) were associated with development of thrombosis. CONCLUSION: In obstetric patients with APS we documented significant changes in T, B, and NK cell homeostasis. Increased levels of CD8+DR+ and CD19+CD27-IgD+ cells might identify obstetric patients with APS at risk of having thrombosis.

A case of ring chromosome 22 with deletion of the 22q13.3 region associated with agenesis of the corpus callosum, fornix and septum pellucidum.

We report a 16-week-gestation foetus obtained by voluntary abortion after prenatal diagnosis, in which a ring chromosome 22 was observed with deletion of the 22q13.3 region. A prenatal study of the amniotic fluid by standard chromosome technique with G bands and FISH (fluorescence in situ hybridisation) was performed. After the abortion, the anatomopathological study of the obtained foetus was carried out. Morphological and histological analysis of the foetus did not reveal severe physical abnormalities, although alterations of the nervous system were observed consisting of corpus callosum, fornix and septum pellucidum agenesia. It could be that the genes in this region that were involved in the development of the central nervous system were responsible for the alterations found in the morphological study. The wide range of manifestations observed in patients with this cytogenetic alteration is probably due to size differences in the deleted region.

Actualización del síndrome antifosfolipídico / Antiphospholipid syndrome updating

El síndrome antifosfolipídico fue definido hace 20 años por Hughes como el estado trombofílico (arterial y/o venoso) y/o de pérdidas fetales recurrentes en presencia de anticuerpos antifosfolípido, como el anticoagulante lúpico o anticardiolipina, acompañado, a veces, de trombocitopenia. Se puede estimar que presenta una prevalencia entre 3-200 casos por 100.000 habitantes; sin embargo, muchos de ellos no son diagnosticados correctamente. Los episodios trombóticos recurrentes pueden presentarse en cualquier parte del árbol vascular, pero las venas profundas y las arterias cerebrales son las afectadas con mayor frecuencia. Su amplio abanico de manifestaciones ha llevado a incluirlo en la práctica totalidad de las especialidades clínicas. A pesar del gran avance efectuado en la comprensión de la etiología y los mecanismos patogénicos del síndrome, su etiología y etiopatogenia todavía no se conocen con certeza. El tratamiento es difícil debido a la ausencia de una orientación clara sustentada en la evidencia científica, pero sigue estando basado en los medicamentos anticoagulantes y antiagregantes, tanto para el tratamiento de los problemas vasculares como de los obstétricos; sin embargo, se detecta una tendencia a reclamar un tratamiento individualizado del paciente acorde con su riesgo trombótico o de pérdida fetal. En esta revisión se pretende, con una exhaustiva búsqueda bibliográfica sobre el tema en las publicaciones de los dos últimos años, dar una visión del estado actual del conocimiento, abordando los requisitos diagnósticos, la etiopatogenia, las principales presentaciones clínicas asociadas, el manejo y el tratamiento de esta enfermedad, que sigue siendo un reto para el médico clínico (AU)