Detection of the Copy Number Variants of Genes in Patients with Familial Cardiac Diseases by Massively Parallel Sequencing.

INTRODUCTION: The implication of copy number variations in familial heart disease is known, although in-depth knowledge is lacking; hence, more studies are needed to further our understanding. Massively parallel sequencing, thanks to its recent surge in use, is emerging as a valid tool for the detection of this type of variant, through the use of appropriate software. METHODS: We conducted a study with 182 patients diagnosed with mendelian cardiovascular diseases who underwent sequencing using a cardiac gene panel and then a specific calling process for copy number variations (CNVs) with ExomeDepth software, which provides us with a Bayes factor (BF), a score of the probability that a CNV detected is true. RESULTS: After a rigorous CNV prioritization process, we confirmed the variants obtained by MLPA or SNP-based array, finding three real CNVs in five individuals in the MYH11, FBN1 and PDMI7 genes. CONCLUSION: The confirmed CNVs present in all cases BF values > 60, thus establishing a threshold to consider real CNVs in the calling process carried out by ExomeDepth on our gene panel.

Interacciones medicamentosas potenciales en pacientes COVID 19 en tratamiento con lopinavir/ritonavir / Potential drug-drug interactions in COVID 19 patients in treatment with lopinavir/ritonavir

OBJETIVOS: Determinar la prevalencia de interacciones potenciales en pacientes COVID-19 en tratamiento con lopinavir/ritonavir (LPV/r). El objetivo secundario fue elaborar recomendaciones e identificar los factores de riesgo asociados a presentar interacciones potenciales con LPV/r. SUJETOS Y MÉTODOS: Estudio transversal y multicéntrico con la participación de 2 hospitales. Se incluyeron pacientes COVID-19 mayores de 18 años, con ingreso hospitalario y en tratamiento con LPV/r. Se realizó un cribado de las interacciones potenciales relacionadas con LPV/r y la medicación domiciliaria y hospitalaria. Se utilizó como base de datos de consulta Lexicomp® (Uptodate), HIV-drug interacctions y COVID-drug interacctions. RESULTADOS: Se incluyeron 361 pacientes con una media de edad de 62,77 ± 14,64 años, donde el 59,6% (n = 215) fueron hombres. El 62,3% (n=225) tuvieron una o más interacciones potenciales y el 26, 87% (n = 97) 2 o más. Las variables independientes asociadas a presentar ≥ 1 interacciones potenciales fueron la edad (> 65) (OR 1,95; IC 95% 1,06-3,59; P = 0,033), el ingreso en UCI (OR 9,22; IC 95% 1,98-42,93; P = 0,005), la enfermedad previa respiratoria (OR 2,90; IC 95% 1,15-7,36; P = 0,024), psiquiátrica (OR 4,14; IC 95% 1,36-12,61; P = 0,013), la dislipemia (OR 3,21; IC 95% 1,63-6,35; P = 0,001) y el número de fármacos prescrito (OR 4,33; IC 95% 2,40-7,81; P = 0,000). CONCLUSIÓN: La prevalencia de interacciones potenciales en paciente COVID-19 en tratamiento con LPV/r es elevada, comportándose como factores de riesgo asociados la edad (> 65), el ingreso en UCI, la enfermedad previa respiratoria, psiquiátrica y la dislipemia y el número de fármacos prescritos

OBJECTIVES: To determine the prevalence of potential interactions in COVID-19 patients receiving lopinavir/ritonavir (LPV/r). The secondary objective was to develop recommendations and identify the risk factors associated with presenting potential interactions with LPV/r. SUBJECTS AND METHODS: Cross-sectional and multicenter study with the participation of 2 hospitals. COVID-19 patients over 18 years of age, admitted to hospital and under treatment with LPV/r were included. A screening of potential interactions related to LPV/r and home and hospital medication was carried out. Lexicomp® (Uptodate), HIV-drug interactions and COVID-drug interactions were used as the query database. RESULTS: 361 patients with a mean age of 62.77 ± 14.64 years were included, where 59.6% (n = 215) were men. 62.3% (n = 225) had 1 or more potential interactions and 26, 87% (n = 97) 2 or more. The independent variables associated with presenting ≥ 1 potential interactions were age (> 65) (OR 1.95; 95% CI 1.06-3.59, P =.033), ICU admission (OR 9.22; CI 95% 1.98-42.93; P = .005), previous respiratory pathology (OR 2.90; 95% CI 1.15-7.36; P =.024), psychiatric (OR 4.14; 95 CI % 1.36-12.61; P =.013), dyslipidemia (OR 3.21; 95% CI 1.63-6.35; P = .001) and the number of drugs prescribed (OR 4.33; 95% CI 2.40-7.81; P =.000). CONCLUSION: The prevalence of potential interactions in COVD-19 patient undergoing treatment with LPV/r is high, with age (> 65), ICU admission, previous respiratory and psychiatric pathology, dyslipidemia and the number of prescribed drugs acting as risk factors

Potential drug-drug interactions in COVID 19 patients in treatment with lopinavir/ritonavir.

OBJECTIVES: To determine the prevalence of potential interactions in COVID19 patients receiving lopinavir/ritonavir (LPV/r). The secondary objective was to develop recommendations and identify the risk factors associated with presenting potential interactions with LPV/r. SUBJECTS AND METHODS: Cross-sectional and multicenter study with the participation of 2 hospitals. COVID 19 patients over 18 years of age, admitted to hospital and under treatment with LPV/r were included. A screening of potential interactions related to LPV/r and home and hospital medication was carried out. Lexicomp® (Uptodate), HIV-drug interactions and COVID-drug interactions were used as the query database. RESULTS: 361 patients with a mean age of 62.77 ±â€¯14.64 years were included, where 59.6% (n = 215) were men. 62.3% (n = 225) had 1 or more potential interactions and 26, 87% (n = 97) 2 or more. The independent variables associated with presenting ≥1 potential interactions were age (>65) (OR 1.95; 95% CI 1.06-3.59, P = .033), ICU admission (OR 9.22; CI 95% 1.98-42.93; P = .005), previous respiratory pathology (OR 2.90; 95% CI 1.15-7.36; P = .024), psychiatric (OR 4.14; 95 CI% 1.36-12.61; P = .013), dyslipidemia (OR 3.21; 95% CI 1.63-6.35; P = .001) and the number of drugs prescribed (OR 4.33; 95% CI 2.40-7.81; P = .000). CONCLUSION: The prevalence of potential interactions in COVD 19 patient undergoing treatment with LPV/r is high, with age (>65), ICU admission, previous respiratory and psychiatric pathology, dyslipidemia and the number of prescribed drugs acting as risk factors.

OBJETIVOS: Determinar la prevalencia de interacciones potenciales en pacientes COVID19 en tratamiento con lopinavir/ritonavir (LPV/r). El objetivo secundario fue elaborar recomendaciones e identificar los factores de riesgo asociados a presentar interacciones potenciales con LPV/r. SUJETOS Y MÉTODOS: Estudio transversal y multicéntrico con la participación 2 hospitales. Se incluyeron pacientes COVID 19 mayores de 18 años, con ingreso hospitalario y en tratamiento con LPV/r. Se realizó un cribado de las interacciones potenciales relacionadas con LPV/r y la medicación domiciliaria y hospitalaria. Se utilizó como base de datos de consulta Lexicomp® (Uptodate), HIV-drug interacctions y COVID-drug interacctions. RESULTADOS: Se incluyeron 361 pacientes con una media de edad de 62,77 ±â€¯14,64 años, donde el 59,6% (n = 215) fueron hombres. El 62,3% (n = 225) tuvieron 1 o más interacciones potenciales y el 26, 87% (n = 97) 2 o más. Las variables independientes asociadas a presentar ≥ 1 interacciones potenciales fueron la edad (> 65) (OR 1,95; IC 95% 1,06­3,59; P = ,033), el ingreso en UCI (OR 9,22; IC 95% 1,98­42,93; P = ,005), la patología previa respiratoria (OR 2,90; IC 95% 1,15­7,36; P = ,024), psiquiátrica (OR 4,14; IC 95% 1,36­12,61; P = ,013), la dislipemia (OR 3,21; IC 95% 1.63­6,35; P = ,001) y el número de fármacos prescrito (OR 4,33; IC 95% 2,40­7,81; P = ,000). CONCLUSIÓN: La prevalencia de interacciones potenciales en paciente COVD 19 en tratamiento con LPV/r es elevada, comportándose como factores de riesgo asociados la edad (>65), el ingreso en UCI, la patología previa respiratoria, psiquiátrica y la dislipemia y el número de fármacos prescritos.

[Potential drug-drug interactions in COVID 19 patients in treatment with lopinavir/ritonavir]. / Interacciones medicamentosas potenciales en pacientes COVID 19 en tratamiento con lopinavir/ritonavir.

OBJECTIVES: To determine the prevalence of potential interactions in COVID-19 patients receiving lopinavir/ritonavir (LPV/r). The secondary objective was to develop recommendations and identify the risk factors associated with presenting potential interactions with LPV/r. SUBJECTS AND METHODS: Cross-sectional and multicenter study with the participation of 2 hospitals. COVID-19 patients over 18 years of age, admitted to hospital and under treatment with LPV/r were included. A screening of potential interactions related to LPV/r and home and hospital medication was carried out. Lexicomp® (Uptodate), HIV-drug interactions and COVID-drug interactions were used as the query database. RESULTS: 361 patients with a mean age of 62.77 ± 14.64 years were included, where 59.6% (n = 215) were men. 62.3% (n = 225) had 1 or more potential interactions and 26, 87% (n = 97) 2 or more. The independent variables associated with presenting ≥1 potential interactions were age (> 65) (OR 1.95; 95% CI 1.06-3.59, P =.033), ICU admission (OR 9.22; CI 95% 1.98-42.93; P =.005), previous respiratory pathology (OR 2.90; 95% CI 1.15-7.36; P =.024), psychiatric (OR 4.14; 95 CI % 1.36-12.61; P =.013), dyslipidemia (OR 3.21; 95% CI 1.63-6.35; P =.001) and the number of drugs prescribed (OR 4.33; 95% CI 2.40-7.81; P =.000). CONCLUSION: The prevalence of potential interactions in COVD-19 patient undergoing treatment with LPV/r is high, with age (> 65), ICU admission, previous respiratory and psychiatric pathology, dyslipidemia and the number of prescribed drugs acting as risk factors.

The economic benefits of increasing breastfeeding rates in Spain.

BACKGROUND: Interventions aimed at promoting breastfeeding rates are among the most effective possible health policies available, with an estimated return of US$35 per dollar invested. Indeed, some authors found that a 10% increase in exclusive breastfeeding rates in the first two years of life led to a reduction in treatment costs of US$312 million in the US, US$7.8 million in the UK, US$30 million in China, and US$1.8 million in Brazil. Among high-income countries, Spain stands out for its low breastfeeding rate. METHODS: We calculated the savings that the Spanish National Health System would have benefited from had breastfeeding rates been higher in Spain, both from the time of hospital discharge and at 6 months postpartum. We followed the methods used in similar studies carried out in the US, Italy, Australia, the Netherlands, and the UK, to conservatively estimate these potential savings by considering only the lower thresholds in all our estimates. Here we approximated the benefits of having increased exclusive breastfeeding rates based on the lower incidence of infantile pathologies among exclusively breastfed infants. Robust evidence indicates that among breastfed infants there is a lower prevalence of otitis media, gastroenteritis, respiratory infections, and necrotising enterocolitis. We obtained the estimated monetary cost of these diseases by combining their prevalences with data about their economic costs for diagnosis-related groups. RESULTS: The estimated effects we calculated imply that the Spanish National Health System could have saved more than €5.6 million for every percentage point increase in exclusive breastfeeding rates in Spain during 2014. CONCLUSIONS: Breastfeeding is essential both for the health of mothers and the health and development of newborns but is rarely considered as an economic issue and remains economically invisible. In addition to the improved wellbeing of mothers and their infants, breastfeeding can positively impact society as a whole and should therefore be better defined in public policies. Thus, strategies aimed at increasing exclusive breastfeeding rates would likely contribute to lowering the fiscal burden of the Spanish National Health System. Moreover, the magnitude of these potential benefits suggests that such policies would likely be socially cost-effective.

Sudden infant death as the most severe phenotype caused by genetic modulation in a family with atrial fibrillation.

AIMS: To assess the functional impact of two combined KCNH2 variants involved in atrial fibrillation, syncope and sudden infant death syndrome. METHODS AND RESULTS: Genetic testing of a 4-month old SIDS victim identified a rare missense heterozygous in KCNH2 variant (V483I) and a missense homozygous polymorphism (K897T) which is often described as a genetic modifier. Electrophysiological characterisation of heterologous HERG channels representing two different KCNH2 genotypes within the family, showed significant differences in both voltage and time dependence of activation and inactivation with a global gain-of-function effect of mutant versus wild type channels and, also, differences between both types of recombinant channels. CONCLUSIONS: The rare variant V483I in combination with K897T produces a gain-of-function effect that represents a pathological substrate for atrial fibrillation, syncope and sudden infant death syndrome events in this family. Ascertaining the genotype-phenotype correlation of genetic variants is imperative for the correct assessment of genetic testing and counselling. TRANSLATIONAL PERSPECTIVE: According to the current guidelines for clinical interpretation of sequence variants, functional studies are an essential tool for the ascertainment of variant pathogenicity. They are especially relevant in the context of sudden infant death syndrome and sudden cardiac death, where individuals cannot be clinically evaluated. The patch-clamp technique is a gold-standard for analysis of the biophysical mechanisms of ion channels.

Genetic susceptibility in pharmacodynamic and pharmacokinetic pathways underlying drug induced arrhythmia and sudden unexplained deaths

Abstract Drug-induced arrhythmia is an adverse drug reaction that can be potentially fatal since it is mostly related to drug-induced QT prolongation, a known risk factor for Torsade de Pointes and sudden cardiac death (SCD). Several risk factors have been described in association to these drug-induced events, such as preexistent cardiac disease and genetic variation. Our objective was to study the genetic susceptibility in pharmacodynamic and pharmacokinetic pathways underlying suspected drug-induced arrhythmias and sudden unexplained deaths in 32 patients. The genetic component in the pharmacodynamic pathway was studied by analyzing 96 genes associated with higher risk of SCD through massive parallel sequencing. Pharmacokinetic mediated genetic susceptibility was investigated by studying the genes encoding cytochrome P450 enzymes using medium-throughput genotyping. Pharmacodynamic analysis showed three probably pathogenic variants and 45 variants of uncertain significance in 28 patients, several of them previously described in relation to mild or late onset cardiomyopathies. These results suggest that genetic variants in cardiomyopathy genes, in addition to those related with channelopathies, could be relevant to drug-induced cardiotoxicity and contribute to the arrhythmogenic phenotype. Pharmacokinetic analysis showed three patients that could have an altered metabolism of the drugs they received involving CYP2C19 and/or CYP2D6, probably contributing to the arrhythmogenic phenotype. The study of genetic variants in both pharmacodynamic and pharmacokinetic pathways may be a useful strategy to understand the multifactorial mechanism of drug-induced events in both clinical practice and forensic field. However, it is necessary to comprehensively study and evaluate the contribution of the genetic susceptibility to drug-induced cardiotoxicity. (AU)

The Use of Insecticide-Treated Curtains for Control of Aedes aegypti and Dengue Virus Transmission in "Fraccionamiento" Style Houses in México.

Dengue, chikungunya, yellow fever, and Zika viruses transmitted by Aedes aegypti mosquitoes are major public health threats in the tropical and subtropical world. In México, construction of large tracts of "fraccionamientos" high density housing to accommodate population growth and urbanization has provided fertile ground for Ae. aegypti-transmitted viruses. We investigated the utility of pyrethroid-treated window curtains to reduce both the abundance of Ae. aegypti and to prevent dengue virus (DENV) transmission in fraccionamiento housing. Windows and doors of fraccionamiento homes in urban/suburban areas, where Ae. aegypti pyrethroid resistance associated with the Ile1016 knock down resistance (kdr) mutation in the voltage gated sodium channel gene was high, and in rural areas, where kdr resistance was low, were fitted with either insecticide-treated curtains (ITCs) or non-treated curtains (NTCs). The homes were monitored for mosquito abundance and DENV infection. ITCs reduced the indoor abundance of Ae. aegypti and the number of DENV-infected mosquitoes in homes in rural but not in urban/suburban study sites. The presence of non-treated screens also was associated with reduced numbers of mosquitoes in homes. "Super-infested" homes, yielding more than 50 mosquitoes, including DENV-infected mosquitoes, provide a significant public health risk to occupants, visitors, and people in neighboring homes.

Broad-based molecular autopsy: a potential tool to investigate the involvement of subtle cardiac conditions in sudden unexpected death in infancy and early childhood.

OBJECTIVES: Sudden unexplained death in children is a tragic and traumatic event, often worsened when the cause of death cannot be determined. This work aimed to investigate the presence of putative pathogenic genetic variants in a broad spectrum of cardiomyopathy, channelopathy and aortic disease associated genes that may have increased these children's vulnerability to sudden cardiac death. DESIGN: We performed molecular autopsy of 41 cases of sudden unexplained death in infants and children through massive parallel sequencing of up to 86 sudden cardiac death-related genes. Multiple in silico analyses were conducted together with a thorough review of the literature in order to prioritise the putative pathogenic variants. RESULTS: A total of 63 variants in 35 cases were validated. The largest proportion of these variants is located within cardiomyopathy genes although this would have been more expected of channelopathy gene variants. Subtle microscopic features of heart tissue may indicate the presence of an early onset cardiomyopathy as a predisposing condition to sudden unexpected death in some individuals. CONCLUSIONS: Next-generation sequencing technologies reveal the existence of a wide spectrum of rare and novel genetic variants in sarcomere genes, compared with that of cardiac ion channels, in sudden unexplained death in infants and children. Our findings encourage further investigation of the role of early onset inherited cardiomyopathies and other diseases involving myocardial dysfunction in these deaths. Early detection of variants in these individuals could help to unmask subtle forms of disease within their relatives, who would eventually benefit from better counselling about their genetic history.

Genotoxicity and cytotoxicity of sevoflurane in two human cell lines in vitro with ionizing radiation.

OBJECTIVE: To determine the in vitro toxicity of different concentrations of sevoflurane in cells exposed to X-ray. METHODS: The genotoxic effects of sevofluorane were studied by means of the micronucleus test in cytokinesis-blocked cells of irradiated human lymphocytes. Subsequently, its cytotoxic effects on PNT2 (normal prostate) cells was determined using the cell viability test (MTT) and compared with those induced by different doses of X-rays. RESULTS: A dose- and time-dependent cytotoxic effect of sevofluorane on PNT2 cells was determined (p >0.001) and a dose-dependent genotoxic effect of sevofluorane was established (p >0.001). However, at volumes lower than 30 µL of sevofluorane at 100%, a non-toxic effect on PNT2 cells was shown. CONCLUSION: Sevofluorane demonstrates a genotoxic capacity as determined in vitro by micronucleus test in cytokinesis-blocked cells of irradiated human lymphocytes.

OBJETIVO: Determinar la capacidad genotóxica del anestésico sevofluorano en en células expuestas a radiación ionizante. MÉTODOS: La genotoxicidad del sevofluorane se determinó mediante el test del bloqueo citocinético de linfocitos humanos irradiados bloqueados con citochalasina. La capacidad citotóxica se determino mediante el test de viabilidad celular e inhibición del crecimiento celular (MTT) en células PNT2 (epiteliales de próstata), comparando sus resultados con los inducidos por diferentes dosis de rayos X. RESULTADOS: Se ha determinado un efecto citotóxico del sevofluorane sobre las células PNT2 que presenta correlación con la dosis administrada y el tiempo de estudio utilizado (p >0.001), así como un efecto genotóxico con características dosis-dependientes (p >0.001). Sin embargo, con volúmenes de sevofluorane puro inferiores a 30 µL no encontramos efecto citotóxico sobre las células PNT2. CONCLUSIÓN: Sevofluorane muestra una significativa capacidad genotóxica in vitro determinada mediante el test de micronúcleos en linfocitos humanos irradiados con bloqueados citocinético mediante citochalsina.

Household use of insecticide consumer products in a dengue-endemic area in México.

OBJECTIVES: To evaluate the household use of insecticide consumer products to kill mosquitoes and other insect pests, as well as the expenditures for using these products, in a dengue-endemic area of México. METHODS: A questionnaire was administered to 441 households in Mérida City and other communities in Yucatán to assess household use of insecticide consumer products. RESULTS: A total of 86.6% of surveyed households took action to kill insect pests with consumer products. The most commonly used product types were insecticide aerosol spray cans (73.6%), electric plug-in insecticide emitters (37.4%) and mosquito coils (28.3%). Mosquitoes were targeted by 89.7% of households using insecticide aerosol spray cans and >99% of households using electric plug-in insecticide emitters or mosquito coils. Products were used daily or every 2 days in most of the households for insecticide aerosol spray cans (61.4%), electric plug-in insecticide emitters (76.2%) and mosquito coils (82.1%). For all products used to kill insect pests, the median annual estimated expenditure per household that took action was 408 Mexican pesos ($MXN), which corresponded to approximately 31 $US. These numbers are suggestive of an annual market in excess of 75 million $MXN (>5.7 million $US) for Mérida City alone. CONCLUSION: Mosquitoes threaten human health and are major nuisances in homes in the study area in México. Households were found to have taken vigorous action to kill mosquitoes and other insect pests and spent substantial amounts of money on insecticide consumer products.

Next generation sequencing challenges in the analysis of cardiac sudden death due to arrhythmogenic disorders.

Inherited arrhythmogenic disorders is a relatively common cause of cardiac sudden death in young people. Diagnosis has been difficult so far due to the genetic heterogeneity of the disease. Next generation sequencing (NGS) is offering a new scenario for diagnosis. The purpose of our study was to validate NGS for the analysis of twenty-eight genes known to be associated with inherited arrhythmogenic disorders and therefore with sudden cardiac death. SureSelect hybridization was used to enrich DNA from 53 samples, prior to be sequenced with the SOLID™ System of Life Technologies. Depth of coverage, consistency of coverage across samples, and location of variants identified were assessed. All the samples showed a depth of coverage over 200×, except one of them discarded because of its coverage below 30×. Average percentage of target bp covered at least 20× was 96.45%. In the remaining samples, following a prioritization process 46 possible variants in 31 samples were found, of which 45 were confirmed by Sanger sequencing. After filtering variants according to their minor allele frequency in the Exome Sequencing Project 27 putative pathogenic variants in 20 samples remained. With the use of in silico tools, 13 variants in 11 samples were classified as likely pathogenic. In conclusion, NGS allowed us to accurately detect arrhythmogenic disease causing mutations in a fast and cost-efficient manner that is suitable for daily clinical and forensic practice of genetic testing of this type of disorders.

Estudio comparativo de la eficacia de distintos protocolos de estimulación en ciclos de donantes de ovocitos / Comparative study of different stimulation protocols efficacy in oocyte donors’ cycles

Introducción: Los estudios clínicos previos que comparan los diferentes tipos de gonadotropinas en estimulación de donantes de ovocitos no son concluyentes. Algunos indican mejores resultados gestacionales por ciclo para pacientes tratadas con FSHr vs. FSHu mientras que otros no observan diferencias significativas. Este estudio compara 3 grupos de donantes de ovocitos que usaron: FSHr+hMGu, FSHu+hMGu y hMGu sin combinación. Materiales y Métodos: Se diseñó un estudio retrospectivo comparativo en 510 estimulaciones de donantes de ovocitos sometidas a estimulación ovárica desde el 1 de enero de 2009 hasta el 1 de enero de 2012. Se empleó un protocolo largo con agonistas de la GnRH y anticonceptivos orales previos. Para el análisis estadístico se usaron test paramétricos (t-student) y no paramétricos (Chi-cuadrado) con límite de significación de p<0,05. Resultados: Las estimulaciones se dividieron en tres grupos: 247 estimulaciones con FSHu y hMGu, 163 estimulaciones con hMGu y 100 estimulaciones con FSHr en combinación con hMGu. Se encontraron diferencias significativas en la tasa de cancelación, número de ovocitos aspirados, maduros y dosis total de gonadotropinas a favor de los grupos de estimulación combinada vs. hMGu. Los resultados gestacionales fueron similares entre los grupos. Conclusiones: Los tres grupos de estimulación evaluados nos proporcionan un resultado similar en cuanto a las tasas de fecundación, calidad embrionaria, gestación clínica e implantación, mientras que hMGu sin combinación con ninguna otra gonadotropina produce mayores tasas de cancelación. Ante estos resultados comparables clínicamente, concluimos que la eficacia de la FSHu y FSHr es similar, por lo que podrán valorarse otros factores como el coste económico a la hora de elegir el protocolo de gonadotropinas en ciclos de donación ovocitaria (AU)

Introduction: The previous clinical studies that have compared the different stimulation protocols in IVF cycles are not conclusive. Some works provide better pregnancy results per cycle in patients stimulated with FSHr vs. FSHu while others do not observe significant differences. This study compares 3 groups of oocyte donors: FSHr+hMGu, FSHu+hMGu, and hMGu. Materials and Methods: We designed a retrospective comparative study of 510 oocyte donors’ cycles undergoing ovarian stimulation from 1 January 2009 until 1 January 2012. A long protocol half-dose with GnRH agonists and contraceptive pills were used in all cases. Comparisons were carried-out using chi-square and t-student test (p<0, 05). Results: Donors were divided into three stimulation’s groups: 247 with FSHu + hMGu, 163 with hMGu, and 100 with FSHr + hMGu. We found significant differences in the cancellation rate, number of retrieved and mature oocytes, and total dose of gonadotropins in favour of combined ovarian stimulation groups. The pregnancy results were comparable in the three groups. Conclusions: All the study stimulation groups provide similar results in terms of fertilization rate, embryo quality, and pregnancy and implantation rates. hMGu group produces higher cancellation rates. With these clinically equivalent results we conclude that the efficacy of treatments with FSHu and FSHr is similar, therefore we could evaluate other factors such as the economic cost to choose the adequate ovarian stimulation protocol in oocyte donors´ cycles (AU)

Effects of post-thaw incubation on motility, acrosomal integrity and in vitro fertilizing capacity of boar spermatozoa cryopreserved in 0.5 and 5ml straws.

The effect of post-thaw incubation (0 vs. 5h at 15°C) and straw size (5 vs. 0.5ml) on motility, acrosomal integrity and in vitro fertilizing (IVF) capacity of cryopreserved boar spermatozoa was studied. In samples assessed immediately after thawing, no differences were found between the two straw sizes. After 5h post-thaw incubation, all parameters, except polyspermy, decreased and, spermatozoa packaged in 5ml straws showed better functional and IVF parameters than these in 0.5ml straws.

Sarcomeric gene mutations in sudden infant death syndrome (SIDS).

In developed countries, sudden infant death syndrome (SIDS) represents the most prevalent cause of death in children between 1 month and 1 year of age. SIDS is a diagnosis of exclusion, a negative autopsy which requires the absence of structural organ disease. Although investigators have confirmed that a significant percentage of SIDS cases are actually channelopathies, no data have been made available as to whether other sudden cardiac death-associated diseases, such as hypertrophic cardiomyopathy (HCM), could be responsible for some cases of SIDS. The presence of a genetic mutation in the sarcomeric protein usually affects the force of contraction of the myocyte, whose weakness is compensated with progressive hypertrophy and disarray. However, it is unclear whether in the most incipient forms, that is, first years of life, the lack of these phenotypes still confers a risk of arrhythmogenesis. The main goal of the present study is to wonder whether genetic defects in the sarcomeric proteins, previously associated with HCM, could be responsible for SIDS. We have analysed 286 SIDS cases for the most common genes implicated in HCM in adults. A total of 680 mutations localised in 16 genes were analysed by semi-automated matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDITOF-MS) using the Sequenom MassARRAY(®) System. Ten subjects with completely normal hearts showed mutated alleles at nine of the genetic variants analysed, and one additional novel mutation was detected by conventional sequencing. Therefore, a genetic mutation associated with HCM may cause sudden cardiac death in the absence of an identifiable phenotype.

Prevalence of HCM and long QT syndrome mutations in young sudden cardiac death-related cases.

Cardiomyopathies and channelopathies are major causes of sudden cardiac death. The genetic study of these diseases is difficult because of their heterogenic nature not only in their genetic traits but also in their phenotypic expression. The purpose of the present study is the analysis of a wide spectrum of previously known genetic mutations in key genes related to hypertrophic cardiomyopathy (HCM), long QT syndrome (LQTS), and Brugada syndrome (BrS) development. The samples studied include cases of sudden cardiac death (SCD) in young adults and their relatives in order to identify the real impact of genetic screening of SCD in forensic cases. Genetic screening of described variation in 16 genes implicated in the development of HCM and three more genes implicated in LQTS and BrS was performed by using MassARRAY technology. In addition, direct sequencing of the two most prevalent genes implicated in the development of SQTL type 1 and 2 was also carried out. Genetic screening allowed us to unmask four possibly pathogenic mutation carriers in the 49 SCD cases considered; carriers of mutation represent 9% (2/23) of the probands with structural anomalies found after autopsy and 7% (1/14) of the probands with structurally normal hearts after in depth autopsy protocol. One mutation was found among 12 of the recovered SCD cases considered. In people with direct family history of sudden cardiac death, but not themselves, 11 additional mutation carriers were found. Three different mutations were found in six of the 19 LQTS patients, representing three families and two different mutations were found among six patients with previous syncope. Genetic analysis in sudden cardiac death cases could help to elucidate the cause of death, but it also can help in the prevention of future deaths in families at risk. The study presented here shows the importance and relevance of genetic screening in patients with signs of cardiac hypertrophy and in family cases with more than one relative affected.

Successful aging: un enfoque holístico / Successful aging: A holistic perspective

Introducción: el envejecimiento creciente de la sociedad ha promovido que exista un mayor interés por comprender el término successful aging. Objetivo: explorar el término successful aging desde una perspectiva holística. Método: se llevó a cabo una revisión de la evidencia disponible en las principales bases de datos, utilizando los términos, ‘envejecimiento’, ‘envejeciminto exitoso’ y ‘enfermería’, durante el período 2000-2010. Resultados:en base al análisis de los 14 artículos seleccionados, se han descrito las principales categorías y subcategorías del término successful aging. Conclusiones: se ofrece una posible definición del término successful aging desde una perspectiva holística, teniendo en cuenta la multidimensionalidad, el dinamismo y la operatividad del concepto.Desde esta revisión se plantea la realización de futuros estudios (AU)

Introduction: the increasing aging of society has promoted the existence of a greater interest in understanding the term Successful aging. Aim: to explore the term Successful aging from a holistic perspective. Method. a review was conducted in the main databases for the period 2000-2010. The search terms ‘aging’, ‘successful aging’ y ‘Nursing’were combined. Results: based on the analysis of the fourteen selected articles the major categories and subcategories of the term Successful aging were described. Conclusion: it offers a possible definition of Successful agingfrom a holistic perspective, taking into account the multidimensionality, the dynamism and viability of the concept.From this review future studies are proposed (AU)

Identification of a novel MYBPC3 gene variant in a patient with hypertrophic cardiomyopathy.

Hypertrophic cardiomyopathy (HCM) is a genetic disorder characterized by cardiac hypertrophy caused by mutations in sarcomere protein genes. MYBPC3 mutations are reported as a frequent cause of HCM. We aimed to identify the gene mutation underlying HCM in an Italian patient and his family composed of 13 relatives. Mutation screening of 658 known mutations was performed using a rapid and efficient mutation detection system based on semiautomated MALDI-TOF mass spectrometry using the Sequenom MassArray System and iPLEX Gold genotyping chemistry. Subsequently, direct sequencing of the coding exons and flanking intronic regions was performed for the most suitable HCM genes (MYBPC3, MYH7, TNNT2, TNNI3, and TPM1) in the index patient. We found a novel MYBPC3 gene mutation: G13999T (Gln689His). No other sarcomere gene mutation was found in this family. This genetic variant, which changes the last amino acid of MYBPC3 exon 21, affects a highly conserved residue. Furthermore, the Gln689His does not appear in public databases and has never been described as a polymorphism. The potential pathogenic role of this novel mutation was underlined by its absence in a sample of healthy subjects (n = 122) from the general Italian population. In summary, a novel MYBPC3 gene mutation has been identified in a patient affected by HCM, whereas it was absent in 244 reference alleles.

A new approach to long QT syndrome mutation detection by Sequenom MassARRAY system.

Congenital long QT syndrome is an inherited cardiac disorder characterized by a prolonged QT interval and polymorphic ventricular arrhythmias that could result in recurrent syncope, seizures or sudden death as the most dramatic event. Until now QT interval mutations have been described in 12 genes, where the majority of mutations reside in three genes KCNQ1, KCNH2, and SCN5A. Diagnosis and prognosis are directly related with the gene and mutation involved. We have developed a diagnostic approach for long QT syndrome and Brugada syndrome based on published mutations and Sequenom MassArray system. Three diagnostic tests have been developed, oriented to each of the three most prevalent genes in the long QT syndrome. A total of 433 mutations are analyzed in 38 multiplex reactions, allowing their detection in about 48 h. Tests were validated on 502 samples from individuals with different clinical conditions and family history. The average call rates obtained for each of the tests were 93, 83, and 73% in KCNQ1, KCNH2, and SCNA, respectively. Sequenom MassARRAY mutation detection is a reliable, highly flexible, and cost-efficient alternative to conventional methods for genetic testing in long QT syndrome and Brugada syndrome, facilitating flexible upgrades of the version of the test presented here with the inclusion of new mutations.

Reservorio intraarterial hepático para quimioterapia. Colocación percutánea en la arteria subclavia izquierda / Intra-arterial hepatic reservoir for chemotherapy. Percutaneous placement in left subclavian artery

Objetivos: Describir la técnica empleada y valorar los resultados, en la implantación percutánea de un catéter en la arteria hepática a través de la subclavia izquierda, para administración de quimioterapia regional. Material y métodos: Entre abril de 1999 y junio de 2002, a 33 pacientes (17 varones y 16 mujeres) de 52,9 años de media y con evidencia de lesión hepática (32 metástasis hepáticas, un hepatocarcinoma fibrolamelar), se les implantaron 36 catéteres intraarteriales. El acceso en todos los casos fue la arteria subclavia izquierda con guía ecográfica. Se estudió la vascularización hepática y se emplazó el catéter distal en arteria hepática. Asimismo, se embolizaron ramas extrahepáticas de la arteria hepática, para evitar fuga del quimioterápico. La cánula se conectó al reservorio (Port-a-cath Titaniumvenous system. Daltec MN. USA) implantado, subcutáneo, próximo al punto de punción. Resultados: El éxito técnico se consiguió en todos los casos (catéter en arteria hepática común en 31 casos, tres en la arteria hepática derecha, uno en la arteria hepática izquierda procedente de la gástrica izquierda y uno en la arteria hepática derecha procedente de la mesentérica superior). La permanencia de los reservorios osciló entre 22 y 740 días (media, 222,4 días). Se registraron complicaciones en 13 casos (36,1%): cinco migraciones de cánula, cuatro vainas de fibrina, tres obstrucciones del vaso cateterizado, una contaminación y una fuga en la conexión. Se solucionaron de modo percutáneo o con retirada del dispositivo. Conclusión: La implantación percutánea de un catéter intraarterial hepático y reservorio, a través de la subclavia izquierda con ayuda ecográfica, es técnicamente factible, y con una tasa de complicaciones aceptable además de solucionables percutáneamente

Objetives: To describe the technique used and to assed about the results obtained in the percutaneous implantation of a catheter in hepatic artery (HA) through left subclavian artery to administer regional chemotherapy. Material and methods: Between April 1999 and June 2002, 33 patients (17 men and 16 women) with a mean age of 52.9 years and with evidence of hepatic lesion (32 hepatic metastases, 1 fibrolamellar hepatocarcinoma (HCC), 36 intra-arterial catheters were implanted. Access in every case was the left subclavian artery with ultrasonographic guide. Hepatic vascularization was studied and distal catheter was located in the HA. Furthermore, extrahepatic branches of the HA were embolized to avoid chemotherapeutic escape. The cannula was connected to the subcutaneous implanted reservoir (Port-a-cath Titaniumvenous system. Daltec. M.N. USA) close to the puncture point. Results: Technical success was achieved in every case (catheter in common HA 31 cases, 3 in right HA, 1 in left HA from left gastric artery and 1 in right HA from upper mesenteric artery). The permanence of the reservoirs ranged from 22 to 740 days (mean 222.4 days). Complications were recorded in 13 cases (36.1%): 5 canula migrations, 4 fibrin sheath, 3 obstruction of catheterized vessel, 12 contamination and 1 escape from the connection. They were solved percutaneously or by withdrawal of the device. Conclusion: Percutaneous implantation of an intra-arterial hepatic catheter and reservoir through the left subclavian artery with ultrasonographic guidance is technically feasible and has an acceptable rate of complications besides being percutaneously solutionable