Adapting an established Ampliseq microhaplotype panel to nanopore sequencing through direct PCR.

We have adapted an established Ampliseq microhaplotype panel for nanopore sequencing with the Oxford Nanopore Technologies (ONT) system, as a cost-effective and highly scalable solution for forensic genetics applications. For this purpose, we designed a protocol combining direct PCR amplification from unextracted DNA with ONT library construction and sequencing using the MinION device and workflow. The analysis of reference samples at input amounts of 5-10 ng of DNA demonstrates stable coverage patterns, allele balance, and strand bias, reaching profile completeness and concordance rates of ∼95%. Similar levels were achieved when using direct-PCR from blood, buccal and semen swabs. Dilution series results indicate sensitivity is maintained down to 250 pg of input DNA, and informative profiles are produced down to 62.5 pg. Finally, we demonstrated the forensic utility of the nanopore workflow by analyzing two third degree pedigrees that showed low likelihood ratio values after the analysis of an extended panel of 38 STRs, achieving likelihood ratios 2-3 orders of magnitude higher when testing with the MinION-based haplotype data.

Actividades deportivas con soporte musical: ¿una nueva forma de aprender reanimación cardiopulmonar? / Sports Activities with Musical Support: A New Way to Learn Cardiopulmonary Resuscitation?

El objetivo del estudio fue analizar la viabilidad de enseñar RCP en actividades deportivas con apoyo musical y evaluar si este nuevo enfoque iguala los resultados de un curso tradicional de reanimación cardiopulmonar (RCP). Participaron 84 estudiantes universitarias sin conocimientos de RCP distribuidas en dos grupos de forma aleatoria. El primero recibió formación de RCP sólo manos integradas en una clase de aeróbic y el otro recibió un curso de RCP de manera convencional. Los resultados en la clase coreografiada comparados con el curso tradicional fueron: Profundidad media de compresiones (41,64 vs 42,92; p = 0,446), re-expansión correcta (65,47% vs 72,47%; p = 0,423), frecuencia de compresiones en un minuto (102,50 vs 138,53; p < 0,001), compresiones totales (202,50 vs 277,95; p < 0,001). Los dos métodos formativos alcanzaron resultados similares por lo que la introducción de la enseñanza de RCP en actividades deportivas puede ser un nuevo método formativo

The aim of the study was to analyse the feasibility of teaching CPR in sports activities with musical support in order to assess if this new methodology of CPR training is as effective as a traditional CPR course. 84 university students with no knowledge of CPR participated. They were distributed in two groups randomly. The first one received an experimental fitness dance class CPR course and the other was given a traditional training CPR course. The following parameters were obtained: fitness dance class group vs traditional CPR course: Average compression depth (41,64 vs 42,92; p = 0,446), Chest Recoil (65,47% vs 72,47%; p = 0,423), average compression rate (102.50 vs 138.53; p<0.001), total number of compressions (202.50 vs 277.95; p< 0.001). The two training methods achieved similar results, so CPR teaching in sports activities with musical support can be a new training method

Fine tuning of magnetite nanoparticle size distribution using dissymmetric potential pulses in the presence of biocompatible surfactants and the electrochemical characterization of the nanoparticles.

The effects of varying the surfactant concentration and the anodic pulse potential on the properties and electrochemical behaviors of magnetite nanoparticles were investigated. The nanoparticles were synthesized with an electrochemical method based on applying dissymmetric potential pulses, which offers the advantage that can be used to tune the particle size distribution very precisely in the range of 10 to 50 nm. Under the conditions studied, the surfactant concentration directly affects the size distribution, with higher concentrations producing narrower distributions. Linear voltammetry was used to characterize the electrochemical behavior of the synthesized nanoparticles in both the anodic and cathodic regions, which are attributed to the oxidation of Fe(2+) and the reduction of Fe(3+); these species are part of the spinel structure of magnetite. Electrochemical impedance spectroscopy data indicated that the reduction and oxidation reactions of the nanoparticles are not controlled by the mass transport step, but by the charge transfer step. The sample with the highest saturation magnetization was that synthesized in the presence of polyethylene glycol.

Differential cross sections and product rotational polarization in A + BC reactions using wave packet methods: H+ + D2 and Li + HF examples.

The state-to-state differential cross sections for some atom + diatom reactions have been calculated using a new wave packet code, MAD-WAVE3, which is described in some detail and uses either reactant or product Jacobi coordinates along the propagation. In order to show the accuracy and efficiency of the coordinate transformation required when using reactant Jacobi coordinates, as recently proposed [ J. Chem. Phys. 2006 , 125 , 054102 ], the method is first applied to the H + D(2) reaction as a benchmark, for which exact time-independent calculations are also performed. It is found that the use of reactant coordinates yields accurate results, with a computational effort slightly lower than that when using product coordinates. The H(+) + D(2) reaction, with the same masses but a much deeper insertion well, is also studied and exhibits a completely different mechanism, a complex-forming one which can be treated by statistical methods. Due to the longer range of the potential, product Jacobi coordinates are more efficient in this case. Differential cross sections for individual final rotational states of the products are obtained based on exact dynamical calculations for some selected total angular momenta, combined with the random phase approximation to save the high computational time required to calculate all partial waves with very long propagations. The results obtained are in excellent agreement with available exact time-independent calculations. Finally, the method is applied to the Li + HF system for which reactant coordinates are very well suited, and quantum differential cross sections are not available. The results are compared with recent quasiclassical simulations and experimental results [J. Chem. Phys. 2005, 122, 244304]. Furthermore, the polarization of the product angular momenta is also analyzed as a function of the scattering angle.

Cirugía de la incontinencia urinaria bajo anestesia general sin ingreso / No disponible

No disponible

Entropic forces in dilute colloidal systems.

Depletion forces can be accounted for by a contraction of the description in the framework of the integral equations theory of simple liquids. This approach includes, in a natural way, the effects of the concentration on the depletion forces, as well as energetic contributions. In this paper we systematically study this approach in a large variety of dilute colloidal systems composed of spherical and nonspherical hard particles, in two and in three dimensions, in the bulk and in front of a hard wall with a relief pattern. We show by this way the form in which concentration and geometry determine the entropic interaction between colloidal particles. The accuracy of our results is corroborated by comparison with computer simulations.

Grado de utilización de los recursos sanitarios y formas de presentación de los trastornos somatomorfos en atención primaria / Level of use of the sanitary services and forms of appearance o famatoform disorders in primary care

El objetivo de nuestro estudio es conocer el grado de utilización de los recursos sanitarios por parte de los pacientes con diagnóstico de trastorno somatomorfo y las formas de presentación del mismo en nuestro medio. Para ello diseñamos un estudio de casos y controles en el Centro de Salud de La Bañeza (León). Se midió la frecuentación de la consulta de atención primaria, estudios complementarios realizados, derivaciones al segundo nivel asistencial, derivaciones a Salud Mental, prescripciones farmacéuticas realizadas y motivos de consulta, observando que los pacientes con trastorno somatomorfo son más frecuentadores y consumen más recursos que los pacientes sin ese diagnóstico y que los síntomas neurológicos son la forma de presentación más frecuente en nuestro medio

The objective of our study is to know the grade of use of the sanitary services by the patients with the diagnosis of somatoform disorder and their forms of presentation. For this purpose we designed a case-control study in the health center of La Bañeza (León). The frequency these patients go to the health center, the complementary studies made, the enquiries made to other specialists, the enquiries made to the mental health centers, the pharmaceutical prescriptions made and the reasons of consultation were measured, observing that the patients with somatoform disorders go more times to the health center and consume more resources that the patients without that diagnosis and that the neurological symptoms is the more frequent form of appearance of these disorders

Perspectivas prometedoras en la enfermedad cerebrovascular isquémica. Parte II: tratamiento / Promising perspectives in the ischemic cerebrovascular disease. Part II: treatment

Se efectúa una revisión sobre la terapéutica de la enfermedad cerebrovascular isquémica, basado en la fisiopatología y el modo de acción de los diferentes medicamentos de que disponemos en el momento actual. Se analizan las modalidades de tratamiento más efectivo utilizados por diferentes investigadores en centros hospitalarios dedicados al estudio de esta enfermedad. (AU)

Perspectivas prometedoras en la enfermedad cerebrovascular isquémica. Parte I: patogenia / Promising perspectives in the ischemic cerebrovascular disease. Part I: pathogenesis

Se realiza una revisión fisiopatológica de los diferentes aspectos que inciden en la enfermedad cerebrovascular isquémica, lo cual permitirá nuevas posibilidades terapéuticas en el futuro de esta enfermedad de gran morbi-mortalidad en el mundo (AU)

MDM2 mediated nuclear exclusion of p53 attenuates etoposide-induced apoptosis in neuroblastoma cells.

The p53 gene in neuroblastoma tumors (NB) is rarely mutated but the protein accumulates in the cytoplasm. Because p53 can mediate the cytotoxic effects of chemotherapeutic agents, it is important to determine whether accumulation of p53 in the cytoplasm impairs p53 function. Data presented here indicate that hyperactive nuclear export of p53 suppresses etoposide-induced apoptosis but does not prevent growth arrest. We compared p53 function in a pair of NB subclones. Our data show etoposide induces complete trans-location of p53 to the nucleus and activation of apoptosis in the neuroblastic NB cell line SH-SY5Y (N-type), which expresses low levels of MDM2. However, in Schwann cell-like SH-EP1 cells (S-type), which have up to 10-fold higher levels of MDM2, p53 accumulates in the cytoplasm and the cells are extremely resistant to etoposide-induced apoptosis. Notably, when MDM2 expression is inhibited in S-type cells, with a phosphorothioated antisense oligonucleotide (AS5), then p53 accumulates in the nucleus and the SH-EP1 cells undergo apoptosis. Surprisingly, induction of p21 and G1-arrest are not attenuated in S-type cells, despite the predominantly cytoplasmic location of p53. Whereas, G1-arrest is attenuated in the SH-SY5Y cells, which have high levels of nuclear p53. Taken together, these findings suggest attenuation of G1-arrest is related to the differentiation status of neuroblastomas and occurs downstream of p53 nuclear accumulation. These results demonstrate for the first time that hyperactive nuclear export of p53 attenuates chemotherapy-induced apoptosis in NB cells, and our findings suggest that inhibitors of MDM2 may enhance the therapeutic efficacy of etoposide by promoting apoptosis rather than trans-differentiation.

[Prevalence of dental caries and associated factors in children at day care centers]. / Prevalencia de caries y factores asociados en niños de estancias infantiles.

UNLABELLED: Currently dental caries is regarded as a public health problem throughout world. Ninety percent of the Latin American population suffers from caries. In Mexico, a research informs of a prevalence of 78% in 5-year-old children from the Southern of the Federal District, while the nearby State of Mexico reports 90% in children the same age that suffers from this same problem. OBJECTIVE: Identifying the prevalence of dental caries, dmf-t and dmf-s indexes, and the relation ship with associated factors in daycare centers of the # 3 Southwest Delegation of the Instituto Mexicano del Seguro Social (IMSS). A total of 934 children from ages 1 to 6 years of age from October 1997 to May 1998, were studied; 52.3% were boys and 47.7%, girls. The presence of caries was examined and determined through WHO criteria. Socioeconomic level was identified the parents questionnaire. Prevalence of caries in the population studied was 22.1%; we also found a decayed, missing, filled-teeth index (dmf-t) for the whole population of 0.71. The average of this index was 3.71.

Increased apoptosis susceptibility in mesangial cells from spontaneously hypertensive rats.

We have examined the susceptibility to apoptosis in mesangial cells from spontaneously hypertensive rats (SHR) or from normotensive rats (WKY) and the possible involvement of nitric oxide in this process. Mesangial cells monolayers from either SHR or normal rats were incubated for 12 h in medium with or without fetal calf serum (FCS) and with or without thapsigargin (Tg, 10(-6) M). A series of cultures from rats of both groups was treated with N(G)-nitro-l-arginine methyl ester (l-NAME, 10(-4) M). We assessed apoptosis by propidium iodide staining, by TUNEL nitrite production (Griess reaction), by inducible nitric oxide synthase (iNOS) and Bcl-2 and Bax by Western blot. Incubated with a FCS-free medium, cells from SHR showed a significantly higher apoptotic rate (10.7 +/- 2.0) than with 10% FCS (10% FCS, 4.7 +/- 0.3), while WKY cells did not show this increment (10% FCS, 4.7 +/- 0.3; 0% FCS, 5.9 +/- 0. 3). Apoptosis in cells from WKY increased when incubated with thapsigargin in FCS-free medium (0% FCS+ Tg, 17.7 +/- 2.9%) and increased even more in SHR cells (0% FCS+ Tg, 19.7 +/- 2.9%). Treatment with l-NAME decreased thapsigargin-induced apoptosis in both SHR (8.2 +/- 2.4%) and WKY cells (9.3 +/- 2.4%). An increase in nitrite production and iNOS expression was detected in groups in which the apoptosis rate was elevated. A high rate of apoptosis was also associated with a decrease in the Bcl-2/Bax ratio. Our results indicate that in SHR cells, short-term serum deprivation and the increase in intracellular free calcium concentration with thapsigargin are able to enhance the apoptosis rate in primary cultures and that the expression of iNOS, and hence NO production, is involved in this effect.

Bcl-2 overexpression results in reciprocal downregulation of Bcl-X(L) and sensitizes human testicular germ cell tumours to chemotherapy-induced apoptosis.

Testicular germ cell tumours are hypersentive to chemotherapy and cell lines derived from these tumours are chemosensitive in vitro. We have previously shown that these cell lines express undetectable levels of the suppressor of apoptosis Bcl-2 and relatively high levels of the apoptosis inducer Bax (Chresta et al., 1996). To determine whether the absence of Bcl-2 in these cell lines makes them highly susceptible to drug-induced apoptosis, Bcl-2 was expressed ectopically in the 833K testicular germ cell tumour cell line. Stable overexpressing clones were isolated and three clones were studied further. Surprisingly, Bcl-2 overexpressing cells were sensitized to chemotherapy-induced apoptosis compared to the parental and vector control cells. Analysis of potential mechanisms of sensitization revealed there was a reciprocal downregulation of the endogenously expressed Bcl-X(L) in the Bcl-2 overexpressing clones. Downregulation of Bcl-X(L) to the same extent using antisense oligonucleotides enhanced etoposide-induced apoptosis by twofold. Our results indicate that Bcl-2 and Bcl-X(L) have different abilities to protect against chemotherapy-induced apoptosis in testicular germ cell tumours. In contrast to findings in some tumour cell types, Bcl-2 did not act as a gatekeeper to prevent entry of p53 to the nucleus.

Nitric oxide is involved in apoptosis induced by thapsigargin in rat mesangial cells.

The purpose of this study was to examine the mechanisms of thapsigargin-induced apoptosis in rat glomerular mesangial cells and the possible involvement of nitric oxide (NO) in this process. In mesangial cell monolayers incubated for 12 h in a medium without growth factors and with 10(-6) M thapsigargin, a known specific inhibitor of endoplasmic reticulum Ca(2+)-ATPase, a high percentage of cells showed typical nuclear features of apoptosis, assessed either by staining with propidium iodide (23 vs. 9% in control conditions) or by terminal desoxynucleotidyl transferase-mediated dUTP biotin nick end labelling (TUNEL; 17 vs. 5% in control conditions). When cells were maintained in a medium containing 10% fetal calf serum (FCS) or in a free-calcium medium, the thapsigargin-induced apoptosis rate was very low. In rat mesangial cells treatment with thapsigargin decreased the expression of BCL-2 protein and bcl-2 mRNA, whereas it did not alter the levels of BAX protein or bax mRNA. When mesangial cells were incubated with thapsigargin in the absence of FCS, we detected a significant increase in nitrite production (3.78 +/- 0.96 vs. 1.76 +/- 0.44 micromol/well). Furthermore, the treatment with the NO synthesis inhibitor L-NAME (10(-4) M) induced a significant decrease in the number of apoptotic cells (9%), whereas incubation with the NO donor SIN-1 (10(-5) M) induced a marked increase in the rate of apoptosis (29%). Western and Northern blot analysis of macrophage-type inducible NO synthase (iNOS) demonstrated that thapsigargin treatment induces the expression of the iNOS protein and iNOS mRNA. Treatment with L-NAME prevented the thapsigargin-induced BCL-2 decrease, whereas incubation with SIN-1 potentiated the effect of thapsigargin on BCL-2. Double labelling by immunohistochemistry for iNOS and TUNEL revealed that the same cells that suffered apoptosis were positive for iNOS. In summary, our results indicate that thapsigargin is able to enhance the apoptosis rate of rat mesangial cells by a mechanism that is mediated by an increase in cytosolic free calcium. Increased iNOS expression, and hence increased NO production, seems to be involved in this effect.

Glomerular cell proliferation and apoptosis in uninephrectomized spontaneously hypertensive rats.

We studied renal function, glomerular cell proliferation and apoptosis for three months after uninephrectomy (UNX) in young, male, spontaneously hypertensive rats (SHR). Apoptosis was assessed by in situ dUTP biotin nick-end labeling method (TUNEL) and by propidium iodide staining. Proliferation rate was determined by immunohistochemistry to proliferating cell nuclear antigen (PCNA). Glomerular bcl-2 expression was assessed by Northern blot analysis. Our results indicate a parallel increase in proliferation and in apoptotic rates in glomerular cells from the first to the second month after UNX. In the third month after UNX, PCNA-labeled cell number continues increasing, whereas TUNEL-labeled cells did not increase. Bcl-2 expression was negative in the first and second months and increased in the third month. Glomerular size and proteinuria increased progressively along the three months of follow-up. Our observations demonstrate a different profile of cell proliferation and apoptosis during the genesis of early glomerular damage in UNX-SHR.

Increased nitric oxide synthesis and inducible nitric oxide synthase expression in patients with alcoholic and non-alcoholic liver cirrhosis.

1. The synthesis and release of nitric oxide may play a role in the pathogenesis of peripheral vasodilatation and hyperdynamic circulation observed in liver cirrhosis. In this work, we analysed the synthesis of nitric oxide by the lympho-mononuclear cells of peripheral blood from patients with chronic alcoholic and non-alcoholic liver disease and we identified the isoform of nitric oxide synthase involved in the increased nitric oxide synthesis. 2. Patients were classified following clinical and histological criteria in non-alcoholic cirrhotic, alcoholic cirrhotic and non-cirrhotic chronic liver disease. We studied clinical and analytical characteristics, haemodynamic parameters and endotoxin levels in these patients. 3. Cirrhotic patients showed an increase of cardiac output and a decrease of peripheral vascular resistance. These patients had higher levels of plasma endotoxin than those observed in the control group. N omega-Nitro-L-arginine methyl ester (L-NAME)-inhibitable nitrite production from mononuclear lymphocyte cells was higher in patients than in the control group, the highest levels being in non-alcoholic cirrhotic patients, and the lowest levels in patients with non-cirrhotic alcoholic liver disease. 4. Immunocytochemistry studies revealed a positive immunoreactivity for the inducible isoform of nitric oxide synthase in lympho-mononuclear cells that was more evident in non-alcoholic than in alcoholic cirrhotic patients. By Northern blot, inducible nitric oxide synthase mRNA expression was observed only in lymphomononuclear cells from non-alcoholic cirrhotic patients. 5. Our patients show a correlation between nitric oxide synthesis, endotoxin levels and haemodynamic parameters. 6. These findings indicate that lympho-mononuclear cell stimulation may play a role in elevated nitric oxide production in hepatic cirrhosis. Thus, this increased nitric oxide synthesis could be implicated in the pathogenesis of the haemodynamic disturbances frequently found in cirrhotic patients. This increase seems to be induced, at least in part, by activation of an inducible isoform of nitric oxide synthase.

Endogenous angiotensin II and cell hypertrophy in vascular smooth muscle cultures from hypertensive Ren-2 transgenic rats.

We investigated the possible role of a tissular renin-angiotensin system in promoting the growth of vascular smooth muscle cells (VSMCs) from hypertensive transgenic rats (TGRs) with the mouse renin gene Ren-2. Mean arterial pressure values were 99.4 +/- 2.8 and 186.7 +/- 5.0 mm Hg for control Sprague-Dawley rats (SDs) and TGRs, respectively (p < 0.05). The tunica media of femoral arteries obtained from hypertensive TGRs was found to be thickened compared to that of age-matched normotensive SDs. Angiotensin II could be detected by dot blot and immunocytochemistry and quantified by radioimmunoassay in transgenic VSMCs, but not in control SD ones. Under serum-free conditions, VSMCs derived from TGRs showed a higher protein content than those derived from SDs (337 +/- 19 vs. 269 +/- 14 pg/cell, p < 0.05, n = 3). Under the same basal conditions, the mean planar cell surface area was significantly higher in TGR VSMCs than in SD ones (4,764 +/- 204 vs. 4,074 +/- 238 micron 2, p < 0.05). In addition, TGR VSMCs showed an enhanced [14C]-leucine uptake but SD VSMCs did not (13,188 +/- 663 vs. 7,633 +/- 713 dpm/well, p < 0.05). VSMCs showed a concentration-dependent proliferative response to fetal calf serum (FCS) that was more marked in TGRs than in SDs. In the absence of FCS, c-fos and c-jun mRNAs were expressed only in transgenic cultures. From the present results, we can hypothesize that cultured TGR VSMCs are able to synthesize angiotensin II that, being almost exclusive into the cells, contributes to produce VSMC growth in the absence of FCS stimulation.

Bacteriuria asintomática en la diabetes mellitus / Asymptomatic bacteriuria in diabetes mellitus

Con el objetivo de investigar la relación entre la bacteriuria y la diabetes mellitus se realizó un estudio descriptivo transversal en 180 pacientes diabéticos e igual número de sujetos controles no diabéticos con correspondencia en edad y sexo, en el Hospital General Docente "Martín Chang Puga" de Nuevitas desde enero de 1994 a enero de 1995. La incidencia de bacteriuria en los pacientes diabéticos fue de 6,1 por ciento significativamente superior a 1,7 por ciento encontrada en los sujetos no diabéticos. Siendo más frecuente en la diabetes mellitus insulino dependiente (19,4 por ciento), en los pacientes de edad superior a los 50 años (71 por ciento), con un tiempo de evolución entre 10 y 19 años y con un mayor deterioro del control metabólico. El residuo vesical constituyó un factor importante (72,7 por ciento) asociado a la bacteriuria en diabéticos. Se recomienda el pesquisaje de la bacteriuria en los diabéticos para prevención de las infecciones urinarias y su posible repercusión en el deterioro de la función renal