Gaining Insights into Inherited Bleeding Disorders of Complex Etiology in Pediatric Patients: Whole-Exome Sequencing as First-Line Investigation Tool.

INTRODUCTION: Investigation of the molecular basis of inherited bleeding disorders (IBD) is mostly performed with gene panel sequencing. However, the continuous discovery of new related genes underlies the limitation of this approach. This study aimed to identify genetic variants responsible for IBD in pediatric patients using whole-exome sequencing (WES), and to provide a detailed description and reclassification of candidate variants. MATERIAL AND METHODS: WES was performed for 18 pediatric patients, and variants were filtered using a first-line list of 290 genes. Variant prioritization was discussed in a multidisciplinary team based on genotype-phenotype correlation, and segregation studies were performed with available family members. RESULTS: The study identified 22 candidate variants in 17 out of 18 patients (94%). Eleven patients had complete genotype-phenotype correlation, resulting in a diagnostic yield of 61%, 5 (28%) were classified as partially solved, and 2 (11%) remained unsolved. Variants were identified in platelet (ACTN1, ANKRD26, CYCS, GATA1, GFI1B, ITGA2, NBEAL2, RUNX1, SRC, TUBB1), bleeding (APOLD1), and coagulation (F7, F8, F11, VWF) genes. Notably, 9 out of 22 (41%) variants were previously unreported. Variant pathogenicity was assessed according to the American College of Medical Genetics and Genomics guidelines and reclassification of three variants based on family segregation evidence, resulting in the identification of 10 pathogenic or likely pathogenic variants, 6 variants of uncertain significance, and 6 benign or likely benign variants. CONCLUSION: This study demonstrated the high potential of WES in identifying rare molecular defects causing IBD in pediatric patients, improving their management, prognosis, and treatment, particularly for patients at risk of malignancy and/or bleeding due to invasive procedures.

Oral amoxicillin/clavulanate for the prevention of bacteremia following dental extractions.

This study aimed to evaluate the effectiveness of oral amoxicillin/clavulanate (AMX-CL) for the prevention of bacteremia following dental extractions. The study group (AMX-CLG) comprised 40 adults requiring dental extractions under general anesthesia who were administered a prophylactic regimen of 1875/125 mg of AMX-CL orally 1-2 h prior to the surgery. Venous blood samples were collected from each patient at baseline and at 30 s and 15 min after dental extractions. Samples were inoculated into BACTEC Plus culture bottles and processed in the BACTEC 9240. Conventional microbiological techniques were used for subcultures and further identification of the isolated bacteria. The results for the AMX-CLG were compared with those of a control group (CG; no prophylaxis) and an amoxicillin group (AMXG; 2 g of amoxicillin orally), consisting of randomly selected patients from among those participating in two clinical trials that we have previously published. The prevalence of bacteremia in the CG, AMXG, and AMX-CLG was 97%, 50%, and 15%, respectively, at 30 s after completing the extractions, and 67%, 10%, and 4% at 15 min, respectively, after the last extraction. The prevalence of bacteremia in the AMXG and the AMX-CLG at 30 s and at 15 min after completing the extractions was significantly lower than that in the CG (p < 0.001 and p < 0.001, respectively; Fisher's exact test). The prevalence of bacteremia in the AMX-CLG at 30 s after completing the extractions was significantly lower than that in the AMXG (p < 0.001; Fisher's exact test). Based in the results of this preliminary study, oral AMX-CL could be an excellent option for preventing bacteremia secondary to dental procedures in patients at risk.

Natural killer cell receptors and ligand variants modulate response to tyrosine kinase inhibitors in patients with chronic myeloid leukemia.

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm treated with tyrosine kinase inhibitors (TKIs). Although survival rates have improved, response to these treatments is highly heterogeneous. Variations in response rates may be due to different causes such as, treatment adherence, mutations in the BCR-ABL1 gene, clonal evolution and amplification of the BCR-ABL1 gene, but innate immune response is also considered to play a very important role and, specifically, NK cell activity through their receptors and ligands, could be determinant. The aim of this retrospective study was to explore the role of different activating and inhibiting KIR genes as well as the activating NKG2D receptor, present in NK cells, and also their respective ligands, HLA-A, -B, -C, -G, -F, MICA and MICB, in the progression of 190 patients with CML and treated at two hospitals from Barcelona between 2000 and 2019. Early molecular response (EMR), major molecular response (MMR) or MR3.0 and deep molecular response (DMR) or MR4.0 were correlated. As control samples, healthy donors from the Barcelona Blood Bank were analyzed. The presence of KIR2DL2/KIR2DS2 was associated with the achievement of EMR, MR3.0, and MR4.0. Carriers of the higher expression NKG2D variant and MICA*009:01 were also likely to achieve molecular response (MR). The most remarkable difference between CML patients and controls was a higher frequency of the lower expression NKG2D variant in CML patients. In summary, our results showed that activating NK receptor phenotypes might help to achieve MR and DMR in CML patients treated with TKIs although confirmatory studies are necessary.

Prevalence and factors associated with SARS-CoV-2 seropositivity in the Spanish HIV Research Network Cohort.

OBJECTIVES: We aimed to assess the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and factors associated with seropositivity and asymptomatic coronavirus disease 2019 (COVID-19) among people with HIV (PWH). METHODS: This was a cross-sectional study carried out within the cohort of the Spanish HIV Research Network. Participants were consecutive PWH with plasma collected from 1st April to 30th September 2020. We determined SARS-CoV-2 antibodies (Abs) in plasma. Illness severity (NIH criteria) was assessed by a review of medical records and, if needed, participant interviews. Multivariable logistic regression analysis was used to identify predictors of seropositivity among the following variables: sex, age, country of birth, education level, comorbidities (hypertension, chronic heart disease, diabetes mellitus, non-AIDS-related cancer, chronic kidney disease, cirrhosis), route of HIV acquisition, prior AIDS, CD4+ cell count, HIV viral load, nucleoside/nucleotide reverse transcriptase inhibitor (N [t]RTI) backbone, type of third antiretroviral drug, and month of sample collection. RESULTS: Of 1076 PWH (88.0% males, median age 43 years, 97.7% on antiretroviral therapy, median CD4+ 688 cells/mm3, 91.4% undetectable HIV viral load), SARS-CoV-2 Abs were detected in 91 PWH, a seroprevalence of 8.5% (95%CI 6.9-10.3%). Forty-five infections (45.0%) were asymptomatic. Variables independently associated with SARS-CoV-2 seropositivity were birth in Latin American countries versus Spain (adjusted odds ratio (aOR) 2.30, 95%CI 1.41-3.76, p 0.001), and therapy with tenofovir disoproxil fumarate plus emtricitabine (TDF/FTC) versus tenofovir alafenamide (TAF)/FTC as the N(t)RTI backbone (aOR 0.49, 95%CI 0.26-0.94, p 0.031). CONCLUSIONS: Many SARS-CoV-2 infections among PWH were asymptomatic, and birth in Latin American countries increased the risk of SARS-CoV-2 seropositivity. Our analysis, adjusted by comorbidities and other variables, suggests that TDF/FTC may prevent SARS-CoV-2 infection among PWH.

Evaluation of the Spanish population coverage of a prospective HLA haplobank of induced pluripotent stem cells.

BACKGROUND: iPSC (induced pluripotent stem cells) banks of iPSC lines with homozygous HLA (human leukocyte antigen) haplotypes (haplobanks) are proposed as an affordable and off-the-shelf approach to allogeneic transplantation of iPSC derived cell therapies. Cord blood banks offer an extensive source of HLA-typed cells suitable for reprogramming to iPSC. Several initiatives worldwide have been undertaken to create national and international iPSC haplobanks that match a significant part of a population. METHODS: To create an iPSC haplobank that serves the Spanish population (IPS-PANIA), we have searched the Spanish Bone Marrow Donor Registry (REDMO) to identify the most frequently estimated haplotypes. From the top ten donors identified, we estimated the population coverage using the criteria of zero mismatches in HLA-A, HLA-B, and HLA-DRB1 with different stringencies: high resolution, low resolution, and beneficial mismatch. RESULTS: We have calculated that ten cord blood units from homozygous donors stored at the Spanish cord blood banks can provide HLA-A, HLA-B, and HLA-DRB1 matching for 28.23% of the population. CONCLUSION: We confirm the feasibility of using banked cord blood units to create an iPSC haplobank that will cover a significant percentage of the Spanish and international population for future advanced therapy replacement strategies.

SARS-CoV-2/COVID-19 pandemic: first wave, impact, response and lessons learnt in a fully integrated Regional Blood and Tissue Bank. A narrative report.

BACKGROUND: The COVID-19 pandemic is placing blood and tissue establishments under unprecedented stress, putting its capacity to provide the adequate care needed at risk. Here we reflect on how our integrated organisational model has faced the first impact of the pandemic and describe what challenges, opportunities and lessons have emerged. MATERIALS AND METHODS: The organisational model of the Catalan Blood and Tissue Bank (Banc de Sang i Teixits, BST) is described. The new scenario was managed by following international recommendations and considering the pandemic in a context of volatility, uncertainty, complexity, and ambiguity (VUCA), allowing rapid measures to be taken. These aimed to: ensure donor safety, promote proper responses to patients' needs, ensure the health and well-being of personnel, and prepare for future scenarios. RESULTS: The BST has adapted its activities to the changes in demand. No shortage of any product or service occurred. Donor acceptance, safety and wellbeing were maintained except for tissue donation, which almost completely stopped. To support the health system, several activities have been promoted: large-scale convalescent plasma (CP) production, clinical trials with CP and mesenchymal stromal cells, massive COVID-19 diagnoses, and participation in co-operative research and publications. Haemovigilance is running smoothly and no adverse effects have been detected among donors or patients. DISCUSSION: Several elements have proven to be critical when addressing the pandemic scenario: a) the early creation of a crisis committee in combination with technical recommendations and the recognition of a VUCA scenario; b) identification of the strategies described; c) the integrated donor-to-patient organisational model; d) active Research and Development (R&D); and e) the flexibility of the staff. It is essential to underline the importance of the need for centralised management, effective contingency strategies, and early collaboration with peers.

High-throughput genotyping of HLA-G, HLA-F, MICA, and MICB and analysis of frequency distributions in healthy blood donors from Catalonia.

Similarly to HLA class I molecules, certain non-classical HLA class I genes and MHC class I polypeptide-related sequences A and B (MICA and MICB) act as ligands for KIR and NKG2D natural killer receptors. Although these genes are less polymorphic than HLA class I, few studies have analyzed their association with diseases. Information on allele frequencies in healthy donors is needed to map their distribution worldwide. This study is the first to analyze high-resolution HLA-G, HLA-F, MICA, and MICB allele frequencies using a novel high-throughput next generation-sequencing method. We analyzed DNA samples from 96 unrelated blood donors resident in Catalonia, Spain, and registered in the Barcelona Blood and Tissue Bank. Using the first two fields of the HLA nomenclature, we detected six HLA-G and two HLA-F alleles. The most frequent alleles were HLA-G*01:01 (77.08%) and HLA-F*01:01(84.90%). When the four fields were analyzed, we detected 16 and 10 alleles, respectively. Nineteen alleles were detected for MICA and 10 for MICB. The most frequent alleles in these cases were MICA*008:01 (16.15%) and MICB*005:02 (46.84%). All frequencies were in Hardy Weinberg equilibrium except MICA. We also estimated maximum-likelihood haplotype frequencies and calculated corresponding linkage disequilibrium (LD) values and found that few allele pairs were in disequilibrium. Strong LD between MICA and HLA-B (using data from a previous study) was observed. Our findings will be useful for guiding further research evaluating the functional role of these genes in different diseases and populations.

Influence of Puerperal Health Literacy on Tobacco Use during Pregnancy among Spanish Women: A Transversal Study.

Background: Despite the fact that tobacco use during pregnancy produces adverse perinatal effects, some women continue to smoke. Health literacy (HL) is essential for health outcomes in adults. However, little is known about HL in pregnant women or postpartum women. The study aimed to analyse the relationship between the degree of HL of women during the early puerperium and tobacco use during pregnancy. METHODS: A multicentre, descriptive, cross-sectional study was carried out with women in the early puerperium in a region of eastern Spain, between November 2017 and May 2018. Their HL level was obtained using the Newest Vital Sign (NVS) tool. Multivariate logistic models were adjusted to estimate the magnitude of association with tobacco use in pregnancy. Odds ratios (OR) were estimated with a 95% confidence interval. RESULTS: 193 were included in the total. 29.5% (57) of pregnant women smoked tobacco during pregnancy, with a smoking cessation rate of 70.1% (40) while pregnant. 42.0% (81) of pregnant women had inadequate or limited HL. A low level of HL was strongly associated with tobacco use, adjusted by catchment area and age of first pregnancy (LRT p < 0.001; ROC curve = 0.71, 95% CI: 0.64-0.79). CONCLUSION: A low HL is associated with tobacco consumption during pregnancy. Whether low HL reflects the wide constellation of already-known socioeconomic, political and commercial determinants of tobacco use, or whether incorporating HL support interventions strengthens tobacco cessation activities in pregnancy, warrants further research. Still, it should be considered as essential to understanding the health disparities related to its consumption.

Ecografía del tercer trimestre combinada con un método clínico para mejorar la predicción del peso del recién nacido a término: un estudio de cohortes en España / Third trimester ultrasound scan combined with a clinical method for accurate birthweight prediction at term: a cohort study in spain

Abstract Objective: To develop and assess an equation based on maternal clinical parameters and third trimester ultrasound biometry (combined method), and compare it with ultrasound estimated foetal weight (EFW) calculated using the Hadlock 2 formula. Material and methods: Cohort study. A total of 1,224 women with singleton pregnancies who had undergone foetal ultrasound scanning (USS) at 34 weeks were recruited. The study was conducted at a reference center in Valencia (Spain) between January and December 2016. A gestation-adjusted projection (GAP) method was applied to estimated foetal-weight-for-gestational-age by foetal gender at delivery (EFWa). A multivariate regression was created to estimate foetal weight at term (EFWmr) using anthropometric, demographic, ultrasonographic and obstetric-neonatal variables. EFWa and EFWmr were calculated and compared with actual birthweight. Results: The proportion for EFWmr within <10% of actual birthweight was greater than EFWa (82% vs. 65%, p<0.001). The mean relative error in foetal-weight predictions by using EFWmr was reduced from 6.7% to 0.9% (difference 5.7% 95% CI: 5.4 to 6.0) paired t-test p<0.001, significantly improving the accuracy attainable with USS. The EFWmr outperformed the GAP method in predicting birthweight, within 1% relative error. For new- borns <2,500 g, the proportion of estimates within <10% of the actual birthweight for the EFWmr was greater than that of the EFWa (20.4 vs. 16.3%, p=0.005). For babies with normal birthweight (2,500-3,999 g), EFWmr was a better predictor of birthweight than EFWa (84.5 vs. 65.7%, p<0.001). Conclusions: Mathematical modelling to predict birthweight improves third trimester routine ultrasound measurement to estimate neonatal weight at term.

RESUMEN Objetivos: desarrollar y evaluar un modelo predictivo de acuerdo con los parámetros clínicos maternos y la biometría de la ecografía del tercer trimestre, que pueda mejorar el poder de predicción del peso al nacer en el recién nacido a término, en comparación con la estimación calculada por ecografía del peso fetal (PFE) usando la fórmula de Hadlock II. Materiales y métodos: revisión de 1224 mujeres con embarazos únicos que se habían sometido a una ecografía fetal a las 34 semanas (EF). El estudio se realizó en un centro de referencia en Valencia (España) entre enero y diciembre de 2016. Se aplicó un método de proyección ajustada de gestación (PAG) para estimar el peso al nacer para la edad gestacional y sexo fetal en el parto (PFEa). Se creó una regresión multivariante para estimar el peso fetal al nacer (PFErm) mediante variables antropométricas, demográficas, ecográficas y obstétrico-neonatales. Los modelos PFErm y PFEa fueron calculados para comparar sus diferencias respecto al peso real al nacer. Resultados: la proporción de PFErm dentro de < 10 % del peso real al nacer fue mayor que la de PFEa (82 % vs. 65 %, p< 0,001). El error relativo medio en las predicciones de peso fetal mediante el uso PFErm pasó de 6, a 0,9 % (Diferencia de proporciones: 5,7 %; IC 95 %: 5,4-6,0); medias pareadas: p < 0,001, siendo significativamente mejor que la precisión que puede ser obtenida con el método ecográfico. El PFErm superó al método PAG y predice el peso al nacer con un error relativo del 1 %. Para recién nacidos con < 2500 g la proporción de estimaciones del peso real < 10 % del PFErm fue mayor que la del PFEa (20,4 % vs. 16,3 %; p = 0,005). En los recién nacidos con peso normal al nacer (2500-3999 g), la capacidad predictiva para estimar el peso al nacer realizada mediante PFErm fue mejor que la realizada mediante PFEa (84,5 % vs. 65,7%; p < 0,001). Conclusiones: el modelo matemático creado para predecir el peso al nacer mejora la medición rutinaria de la ecografía en el tercer trimestre del embarazo para estimar el peso del recién nacido a término.

Analysis of the Spanish CCR5-∆32 inventory of cord blood units: lower cell counts in homozygous donors.

The possibility to use CCR5-∆32 umbilical cord blood to cure HIV infection in patients in need of a hematopoietic transplant has been suggested. The less stringent HLA compatibility needed in this type of transplant facilitates the search of a suitable donor having the CCR5-∆32 mutation. To achieve an inventory of CCR5-∆32 cord blood units, the 20,236 best cell quality units of the Spanish Registry were genotyped. Furthermore, their CD34+ and total nucleated cells counts, blood type, gender, HLA and donor's geographical and ancestral origin were analyzed. The results showed 130 (0.64%) units homozygous for the deletion, 2,646 (13.08%) heterozygous and 17,460 (86.28%) did not present the mutation. Interestingly, a significant lower amount of CD34+ cells was found in the CCR5-∆32 homozygous units. In addition, a significant association was found among donor's ancestral origin and the mutation, with a higher percentage of CCR5-∆32 units with a European ancestry. In summary, identification of a relatively high number of CCR5-∆32 units is feasible and will facilitate the development of clinical trials for HIV cure in patients requiring hematopoietic transplantation. Further studies are required to understand the significance of lower cell counts within the CCR5-∆32 homozygous group and its clinical impact.

Design and Validation of a Multiplex KIR and HLA Class I Genotyping Method Using Next Generation Sequencing.

Killer cell immunoglobulin-like receptors (KIR), considered the most polymorphic natural killer (NK) cell regulators, bind HLA class-I molecules or still unknown ligands. Interest in KIR genotyping is increasing because of the importance of these receptors for identifying the best possible donor in hematopoietic stem cell transplantation to obtain a graft-versus-leukemia effect. Currently, routine protocols to determine the gene content of the KIR cluster are exclusively performed by PCR-SSO and PCR-SSP. To improve the study of these genes, we developed a multiplex, long-range PCR strategy suitable for simultaneous, high-resolution HLA class I and KIR genotyping by next generation sequencing (NGS). This protocol allows amplification of the 14 KIR genes, 2 KIR pseudogenes, and HLA class I genes, with subsequent sequencing on an Illumina platform. The bioinformatics analysis for KIR genotyping was performed by virtual hybridization of gene-specific probes, and HLA genotyping was done by GenDx NGSengine software. To validate the method reliability, 192 genomic DNA samples previously characterized by PCR-SSO were used. When a specific KIR gene was present, a large number of gene-specific virtual probes were detected, whereas when it was absent, very few or none were found, enabling cutoff establishment. Concordance for both the KIR and HLA assignments as compared with the previous characterization was 100%. In conclusion, the multiplex PCR NGS-based strategy presented could provide an efficient, less costly method for KIR-ligand genotyping by gene presence/absence. Furthermore, allele resolution will be possible when KIR-specific software becomes available.

Tobacco use in the third trimester of pregnancy and its relationship to birth weight. A prospective study in Spain.

BACKGROUND: Few studies have been carried out in Spain examining the use of tobacco amongst expectant mothers and its effect on birth weight. AIMS: To observe the proportion of expectant mothers who smoke during their pregnancy, and the impact of tobacco consumption on maternal and birth weight. We also aimed to identify the trimester of pregnancy in which tobacco use produced the greatest reduction in birth weight. METHODS: Prospective observational study in Spain. A random sampling strategy was used to select health centres and participant women. A total of 137 individuals were enrolled in the study. Exposure to tobacco was measured through a self-reported questionnaire. Regressions were performed to obtain a predictive model for birth weight related to smoking. FINDINGS: Overall, 35% of study participants were smokers during the pre-gestational period (27% in the first trimester, 21.9% in the second and 21.2% in the third). 38.7% of smoking cessation attempts took place in the third-trimester. Pregnant women who smoked up to the third trimester had a higher risk of giving birth to a baby under 3000g, compared to non-smokers (OR=5.94, CI 95%: 1.94-18.16). Each additional unit of tobacco consumed daily in the 3rd trimester led to a 32g reduction in birth weight. CONCLUSION: An important proportion of pregnant women in Spain smoke during pregnancy. Pregnant women exposed to tobacco have newborns with lower birth weight. Smoking during the 3rd trimester of pregnancy is associated with the greatest risk of lower birth weight.

miR-146a targets Fos expression in human cardiac cells.

miR-146a is a microRNA whose transcript levels are induced in the heart upon activation of NF-κB, a transcription factor induced by pro-inflammatory molecules (such as TNF-α) that is strongly related to the pathogenesis of cardiac disorders. The main goal of this study consisted of studying new roles of miR-146a in cardiac pathological processes caused by the pro-inflammatory cytokine TNF-α. Our results demonstrate that miR-146a transcript levels were sharply increased in cardiac ventricular tissue of transgenic mice with specific overexpression of TNF-α in the heart, and also in a cardiomyocyte cell line of human origin (AC16) exposed to TNF-α. Among all the in silico predicted miR-146a target genes, Fos mRNA and protein levels notably decreased after TNF-α treatment or miR-146a overexpression. These changes correlated with a diminution in the DNA-binding activity of AP-1, the Fos-containing transcription factor complex. Interestingly, AP-1 inhibition was accompanied by a reduction in matrix metalloproteinase (MMP)-9 mRNA levels in human cardiac cells. The specific regulation of this MMP by miR-146a was further confirmed at the secretion and enzymatic activity levels, as well as after anti-miR-mediated miR-146a inhibition. The results reported here demonstrate that Fos is a direct target of miR-146a activity and that downregulation of the Fos-AP-1 pathway by miR-146a has the capacity to inhibit MMP-9 activity. Given that MMP-9 is an AP-1 target gene involved in cardiac remodeling, myocardial dysfunction and progression of heart failure, these findings suggest that miR-146a might be a new and promising therapeutic tool for treating cardiac disorders associated with enhanced inflammation in the heart.

[Relationship between maternal body mass index, gestational weight gain and birth weight; prospective study in a health department]. / Asociación entre el índice de masa corporal materno, la ganancia de peso gestacional y el peso al nacer; estudio prospectivo en un departamento de salud.

OBJECTIVES: To ascertain the relationship between maternal weight gain and birth weight, in every pre-gestational body mass index (BMI) category. MATERIAL AND METHODS: A two-stage sampling observational and descriptive study was carried out in the health department of La Ribera (Valencia, Spain). The sample was divided into four groups according to pre-gestational BMI. FINDINGS: 140 pregnant women were studied. We observed rising pre-gestational weight gain (PWG) and trimestral gradients. There was a higher increase from the first to the second trimester than from the second to the third trimester in every pre-gestational BMI category. According to the international recommendations of Institute of Medicine, 16.4% of women had an inferior gestational weight gain (GWG), 38.6% were within the recommendations and 45% were above them. The pre-gestational BMI, categorized by the WHO, is related to the birth weight, showing a statistical significance (F=6.636 and and p<0.001). Obese mothers with a higher weight gain than the recommended have newborns with higher birth weight (4,353 ± 821.924 g) and, underweight mothers with a lower weight gain than the recommended, have newborns with lower birth weights (2,900 ± 381.83 g) than the rest of the groups. CONCLUSIONS: The absolute gestational weight gain did not show a statistical significance compared to the birthweight in any of the pre-gestational BMI categories and, as an isolated indicator, is not an added value to the prenatal quality control.

Objetivo: Contrastar la relación que existe entre la ganancia de peso gestacional y el peso del recién nacido, entre cada categoría de IMC pregestacional. Material y Métodos: Estudio observacional y descriptivo con muestreo bietápico en el Departamento de Salud de la Ribera (Valencia, España). Se clasificaron en cuatro grupos dependiendo del índice de masa corporal (IMC) pregestacional. Resultados: Se estudiaron 140 gestantes. La evolución de la ganancia de peso gestacional (GPG) y de sus gradientes trimestrales fue ascendente. Se produjo un mayor incremento del primer al segundo trimestre que del segundo al tercero para todas las categorías de IMC pregestacional. Según las recomendaciones internacionales de GPG el 16,4% de la muestra tuvo una ganancia de peso inferior a la recomendada, el 38,6% una ganancia de peso igual y el 45% una ganancia de peso superior. El IMC pregestacional categorizado por la OMS, está relacionado con el peso al nacer, mostrando significación estadística (F=6,636 y p<0,001). Las obesas con una ganancia de peso mayor de la recomendada tienen recién nacidos con mayor peso (4.353±821,924 g) y las de bajo-peso con ganancia menor de la recomendada, tienen recién nacidos con pesos menores (2.900±381,83 g) que el resto de grupos. Conclusiones: La GPG de forma absoluta no mostró significación estadística con el peso al nacer con ninguna categoría materna de IMC pregestacional y, como indicador aislado, no es un valor añadido a la calidad del control prenatal.

miR-125b acts as a tumor suppressor in breast tumorigenesis via its novel direct targets ENPEP, CK2-α, CCNJ, and MEGF9.

MicroRNAs (miRNAs) play important roles in diverse biological processes and are emerging as key regulators of tumorigenesis and tumor progression. To explore the dysregulation of miRNAs in breast cancer, a genome-wide expression profiling of 939 miRNAs was performed in 50 breast cancer patients. A total of 35 miRNAs were aberrantly expressed between breast cancer tissue and adjacent normal breast tissue and several novel miRNAs were identified as potential oncogenes or tumor suppressor miRNAs in breast tumorigenesis. miR-125b exhibited the largest decrease in expression. Enforced miR-125b expression in mammary cells decreased cell proliferation by inducing G2/M cell cycle arrest and reduced anchorage-independent cell growth of cells of mammary origin. miR-125b was found to perform its tumor suppressor function via the direct targeting of the 3'-UTRs of ENPEP, CK2-α, CCNJ, and MEGF9 mRNAs. Silencing these miR-125b targets mimicked the biological effects of miR-125b overexpression, confirming that they are modulated by miR-125b. Analysis of ENPEP, CK2-α, CCNJ, and MEGF9 protein expression in breast cancer patients revealed that they were overexpressed in 56%, 40-56%, 20%, and 32% of the tumors, respectively. The expression of ENPEP and CK2-α was inversely correlated with miR-125b expression in breast tumors, indicating the relevance of these potential oncogenic proteins in breast cancer patients. Our results support a prognostic role for CK2-α, whose expression may help clinicians predict breast tumor aggressiveness. In particular, our results show that restoration of miR-125b expression or knockdown of ENPEP, CK2-α, CCNJ, or MEGF9 may provide novel approaches for the treatment of breast cancer.

Estrutura fatorial do WISC-III em crianças com dificuldades de aprendizagem / WISC-III factor structure for children with learning disabilities / Estructura factorial del WISC-III en niños con problemas de aprendizaje

O presente trabalho busca verificar qual o modelo fatorial mais adequado para o WISC-III no grupo clínico das crianças com dificuldades de aprendizagem. Foram analisados 263 protocolos de testes aplicados em alunos de escolas públicas, encaminhados por seus professores para avaliação psicológica. Foram utilizadas as técnicas estatísticas da análise fatorial exploratória e confirmatória. O presente estudo, além de corroborar a estrutura fatorial definida na padronização brasileira, vai ao encontro dos resultados da pesquisa internacional quanto à definição do modelo de quatro fatores como o de melhor ajuste para esse grupo clínico. Apesar de também ter identificado dois modelos trifatoriais como vantajosos quanto ao ajuste, parcimônia e interpretabilidade teórica, a estrutura quadrifatorial é a mais indicada para interpretar clinicamente as pontuações que expressam as habilidades cognitivas do grupo estudado, uma vez que permite aproveitar as normas existentes do WISC-III para a população geral.

This work contributes to the investigation of a factorial model that is more appropriate for Brazilian children with learning disabilities (LD). 263 WISC-III test protocols of public school students with academic difficulties referred by their teachers for psychological evaluation were analyzed. Statistical techniques of exploratory factor analysis and confirmatory factor analysis were performed. This study, besides corroborating the factor structure defined in the Brazilian standardization, meets the results of the international research for the definition of four-factor model as the best adjusting for the LD population. Although we have also identified two three-factor models as advantageous as to the fit, parsimony and theoretical interpretability, the four-factor structure is the most suitable for clinical interpretation of the scores that express the LD group cognitive abilities, since it allows us to leverage existing standards of WISC-III for the general population.

El presente trabajo busca verificar cual el modelo factorial más adecuado para el WISC-III en el grupo clínico de niños con dificultades de aprendizaje. Se analizaron 263 testes aplicados en alumnos de escuelas públicas, encaminados por sus maestros para evaluación psicológica. Se utilizaron las técnicas estadísticas del análisis factorial exploratorio y confirmatorio. Este estudio, además de corroborar la estructura factorial definida en la estandarización brasileña, coincide con los resultados de la investigación internacional en cuanto a la definición del modelo de cuatro factores como de mejor ajuste para ese grupo clínico. Además de haber también identificado dos modelos trifactoriales como ventajosos en cuanto al ajuste, a la economía factorial y a la interpretabilidad teórica, la estructura de cuatro factores es la más indicada para interpretar clinicamente los puntajes que expresan las habilidades cognitivas del grupo estudiado, pues permite aprovechar las normas ya existentes del WISC-III para la población general.

Controlling terahertz radiation with nanoscale metal barriers embedded in nano slot antennas.

Nanoscale metallic barriers embedded in terahertz (THz) slot antennas are shown to provide unprecedented control of the transition state arising at the crossover between the full- and half-wavelength resonant modes of such antennas. We demonstrate strong near-field coupling between two paired THz slot antennas separated by a 5 nm wide nanobarrier, almost fully inducing the shift to the resonance of the double-length slot antenna. This increases by a factor of 50 the length-scale needed to observe similar coupling strengths in conventional air-gap antennas (around 0.1 nm), making the transition state readily accessible to experiment. Our measurements are in good agreement with a quantitative theoretical modeling, which also provides a simple physical picture of our observations.

Tolerance induction in experimental autoimmune encephalomyelitis using non-myeloablative hematopoietic gene therapy with autoantigen.

Experimental autoimmune encephalomyelitis (EAE) constitutes a paradigm of antigen (Ag)-specific T cell driven autoimmune diseases. In this study, we transferred bone marrow cells (BMCs) expressing an autoantigen (autoAg), the peptide 40-55 of the myelin oligodendrocytic glycoprotein (MOG(40-55)), to induce preventive and therapeutic immune tolerance in a murine EAE model. Transfer of BMC expressing MOG(40-55) (IiMOG-BMC) into partially myeloablated mice resulted in molecular chimerism and in robust protection from the experimental disease. In addition, in mice with established EAE, transfer of transduced BMC with or without partial myeloablation reduced the clinical and histopathological severity of the disease. In these experiments, improvement was observed even in the absence of engraftment of the transduced hematopoietic cells, probably rejected due to the previous immunization with the autoAg. Splenocytes from mice transplanted with IiMOG-BMC produced significantly higher amounts of interleukin (IL)-5 and IL-10 upon autoAg challenge than those of control animals, suggesting the participation of regulatory cells. Altogether, these results suggest that different tolerogenic mechanisms may be mediating the preventive and the therapeutic effects. In conclusion, this study demonstrates that a cell therapy using BMC expressing an autoAg can induce Ag-specific tolerance and ameliorate established EAE even in a nonmyeloablative setting.