Genome-wide association study for yield-related traits in faba bean (Vicia faba L.).

Yield is the most complex trait to improve crop production, and identifying the genetic determinants for high yield is a major issue in breeding new varieties. In faba bean (Vicia faba L.), quantitative trait loci (QTLs) have previously been detected in studies of biparental mapping populations, but the genes controlling the main trait components remain largely unknown. In this study, we investigated for the first time the genetic control of six faba bean yield-related traits: shattering (SH), pods per plant (PP), seeds per pod (SP), seeds per plant (SPL), 100-seed weight (HSW), and plot yield (PY), using a genome-wide association study (GWAS) on a worldwide collection of 352 homozygous faba bean accessions with the aim of identifying markers associated with them. Phenotyping was carried out in field trials at three locations (Spain, United Kingdom, and Serbia) over 2 years. The faba bean panel was genotyped with the Affymetrix faba bean SNP-chip yielding 22,867 SNP markers. The GWAS analysis identified 112 marker-trait associations (MTAs) in 97 candidate genes, distributed over the six faba bean chromosomes. Eight MTAs were detected in at least two environments, and five were associated with multiple traits. The next step will be to validate these candidates in different genetic backgrounds to provide resources for marker-assisted breeding of faba bean yield.

A Clear Cell Sarcoma Case: A Diagnostic and Treatment Challenge, with a Promising Response to Trabectedin.

Introduction: Clear cell sarcoma (CCS) is a rare and aggressive soft tissue sarcoma. CCS is characterized by the translocation t(12;22) (q13;q12), involving the fusion of EWSR1 and ATF1 genes, and less frequently the fusion gene EWSR1-CREB1. Usually, CCSs are considered poorly responsive to conventional chemotherapy. However, trabectedin has shown activity against translocation-related sarcomas. Furthermore, preclinical results suggest that trabectedin is a promising antitumor agent for CCS, potentially inducing melanocytic differentiation. Case Presentation: We report the case of a challenging anatomopathological diagnosis in a patient with an aggressive metastatic CCS. Following the diagnosis of CCS, the patient experienced a clinical and radiological tumor response to trabectedin after four lines of treatment. Conclusion: This is a novel report of CCS treated with trabectedin that resulted in a partial response and suggests the need for further research on trabectedin as a therapeutic option for CCS.

Variability in drought stress response in a panel of 100 faba bean genotypes.

Faba bean is an important protein crop for food and feed worldwide and provides a range of advantages in crop rotations. Its limited use in modern agriculture is mainly due to the high fluctuations in yield. A well known limiting factor in most legumes, and particularly in faba bean, is the high sensitivity to water shortage, which is further aggravated by climate change. The present study was undertaken to exploit the genetic variation in drought stress response in a faba bean collection of 100 accessions with diverse origins and to assess selection criteria for identifying drought tolerant genotypes. Physiological, phenological and yield related traits evaluated under drought or water-sufficient conditions responded significantly to the end-terminated drought stress. Comparison of yield relations showed the advantage of using a stress tolerance index (STI) to identify genotypes combining high yield potential with high stress yield. With regard to physiological traits, SPAD (chlorophyll content) values were significantly related to yield as well as to STI, while the other traits also contributed to different extents to variation in yield formation. Among the yield related traits, seeds per plant proved to be the most important trait followed by pods per plant. Interestingly, the eight genotypes with the best STI performance use different strategies to cope with drought stress.

Genetic analysis of global faba bean diversity, agronomic traits and selection signatures.

KEY MESSAGE: We identified marker-trait associations for key faba bean agronomic traits and genomic signatures of selection within a global germplasm collection. Faba bean (Vicia faba L.) is a high-protein grain legume crop with great potential for sustainable protein production. However, little is known about the genetics underlying trait diversity. In this study, we used 21,345 high-quality SNP markers to genetically characterize 2678 faba bean genotypes. We performed genome-wide association studies of key agronomic traits using a seven-parent-MAGIC population and detected 238 significant marker-trait associations linked to 12 traits of agronomic importance. Sixty-five of these were stable across multiple environments. Using a non-redundant diversity panel of 685 accessions from 52 countries, we identified three subpopulations differentiated by geographical origin and 33 genomic regions subjected to strong diversifying selection between subpopulations. We found that SNP markers associated with the differentiation of northern and southern accessions explained a significant proportion of agronomic trait variance in the seven-parent-MAGIC population, suggesting that some of these traits were targets of selection during breeding. Our findings point to genomic regions associated with important agronomic traits and selection, facilitating faba bean genomics-based breeding.

Genome-wide association analysis for drought tolerance and associated traits in faba bean (Vicia faba L.).

Faba bean (Vicia faba L.) is an important high protein legume adapted to diverse climatic conditions with multiple benefits for the overall sustainability of the cropping systems. Plant-based protein demand is being expanded and faba bean is a good candidate to cover this need. However, the crop is very sensitive to abiotic stresses, especially drought, which severely affects faba bean yield and development worldwide. Therefore, identifying genes associated with drought stress tolerance is a major challenge in faba bean breeding. Although the faba bean response to drought stress has been widely studied, the molecular approaches to improve drought tolerance in this crop are still limited. Here we built on recent genomic advances such as the development of the first high-density SNP genotyping array, to conduct a genome-wide association study (GWAS) using thousands of genetic polymorphisms throughout the entire faba bean genome. A worldwide collection of 100 faba bean accessions was grown under control and drought conditions and 10 morphological, phenological and physiological traits were evaluated to identify single nucleotide polymorphism (SNP) markers associated with drought tolerance. We identified 29 SNP markers significantly correlated with these traits under drought stress conditions. The flanking sequences were blasted to the Medicago truncatula reference genomes in order to annotate potential candidate genes underlying the causal variants. Three of the SNPs for chlorophyll content after the stress, correspond to uncharacterized proteins indicating the presence of novel genes associated with drought tolerance in faba bean. The significance of stress-inducible signal transducers provides valuable information on the possible mechanisms underlying the faba bean response to drought stress, thus providing a foundation for future marker-assisted breeding in the crop.

Antioxidant and Chemopreventive Activity of Protein Hydrolysates from Raw and Germinated Flour of Legumes with Commercial Interest in Colorectal Cancer.

Legumes are a highly nutritious source of plant protein, fiber, minerals and vitamins. However, they also contain several bioactive compounds with significant potential benefits for human health. The objectives of this study were to evaluate the antioxidant, antitumor and chemopreventive activity of functional extracts from legumes using raw and germinated flours of six legume species of commercial interest. The methodology carried out consisted on the development of protein hydrolysates, assessment of their antioxidant capacity and in vitro tests on T84, HCT15 and SW480 colorectal cancer (CRC) cell lines. Our results showed a high antitumor activity of protein hydrolysate from M. sativa. Likewise, when combined with 5-Fluorouracile (5-Fu), there was a synergistic effect using extract concentrations from 50 to 175 µg/mL and 5-Fu concentrations from 1.5 to 5 µM. Similarly, the induction effect on detoxifying enzymes by the extracts of M. sativa, germinated V. faba Baraca × LVzt1 and V. narbonensis, which produced a higher induction rate than the positive control sulforaphane (10 µM), should be highlighted. Therefore, incorporating these enzymes into the diet could provide nutritional effects, as well as play an effective role in cancer chemoprevention and therapy.

Identification of Leukemia-Associated Immunophenotypes by Databaseguided Flow Cytometry Provides a Highly Sensitive and Reproducible Strategy for the Study of Measurable Residual Disease in Acute Myeloblastic Leukemia.

Background: Multiparametric Flow Cytometry (MFC) is an essential tool to study the involved cell lineages, the aberrant differentiation/maturation patterns and the expression of aberrant antigens in acute myeloid leukemia (AML). The characterization of leukemia-associated immunophenotypes (LAIPs) at the moment of diagnosis is critical to establish reproducible strategies for the study of measurable residual disease using MFC (MFC-MRD). Methods: In this study, we identify and characterize LAIPs by comparing the leukemic populations of 145 AML patients, using the EuroFlow AML/ MDS MFC panel, with six databases of normal myeloid progenitors (MPCs). Principal component analysis was used to identify and characterize the LAIPs, which were then used to generate individual profiles for MFC-MRD monitoring. Furthermore, we investigated the relationship between the expression patterns of LAIPs and the different subtypes of AML. The MFC-MRD study was performed by identifying residual AML populations that matched with the LAIPs at diagnosis. To further validate this approach, the presence of MRD was also assessed by qPCR (qPCR-MRD). Finally, we studied the association between MFC-MRD and progression-free survival (PFS). Results: The strategy used in this study allowed us to describe more than 300 different LAIPs and facilitated the association of specific phenotypes with certain subtypes of AML. The MFC-MRD monitoring based on LAIPs with good/strong specificity was applicable to virtually all patients and showed a good correlation with qPCR-MRD and PFS. Conclusions: The described methodology provides an objective method to identify and characterize LAIPs. Furthermore, it provides a theoretical basis to develop highly sensitive MFC-MRD strategies.

Asociación entre estado nutricional y la cantidad de dientes permanentes en niños escolares en Costa Rica / Associação entre o estado nutricional e o número de dentes permanentes em crianças em idade escolar na Costa Rica / Association between nutritional status and the number of permanent teeth in a school child in Costa Rica

Resumen: El sobrepeso y la obesidad infantil se han convertido en uno de los desafíos de la salud pública a nivel mundial. Estas condiciones pueden afectar varios sistemas, asociándose con un inicio temprano de la pubertad y erupción dental. Objetivo: evaluar si existe una asociación entre el estado nutricional y la cantidad de dientes permanentes erupcionados en una población de niños escolares en Costa Rica. Material y métodos: Estudio transversal con una muestra de 753 niños entre los 6 y 12 años. Se realizó un examen clínico donde se registró la presencia o ausencia de dientes permanentes y se tomaron medidas de peso y talla. El estado antropométrico fue evaluado según los criterios de la Organización Mundial de la Salud. Se efectuó análisis descriptivo para todas las variables, comparación de medias entre los grupos (sobrepeso/obesidad vs peso normal) con la prueba U de Mann-Whitney. Se utilizó un análisis de regresión lineal con la edad y el sexo como covariantes. Resultados: Un 69.20% niños presentaba un peso normal y 30.80 % sobrepeso/obesidad. El promedio de dientes erupcionados era mayor en los niños con sobrepreso/obesidad que en aquellos con un peso normal (p=0.001). Al realizar la comparación según el sexo, la diferencia se mantiene (hombres, p=0.001; mujeres, p=0.018). El análisis de regresión lineal demostró que el estado nutricional (p=0.001) y la edad (p=0.001) están asociadas con el número de dientes permanentes erupcionados. Conclusión: los niños en edad escolar con sobrepeso/obesidad presentan mayor cantidad de dientes erupcionados que sus pares con un peso saludable.

Resumo: O sobrepeso e a obesidade na infância tornaram-se um dos desafios da saúde pública no mundo. Essas condições podem afetar vários sistemas, estando associadas ao início precoce da puberdade e a erupção dentária. Objetivo: Avaliar se há associação entre o estado nutricional e o número de dentes permanentes erupcionados em uma população de escolares da Costa Rica. Materiais e método: Estudo transversal com amostra de 753 crianças entre 6 e 12 anos. Foi realizado un exame clínico onde foi registrada a presença ou ausência dos dentes permanentes e as medidas do peso e altura. O estado antropométrico foi avaliado de acordo com os critérios da Organização Mundial da Saúde. Fio realizada una análise descritiva para todas as variáveis, comparação das médias entre os grupos (sobrepeso/obesidade vs peso normal) com o teste U de Mann-Whitney. Uma análise de regressão linear foi utilizada como a ideade e o sexo como covariáveis. Resultados: 69,20% das crianças apresentavam peso normal e 30,80% sobrepeso/obesidade. O número médio de dentes erupcionados foi maior em crianças com sobrepeso/obesidades (p=0.001). Ao fazer a comparação pelo sexo, a diferença permanece (homens, p=0.001; mulheres, p=0.018). A análise de regressão linear demostrou que o estado nutricional (p=0.001) e a idade (p=0.001) estão associados ao número de dentes permanentes erupcionados. Conclusão: Crianças em idade escolar com sobrepeso/obesas tam mais dentes erupcionados do que seus pares com peso saudável.

Abstract: Childhood overweight and obesity have become one of the public health challenges worldwide. These conditions can affect various systems, they are associated with an early onset of puberty and tooth eruption. Objective: to evaluate whether there is an association between nutritional status and the number of permanent teeth erupted in a population of school children in Costa Rica. Materials and methods: Cross-sectional study with a sample of 753 children between 6 and 12 years old. A clinical examination was performed the presence or absence of permanent teeth weight and height measurements were recorded. The anthropometric status was evaluated according to the criteria of the World Health Organization. Descriptive analysis was carried out for all variables, comparison of means between groups (overweight/obesity versus normal weight) with the Mann-Whitney U test. A linear regression analysis was used with age and sex as covariates. Results: 69,20% children had a normal weight and 30,80% were overweight/obese. The average number or erupted teeth as higher in children with overweight/obesity than in those with normal weight (p=0.001). This difference remains in the analysis y sex (men, p=0.001; women, p=0.018). Linear regression analysis demonstrated that nutritional status (p=0.001) and age (p=0.001) are associated with the number of permanent teeth erupted. Conclusion: Overweight/obese school-age children have more erupted teeth than their healthy-weight peers.

QTL dissection and mining of candidate genes for Ascochyta fabae and Orobanche crenata resistance in faba bean (Vicia faba L.).

BACKGROUND: Ascochyta blight caused by Ascochyta fabae Speg. and broomrape (Orobanche crenata) are among the economically most significant pathogens of faba bean. Several QTLs conferring resistance against the two pathogens have been identified and validated in different genetic backgrounds. The aim of this study was to saturate the most stable QTLs for ascochyta and broomrape resistance in two Recombinant Inbred Line (RIL) populations, 29H x Vf136 and Vf6 x Vf136, to identify candidate genes conferring resistance against these two pathogens. RESULTS: We exploited the synteny between faba bean and the model species Medicago truncatula by selecting a set of 219 genes encoding putative WRKY transcription factors and defense related proteins falling within the target QTL intervals, for genotyping and marker saturation in the two RIL populations. Seventy and 50 of the candidate genes could be mapped in 29H x Vf136 and Vf6 x Vf136, respectively. Besides the strong reduction of the QTL intervals, the mapping process allowed replacing previous dominant and pedigree-specific RAPD flanking markers with robust and transferrable SNP markers, revealing promising candidates for resistance against the two pathogens. CONCLUSIONS: Although further efforts in association mapping and expression studies will be required to corroborate the candidate genes for resistance, the fine-mapping approach proposed here increases the genetic resolution of relevant QTL regions and paves the way for an efficient deployment of useful alleles for faba bean ascochyta and broomrape resistance through marker-assisted breeding.

Efectividad de una unidad de dolor torácico en el primer nivel de atención en salud con telecardiología: un estudio cuasiexperimental de series de tiempo interrumpidas / Effectiveness of the chest pain unit at primary level of health care with telecardiology: a quasi-experimental study of interrupted time series

Resumen Introducción: Las unidades de dolor torácico disminuyen la morbimortalidad de los pacientes con síndrome coronario agudo. No obstante, se desconoce su efectividad en el primer nivel de atención con el apoyo de la telecardiología. Objetivo: Evaluar la efectividad de las unidades de dolor torácico sobre los tiempos de atención, la concordancia diagnóstica y la frecuencia de reconsultas a urgencias y hospitalización. Método: Estudio cuasiexperimental de series de tiempo interrumpidas, robusto, que incluyó 20,412 pacientes que consultaron al servicio de urgencias por dolor torácico, antes y después de implementar una unidad de dolor torácico. Se analizaron los cambios en la pendiente, el nivel, la autocorrelación y la varianza de los desenlaces estudiados entre ambos periodos (previo y posterior a las unidades de dolor torácico) a 30 días. Resultados: El promedio de edad fue de 44.9 ± 17.6 años y el 45.8% fueron hombres. La proporción global de pacientes remitidos para hospitalización fue del 9.0%. La pendiente de las tasas de reconsultas a urgencias disminuyó (diferencia: −1.23; intervalo de confianza del 95% [IC95%]: −2.46 a −0.01; p = 0.049) al comparar los dos periodos de observación. Igualmente, la proporción de pacientes remitidos sin síndrome coronario agudo y que finalmente tuvieron este diagnóstico en el tercer nivel de atención disminuyó en el periodo posterior a las unidades de dolor torácico con relación al previo (diferencia: −8.31; IC95%: −15.52 a −1.11; p = 0.020). Conclusiones: Las unidades de dolor torácico incrementaron los egresos de forma segura, con disminución de las reconsultas a urgencias en los siguientes 30 días por la misma causa. Además, mejoró la concordancia diagnóstica del síndrome coronario agudo, sin modificar los tiempos de atención ni la frecuencia de rehospitalización por enfermedad cardiovascular en el seguimiento.

Abstract Introduction: Chest pain units (CPU) decrease morbi-mortality in patients with acute coronary syndrome (ACS). Nevertheless, its effectiveness at primary level of health care with telecardiology support is unknown. Objective: To evaluate effectiveness of CPU on times of observation, diagnostic agreement and emergency department re-admission and hospitalizations. Method: Quasi-experimental study of robust interrupted time series, which included 20,412 patients admitted to the emergency department for chest pain, before and after the implementation of a CPU. Changes in slope, level, autocorrelation and, variance between both periods (before-CPU and after-CPU) in outcomes at 30 day follow-up were analyzed. Results: Subjects had a mean age of 44.9 ± 17.6 years-old and 45.8% were men. The overall rate of hospital admission was 9.0%. The slope of emergency re-consultation rates decreased (difference: −1.23; 95% CI: −2.46 to −0.01; p = 0.049), when comparing the two observation periods. Also, the level of proportion of patients admitted without ACS who finally had an inpatient diagnosis of ACS decreased after-CPU implementation (difference: −8.31; 95% CI: −15,52 to −1.11; p = 0.020). Conclusions: The CPU increased patient discharge safely with a reduction of 30-day re-admissions. In addition, an improvement in the ACS diagnostic agreement without affecting the time of observation or the frequency of re-hospitalization for cardiovascular disease was obtained during the follow-up.

Efficacy of fluoride varnishes with different compositions on white spot lesions remineralization / Eficácia de vernizes fluoretados com diferentes composições na remineralização de manchas brancas

Objective: To evaluate the efficacy of different fluoride varnishes on white spot lesions (WSL) remineralization. Material and Methods: Polished bovine enamel specimens were obtained (n = 60) and had their initial surface Knoop microhardness (SMH) determined. WSL were created and the SMH was measured again. Then, specimens were allocated into six groups: C ­ Control (without varnish); BF ­ Bifluorid 12 (6% NaF + 6% CaF2); DP ­ Duraphat (5% NaF); PF ­ Profluorid (5% NaF); FP - Fluor Protector (0.2% NaF + 0.9% difluorsilane); CW - Clinpro White Varnish (5% NaF + 5% TCP). After varnishes application, specimens were immersed in artificial saliva for 24 h. Then, pH-cycling was performed for 8 days and SMH was measured. Data were analyzed by one-way ANOVA and Tukey's test. Results: Non-significant differences were observed among the groups at baseline (p = 0.187) and after WSL formation (p = 0.999). After treatments, significant differences were observed among the groups (p = 0.001). Mean % of alteration (SD) and results of Tukey test were: C- 92.40 (12.10)a; PF- 88.66 (10.66)a; FP- 85.90 (14.49)ab; BF- 67.85 (17.86)bc; CW- 66.60 (18.48)c; DP- 58.62 (8.69)c. Conclusion: Bifluorid 12, Clinpro White Varnish, and Duraphat showed higher efficacy than artificial saliva in promoting the remineralization of WSL, nevertheless, none of the treatments were able to recover sound enamel baseline microhardness (AU)

Objetivo: Avaliar a eficácia de diferentes vernizes fluoretados na remineralização de lesões de mancha branca (LMB). Material e métodos: Espécimes de esmalte bovino polido (n = 60) foram submetidos à análise de microdureza superficial Knoop (KMH) inicial. Foram então criadas LMB artificialmente e os espécimes foram alocados em seis grupos: C ­ Controle (sem aplicação de verniz); BF ­ Bifluorid 12 (6% NaF + 6% CaF2); DP ­ Duraphat (5% NaF); PF ­ Profluorid (5% NaF); FP - Fluor Protector (0.2% NaF + 0.9% difluorsilano); CW - Clinpro White Varnish (5% NaF + 5% TCP). Após a aplicação dos vernizes, os espécimes ficaram imersos em saliva artificial por 24h e uma ciclagem de pH foi realizada por 8 dias. Após a ciclagem, KMH final foi realizada. Os dados foram analisados por ANOVA e teste de Tukey (5%). Resultados: Não foi observada diferença significante para os grupos após a KHM inicial (p = 0.187) e após a formação de LMB (p = 0.999). Após os tratamentos, diferenças significativas foram observadas entre os grupos (p = 0.001). Valores de média de % de alteração superficial (desvio-padrão) e resultados do teste de Tukey foram: C- 92.40 (12.10)a; PF- 88.66 (10.66)a; FP- 85.90 (14.49)ab; BF- 67.85 (17.86)bc; CW- 66.60 (18.48)c; DP- 58.62 (8.69)c. Conclusão: Os vernizes Bifluorid 12, Clinpro White Varnish e Duraphat apresentaram maior eficácia na remineralização das LMB quando comparados à saliva artificial, entretanto, nenhum dos produtos testados foi capaz de recuperar os valores iniciais de microdureza. (AU)

Outcomes of COVID-19 in Patients With Lung Cancer Treated in a Tertiary Hospital in Madrid.

Background: Cancer patients represent a vulnerable population for COVID-19 illness. We aimed to analyze outcomes of lung cancer patients affected by COVID-19 in a tertiary hospital of a high-incidence region during the pandemic. Methods: We annotated 23 lung cancer patients consecutively diagnosed with COVID-19 at our institution (HGUGM; Madrid, Spain) between March 4th, 2020 and May 12th, 2020. Only patients with a confirmatory SARS-CoV-2 RT-PCR were included in the study. Results: All patients had at least 1 COVID-19 related symptom; cough (48%), shortness of breath (48%), fever (39%), and low-grade fever (30%) were the most common. Time from symptoms onset to first positive SARS-CoV-2 PCR was 5.5 days (range 1-17), with 13% of cases needed from a 2nd PCR to confirm diagnosis. There was a high variability on thoracic imaging findings, with multilobar pneumonia as the most commonly found pattern (74%). Main lab test abnormalities were low lymphocytes count (87%), high neutrophil to lymphocyte ratio -NLR- (78%), and elevated inflammatory markers: fibrinogen (91%), c-reactive protein -CRP- (87%), and D-dimer (70%). In our series, hospitalization rate was 74%, 39% of patients developed acute respiratory distress syndrome (ARDS), and the case-fatality rate was 35% (8/23). 87% of patients received anti-viral treatment (87% hydroxychloroquine, 74% lopinavir/ritonavir, 13% azithromycin), 43% corticosteroids, 26% interferon-ß, 4% tocilizumab, and 82% of hospitalized patients received anticoagulation. High-oxygen requirements were needed in 39% of patients, but only 1 pt was admitted for invasive MV and was discharged 42 days after admission. Multiple variables related to tumor status, clinical baseline conditions, and inflammation markers were associated with mortality but did not remain statistically significant in a multivariate model. In patients with lung cancer receiving systemic therapy (n = 242) incidence and mortality from COVID-19 were 4.5, and 2.1%, respectively, with no differences found by type of treatment. Conclusions: Lung cancer patients represent a vulnerable population for COVID-19, according to the high rate of hospitalization, onset of ARDS, and high mortality rate. Although larger series are needed, no differences in mortality were found by type of cancer treatment. Measures to minimize the risk of SARS-CoV-2 infection remain key to protect lung cancer patients.

The bHLH transcription factor VfTT8 underlies zt2, the locus determining zero tannin content in faba bean (Vicia faba L.).

Faba bean (Vicia faba L.) is an important protein-rich fodder crop, which is widely cultivated in temperate areas. However, antinutritional compounds such as condensed tannins, limit the use of this protein source in monogastric feed formulations. Previous studies demonstrated that two recessive and complementary genes, zt1 and zt2, control absence of tannin and white flower colour in faba bean. An ortholog of the Medicago WD40 transcription factor TTG1 was reported to encode the zt1 phenotype, but the responsible gene for zt2 is still unknown. Here we used a candidate gene approach combined with linkage mapping, comparative genomics and gene expression to fine map the zt2 genomic region and to identify the regulatory gene controlling both traits. Seventy-two genes, including 23 MYB and bHLH regulatory genes predicted to be associated with anthocyanin expression together with WRKY proteins, were screened and genotyped in three mapping populations. The linkage groups constructed identified the regulatory gene, TRANSPARENT TESTA8 (TT8), encoding a basic helix-loop-helix (bHLH) transcription factor, as the candidate for zt2. This finding was supported by qPCR analysis and further validated in different genetic backgrounds. Accordingly, VfTT8 was downregulated in white flowered types while showing high levels of expression in wild genotypes. Our results provide new insights on the regulatory mechanisms of tannin biosynthesis in faba bean and will facilitate the development of an ultimate zt2 diagnostic marker for the fast generation of new value-added cultivars free of tannins and with improved nutritional value.

Bond Strength of Composite Resin Restoration Repair: Influence of Silane and Adhesive Systems.

AIM: The aim of this study is to evaluate the effect of silane (Si) application and different adhesive systems on the bond strength of composite resin repair. MATERIALS AND METHODS: One hundred composite truncated cone-shaped specimens were prepared and submitted to 5,000 thermal cycles to simulate existing restorations. Their top surfaces were airborne particle abraded with aluminum oxide, etched with phosphoric acid, and divided into two groups (n = 50) with or without Si application. Each group was divided into five subgroups (n = 10) according to the adhesive system applied: Solobond Plus Primer and Adhesive (SPA)-two-bottle, Solobond Plus adhesive (SA), Admira Bond (A)-one bottle, Futurabond DC (FDC)-self-etch, and Futurabond M (FM)-self-etch. New composite resin was applied over the bonded area. A control group was prepared to evaluate the cohesive strength of the composite resin. Specimens were submitted to tensile stress. Data were analyzed with two-way analysis of variance (ANOVA), and the Tukey and Dunnett tests. RESULTS: Si application reduced the bond strength of all adhesives (p = 0.001). Groups SA and SPA showed higher bond strengths in relation to other groups (p = 0.01). Groups FDC + Si, FM, FM + Si, and A + Si showed smaller mean bond strength values than that of the control group (p < 0.05). CONCLUSION: Previous Si application reduced bond strength values. The two-bottle adhesive showed better results than one-bottle or self-etching systems for composite resin repairs. CLINICAL SIGNIFICANCE: The kind of adhesive system applied for composite resin repairs has a great influence on bond strength values. The use of Si in this situation is not recommended.

Characterization and diagnostic marker for TTG1 regulating tannin and anthocyanin biosynthesis in faba bean.

Condensed tannins, found in coloured-flowering varieties of faba bean (Vicia faba L) are, after vicine and convicine, one of the major anti-nutritional factors for monogastric animals. The development of tannin-free cultivars is a key goal in breeding to broaden the use of this legume in the animal feed industry. Two recessive genes, zt-1 and zt-2, control the zero-tannin content and promote white-flowered plants. Previous studies exploiting synteny with the model Medicago truncatula reported a mutation in TTG1, a gene encoding a WD40 transcription factor located in chromosome II, as the responsible for the zt-1 phenotypes. Here a comprehensive analysis of VfTTG1 (including phylogenetic relationships, gene structure and gene expression) has been conducted to confirm the identity of the gene and to reveal structural changes that may result in different functional alleles. The results confirmed the identity of the candidate and revealed the existence of two different alleles responsible for the phenotype: ttg1-a, probably due to a mutation in the promoter region, and ttg1-b caused by a deletion at the 5'end of VfTTG1. Based on the sequencing results, an allele-specific diagnostic marker was designed that differentiate zt-1 from wild and zt-2 genotypes and facilitates its deployment in faba bean breeding programs.

Species Deletions from Microbiome Consortia Reveal Key Metabolic Interactions between Gut Microbes.

The gut microbiome is a complex microbial community that plays a key role in human health. Diet is an important factor dictating gut microbiome composition. This is mediated by multiple microbe-microbe interactions that result in the fermentation of nondigestible carbohydrates and the production of short-chain fatty acids. Certain species play key metabolic roles in the microbiome, and their disappearance could result in dysbiosis. In this work, a synthetic consortium of 14 gut microbes was studied during the utilization of prebiotic inulin in batch bioreactors. Fermentations were repeated leaving one species out every time, in order to evaluate the impact of their elimination on the system. Substrate consumption, microbial composition, and metabolite production were determined. Single deletions never resulted in a complete loss of bacterial growth or inulin consumption, suggesting functional redundancy. Deletions of Bacteroides dorei and Lachnoclostridium clostridioforme resulted in lower biomass and higher residual inulin. The absence of B. dorei impacted the abundance of the other 10 species negatively. Lachnoclostridium symbiosum, a butyrate producer, appeared to be the most sensitive species to deletions, being stimulated by the presence of Escherichia coli, Bifidobacterium adolescentis, B. dorei, and Lactobacillus plantarum Conversely, bioreactors without these species did not show butyrate production. L. clostridioforme was observed to be essential for propionate production, and B. dorei for lactate production. Our analysis identified specific members that were essential for the function of the consortium. In conclusion, species deletions from microbial consortia could be a useful approach to identify relevant interactions between microorganisms and defining metabolic roles in the gut microbiome.IMPORTANCE Gut microbes associate, compete for, and specialize in specific metabolic tasks. These interactions are dictated by the cross-feeding of degradation or fermentation products. However, the individual contribution of microbes to the function of the gut microbiome is difficult to evaluate. It is essential to understand the complexity of microbial interactions and how the presence or absence of specific microorganisms affects the stability and functioning of the gut microbiome. The experimental approach of this study could be used for identifying keystone species, in addition to redundant functions and conditions that contribute to community stability. Redundancy is an important feature of the microbiome, and its reduction could be useful for the design of microbial consortia with desired metabolic properties enhancing the tasks of the keystone species.

Minimally invasive plate osteosynthesis in humerus nonunion after intramedullary nailing.

INTRODUCTION: The purpose of this study was to evaluate the results achieved after the use of lateral minimally invasive plate osteosynthesis (MIPO) in oligotrophic humerus nonunions resulting from failed intramedullary nailing (IM). METHOD: We evaluated nine patients with humerus nonunion after failed locked intramedullary nailing, all treated using 3.5-mm locked compression plates (LCP) placed through lateral minimally invasive approaches, between 2010 and 2016. Patient's age averaged 39.7 years. All nonunions were diaphyseal and oligotrophic. All nonunions had previous surgical treatment with static locked nails (seven antegrade and two retrograde). The IM nails were all well inserted in the humerus (none of them protruded or had rotator cuff lesions associated). Pre-operative Disabilities of the Arm, Shoulder and Hand (DASH) score averaged 25.5 points. Pre-operative Constant's score averaged 80.2. Pre-operative visual analog scale of pain averaged 2.4 points. RESULTS: Follow-up averaged 17.7 months. Time between initial surgery and revision procedure averaged 11.7 months. Union was achieved in all cases, after an average of 4.8 months. DASH score at last follow-up averaged 5.1 points, and final Constant's score averaged 93.7 points. The analog scale of pain averaged 0.7 points. Time from definitive surgery to work return averaged 3.9 months. Long 3.5-mm LCPs were used (plate length averaged 16.9 screw holes). In two cases, a third 4-cm incision at the nonunion site was performed and cancellous autologous iliac crest bone graft was associated. CONCLUSION: In our series of nine patients, we achieved union and good objective and subjective results, with high patient satisfaction, using a lateral MIPO technique and placing long 3.5-mm LCPs in selected oligotrophic humerus nonunions after failed IM nailing.

Prognostic Factors in Severe Chagasic Heart Failure.

BACKGROUND: Prognostic factors are extensively studied in heart failure; however, their role in severe Chagasic heart failure have not been established. OBJECTIVES: To identify the association of clinical and laboratory factors with the prognosis of severe Chagasic heart failure, as well as the association of these factors with mortality and survival in a 7.5-year follow-up. METHODS: 60 patients with severe Chagasic heart failure were evaluated regarding the following variables: age, blood pressure, ejection fraction, serum sodium, creatinine, 6-minute walk test, non-sustained ventricular tachycardia, QRS width, indexed left atrial volume, and functional class. RESULTS: 53 (88.3%) patients died during follow-up, and 7 (11.7%) remained alive. Cumulative overall survival probability was approximately 11%. Non-sustained ventricular tachycardia (HR = 2.11; 95% CI: 1.04 - 4.31; p<0.05) and indexed left atrial volume ≥ 72 mL/m2 (HR = 3.51; 95% CI: 1.63 - 7.52; p<0.05) were the only variables that remained as independent predictors of mortality. CONCLUSIONS: The presence of non-sustained ventricular tachycardia on Holter and indexed left atrial volume > 72 mL/m2 are independent predictors of mortality in severe Chagasic heart failure, with cumulative survival probability of only 11% in 7.5 years.

Prognostic Factors in Severe Chagasic Heart Failure / Fatores Prognósticos na Insuficiência Cardíaca Grave de Etiologia Chagásica

Abstract Background: Prognostic factors are extensively studied in heart failure; however, their role in severe Chagasic heart failure have not been established. Objectives: To identify the association of clinical and laboratory factors with the prognosis of severe Chagasic heart failure, as well as the association of these factors with mortality and survival in a 7.5-year follow-up. Methods: 60 patients with severe Chagasic heart failure were evaluated regarding the following variables: age, blood pressure, ejection fraction, serum sodium, creatinine, 6-minute walk test, non-sustained ventricular tachycardia, QRS width, indexed left atrial volume, and functional class. Results: 53 (88.3%) patients died during follow-up, and 7 (11.7%) remained alive. Cumulative overall survival probability was approximately 11%. Non-sustained ventricular tachycardia (HR = 2.11; 95% CI: 1.04 - 4.31; p<0.05) and indexed left atrial volume ≥ 72 mL/m2 (HR = 3.51; 95% CI: 1.63 - 7.52; p<0.05) were the only variables that remained as independent predictors of mortality. Conclusions: The presence of non-sustained ventricular tachycardia on Holter and indexed left atrial volume > 72 mL/m2 are independent predictors of mortality in severe Chagasic heart failure, with cumulative survival probability of only 11% in 7.5 years.

Resumo Fundamento: Fatores prognósticos são bastante estudados na insuficiência cardíaca (IC), mas ainda não possuem um papel estabelecido na IC grave de etiologia chagásica. Objetivo: Identificar a associação de fatores clínicos e laboratoriais com o prognóstico da IC grave de etiologia chagásica, bem como a associação desses fatores com a taxa de mortalidade e a sobrevida em um seguimento de 7,5 anos. Métodos: 60 pacientes portadores de IC grave de etiologia chagásica foram avaliados com relação às seguintes variáveis: idade, pressão arterial, fração de ejeção, sódio plasmático, creatinina, teste de caminhada de 6 minutos, taquicardia ventricular não sustentada, largura do QRS, volume do átrio esquerdo indexado e classe funcional. Resultados: 53 (88,3%) pacientes foram a óbito durante o período de seguimento e 7 (11,7%) permaneceram vivos. A probabilidade de sobrevida geral acumulada foi de aproximadamente 11%. Taquicardia ventricular não sustentada (HR = 2,11; IC 95%: 1,04 - 4,31; p<0,05) e volume do átrio esquerdo indexado ≥ 72 ml/m2 (HR = 3,51; IC 95%: 1,63 - 7,52; p<0,05) foram as únicas variáveis que permaneceram como preditores independentes de mortalidade. Conclusão: A presença de taquicardia ventricular não sustentada ao Holter e o volume do átrio esquerdo indexado > 72 ml/m2 são preditores independentes de mortalidade na IC chagásica grave, com probabilidade de sobrevida acumulada de apenas 11% em 7,5 anos.

Análise da efetividade de polimerização, adaptação marginal e interna de restaurações Classe II de resinas compostas de inserção em bloco utilizando diferentes matrizes / Effectiveness of cure, marginal and internal adaptation of Class II bulkfill composites restorations made with different matrices bands

O objetivo deste estudo foi analisar a efetividade de polimerização, a adaptação marginal e interna de resinas compostas para dentes posteriores, empregando diferentes matrizes associadas ao envelhecimento artificial. Cento e vinte incisivos bovinos foram cortados e desgastados para simular dentes posteriores, nos quais foram realizados preparados de Classe II, com o ângulo cavo superficial gengival em dentina. As cavidades tinham 6 mm de altura (ocluso-cervical), 3 de largura(vestibulo-lingual) e 1,5 de profundidade (mesio-distal). Os espécimes foram divididos em seis grupos de acordo com o material restaurador utilizado: GrandioSO (Voco)– convencional incremental, G-aenial universal flo (GC)– fluída incremental, Xtrafil (Voco)– convencional com inserção em bloco, X-tra base (Voco)– fluída com inserção em bloco, Venus Bulk Fill (Kulzer)– fluída com inserção em bloco e SureFil SDR (Dentsply)– fluída com inserção em bloco. As resinas fluídas com inserção em bloco foram inseridas em incremento de 4 mm e receberam uma cobertura oclusal de 2 mm utilizando a resina GrandioSO, com exceção da X-tra fil que foi coberta com uma camada adicional do mesmo material. Nos demais grupos foi utilizada uma técnica incremental de 2 mm. Metade dos espécimes foi restaurada com matriz metálica e a outra metade com matriz de poliéster. O percentual de polimerização foi mensurado através da dureza Knoop e da relação dureza base/topo. A média da largura das fendas marginais presentes nas margens de cada restauração foi calculada após medição, em micrometros, no microscópio óptico (100x). Os espécimes foram submetidos à ciclagem mecânica (500,000 ciclos) e térmica (1,000 ciclos) e a fenda marginal foi avaliada novamente. Para fenda interna, os espécimes foram seccionados em uma cortadeira de precisão e o comprimento, em milímetros, da parede gengival que apresentava fenda foi mensurado no microscópio óptico. Foi calculada então a porcentagem de fenda em relação ao comprimento total...

The aim of this study was to analyze the effectiveness of cure, marginal and internal adaptation of composite resins for posterior teeth using different matrices bands associated with artificial aging. One hundred and twenty bovine incisors were cut and flattened in order to simulate posterior teeth, and then were prepared for class II cavities with gingival cavosurface margin in dentin. Cavities had 6 mm height(occluso-cervical), 3 mm width (bucco-lingual) and 1.5 mm deep (mesio-distal).Specimens were divided into six groups according to the restorative material:GrandioSO (Voco)- conventional, G-aenial universal flo (GC)- flowable, X-tra fil (Voco)- conventional bulk, X-tra base (Voco)- flowable bulk, Venus Bulk Fill (Kulzer)- flowable bulk, and SureFil SDR (Dentsply)- flowable bulk. Bulk-fill composites were placed in single increment of 4mm and received a 2mm occlusal coverage of GrandioSO, with the exception of X-tra fil which was covered with a layer of the same material. The remmaing groups were restored in 2 mm incremental technique. Half ofthe specimens were restored with metallic and the other half with polyester matrix. of cure was measured by Knoop hardness and the bottom/tophardness ratio was evaluated. The average width of marginal gaps present in the restoration was calculated after optical microscope measuring (100x), in microns. The specimens were subjected to mechanical (500.000 cycles) and thermal cycling (1.000 cycles) and marginal gap was evaluated again. For internal adaptation, the specimens were sectioned and the gap length, in millimeters, present on the gingivalwall was measured in an optical microscope. The percentage of gap length in relation to the total length of the wall was also calculated. Data were analyzed with three andtwo-way ANOVA and Tukey´s test. Regarding effectiveness of cure, significant differences were observed for resin factor. Composites G-aenial universal flo (82.08%) and GrandioSO(84.62%) had the highest cure...