The Role of a WRKY Transcription Factor in Mediating Hormone Response During Rice Seed Germination

This dissertation provides genetic evidence that addresses the biological function of OsWRKY71, a transcription factor expressed abundantly in rice seed tissues. This work is significant as it is the first report on oswrky71 mutants, which display an early seed germination phenotype hyposensitive to ABA. This dissertation first focused on providing the most thorough-to-date RNA-seq time course analysis of rice embryos during germination. Since embryo and aleurone layers communication during germination is a crucial process, we then studied the aleurone transcriptomics, which provided further insight into the regulatory mechanisms involved in the germination of oswrky71 seeds. By profiling the transcriptomes, we identified key genes regulated by OsWRKY71 in both embryos and aleurone cells that are involved in the control of the germination process. We provide a working model where OsWRKY71 negatively regulates seed germination by positively regulating the abundance of a critical ABA signaling node and negatively regulating seed storage mobilization, cell wall loosening, water transport, and epiblast tissue weakening. Association studies revealed an OsWRKY71-containing QTL, qLTG-2, which is known to be associated with seed germination speed at low temperatures. Indeed, oswrky71 mutants germinate early at low temperatures, suggesting that WRKY71 is the primary gene of qLTG-2. As such, this dissertation provides new knowledge on the WRKY-dependent regulation of ABA signaling during rice germination. This information is valuable in developing strategies to improve seed germination rates and ensure optimal crop yields to alleviate food insecurity. Rice germinating faster is needed in water-limited growing regions and will allow the production of pregerminated rice for food processing applications to be created using less energy. Lastly, we performed a comprehensive and high-throughput characterization of the metabolomes of wild type and oswrky71 threshed grains in conjunction with standard analytical methods for grain nutrient contents. This work allowed us to identify differential metabolites between the wild type and oswrky71 mutant for risk assessment if we position this mutant for human consumption. Overall, our study provides a deeper understanding of the molecular mechanisms of rice seed germination that is crucial to improving rice crops\u27 yield and resilience

Comparative Nutritional Assessment and Metabolomics of a WRKY Rice Mutant with Enhanced Germination Rates

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Comparative Nutritional Assessment and Metabolomics of a WRKY Rice Mutant with Enhanced Germination Rates

Rice is the primary staple food for half the world’s population. Climate change challenges and food insecurity supports the need for rice with agronomically advantageous traits. We report on a transposon insertional rice mutant with enhanced germination rates. This trait is advantageous for rice growth in limited water regions and to reduce yield constraints caused by weed and bird competition. Evaluations of vital nutritional components, compositional analysis, and comparative metabolomics on threshed grain samples are performed, as these assays are those used to assess the safety of foods from genetically modified crops. Compared with the wild type (cv. Nipponbare), oswrky71 mutant grains have a similar size, shape, amount of crude fiber, crude fat, and ash content but higher crude protein. Mineral analyses reveal higher contents of phosphorus and zinc but lower calcium, potassium, sodium, and manganese in the mutant. Analysis of B vitamins reveals significantly higher riboflavin concentrations but lower choline chloride, calcium pantothenate, and thiamine. In addition, untargeted metabolomics analyses identify approximately 50 metabolites whose levels differed between the mutant and its wild type. Physical traits and compositional parameters analyzed are mostly similar and within the range or very close to being considered safe for consumption by the International Life Sciences Institute Crop Composition Database. Further agronomic evaluation and cooked rice sensory properties assessment are needed before positioning this mutant for human consumption

Selective IgA Deficiency and Blood Component Transfusion: In Search of the Lost Evidence

Background: Selective IgA deficiency (IgA-D) has been historically considered a high-risk entity for developing allergic/anaphylactic reactions after blood transfusion (AATRs). However, it has been suggested that the IgA-D-related anaphylactic transfusion reaction is not evidence-based. Methods: We conducted three different approaches to collect evidence about epidemiology, AATRs, and transfusion management of patients with IgA-D at La Fe University Hospital. Firstly, we analysed the prevalence of IgA-D in a population of patients diagnosed with acute leukaemia, The second approach consisted of collecting transfusion data from IgA-D patients. Finally, we reviewed the IgA levels of patients recorded in the hemovigilance system suffering an AATR. Results: IgA-D prevalence was 1 in 334 patients. At least one blood component was transfused to 23 patients diagnosed with IgA-D. Plasma was transfused to eight IgA-D patients, while six patients received red blood cells, platelets, and plasma. No adverse reactions were reported in any patient. AATRs occurred in 325 men and 264 women with a median age of 52 years. Severe reactions occurred in 56 patients (1/14,520 components). Mean IgA levels were 215 mg/dL (4–5570) for mild reactions and 214 mg/dL (14–824) for severe reactions (p = ns). Washed platelets were administered to two patients who developed severe and repeated AATRs. Both had normal IgA levels. Conclusions: Since the AATRs related to IgA-D are extremely low, as reported in current hemovigilance systems, IgA-D should not be considered a high-risk entity to develop AATRs. On the contrary, our findings support standard transfusion management of IgA-D patients

Methodological Approach of the Iron and Muscular Damage: Female Metabolism and Menstrual Cycle during Exercise Project (IronFEMME Study)

Background: The increase in exercise levels in the last few years among professional and recreational female athletes has led to an increased scientific interest about sports health and performance in the female athlete population. The purpose of the IronFEMME Study described in this protocol article is to determine the influence of different hormonal profiles on iron metabolism in response to endurance exercise, and the main markers of muscle damage in response to resistance exercise; both in eumenorrheic, oral contraceptive (OC) users and postmenopausal well-trained women. Methods: This project is an observational controlled randomized counterbalanced study. One hundered and four (104) active and healthy women were selected to participate in the IronFEMME Study, 57 of which were eumenorrheic, 31 OC users and 16 postmenopausal. The project consisted of two sections carried out at the same time: iron metabolism (study I) and muscle damage (study II). For the study I, the exercise protocol consisted of an interval running test (eight bouts of 3 min at 85% of the maximal aerobic speed), whereas the study II protocol was an eccentric-based resistance exercise protocol (10 sets of 10 repetitions of plate-loaded barbell parallel back squats at 60% of their one repetition maximum (1RM) with 2 min of recovery between sets). In both studies, eumenorrheic participants were evaluated at three specific moments of the menstrual cycle: early-follicular phase, late-follicular phase and mid-luteal phase; OC users performed the trial at two moments: withdrawal phase and active pill phase. Lastly, postmenopausal women were only tested once, since their hormonal status does not fluctuate. The three-step method was used to verify the menstrual cycle phase: calendar counting, blood test confirmation, and urine-based ovulation kits. Blood samples were obtained to measure sex hormones, iron metabolism parameters, and muscle damage related markers. Discussion: IronFEMME Study has been designed to increase the knowledge regarding the influence of sex hormones on some aspects of the exercise-related female physiology. Iron metabolism and exercise-induced muscle damage will be studied considering the different reproductive status present throughout well-trained females' lifespan

Enfermedad hepática por alcohol. Guías de práctica clínica. Documento de consenso auspiciado por la AEEH

[EN]: Alcohol-related liver disease (ARLD) is the most prevalent cause of advanced liver disease and liver cirrhosis in Europe, including Spain. According to the World Health Organization the fraction of liver cirrhosis attributable to alcohol use in Spain is 73.8% among men and 56.3% among women. ARLD includes various stages such as steatohepatitis, cirrhosis and hepatocellular cancer. In addition, patients with underlying ARLD and heavy alcohol intake may develop alcoholic hepatitis, which is associated with high mortality. To date, the only effective treatment to treat ARLD is prolonged withdrawal. There are no specific treatments, and the only treatment that increases life expectancy in alcoholic hepatitis is prednisolone. For patients with alcoholic hepatitis who do not respond to treatment, some centres offer the possibility of an early transplant. These clinical practice guidelines aim to propose recommendations on ARLD taking into account their relevance as a cause of advanced chronic liver disease and liver cirrhosis in our setting. This paper aims to answer the key questions for the clinical practice of Gastroenterology, Hepatology, as well as Internal Medicine and Primary Health Centres, making the most up-to-date information regarding the management and treatment of ARLD available to health professionals. These guidelines provide evidence-based recommendations for the clinical management of this disease.[ES]: La enfermedad hepática alcohólica (EHA) es la causa más prevalente de enfermedad hepática avanzada y cirrosis hepática en Europa incluyendo a España. De acuerdo con la Organización Mundial de la Salud la fracción de cirrosis hepática atribuible al uso de alcohol en España es del 73,8% entre varones y del 56,3% entre mujeres. La EHA incluye diversos estadios como la esteatohepatitis, la cirrosis y el cáncer hepatocelular. Además, enfermos con EHA de base e ingesta abundante de alcohol pueden desarrollar hepatitis alcohólica, que cursa con una elevada mortalidad. Hasta la fecha, el único tratamiento efectivo para tratar la EHA es la abstinencia prolongada. No existen tratamientos específicos, y el único tratamiento que aumenta la esperanza de vida en la hepatitis alcohólica es la prednisolona. Para enfermos con hepatitis alcohólica que no responden al tratamiento, algunos centros ofrecen la posibilidad de un trasplante precoz. Estas guías de práctica clínica tienen como objetivo proponer recomendaciones sobre la EHA teniendo en cuenta su relevancia como causa de hepatopatía crónica avanzada y cirrosis hepática en nuestro medio. En el presente trabajo se propone como objetivo responder las preguntas claves para la práctica clínica de Gastroenterología, Hepatología, así como de Medicina Interna y centros de salud primaria, poniendo al servicio del profesional de la salud la información más actualizada respecto al manejo y tratamiento de la EHA. Estas guías proporcionan recomendaciones basadas en la evidencia para el manejo clínico de esta enfermedad

Novel Molecular Mechanisms in the Development of Non-Alcoholic Steatohepatitis

Non-alcoholic fatty liver disease (NAFLD) is one of the most common causes of chronic liver disease in adults and children worldwide. NAFLD has become a severe health issue and it can progress towards a more severe form of the disease, the non-alcoholic steatohepatitis (NASH). A combination of environmental factors, host genetics, and gut microbiota leads to excessive accumulation of lipids in the liver (steatosis), which may result in lipotoxicity and trigger hepatocyte cell death, liver inflammation, fibrosis, and pathological angiogenesis. NASH can further progress towards liver cirrhosis and cancer. Over the last few years, cell-derived extracellular vesicles (EVs) have been identified as effective cell-to-cell messengers that transfer several bioactive molecules in target cells, modulating the pathogenesis and progression of NASH. In this review, we focused on recently highlighted aspects of molecular pathogenesis of NASH, mediated by EVs via their bioactive components. The studies included in this review summarize the state of art regarding the role of EVs during the progression of NASH and bring novel insight about the potential use of EVs for diagnosis and therapeutic strategies for patients with this disease

Bacteriophage targeting of gut bacterium attenuates alcoholic liver disease

Chronic liver disease due to alcohol-use disorder contributes markedly to the global burden of disease and mortality. Alcoholic hepatitis is a severe and life-threatening form of alcohol-associated liver disease. The gut microbiota promotes ethanol-induced liver disease in mice, but little is known about the microbial factors that are responsible for this process. Here we identify cytolysin-a two-subunit exotoxin that is secreted by Enterococcus faecalis-as a cause of hepatocyte death and liver injury. Compared with non-alcoholic individuals or patients with alcohol-use disorder, patients with alcoholic hepatitis have increased faecal numbers of E. faecalis. The presence of cytolysin-positive (cytolytic) E. faecalis correlated with the severity of liver disease and with mortality in patients with alcoholic hepatitis. Using humanized mice that were colonized with bacteria from the faeces of patients with alcoholic hepatitis, we investigated the therapeutic effects of bacteriophages that target cytolytic E. faecalis. We found that these bacteriophages decrease cytolysin in the liver and abolish ethanol-induced liver disease in humanized mice. Our findings link cytolytic E. faecalis with more severe clinical outcomes and increased mortality in patients with alcoholic hepatitis. We show that bacteriophages can specifically target cytolytic E. faecalis, which provides a method for precisely editing the intestinal microbiota. A clinical trial with a larger cohort is required to validate the relevance of our findings in humans, and to test whether this therapeutic approach is effective for patients with alcoholic hepatitis