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Abstract Subcellular compartmentalization of metabolic enzymes establishes a unique metabolic environment that elicits specific cellular functions. Indeed, the nuclear translocation of certain metabolic enzymes is required for epigenetic regulation and gene expression control. Here, we show that the nuclear localization of the mitochondrial enzyme methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) ensures mitosis progression. Nuclear MTHFD2 interacts with proteins involved in mitosis regulation and centromere stability, including the methyltransferases KMT5A and DNMT3B. Loss of MTHFD2 induces severe methylation defects and impedes correct mitosis completion. MTHFD2 deficient cells display chromosome congression and segregation defects and accumulate chromosomal aberrations. Blocking the catalytic nuclear function of MTHFD2 recapitulates the phenotype observed in MTHFD2 deficient cells, whereas restricting MTHFD2 to the nucleus is sufficient to ensure correct mitotic progression. Our discovery uncovers a nuclear role for MTHFD2, supporting the notion that translocation of metabolic enzymes to the nucleus is required to meet precise chromatin needs.

Abstract Nuclear metabolism and DNA damage response are intertwined processes, but the precise molecular links remain elusive. Here, we explore this crosstalk using triple-negative breast cancer (TNBC) as a model, a subtype often prone to DNA damage accumulation. We show that the de novo purine synthesis enzyme IMPDH2 is enriched on chromatin in TNBC compared to other subtypes. IMPDH2 chromatin localization is DNA damage dependent, and IMPDH2 repression leads to DNA damage accumulation. On chromatin, IMPDH2 interacts with and modulates PARP1 activity by controlling the nuclear availability of NAD+ to fine-tune the DNA damage response. However, when IMPDH2 is restricted to the nucleus, it depletes nuclear NAD+, leading to PARP1 cleavage and cell death. Our study identifies a non-canonical nuclear role for IMPDH2, acting as a convergence point of nuclear metabolism and DNA damage response.

Societal Impact Statement Table grape production is a traditional practice in southeastern Spain, where locals have produced fresh grapes and raisins for centuries. Many of these vines are now centenary, and they represent a useful source of diversity for developing future table grape varieties with improved traits. Genetic analysis showed that many of the local varieties identified in this study were traditional varieties from Western Mediterranean countries. Others were not identified, and they might be old grape varieties of previous importance in the region. The conservation and characterization of these varieties could be key to ensuring current and future vineyard sustainability. Summary Current worldwide table grape production focuses on a reduced number of Vitis vinifera L. varieties. However, traditional farmers have grown many table grape varieties for centuries, as they provided a steady source of fresh fruit and raisins. These ancient living genotypes potentially store a genetic diversity that can be used now to ensure future grape production. Here we focused on the study of grapevines found across Almería, one of the Spanish regions with longer tradition in table grape production. After an exhaustive inspection, we located 220 old (some centenary) vines producing grapes consumed by owners or in abandoned areas no longer devoted to agriculture. Some of these vines were identified by comparing their simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) genetic profiles with available data from international databases. We found that, while grape growers' efforts focused on the cultivation of traditional grape varieties from Western Mediterranean regions, they also cultivated few exogenous varieties if they provided additional fruit features. Other vines were found to have genetic profiles that did not match reference datasets. Interestingly, some of them were found in multiple locations, suggesting they are endangered varieties with some previous relevance in the region. Besides, first‐degree relationships support the autochthonous origin of many of these unidentified genotypes. Locals kept a high number of different grapevine varieties, now considered reservoirs of genetic diversity. Traditional farming practices have been useful to prevent the loss of this diversity, which now needs to be preserved and further studied to contribute to the sustainability of viticultural systems.

Abstract Background Some evidence suggests that fluid resuscitation with lactated Ringer’s solution (LR) may have an anti-inflammatory effect on acute pancreatitis (AP) when compared to normal saline (NS) and may be associated with a decrease in severity, but existing single-center randomized controlled trials showed conflicting results. The WATERLAND trial aims to investigate the efficacy and safety of fluid resuscitation using LR compared to NS in patients with AP. Methods The WATERLAND trial is an international multicenter, open-label, parallel-group, randomized, controlled, superiority trial. Patients will be randomly assigned in a 1:1 ratio to receive LR versus NS-based fluid resuscitation for at least 48 h. The primary outcome will be moderately severe or severe AP, according to the revision of the Atlanta classification. The secondary objectives of the WATERLAND trial are to determine the effect of LR versus NS fluid resuscitation on several efficacy and safety outcomes in patients with AP. A total sample of 720 patients, 360 in the LR group and 360 in the NS group, will achieve 90% power to detect a difference between the group proportions of 10%, assuming that the frequency of moderately severe or severe AP in the LR group will be 17%. A loss to follow-up of 10% of patients is expected, so the total sample size will be 396 patients in each treatment arm (792 patients overall). The test statistic used is the two-sided Z test with pooled variance set at a 0.05 significance level. Discussion The WATERLAND study aims to improve the early management of AP. Fluid resuscitation is an inexpensive treatment available in any hospital center worldwide. If a better evolution of pancreatitis is demonstrated in one of the treatment arms, it would have important repercussions in the management of this frequent disease. Trial registration ClinicalTrials.gov, NCT05781243. Registration date on January 4, 2023. EudraCT number 2023–000010-18, first posted March 23, 2023.

Montgomery’s medialization thyroplasty is a laryngeal frame surgery to treat patients with unilateral vocal cord paralysis. This procedure uses a silicone implant in six sizes each for male and female patients. The results of this technique are generally good; however, the results can be suboptimal in some female patients because of differences in female and male laryngeal anatomy. The angle formed by the thyroid cartilage is greater in women than in men, meaning that the posterior portion of this cartilage is further away from the midline in women. We describe our results using male prostheses to improve suboptimal results in some of our female patients. This study is a retrospective case series study. We included female patients who underwent thyroplasty reintervention from 2014 to 2021 using male Montgomery thyroplasty prostheses. Functional results were studied using maximum phonation time (MPT), the GRBAS scales, and two questionnaires: Voice-Handicap Index 30 and Eating Assessment Tool 10. The functional results for all of these women improved after secondary thyroplasty. In one case, the MPT parameter did not change but patient satisfaction improved. The use of male Montgomery implants in women can be a useful tool to achieve improvement in female thyroplasty failure.

This article focuses on two facets of young people's active citizenship: their experience of being impacted by socio-political events and their participation behaviour. The idea that underlies the study is that to better understand the relationship between young people's perception of socio-political reality and their multiple ways of exercising active citizenship, we need to conceptualize more broadly what constitutes "the political". Since today's youth engagement includes many different forms, research should focus on what young citizens themselves experience as impacting, going beyond traditional measures of political interest. We conducted a survey among university students (N = 969, 72.7% female) in Spain that included an open-ended question about events that had particularly impacted them in the past year and measures of their experience and willingness to engage in conventional and unconventional political and civic participation. The content analysis established four categories of impacting events: "national politics" and "international politics", which correlated with conventional political participation; events categorized as "social life" and "social justice" were associated with unconventional/civic participation. The results also suggest that most of the participants are, in fact, active citizens, which challenges the view of young people as "disaffected citizens". We conclude that the analysis of their specific socio-political sensitivities helps to understand the intensity and concrete orientation of their actions.

Serbia preserves a high number of local grape varieties, which have been cultivated across the country for centuries. Now, these ancient varieties are in the spotlight, and there is a global trend towards their recovery and characterization because they can revitalize regional, national and international grape and wine sectors. In addition, their genetic study can be useful to find new pedigree relationships to reveal how local varietal assortment evolved over time. Here, the genetic characterization of 138 grapevines from old Serbian vineyards revealed 59 different genetic profiles, 49 of which were identified as grapevine varieties whose origin in the country could be linked to some major Serbian historical periods. Most of the genetic profiles found in this work arranged in a complex pedigree network that integrates numerous grapevine varieties from diverse Balkan countries, agreeing with an intense exchange of plant material among Balkan regions for centuries. This analysis identified some varieties as important founders of Balkan genetic resources, like ‘Alba Imputotato’, ‘Braghina Rosie’, ‘Coarna Alba’, and ‘Vulpea’. After deepening into their genealogy, these major direct founders might have ultimately derived from ‘Visparola’, an ancient variety of likely Balkan origin with a major founding role in some European regions. Our results also indicated the genetic singularity of the grapevine resources from the Balkans when compared to those from other relevant winemaking regions, supporting the interest of their detailed study to evaluate their oenological potential and for the eventual identification of useful traits to counteract current viticulture challenges.

BackgroundSeveral studies suggest that women with Crohn disease (CD) have reduced fertility due to decreased ovarian reserve, among other causes. On the other hand, male CD patients could have difficulties conceiving. The present study aimed to test the effect of CD on both male and female fertility potential, Sertoli cell function and ovarian reserve, assessed by inhibin-B (IB) plus IB:FSH ratio (IFR) and antiMüllerian hormone (AMH), respectively. Sexual dysfunction (SD) was studied as secondary endpoint.MethodsWe performed a cross-sectional, case–control study. Serum IB levels plus IFR were measured in 58 men with CD and compared to 25 age-matched healthy controls (HC). Serum AMH levels were measured in 50 women with CD and in 30 HC matched by age. SD was assessed by means of the International Index of Erectile Function (IIFE-15) in males and the Index of Female Sexual Function (IFSF) in women.ResultsA total of 108 CD patients and 55 HC were included. IB serum levels were significantly lower in CD men than in HC (177 ± 58 vs. 234 ± 75 pg./mL, p = 0.001). IFR was also decreased in CD patients compared to HC (58.27 ± 59.5 vs. 91.35 ± 60.04, p = 0.014). Women with CD > 30 years had lower serum AMH levels compared to HC (1.15 ± 0.74 vs. 2.14 ± 1.68 ng/mL, p = 0.033). In addition, CD women >30 years presented a serum AMH 30 years is the single independent predictor of reduced ovarian reserve in women with CD. These results should be confirmed in further studies in order to properly counsel patients with CD and desire for offspring.

Abstract Background and Study Aims Our aim was to determine the impact of the SARS‐CoV‐2 pandemic on the diagnosis and prognosis of colorectal cancer (CRC). Patients and Methods This prospective cohort study included individuals diagnosed with CRC between March 13, 2019 and June 20, 2021 across 21 Spanish hospitals. Two time periods were compared: prepandemic (from March 13, 2019 to March 13, 2020) and pandemic (from March 14, 2020 to June 20, 2021, lockdown period and 1 year after lockdown). Results We observed a 46.9% decrease in the number of CRC diagnoses (95% confidence interval (CI): 45.1%–48.7%) during the lockdown and 29.7% decrease (95% CI: 28.1%–31.4%) in the year after the lockdown. The proportion of patients diagnosed at stage I significantly decreased during the pandemic (21.7% vs. 19.0%; p = 0.025). Centers that applied universal preprocedure SARS‐CoV‐2 PCR testing experienced a higher reduction in the number of colonoscopies performed during the pandemic post‐lockdown (34.0% reduction; 95% CI: 33.6%–34.4% vs. 13.7; 95% CI: 13.4%–13.9%) and in the number of CRCs diagnosed (34.1% reduction; 95% CI: 31.4%–36.8% vs. 26.7%; 95% CI: 24.6%–28.8%). Curative treatment was received by 87.5% of patients diagnosed with rectal cancer prepandemic and 80.7% of patients during the pandemic post‐lockdown period (p = 0.002). Conclusions The COVID‐19 pandemic has led to a decrease in the number of diagnosed CRC cases and in the proportion of stage I CRC. The reduction in the number of colonoscopies and CRC diagnoses was higher in centers that applied universal SARS‐CoV‐2 PCR screening before colonoscopy. In addition, the COVID‐19 pandemic has affected curative treatment of rectal cancers.

The epithelium is a dynamic barrier and the damage to this epithelial layer governs a variety of complex mechanisms involving not only epithelial cells but all resident tissue constituents, including immune and stroma cells. Traditionally, diseases characterized by a damaged epithelium have been considered “immunological diseases,” and research efforts aimed at preventing and treating these diseases have primarily focused on immuno-centric therapeutic strategies, that often fail to halt or reverse the natural progression of the disease. In this review, we intend to focus on specific mechanisms driven by the epithelium that ensure barrier function. We will bring asthma and Inflammatory Bowel Diseases into the spotlight, as we believe that these two diseases serve as pertinent examples of epithelium derived pathologies. Finally, we will argue how targeting the epithelium is emerging as a novel therapeutic strategy that holds promise for addressing these chronic diseases.

Abstract Triple‐negative breast cancer (TNBC) often develops resistance to single‐agent treatment, which can be circumvented using targeted combinatorial approaches. Here, we demonstrate that the simultaneous inhibition of LOXL2 and BRD4 synergistically limits TNBC proliferation in vitro and in vivo. Mechanistically, LOXL2 interacts in the nucleus with the short isoform of BRD4 (BRD4S), MED1, and the cell cycle transcriptional regulator B‐MyB. These interactions sustain the formation of BRD4 and MED1 nuclear transcriptional foci and control cell cycle progression at the gene expression level. The pharmacological co‐inhibition of LOXL2 and BRD4 reduces BRD4 nuclear foci, BRD4‐MED1 colocalization, and the transcription of cell cycle genes, thus suppressing TNBC cell proliferation. Targeting the interaction between BRD4S and LOXL2 could be a starting point for the development of new anticancer strategies for the treatment of TNBC.

Abstract While cellular metabolism impacts the DNA damage response, a systematic understanding of the metabolic requirements that are crucial for DNA damage repair has yet to be achieved. Here, we investigate the metabolic enzymes and processes that are essential for the resolution of DNA damage. By integrating functional genomics with chromatin proteomics and metabolomics, we provide a detailed description of the interplay between cellular metabolism and the DNA damage response. Further analysis identified that Peroxiredoxin 1, PRDX1, contributes to the DNA damage repair. During the DNA damage response, PRDX1 translocates to the nucleus where it reduces DNA damage‐induced nuclear reactive oxygen species. Moreover, PRDX1 loss lowers aspartate availability, which is required for the DNA damage‐induced upregulation of de novo nucleotide synthesis. In the absence of PRDX1, cells accumulate replication stress and DNA damage, leading to proliferation defects that are exacerbated in the presence of etoposide, thus revealing a role for PRDX1 as a DNA damage surveillance factor.

ObjectivePsychological factors, such as stress, anxiety, and depression, are frequently related to inflammatory bowel disease (IBD). However, few studies have examined these factors in patients newly diagnosed with IBD. The aim of the present study was to test the psychological burden in patients with a recent diagnosis of IBD and the factors related to this psychological burden.MethodsWe performed a prospective, multi-center, observational study in patients with a new diagnosis of IBD (≤6 months). The patients were recruited from four different Spanish hospitals. Clinical and demographic characteristics were collected. Patients were evaluated using the Hospital Anxiety and Depression Scale and quality of life questionnaire for patients with inflammatory bowel disease (IBDQ-32). The Scale of Stress Perceived by the Disease was used to assess stressful life events.ResultsWe included 156 patients newly diagnosed with IBD [69 women; 80 Crohn’s disease (CD) and 76 ulcerative colitis (UC)], with a mean age of 42.3 (SD 16.21) years. A total of 37.2% of patients had symptoms of anxiety and 17.3% had symptoms of depression. Quality of life was affected in 30.1% of patients. Factors related to anxiety in early IBD were being a woman and having CD. The only factor related to depression was the presence of comorbidity. Being a woman and having suffered previous stressful life events were factors related to impaired quality of life.ConclusionAnxiety, depression, and impaired quality of life are frequent in patients with a recent diagnosis of IBD. This psychological burden is greater in women.

Abstract A primary role of the liver is to regulate whole body glucose homeostasis. Glucokinase (GCK) is the main hexokinase (HK) expressed in hepatocytes and functions to phosphorylate the glucose that enters via GLUT transporters to become glucose-6-phosphate (G6P), which subsequently commits glucose to enter downstream anabolic and catabolic pathways. In the recent years, hexokinase domain-containing-1 (HKDC1), a novel 5th HK, has been characterized by our group and others. Its expression profile varies but has been identified to have low basal expression in normal liver but increases during states of stress including pregnancy, nonalcoholic fatty liver disease (NAFLD), and liver cancer. Here, we have developed a stable overexpression model of hepatic HKDC1 in mice to examine its effect on metabolic regulation. We found that HKDC1 overexpression, over time, causes impaired glucose homeostasis in male mice and shifts glucose metabolism towards anabolic pathways with an increase in nucleotide synthesis. Furthermore, we observed these mice to have larger liver sizes due to greater hepatocyte proliferative potential and cell size, which in part, is mediated via yes-associated protein (YAP) signaling.

Objective: To study the impact of age and the interval between disease diagnosis and death on the organotropism of SARS-CoV-2. Method: Patients underwent post-mortem biopsies from lungs, Waldeyer ring, heart, liver, kidneys and bone marrow between 2020‒2021. SARS-CoV-2 organotropism was mapped by using molecular RT-PCR analyses for SARS-CoV2 targeting the Envelope gene (E), the RNA Polymerase Gene (RdRp), and the Nucleocapsid gene (N). Statistical and linear regression analysis was performed to study the impact of age and illness duration in SARS-CoV-2 organotropism. Results: We performed 158 postmortem biopsies in 21 patients, with a mean age of 76 years old. The mean interval between the diagnosis of the infection to the death was 23 days. The RNA of the SARS-CoV-2 was detected in 100% of lung biopsies, 76%‒82% of Waldeyer’s ring biopsies, 55% of heart biopsies, 40% of kidney biopsies, 33% of liver and 25% of bone marrow biopsies. Patients who died before the day 9, presented extensive visceral dissemination of SARS-CoV-2 RNA. Most of the patients older than 80 years (90%) presented visceral dissemination of SARS-CoV-2 RNA, while among younger patients, only 3/11 patients presented visceral dissemination of the virus. The relationship between “age” and “illness duration” and multitropism of the virus was statistically significant (p

Introduction: Asthma is the most common chronic inflammatory disease of the airways. The airway epithelium is a key driver of the disease, and numerous studies have established genome-wide differences in mRNA expression between health and asthma. However, the underlying molecular mechanisms for such differences remain poorly understood. The human TTP family is comprised of ZFP36, ZFP36L1 and ZFP36L2, and has essential roles in immune regulation by determining the stability and translation of myriad mRNAs encoding for inflammatory mediators. We investigated the expression and possible role of the tristetraprolin (TTP) family of RNA binding proteins (RBPs), poorly understood in asthma.Methods: We analysed the levels of ZFP36, ZFP36L1 and ZFP36L2 mRNA in several publicly available asthma datasets, including single cell RNA-sequencing. We also interrogated the expression of known targets of these RBPs in asthma. We assessed the lung mRNA expression and cellular localization of Zfp36l1 and Zfp36l2 in precision cut lung slices in murine asthma models. Finally, we determined the expression in airway epithelium of ZFP36L1 and ZFP36L2 in human bronchial biopsies and performed rescue experiments in primary bronchial epithelium from patients with severe asthma.Results: We found ZFP36L1 and ZFP36L2 mRNA levels significantly downregulated in the airway epithelium of patients with very severe asthma in different cohorts (5 healthy vs. 8 severe asthma; 36 moderate asthma vs. 37 severe asthma on inhaled steroids vs. 26 severe asthma on oral corticoids). Integrating several datasets allowed us to infer that mRNAs potentially targeted by these RBPs are increased in severe asthma. Zfp36l1 was downregulated in the lung of a mouse model of asthma, and immunostaining of ex vivo lung slices with a dual antibody demonstrated that Zfp36l1/l2 nuclear localization was increased in the airway epithelium of an acute asthma mouse model, which was further enhanced in a chronic model. Immunostaining of human bronchial biopsies showed that airway epithelial cell staining of ZFP36L1 was decreased in severe asthma as compared with mild, while ZFP36L2 was upregulated. Restoring the levels of ZFP36L1 and ZFP36L2 in primary bronchial epithelial cells from patients with severe asthma decreased the mRNA expression of IL6, IL8 and CSF2.Discussion: We propose that the dysregulation of ZFP36L1/L2 levels as well as their subcellular mislocalization contributes to changes in mRNA expression and cytoplasmic fate in asthma.