IκBα controls dormancy in hematopoietic stem cells via retinoic acid during embryonic development.

Recent findings suggest that Hematopoietic Stem Cells (HSC) and progenitors arise simultaneously and independently of each other already in the embryonic aorta-gonad mesonephros region, but it is still unknown how their different features are established. Here, we uncover IκBα (Nfkbia, the inhibitor of NF-κB) as a critical regulator of HSC proliferation throughout development. IκBα balances retinoic acid signaling levels together with the epigenetic silencer, PRC2, specifically in HSCs. Loss of IκBα decreases proliferation of HSC and induces a dormancy related gene expression signature instead. Also, IκBα deficient HSCs respond with superior activation to in vitro culture and in serial transplantation. At the molecular level, chromatin regions harboring binding motifs for retinoic acid signaling are hypo-methylated for the PRC2 dependent H3K27me3 mark in IκBα deficient HSCs. Overall, we show that the proliferation index in the developing HSCs is regulated by a IκBα-PRC2 axis, which controls retinoic acid signaling.

Single-cell Multiomics Analysis of Myelodysplastic Syndromes and Clinical Response to Hypomethylating Therapy.

Alterations in epigenetic marks, such as DNA methylation, represent a hallmark of cancer that has been successfully exploited for therapy in myeloid malignancies. Hypomethylating agents (HMA), such as azacitidine, have become standard-of-care therapy to treat myelodysplastic syndromes (MDS), myeloid neoplasms that can evolve into acute myeloid leukemia. However, our capacity to identify who will respond to HMAs, and the duration of response, remains limited. To shed light on this question, we have leveraged the unprecedented analytic power of single-cell technologies to simultaneously map the genome and immunoproteome of MDS samples throughout clinical evolution. We were able to chart the architecture and evolution of molecular clones in precious paired bone marrow MDS samples at diagnosis and posttreatment to show that a combined imbalance of specific cell lineages with diverse mutational profiles is associated with the clinical response of patients with MDS to hypomethylating therapy. SIGNIFICANCE: MDS are myeloid clonal hemopathies with a low 5-year survival rate, and approximately half of the cases do not respond to standard HMA therapy. Our innovative single-cell multiomics approach offers valuable biological insights and potential biomarkers associated with the demethylating agent efficacy. It also identifies vulnerabilities that can be targeted using personalized combinations of small drugs and antibodies.

Targeting lymphoid-derived IL-17 signaling to delay skin aging.

Skin aging is characterized by structural and functional changes that contribute to age-associated frailty. This probably depends on synergy between alterations in the local niche and stem cell-intrinsic changes, underscored by proinflammatory microenvironments that drive pleotropic changes. The nature of these age-associated inflammatory cues, or how they affect tissue aging, is unknown. Based on single-cell RNA sequencing of the dermal compartment of mouse skin, we show a skew towards an IL-17-expressing phenotype of T helper cells, γδ T cells and innate lymphoid cells in aged skin. Importantly, in vivo blockade of IL-17 signaling during aging reduces the proinflammatory state of the skin, delaying the appearance of age-related traits. Mechanistically, aberrant IL-17 signals through NF-κB in epidermal cells to impair homeostatic functions while promoting an inflammatory state. Our results indicate that aged skin shows signs of chronic inflammation and that increased IL-17 signaling could be targeted to prevent age-associated skin ailments.

Analysis of the Structural Characteristics and Psychometric Properties of the Pelvic Floor Bother Questionnaire (PFBQ): A Systematic Review.

BACKGROUND: The Pelvic Floor Bother Questionnaire is a validated and reliable questionnaire that studies the presence and degree of pelvic floor discomfort, providing a global vision of pelvic floor dysfunction. This questionnaire assesses urinary stress incontinence, urinary urgency, urinary frequency, urge urinary incontinence, pelvic organ prolapses, dysuria, dyspareunia, defecatory dysfunction, fecal incontinence, and the disability it causes to the respondent. AIM: The aim of the present study was to analyze the structural characteristics and psychometric properties of the different versions of the pelvic floor bother questionnaire, as well as the methodological quality, the quality of evidence, and the criteria used for good measurement properties. METHODS: A systematic review was carried out in different databases, such as PubMed, SCOPUS, Web of Science, Dialnet, ScienceDirect, and CINAHL, on studies adapting and validating the pelvic floor bother questionnaire in other languages. The data were analyzed taking into account the guidelines of the preferred reporting item statement for systematic reviews and meta-analyses (PRISMA) and following the COSMIN guidelines, considering articles published up to 28 February 2022, and registered in the PROSPERO database. RESULTS: Initially, a total of 443 studies were found, from which a total of four studies were analyzed with regard to structural characteristics and psychometric properties, such as reliability, internal consistency, construct validity, and criterion validity. CONCLUSIONS: The different versions of the questionnaires show basic structural characteristics and psychometric properties for the evaluation of patients with pelvic floor dysfunctions. Most of the analyzed versions present criteria for good measurement properties qualified as sufficient, inadequate-adequate methodological quality, and low-moderate quality of evidence.

Targeting DNA damage response components induces enhanced STING-dependent type-I IFN response in ATM deficient cancer cells and drives dendritic cell activation.

The concept of exploiting tumor intrinsic deficiencies in DNA damage repair mechanisms by inhibiting compensatory DNA repair pathways is well established. For example, ATM-deficient cells show increased sensitivity to the ATR inhibitor ceralasertib. DNA damage response (DDR)-deficient cells are also more sensitive to DNA damaging agents like the DNA crosslinker pyrrolobenzodiazepine (PBD) SG-3199. However, additional antitumor benefits from targeting the DDR pathways, which could operate through the activation of the innate immune system are less well studied. DNA accumulation in the cytosol acts as an immunogenic danger signal, inducing the expression of type-I interferon (IFN) stimulated genes (ISGs) by the activation of the cGAS-STING pathway. Here, we demonstrate that ATM -/- FaDu tumor cells have higher basal expression of ISGs when compared to WT cells and respond to ceralasertib and PBD SG-3199 by inducing higher levels of ISGs in a cGAS-STING-dependent manner. We show that sensitive tumor cells treated with ceralasertib and PBD SG-3199 activate dendritic cells (DCs) via a type-I IFN-dependent mechanism. However, STING deficiency in tumor cells does not prevent DC activation, suggesting that transactivation of the STING pathway occurs within DCs. Furthermore, depletion of the cytosolic DNA exonuclease TREX1 in tumor cells increases DC activation in response to PBD SG-3199-treated tumor cells, indicating that an increase in tumor-derived cytosolic DNA may further enhance DC activation. In summary, in this study, we show that ceralasertib and PBD SG-3199 treatment not only intrinsically target tumor cells but also extrinsically increase tumor cell immunogenicity by inducing DC activation, which is enhanced in ATM-deficient cells.

Single cell cancer epigenetics.

Bulk sequencing methodologies have allowed us to make great progress in cancer research. Unfortunately, these techniques lack the resolution to fully unravel the epigenetic mechanisms that govern tumor heterogeneity. Consequently, many novel single cell-sequencing methodologies have been developed over the past decade, allowing us to explore the epigenetic components that regulate different aspects of cancer heterogeneity, namely: clonal heterogeneity, tumor microenvironment (TME), spatial organization, intratumoral differentiation programs, metastasis, and resistance mechanisms. In this review, we explore the different sequencing techniques that enable researchers to study different aspects of epigenetics (DNA methylation, chromatin accessibility, histone modifications, DNA-protein interactions, and chromatin 3D architecture) at the single cell level, their potential applications in cancer, and their current technical limitations.

Seroprevalencia de infección frente a SARS-CoV-2 en trabajadores de la salud en un hospital monográfico pediátrico en Madrid (España) / Seroprevalence of antibodies against SARS-CoV-2 among health care workers in a pediatric monographic hospital in Madrid (Spain)

Introducción: A 30 de abril de 2020, se habían notificado 203.715 infecciones SARS-CoV-2 en España, 54.486 en Madrid, y el 21,4% eran trabajadores de la salud. El objetivo del estudio es determinar la prevalencia serológica de infección SARS-CoV-2 en trabajadores de un hospital monográfico pediátrico. Método: Del 13 al 30 de abril, 1.523 trabajadores fueron convocados a realizar un test serológico (All Test®) frente a SARS-CoV-2 y respondieron un cuestionario con información demográfica, clínico-epidemiológica y de exposición a COVID-19. Resultados: Mil doscientos noventa y dos (84,8%) fueron estudiados. La prevalencia serológica (IgM y/o IgG+) a SARS-CoV-2 fue del 17,2% (222/1.292) y del 15,5% (201/1.292) considerando IgG positiva. La edad media fue 44±13 años, el 73% eran mujeres. El 33,8% (75/222) fueron asintomáticos. Tenían rRT-PCR positiva previa 81. El 14% (32/222) contacto familiar. Conclusión: La prevalencia serológica SARS-CoV-2 en los trabajadores de un hospital pediátrico fue mayor que en la población general. Muchos pasaron una infección inadvertida.(AU)

Introduction: As of 30 April 2020, 203.715 SARS-CoV-2 infections had been reported in Spain, 54.486 in Madrid, 21.4% were health care workers. Our objective is to determine seroprevalence of COVID-19 among workers in a monographic pediatric hospital. Methods: Between April13th and 30th, 1.523 health workers were recruited to be tested for SARS-CoV-2 serology screening (All Test®) and they answered a questionnaire with demographic, epidemiological and clinical information and previous exposure to COVID-19. Findings: One thousand two hundred ninety two (84.8%) were tested. Positive serology (IgM and/or IgG) to SARS-CoV-2 was found in 17.2% (222/1.292), in 15.5% (201/1.292) if only IgG was considered. Median age was 44±13 years, 73% were female. The 33.8% (75/222) were asymptomatic. Eighty one had a previous positive rRT-PCR. The 14% (32/222) referred a family contact. Conclusion: Serology prevalence for SARS-CoV-2 in workers of a pediatric hospital was higher than in general population. Many of them had an unnoticed infection.(AU)

Seroprevalence of antibodies against SARS-CoV-2 among health care workers in a pediatric monographic hospital in Madrid (Spain).

INTRODUCTION: As of 30 April 2020, 203.715 SARS-CoV-2 infections had been reported in Spain, 54.486 in Madrid, 21.4% were health care workers. Our objective is to determine seroprevalence of COVID-19 among workers in a monographic pediatric hospital. METHODS: Between April13th and 30th, 1.523 health workers were recruited to be tested for SARS-CoV-2 serology screening (All Test®) and they answered a questionnaire with demographic, epidemiological and clinical information and previous exposure to COVID-19. FINDINGS: One thousand two hundred ninety two (84.8%) were tested. Positive serology (IgM and/or IgG) to SARS-CoV-2 was found in 17.2% (222/1.292), in 15.5% (201/1.292) if only IgG was considered. Median age was 44±13 years, 73% were female. The 33.8% (75/222) were asymptomatic. Eighty one had a previous positive rRT-PCR. The 14% (32/222) referred a family contact. CONCLUSION: Serology prevalence for SARS-CoV-2 in workers of a pediatric hospital was higher than in general population. Many of them had an unnoticed infection.

Using retrospective life tables to assess the effect of extreme climatic conditions on ungulate demography.

In Mediterranean areas, severe drought events are expected to intensify in forthcoming years as a consequence of climate change. These events may increase physiological and reproductive stress of wild populations producing demographic changes and distribution shifts. We used retrospective life tables to understand demographic changes on a wild population after severe drought events. We studied the impact of two extreme events (2003 and 2005) on the population dynamics of our model species, the red deer (Cervus elaphus). During both years, population density was high (40 and 36 ind/100 ha, respectively). Thus, we reconstructed retrospectively the age structure of the female part of the population for the period 2000-2010 by using data of known-age individuals culled during the period 2000-2019 (n = 4176). Also, based on previous study results, we aimed to validate this methodology. Both extremely dry years, 2003 and 2005, produced marked and lasting cohort effects on population demography. Age pyramid the following years (2004 and 2006) revealed that the extreme drought caused the female fawn cohort to be similar or even smaller than the yearling cohort. Furthermore, these cohort effects were still perceptible 3 years after these severe events. Results agree with previous findings that showed a negative effect of severe drought events on female pregnancy rates and conception dates. Although simple, this study provides an empirical quantification of the demographic effects of severe drought events for a wild population which might be useful to understand future demographic changes under the context of climate change.

Early life investment in antlers and body growth reflects adult performance in roe deer population under supplementary feeding conditions.

Recent research has challenged the idea that cervid antlers are such costly traits, supporting the assertion early-life antler investment is an honest signal of adult phenotypic quality. However, inferences were made based on antler measurements while growing (velvet) and thus, studies on fully-formed clean antlers are needed to avoid possible bias caused by the inter-individual variation in antler growth phenology. We studied a semi-captive population of European roe deer inhabiting a sub-Mediterranean area (Valsemana research station) and living under optimal conditions (ad libitum food supply and veterinary care). Based on repeated measurements taken from 146 individuals, we assessed whether allocation to secondary sexual traits during early life affected body mass or antler development during adulthood. Furthermore, we evaluated which body measurements better predicted future male quality. Additionally, using 488 individuals, we described age-class-specific variation in male body measurements and assessed the decline in antler size between adult and senescent stages (i.e. magnitude of senescence). Results agree with the assertion that there is no negative effect of a high investment in antler during early life on adult antler size or body mass, but we suggest the use of clean antlers as a more robust and reliable measure. The variables that better predicted body mass during adulthood were yearling body mass and body size at 6 months. Antler score between 10 and 18 months resulted in the best indicator of adult antler size. Finally, we support the idea that the magnitude of senescence in antler size is smaller in environments with higher resource availability during winter.

Effect of Myofascial Therapy on Pain and Functionality of the Upper Extremities in Breast Cancer Survivors: A Systematic Review and Meta-Analysis.

(1) Objective: The purpose was to analyze the effectiveness of myofascial therapy on musculoskeletal pain and functionality of the upper extremities in female breast cancer survivors, and to evaluate the changes in range of motion, quality of life, and mood state of these patients. (2) Methods: Systematic searches were performed on the MEDLINE/PubMed, Web of Science, Scopus, and Physiotherapy Evidence Databases for articles published until October 2020, in order to identify randomized controlled trials which analyzed the effectiveness of myofascial therapy as compared to a control group, passive treatment, placebo, or another intervention, and allowed co-interventions on female breast cancer survivors. Two reviewers examined the sources individually, calculated the risk of bias and extracted the data (PROSPERO number CRD42020215823). (3) Results: A total of eight RCTs were included. The results suggested that myofascial therapy does not have a greater statistically significant immediate effect on pain intensity (SMD: -0.15; 95% CI -0.48, 0.19), functionality (SMD: -0.17; 95% CI -0.43, 0.09) and range of motion in flexion (SMD: 0.30; 95% CI -0.13, 0.74) than an inactive, passive treatment or another intervention. However, a statistically significant result was observed for the abduction shoulder in favor of the experimental group (SMD: 0.46; 95% CI 0.05, 0.87; p = 0.03). (4) Conclusion: In general, although we found greater overall effects in support of the intervention with myofascial therapy than other control groups/types of interventions, the subgroup analysis revealed inconsistent results supporting myofascial therapy applied to breast cancer survivors.

Seroprevalence of antibodies against SARS-CoV-2 among health care workers in a pediatric monographic hospital in Madrid (Spain). / Seroprevalencia de infección frente a SARS-CoV-2 en trabajadores de la salud en un hospital monográfico pediátrico en Madrid (España).

INTRODUCTION: As of 30 April 2020, 203.715 SARS-CoV-2 infections had been reported in Spain, 54.486 in Madrid, 21.4% were health care workers. Our objective is to determine seroprevalence of COVID-19 among workers in a monographic pediatric hospital. METHODS: Between April13th and 30th, 1.523 health workers were recruited to be tested for SARS-CoV-2 serology screening (All Test®) and they answered a questionnaire with demographic, epidemiological and clinical information and previous exposure to COVID-19. FINDINGS: One thousand two hundred ninety two (84.8%) were tested. Positive serology (IgM and/or IgG) to SARS-CoV-2 was found in 17.2% (222/1.292), in 15.5% (201/1.292) if only IgG was considered. Median age was 44±13 years, 73% were female. The 33.8% (75/222) were asymptomatic. Eighty one had a previous positive rRT-PCR. The 14% (32/222) referred a family contact. CONCLUSION: Serology prevalence for SARS-CoV-2 in workers of a pediatric hospital was higher than in general population. Many of them had an unnoticed infection.

Large-scale variation in birth timing and synchrony of a large herbivore along the latitudinal and altitudinal gradients.

Hopkins' Bioclimatic Law predicts geographical patterns in phenological timing by establishing a correspondence between latitudinal and altitudinal gradients. First proposed for key phenological events of plants, such as leaf sprouting or flowering dates, this law has rarely been used to assess the geographical equivalence of key life-history traits of mammals. We hypothesize that (H1) parturition dates of European roe deer Capreolus capreolus are delayed and more synchronized at higher latitudes and altitudes, (H2) parturition timing varies along latitudinal and altitudinal gradients in a way that matches the Hopkins' Bioclimatic Law and (H3) females adjust parturition timing to match the period of high energy demand with peak resource availability. We used parturition dates of 7,444 European roe deer from Switzerland to assess altitudinal variation in birth timing and synchrony from 288 to 2,366 m a.s.l. We then performed a literature survey to compare altitudinal results with those from different populations along the species' latitudinal range of distribution. Finally, we performed spatial analysis combining our highly resolved altitudinal data on parturition dates with plant phenology data. As expected, parturition dates were delayed with increasing latitude and altitude. This delay matched the Bioclimatic Law, as the effect of 1º increase in latitude was similar to 120 m increase in altitude. However, while parturitions were more synchronized with increasing altitude, we did not detect any trend along the latitudinal gradient. Finally, plant phenology explained altitudinal variation in parturition timing better than a linear effect of altitude. Our findings clearly demonstrate the ability of a large herbivore to match parturition timing with phenological conditions across the altitudinal gradient, even at the smallest spatial scales.

Remote mapping of foodscapes using sUAS and a low cost BG-NIR sensor.

The assessment of landscape condition for large herbivores, also known as foodscapes, is fast gaining interest in conservation and landscape management programs worldwide. Although traditional approaches are now being replaced by satellite imagery, several technical issues still need to be addressed before full standardization of remote sensing methods for these purposes. We present a low-cost method, based on the use of a modified blue/green/near-infrared (BG-NIR) camera housed on a small-Unmanned Aircraft System (sUAS), to create foodscapes for a generalist Mediterranean ungulate: the Iberian Ibex (Capra pyrenaica) in Northeast Spain. Faecal cuticle micro-histological analyses were used to assess the dietary preferences of ibexes and then individuals of the most common plant species (n = 19) were georeferenced to use as test samples. Because of the seasonal pattern in vegetation activity, based on the NDVI (Smooth term Month = 21.5, p-value < .01, R2 = 43%, from a GAM), images were recorded in winter and spring to represent contrasting vegetation phenology using two flight heights above ground level (30 and 60 m). Additionally, the range of image pixel sizes was 3.5-30 cm with the smallest pixel size representing the highest resolution. Boosted Trees were used to classify plant taxa based on spectral reflectance and create a foodscape of the study area. The number of target species, the sampling season, the height of flight and the image resolution were analysed to determine the accuracy of mapping the foodscape. The highest classification error (70.66%) was present when classifying all plant species using a 30 cm pixel size from acquisitions at 30 m height. The lowest error (18.7%), however, was present when predicting plants preferred by ibexes, at 3.5 cm pixel size acquired at 60 m height. This methodology can help to successfully monitor food availability and seasonality and to identify individual species.

A cell-engineered system to assess tumor cell sensitivity to CD8+ T cell-mediated cytotoxicity.

In vitro assays that evaluate CD8+ T cell-mediated cytotoxicity are important to aid in the development of novel therapeutic approaches to enhance anti-tumor immune responses. Here, we describe a novel cytotoxicity co-culture assay that circumvents the problem of highly variable allogeneic responses and obviates the constraints of HLA-restriction between effector and target cells. We show that this assay can be easily applied to a panel of tumor cell lines to provide additional insights into intrinsic drivers of sensitivity/resistance to T cell-mediated killing, and to evaluate the impact of targeted therapies on both tumor and T cell compartments.

Falso positivo a morfina en orina por reactividad cruzada con alimemazina / False positive result to morphine in urine due to cross-reactivity to alimemazine

La alimemazina (Variargil(R)) es un antihistamínico anti-H1 reversible inespecífico que atraviesa la barrera hematoencefálica. Actúa como anticolinérgico. Su forma de presentación es en gotas en suspensión oral. Su uso clínico está extendido a rinitis alérgica estacional, angioedema y urticaria leve a partir de los 2 años de edad, conjuntivitis alérgica e insomnio de conciliación en la infancia. Presenta una farmacocinética muy variable, somnolencia excesiva conocida como «heavy hangover» y efecto de rebote tras su retirada. No está autorizado su uso en menores de 2 años. Presentamos el caso de un paciente de 19 meses que acude a Urgencias por referir somnolencia y falta de respuesta a estímulos tras la administración de alimemazina por dificultad para conciliar el sueño

Alimemazine (Variargil(R)) is a non-specific reversible anti-H1 antihistamine that crosses the blood-brain barrier. It acts as anticholinergic drug. It is marketed in drop form (oral suspension). It is used to relieve seasonal allergic rhinitis, angioedema, mild urticaria, allergic conjunctivitis, and for difficulty in falling asleep in children. It has a very variable pharmacokinetic, excessive "heavy hangover drowsiness" and rebound effect after withdrawal. It is not authorised for children under 2 years of age. The case is presented of a patient seen in the emergency room due to drowsiness and lack of response to stimuli after administration of alimemazine due to difficulties in falling asleep

Climate, female traits and population features as drivers of breeding timing in Mediterranean red deer populations.

Understanding the factors that lead to variation in the timing of breeding in widespread species such as red deer (Cervus elaphus) is crucial to predict possible responses of wild populations to different climate scenarios. Here, we sought to further understand the causes of inter-annual variation in the reproduction timing of female deer in Mediterranean environments. An integrative approach was used to identify the relative importance of individual, population and climate traits in the date of conception of free-ranging deer, based on a dataset of 829 hinds culled during 12 years. We found that a population trait, density, was the most important factor explaining the variation in conception dates, with greater densities causing later conception dates. Body mass was the second in importance, with heavier females conceiving earlier than lighter ones. Almost equally important was the spring real bioclimatic index, a measure of plant productivity, causing later conception dates in the least productive springs (drier and hotter). Another climatic component, the end of summer drought, showed that the sooner the autumn arrives (greater rainfalls and cooler temperatures) the earlier the conception dates. Interestingly, age class was found to be a minor factor in determining conception date. Only older females (≥10 years old) conceived significantly later, suggesting reproductive senescence. This study highlights not only the importance of population and individual traits but also the influence of climatic parameters on the deer reproductive cycle in Mediterranean environments, giving valuable insight into how reproductive phenology may respond to seasonality and global climate changes.

Suppression of IRAK1 or IRAK4 Catalytic Activity, but Not Type 1 IFN Signaling, Prevents Lupus Nephritis in Mice Expressing a Ubiquitin Binding-Defective Mutant of ABIN1.

Polymorphisms in the TNIP1 gene encoding A20-binding inhibitor of NF-κB1 (ABIN1) predispose to lupus and other autoimmune diseases in at least eight human populations. We found previously that knock-in mice expressing a ubiquitin-binding-defective mutant of ABIN1 (ABIN1[D485N]) develop autoimmunity as they age and succumb to a disease resembling lupus nephritis in humans. In this article, we report that Flt3-derived dendritic cells from these mice overproduced type 1 IFNs upon stimulation with ligands that activate TLR7 or TLR9. However, crossing ABIN1[D485N] mice to IFNAR1-knockout mice that do not express the α-subunit of the type 1 IFNR did not prevent splenomegaly, the appearance of high serum levels of autoantibodies and other Igs, or liver inflammation and only reduced kidney inflammation modestly. In contrast, crossing ABIN1[D485N] mice to knock-in mice expressing catalytically inactive mutants of IRAK1 or IRAK4 prevented splenomegaly, autoimmunity, and liver and kidney inflammation. Our results support the notion that IRAK1 and/or IRAK4 are attractive targets for the development of drugs to prevent, and perhaps treat, lupus nephritis and other autoinflammatory diseases caused by the decreased ability of ABIN1 or other proteins to restrict the strength of MyD88 signaling.