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Abstract Obesity is a major cause of metabolic dysfunction-associated steatohepatitis (MASH) and is characterized by inflammation and insulin resistance. Interferon-γ (IFNγ) is a pro-inflammatory cytokine elevated in obesity and modulating macrophage functions. Here, we show that male mice with loss of IFNγ signaling in myeloid cells (Lyz-IFNγR2−/−) are protected from diet-induced insulin resistance despite fatty liver. Obesity-mediated liver inflammation is also attenuated with reduced interleukin (IL)−12, a cytokine primarily released by macrophages, and IL-12 treatment in vivo causes insulin resistance by impairing hepatic insulin signaling. Following MASH diets, Lyz-IFNγR2−/− mice are rescued from developing liver fibrosis, which is associated with reduced fibroblast growth factor (FGF) 21 levels. These results indicate critical roles for IFNγ signaling in macrophages and their release of IL-12 in modulating obesity-mediated insulin resistance and fatty liver progression to MASH. In this work, we identify the IFNγ-IL12 axis in regulating intercellular crosstalk in the liver and as potential therapeutic targets to treat MASH.

There is a wide variety of drought indices, yet a consensus on suitable indices and temporal scales for monitoring streamflow drought remains elusive across diverse hydrological settings. Considering the growing interest in spatially distributed indices for ungauged areas, this study addresses the following questions: (i) What temporal scales of precipitation-based indices are most suitable to assess streamflow drought in catchments with different hydrological regimes? (ii) Do soil moisture indices outperform meteorological indices as proxies for streamflow drought? (iii) Are snow indices more effective than meteorological indices for assessing streamflow drought in snow-influenced catchments? To answer these questions, we examined 100 near-natural catchments in Chile with four hydrological regimes, using the standardised precipitation index (SPI), standardised precipitation evapotranspiration index (SPEI), empirical standardised soil moisture index (ESSMI), and standardised snow water equivalent index (SWEI), aggregated across various temporal scales. Cross-correlation and event coincidence analysis were applied between these indices and the standardised streamflow index at a temporal scale of 1 month (SSI-1), as representative of streamflow drought events. Our results underscore that there is not a single drought index and temporal scale best suited to characterise all streamflow droughts in Chile, and their suitability largely depends on catchment memory. Specifically, in snowmelt-driven catchments characterised by a slow streamflow response to precipitation, the SPI at accumulation periods of 12–24 months serves as the best proxy for characterising streamflow droughts, with median correlation and coincidence rates of approximately 0.70–0.75 and 0.58–0.75, respectively. In contrast, the SPI at a 3-month accumulation period is the best proxy over faster-response rainfall-driven catchments, with median coincidence rates of around 0.55. Despite soil moisture and snowpack being key variables that modulate the propagation of meteorological deficits into hydrological ones, meteorological indices are better proxies for streamflow drought. Finally, to exclude the influence of non-drought periods, we recommend using the event coincidence analysis, a method that helps assessing the suitability of meteorological, soil moisture, and/or snow drought indices as proxies for streamflow drought events.

Abstract The electrochemical reduction of carbon dioxide to formic acid is a promising pathway to improve CO2 utilization and has potential applications as a hydrogen storage medium. In this work, a zero-gap membrane electrode assembly architecture is developed for the direct electrochemical synthesis of formic acid from carbon dioxide. The key technological advancement is a perforated cation exchange membrane, which, when utilized in a forward bias bipolar membrane configuration, allows formic acid generated at the membrane interface to exit through the anode flow field at concentrations up to 0.25 M. Having no additional interlayer components between the anode and cathode this concept is positioned to leverage currently available materials and stack designs ubiquitous in fuel cell and H2 electrolysis, enabling a more rapid transition to scale and commercialization. The perforated cation exchange membrane configuration can achieve >75% Faradaic efficiency to formic acid at

As climate change brings about hotter and often drier summers, an improved understanding of how irrigation requirements vary according to climatic conditions is of increasing importance. Within Germany, temperate conditions have historically enabled most agriculture to be supplied solely by green water, but recent crop yield reductions and crop failures have demonstrated its increased vulnerability to climatic conditions. The raster-based mGROWA hydrological water balance model was implemented over all agricultural areas in Germany for the period 1961–2020 at a high spatial (200 m) and temporal (daily) resolution. Grid-cells were each assigned one of 10 major crop classes, which account for 86.7 % of all agricultural areas in Germany, and effectively all irrigated areas. Using crop-specific irrigation rules that reflect actual practices, irrigation requirements were simulated for all crop areas. To investigate the relationship between climatic water balance over the crop growing season and irrigation requirements, the simulated annual irrigation requirements were compared with the standardised precipitation-evapotranspiration index (SPEI-6), calculated at the end of September. Through this comparison, irrigation requirements could be characterised for near-normal and dry conditions, and results were aggregated to the district level. Additionally, using district-level data on the areas with irrigation infrastructure, the actual water used for irrigation was estimated. The results highlight marked increases in irrigation requirements in dry conditions compared to near-normal conditions (median increase of 72 %), which are more pronounced over crops in silty soils than in sandy soils. The results also demonstrate how the increased irrigation requirements in dry years are in many cases higher than what is suggested by guidelines for irrigation management in Germany. This study provides important information for actors related to the agricultural sector and water management and is based on a robust and transferable framework to quantify how irrigation requirements vary according to climatic variability and local soil conditions.

Over the past decades, novel parameter regionalisation techniques have been developed to predict streamflow in data-scarce regions. In this paper, we examined how the choice of gridded daily precipitation (P) products affects the relative performance of three well-known parameter regionalisation techniques (spatial proximity, feature similarity, and parameter regression) over 100 near-natural catchments with diverse hydrological regimes across Chile. We set up and calibrated a conceptual semi-distributed HBV-like hydrological model (TUWmodel) for each catchment, using four P products (CR2MET, RF-MEP, ERA5, and MSWEPv2.8). We assessed the ability of these regionalisation techniques to transfer the parameters of a rainfall-runoff model, implementing a leave-one-out cross-validation procedure for each P product. Despite differences in the spatio-temporal distribution of P, all products provided good performance during calibration (median Kling–Gupta efficiencies (KGE′s) > 0.77), two independent verification periods (median KGE′s >0.70 and 0.61, for near-normal and dry conditions, respectively), and regionalisation (median KGE′s for the best method ranging from 0.56 to 0.63). We show how model calibration is able to compensate, to some extent, differences between P forcings by adjusting model parameters and thus the water balance components. Overall, feature similarity provided the best results, followed by spatial proximity, while parameter regression resulted in the worst performance, reinforcing the importance of transferring complete model parameter sets to ungauged catchments. Our results suggest that (i) merging P products and ground-based measurements does not necessarily translate into an improved hydrologic model performance; (ii) the spatial resolution of P products does not substantially affect the regionalisation performance; (iii) a P product that provides the best individual model performance during calibration and verification does not necessarily yield the best performance in terms of parameter regionalisation; and (iv) the model parameters and the performance of regionalisation methods are affected by the hydrological regime, with the best results for spatial proximity and feature similarity obtained for rain-dominated catchments with a minor snowmelt component.

Study region: Nile Basin, Africa. Study focus: The accurate representation of precipitation (P) and actual evapotranspiration (ETa) patterns is crucial for water resources management, yet there remains a high spatial and temporal variability among gridded products, particularly over data-scarce regions. We evaluated the performance of eleven state-of-the-art P products and seven ETa products over the Nile Basin using a four-step procedure: (i) P products were evaluated at the monthly scale through a point-to-pixel approach; (ii) streamflow was modelled using the Random Forest machine learning technique, and simulated for well-performing catchments for 2009–2018 (to correspond with ETa product availability); (iii) ETa products were evaluated at the multiannual scale using the water balance method; and (iv) the ability of the best-performing P and ETa products to represent monthly variations in terrestrial water storage (ΔTWS) was assessed through a comparison with GRACE Level-3 data. New hydrological insights for the region: CHIRPSv2 was the best-performing P product (median monthly KGE’ of 0.80) and PMLv2 and WaPORv2.1 the best-performing ETa products over the majority of the evaluated catchments. The application of the water balance using these best-performing products captures the seasonality of ΔTWS well over the White Nile Basin, but overestimates seasonality over the Blue Nile Basin. Our study demonstrates how gridded P and ETa products can be evaluated over extremely data-scarce conditions using an easily transferable methodology.

Phosphatidylcholine transfer protein (PC-TP) is a cytosolic phospholipid binding protein and a member of the steroidogenic acute regulatory-related transfer domain superfamily. Its tissue distribution includes liver and macrophages. PC-TP regulates hepatic lipid metabolism, and its absence in cholesterol-loaded macrophages is associated with reduced ATP binding cassette transporter A1-mediated lipid efflux and increased susceptibility to apoptosis induced by unesterified cholesterol. To explore a role for PC-TP in atherosclerosis, we prepared PC-TP-deficient/apolipoprotein E-deficient (Pctp−/−/Apoe−/−) mice and littermate Apoe−/− controls. At 16 weeks, atherosclerosis was increased in chow-fed male, but not female, Pctp−/−/Apoe−/− mice. This effect was associated with increases in plasma lipid concentrations. By contrast, no differences in atherosclerosis were observed between male or female Pctp−/−/Apoe−/− mice and Apoe−/− controls fed a Western-type diet for 16 weeks. At 24 weeks, atherosclerosis in chow-fed male Pctp−/−/Apoe−/− mice tended to be reduced in proportion to plasma cholesterol. The attenuation of atherosclerosis in female Pctp−/−/Apoe−/− mice fed chow or the Western-type diet for 24 weeks was not attributable to changes in plasma cholesterol or triglyceride concentrations. These findings suggest that PC-TP modulates the development of atherosclerosis, in part by regulating plasma lipid concentrations.

Severe acute respiratory syndrome coronavirus (SARS-CoV) encodes a papain-like protease (PLpro) with both deubiquitinating (DUB) and deISGylating activities that are proposed to counteract the post-translational modification of signaling molecules that activate the innate immune response. Here we examine the structural basis for PLpro's ubiquitin chain and interferon stimulated gene 15 (ISG15) specificity. We present the X-ray crystal structure of PLpro in complex with ubiquitin-aldehyde and model the interaction of PLpro with other ubiquitin-chain and ISG15 substrates. We show that PLpro greatly prefers K48- to K63-linked ubiquitin chains, and ISG15-based substrates to those that are mono-ubiquitinated. We propose that PLpro's higher affinity for K48-linked ubiquitin chains and ISG15 stems from a bivalent mechanism of binding, where two ubiquitin-like domains prefer to bind in the palm domain of PLpro with the most distal ubiquitin domain interacting with a "ridge" region of the thumb domain. Mutagenesis of residues within this ridge region revealed that these mutants retain viral protease activity and the ability to catalyze hydrolysis of mono-ubiquitin. However, a select number of these mutants have a significantly reduced ability to hydrolyze the substrate ISG15-AMC, or be inhibited by K48-linked diubuiquitin. For these latter residues, we found that PLpro antagonism of the nuclear factor kappa-light-chain-enhancer of activated B-cells (NFκB) signaling pathway is abrogated. This identification of key and unique sites in PLpro required for recognition and processing of diubiquitin and ISG15 versus mono-ubiquitin and protease activity provides new insight into ubiquitin-chain and ISG15 recognition and highlights a role for PLpro DUB and deISGylase activity in antagonism of the innate immune response.

Resumen: Introducción: Los cuerpos amiláceos (CoA) se presentan en aproximadamente el 60% de los hipocampos atróficos resecados de pacientes con epilepsia del lóbulo temporal farmacorresistente (ELTFR). Su presencia en la neocorteza temporal lateral ha sido observada con menor frecuencia. Objetivo: El objetivo es evaluar la presencia, la distribución y la densidad de CoA en el lóbulo temporal lateral de pacientes con ELTFR y displasia cortical focal (DCF) y la relación de su densidad con variables demográficas y clínicas. Métodos: Analizamos histológicamente el tejido resecado de 35 pacientes con ELTFR. La densidad de los CoA fue evaluada con una escala semicuantitativa según los criterios de Cherian et al. Resultados: La presencia de CoA en la neocorteza de 9 pacientes estuvo asociada a esclerosis hipocampal (DCF tipo iiia, 7 casos), tumor neuroepitelial disembrioplásico (DCF tipo iiib, un caso) y angioma cavernoso (DCF tipo iiic, un caso). Todos los pacientes tuvieron afectación de la superficie meníngea (SM) y en 8 casos se localizaron en el parénquima cerebral (sustancia blanca) y alrededor de los vasos sanguíneos. La densidad de los CoA en SM tuvo una correlación negativa con la edad de inicio de las crisis (r = −0,828, p

Introduction: Corpora amylacea (CoA) are present in about 60% of atrophic hippocampi resected from patients with drug resistant temporal lobe epilepsy (DRTLE). They have also been described in the lateral temporal neocortex, although less frequently. Objective: The objective is to measure the presence, distribution and density of CoA in the lateral temporal lobes of patients with DRTLE and focal cortical dysplasia (FCD), also examining how CoA density may be linked to demographic and clinical traits. Methods: Resected tissue from 35 patients was analysed. CoA density was assessed with a semi-quantitative scale according to the criteria established by Cherian et al. Results: Presence of CoA in the neocortex of nine patients was associated with hippocampal sclerosis (FCD type IIIa, seven cases), dysembryoplastic neuroepithelial tumour (FCD type IIIb, one case), and cavernous angioma (FCD type IIIC, 1 case). The meningeal surface (MS) was involved in all cases, and eight cases displayed CoA in the cerebral parenchyma (white matter) and around blood vessels. CoA density on the MS showed a negative correlation with age at seizure onset (r = −0.828, P

The objectives of this study were to evaluate the effects of daily top-dressing (individually feeding on the top of the total mixed ration) with rumen-protected methionine (RPM) from 30 ± 3 until 126 ± 3 Days in milk on productive and reproductive performance in lactating dairy cows. A total of 309 lactating dairy Holstein cows (138 primiparous and 171 multiparous) were randomly assigned to treatment diets containing either RPM (21.2 g of RPM + 38.8 g of dried distillers grain; 2.34% Methionine [Met] of metabolizable protein [MP]) or Control (CON; 60 g of dried distillers grain; 1.87% Met of MP). Plasma amino acids were evaluated at the time of artificial insemination (AI) and near pregnancy diagnosis. Milk production and milk composition were evaluated monthly. Pregnancy was diagnosed on Day 28 (by Pregnancy-specific protein B [PSPB]), 32, 47, and 61 (by ultrasound) and sizes of embryonic and amniotic vesicle were determined by ultrasound on Day 33 after AI. Feeding RPM increased plasma Met at 6, 9, 12, and 18 hours after top-dressing with a peak at 12 hours (52.4 vs 26.0 μM; P < 0.001) and returned to basal by 24 hours. Cows fed RPM had a small increase in milk protein percentage (3.08 vs 3.00%; P = 0.04) with no differences on milk yield and milk protein yield. Additionally, in multiparous cows, RPM feeding increased milk protein (3.03 vs 2.95%; P = 0.05) and fat (3.45 vs 3.14%; P = 0.01) percentages, although no effects were observed in primiparous cows. In multiparous cows fed RPM, pregnancy loss was lower between Days 28 to 61 (19.6 [10/51] vs. 6.1% [3/49]; P = 0.03) or between Days 32 to 61 (8.9 [4/45] vs. 0 [0/0] %; P = 0.03), although, there was no effect of treatment on pregnancy loss in primiparous cows. Consistent with data on pregnancy loss, RPM feeding increased embryonic abdominal diameter (P = 0.01) and volume (P = 0.009) and amniotic vesicle volume (P = 0.04) on Day 33 of pregnancy in multiparous cows but had no effect on embryonic size in primiparous cows. Thus, the increase in plasma Met concentrations after feeding RPM was sufficient to produce a small increase in milk protein percentage and to improve embryonic size and pregnancy maintenance in multiparous cows. Further studies are needed to confirm these responses and understand the biological mechanisms that underlie these responses as well as the timing and concentrations of circulating Met that are needed to produce this effect.

Introducción: A pesar del uso extendido del formulario de Zung para ansiedad no se encontraron artículos sobre la invarianza de la prueba. Objetivo: Establecer la invarianza por género del formulario para ansiedad autodiligenciado de Zung desde la Teoría Clásica. Métodos: Muestra no aleaotoria de 336 mujeres y 205 hombres estudiantes de varias universidades colombianas que diligenciaron el formulario. El análisis se realizo con análisis confimatorio de factores. Se investigó la invarianza dimensional, de configuración y métrica. Resultados: Se encontró que las invarianzas dimensional y de configuración se cumplen al demostrarse que el constructo es unidimensional en los 2 géneros, seleccionando los modelos con el Criterio de información Bayesiano y que la carga de los ítems es en general alta. No así la invarianza métrica para la cual debieron eliminarse 9 ítems para lograr invarianza en las cargas. Conclusión: El formulario de Zung presenta invarianzas dimensional, de configuración y métrica con 11 ítems. Salud UIS 2011; 43 (2): 159-166Introduction: There is not a published report of the invariance of anxiety Zung test, although is widely used. Objective: To stablish gender invariance of the Zung self-rating anxiety scale using Classical Test Theory. Methods: A non random sample of 336 women and 205 men students from several colombian universities that answered the scale. Confirmatory factor analysis was performed. Dimensional, configurational and metric invariance were investigated. Results: Dimensional and configurational invariance are met because for both genders theres is one dimension, using Bayesian Information Crietria as the model selection strategy, and the loadings are generally high. The metric invariance was obtained after deleting 9 items. Conclusion: Eleven items of The Zung self-rating anxiety scale show dimensional, configurational and metric invariance. Salud UIS 2011; 43 (2): 159-166

The purpose of this paper is to present a long- term electroclinical and employment follow up in temporal lobe epilepsy (TLE) patients in a comprehensive epilepsy surgery program. Forty adult patients with pharmacoresistant TLE underwent detailed presurgical evaluation. Electroencephalogram (EEG) and clinical follow up assessment for each patient were carried out. The occurrence of interictal epileptiform activity (IEA) and absolute spike frequency (ASF) were tabulated before and after 1, 6, 12, 24 and 72 months surgical treatment. Employment status pre- to post-surgery at the last evaluated period was also examined. Engel scores follow-up was described as follows: at 12 months 70% (28) class I, 10% (4) class II and 19% (8) class III-IV; at 24 months after surgery 55.2% (21) of the patients were class I, 28.9% (11) class II and 15.1% (6) class III-IV. After one- year follow up 23 (57.7%) patients were seizure and aura-free (Engel class IA). These figures changed to 47.3%, and 48.6% respectively two and five years following surgery whereas 50% maintained this condition in the last follow up period. A decline in the ASF was observed from the first year until the sixth year after surgery in relation to the preoperative EEG. The ASF one year after surgery allowed to distinguish “satisfactory” from “unsatisfactory” seizure relief outcome at the last follow up. An adequate social functioning in terms of education and employment in more than 50% of the patients was also found. Results revealed the feasibility of conducting a successful epilepsy surgery program with favorable long term electroclinical and psychosocial functioning outcomes in a developing country as well.

The mammalian high mobility group protein AT-hook 2 (HMGA2) is a chromosomal architectural transcription factor involved in cell transformation and oncogenesis. It consists of three positively charged "AT-hooks" and a negatively charged C-terminus. Sequence analyses, circular dichroism experiments, and gel-filtration studies showed that HMGA2, in the native state, does not have a defined secondary or tertiary structure. Surprisingly, using combined approaches of 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) chemical cross-linking, analytical ultracentrifugation, fluorescence resonance energy transfer (FRET), and mass spectrometry, we discovered that HMGA2 is capable of self-associating into homodimers in aqueous buffer solution. Our results showed that electrostatic interactions between the positively charged "AT-hooks" and the negatively charged C-terminus greatly contribute to the homodimer formation.

Root depth is a major determinant of plant performance during drought and a key trait for strategies to improve soil carbon sequestration to mitigate climate change. While the model Arabidopsis thaliana offers numerous advantages for studies of root system architecture and root depth, its small and fragile roots severely limit the use of the methods and techniques currently available for such studies in soils. To overcome this, we have developed ClearDepth, a conceptually simple, non-destructive, sensitive, and low-cost method to estimate the root depth of Arabidopsis in relatively small pots that are amenable to mid- and large-scale studies. In our method, the root system develops naturally inside of the soil, without considerable space constraints. The ClearDepth parameter wall root shallowness (WRS) quantifies the shallowness of the root system by measuring the depth of roots that reach the transparent walls of clear pots. We show that WRS is a robust and sensitive parameter that distinguishes deep root systems from shallower ones while also capturing relatively smaller differences in root depth caused by the influence of an environmental factor. In addition, we leveraged ClearDepth to study the relation between lateral root angles measured in non-soil systems and root depth in soil. We found that Arabidopsis genotypes characterized by steep lateral roots in transparent growth media produce deeper root systems in the ClearDepth pots. Finally, we show that ClearDepth can also be used to study root depth in crop species like rice., (© 2024 The Author(s). The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

Glioblastoma multiforme (GBM) is a deadly form of glioma notable for its significant intratumoral heterogeneity, which is believed to drive therapy resistance. GBM has been observed to mimic a neural stem cell hierarchy reminiscent of normal brain development. However, it is still unclear how cell-of-origin shapes intratumoral heterogeneity. Here, we develop a model of glioma initiation using neural stem and progenitor cells (NSPCs) purified from fetal human brain tissue. We previously described a method to prospectively isolate and culture tripotent neural stem cells (NSCs), bipotent glial progenitor cells (GPCs), and unipotent oligodendrocyte precursor cells (OPCs). We transduced these isogenic lines with dominant-negative TP53 R175H and NF1 knockdown, a commonly-used genetic model of GBM in mice. These reprogrammed lines robustly engrafted when transplanted into the brains of immunodeficient mice, and showed significant expansion over time. Engrafted cells were reextracted from the mouse brain for single cell RNA sequencing (scRNA-seq), in order to quantify how the cell-of-origin modulates the cellular subtypes found in the resulting tumor. This result revealed the strong influence the cell-of-origin plays in glioma heterogeneity. Our platform is highly adaptable and allows for modular and systematic interrogation of how cell-of-origin shape the tumor landscape., Competing Interests: COMPETING INTERESTS D.D.L. and I.L.W. are listed as inventors on a patent (WO2023225293A1) related to the isolation of neural stem and progenitor cell types from human brain tissue. I.L.W. is a cofounder of Bitterroot Bio, Inc. and Pheast, Inc., neither of which are related to the current study.

Objective: We aim to evaluate whether a wide range of baseline mental disorders predict Internet Gaming Disorder (IGD) one to three years later, among university students., Methods: Prospective cohort study with a follow-up period of one to three years (September 2018-June 2022) in 6 Mexican universities. Participants were first-year university students (N=2,144) free of symptoms indicative of IGD at entry (baseline). Ten mental disorders (bipolar, major depressive disorder, generalized anxiety disorder, panic disorder, alcohol use disorder, drug use disorder, binging and/or purging, intermittent explosive disorder, psychotic experiences, attention deficit hyperactivity disorder) at baseline were the main risk factors for IGD at the end of the follow-up. We used Cox regression to model the IGD incidence rate., Results: Any mental disorder at baseline was associated with an increase in 2.33 times (1.26-4.31) the rate of IGD 1 to 3 years later. Several individual disorders were associated with rates of IGD in multiple models, with comorbid conditions diminishing most of these associations., Conclusions: Only major depressive disorder and bipolar disorder remained associated with a new case of IGD. Discrepant results from available longitudinal studies on the role of specific mental disorders in the development of IGD needs to be further investigated., Competing Interests: The authors report no conflicts of interest.

While CD40 agonism is an attractive approach for activating antigen-presenting cells and initiating antitumor responses, previous attempts have encountered limited clinical efficacy coupled with toxicity. We previously demonstrated that interactions between the antibody Fc domain and the inhibitory receptor FcγRIIB are critical for enhanced antitumor activity. Here, we present the results of a phase 1 study on intratumoral administration of an anti-CD40 agonistic antibody (2141-V11) Fc-engineered to enhance FcγRIIB binding. Primary endpoints included safety, maximum tolerated dose (MTD), and recommended phase 2 dose. Secondary objectives included preliminary clinical activity and correlative studies from biospecimens. 2141-V11 was well-tolerated without dose-limiting toxicities and MTD was not reached. In ten evaluable patients with metastatic cancer, the overall response rate was 20%, with complete responses in two patients (melanoma and breast carcinoma) and stable disease in six patients. 2141-V11 induced tumor regression in injected and non-injected lesions, with increased leukocyte infiltration and tertiary lymphoid structures (TLS) formation in post-treatment biopsies. In a humanized mouse model for CD40 and FcγRs, 2141-V11 induced TLS formation in mice bearing orthotopic breast carcinoma, correlating with local and abscopal antitumor effects, systemic immune activation, and immune memory. These findings support the safety and efficacy of 2141-V11, warranting phase 2 studies and suggesting a unique mechanism of action for this Fc-enhanced immunotherapy (NCT04059588).

Preclinical murine data indicate that fragment crystallizable (Fc)-dependent depletion of intratumoral regulatory T cells (Treg) is a major mechanism of action of anti-CTLA-4. However, the two main antibodies administered to patients (ipilimumab and tremelimumab) do not recapitulate these effects. Here, we investigate the underlying mechanisms responsible for the limited Treg depletion observed with these therapies. Using an immunocompetent murine model humanized for CTLA-4 and Fcγ receptors (FcγR), we show that ipilimumab and tremelimumab exhibit limited Treg depletion in tumors. Immune profiling of the tumor microenvironment (TME) in both humanized mice and humans revealed high expression of the inhibitory Fc receptor, FcγRIIB, which limits antibody-dependent cellular cytotoxicity/phagocytosis. Blocking FcγRIIB in humanized mice rescued the Treg-depleting capacity and antitumor activity of ipilimumab. Furthermore, Fc engineering of antibodies targeting Treg-associated targets (CTLA-4 or CCR8) to minimize FcγRIIB binding significantly enhanced Treg depletion, resulting in increased antitumor activity across various tumor models. Our results define the inhibitory FcγRIIB as an immune checkpoint limiting antibody-mediated Treg depletion in the TME, and demonstrate Fc engineering as an effective strategy to overcome this limitation and improve the efficacy of Treg-targeting antibodies., (©2023 The Authors; Published by the American Association for Cancer Research.)

The use of colony-stimulating factor-1 receptor (CSF1R) inhibitors has been widely explored as a strategy for cancer immunotherapy due to their robust depletion of tumor-associated macrophages (TAMs). While CSF1R blockade effectively eliminates TAMs from the solid tumor microenvironment, its clinical efficacy is limited. Here, we use an inducible CSF1R knockout model to investigate the persistence of tumor progression in the absence of TAMs. We find increased frequencies of granulocytic myeloid-derived suppressor cells (G-MDSCs) in the bone marrow, throughout circulation, and in the tumor following CSF1R deletion and loss of TAMs. We find that G-MDSCs are capable of suppressing macrophage phagocytosis, and the elimination of G-MDSCs through CXCR2 inhibition increases macrophage capacity for tumor cell clearance. Further, we find that combination therapy of CXCR2 inhibition and CD47 blockade synergize to elicit a significant anti-tumor response. These findings reveal G-MDSCs as key drivers of tumor immunosuppression and demonstrate their inhibition as a potent strategy to increase macrophage phagocytosis and enhance the anti-tumor efficacy of CD47 blockade in B16-F10 melanoma., Competing Interests: Competing interests statement:Patent was filed on combination therapy of CXCR2 inhibition and CD47 blockade.

Ductular reactive (DR) cells exacerbate cholestatic liver injury and fibrosis. Herein, we posit that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) emanates from recruited macrophages and restrains DR cell expansion, thereby limiting cholestatic liver injury. Wild type (WT), Trail fl/fl and myeloid-specific Trail deleted (Trail Δmye ) C57BL/6 mice were exposed to DDC diet-induced cholestatic liver injury, which induced hepatomegaly and liver injury as compared to control diet-fed mice. However, parameters of liver injury, fibrosis, and inflammation were all increased in the Trail Δmye mice as compared to the WT and Trail fl/fl mice. High dimensional mass cytometry indicated that cholestasis resulted in increased hepatic recruitment of subsets of macrophages and neutrophils in the Trail Δmye mice. Spatial transcriptomics analysis revealed that the PanCK + cholangiocytes from Trail Δmye mice had increased expression of the known myeloid attractants S100a8, Cxcl5, Cx3cl1, and Cxcl1. Additionally, in situ hybridization of Cxcl1, a potent neutrophil chemoattractant, demonstrated an increased expression in CK19 + cholangiocytes of Trail Δmye mice. Collectively, these data suggest that TRAIL from myeloid cells, particularly macrophages, restrains a subset of DR cells (i.e., Cxcl1 positive cells), limiting liver inflammation and fibrosis. Reprogramming macrophages to express TRAIL may be salutary in cholestasis., (© 2024. The Author(s).)

Despite recent advances in the treatment of acute myeloid leukemia (AML), there has been limited success in targeting surface antigens in AML, in part due to shared expression across malignant and normal cells. Here, high-density immunophenotyping of AML coupled with proteogenomics identified unique expression of a variety of antigens, including the RNA helicase U5 snRNP200, on the surface of AML cells but not on normal hematopoietic precursors and skewed Fc receptor distribution in the AML immune microenvironment. Cell membrane localization of U5 snRNP200 was linked to surface expression of the Fcγ receptor IIIA (FcγIIIA, also known as CD32A) and correlated with expression of interferon-regulated immune response genes. Anti-U5 snRNP200 antibodies engaging activating Fcγ receptors were efficacious across immunocompetent AML models and were augmented by combination with azacitidine. These data provide a roadmap of AML-associated antigens with Fc receptor distribution in AML and highlight the potential for targeting the AML cell surface using Fc-optimized therapeutics., (© 2023. The Author(s).)

BiP (immunoglobulin heavy-chain binding protein) is a Hsp70 monomeric ATPase motor that plays broad and crucial roles in maintaining proteostasis inside the cell. Structurally, BiP is formed by two domains, a nucleotide-binding domain (NBD) with ATPase activity connected by a flexible hydrophobic linker to the substrate-binding domain. While the ATPase and substrate binding activities of BiP are allosterically coupled, the latter is also dependent on nucleotide binding. Recent structural studies have provided new insights into BiP's allostery; however, the influence of temperature on the coupling between substrate and nucleotide binding to BiP remains unexplored. Here, we study BiP's binding to its substrate at the single molecule level using thermo-regulated optical tweezers which allows us to mechanically unfold the client protein and explore the effect of temperature and different nucleotides on BiP binding. Our results confirm that the affinity of BiP for its protein substrate relies on nucleotide binding, by mainly regulating the binding kinetics between BiP and its substrate. Interestingly, our findings also showed that the apparent affinity of BiP for its protein substrate in the presence of nucleotides remains invariable over a wide range of temperatures, suggesting that BiP may interact with its client proteins with similar affinities even when the temperature is not optimal. Thus, BiP could play a role as a "thermal buffer" in proteostasis., (© 2023 The Protein Society.)

Objective: To describe the clinical and epidemiological characteristics of newborn infants with SARS-CoV-2 infection notified in the Colombian Public Health Surveillance System., Design: This epidemiological descriptive analysis was conducted using the data of all cases of newborn infants with confirmed SARS-CoV-2 infection notified in the surveillance system. Absolute frequencies and central tendency measures were calculated and a bivariate analysis comparing variables of interest with symptomatic and asymptomatic disease was performed., Setting: Population-based descriptive analysis., Patients: Laboratory-confirmed COVID-19 cases in newborn infants (aged ≤28 days of life) reported to the surveillance system from 1 March 2020 to 28 February 2021., Results: 879 newborns were identified, corresponding to 0.04% of all reported cases in the country. The mean age at diagnosis was 13 days (range 0-28 days), 55.1% were male and most (57.6%) were classified as symptomatic. Preterm birth and low birth weight were identified in 24.0% and 24.4% of the cases, respectively. Common symptoms were fever (58.3%), cough (48.3%) and respiratory distress (34.9%). A higher prevalence of symptomatic newborns was seen in individuals with low birth weight for gestational age (prevalence ratio (PR): 1.51, 95% CI: 1.44 to 1.59) and newborns with underlying conditions (PR: 1.33, 95% CI: 1.13 to 1.55)., Conclusions: There were a low proportion of confirmed COVID-19 cases in the newborn population. A substantial number of newborns were classified as symptomatic, having low birth weight and being preterm. Clinicians caring for COVID-19-infected newborns should be aware of population characteristics that potentially contribute to disease manifestations and severity., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Temperature is a useful system variable to gather kinetic and thermodynamic information from proteins. Usually, free energy and the associated entropic and enthalpic contributions are obtained by quantifying the conformational equilibrium based on melting experiments performed in bulk conditions. Such experiments are suitable only for those small single-domain proteins whose side reactions of irreversible aggregation are unlikely to occur. Here, we avoid aggregation by pulling single-protein molecules in a thermo-regulated optical tweezers. Thus, we are able to explore the temperature dependence of the thermodynamic and kinetic parameters of MJ0366 from Methanocaldococcus jannaschii at the single-molecule level. By performing force-ramp experiments between 2°C and 40°C, we found that MJ0366 has a nonlinear dependence of free energy with temperature and a specific heat change of 2.3 ± 1.2 kcal/mol ∗ K. These thermodynamic parameters are compatible with a two-state unfolding/refolding mechanism for MJ0366. However, the kinetics measured as a function of the temperature show a complex behavior, suggesting a three-state folding mechanism comprising a high-energy intermediate state. The combination of two perturbations, temperature and force, reveals a high-energy species in the folding mechanism of MJ0366 not detected in force-ramp experiments at constant temperature., Competing Interests: Declaration of interests S.B.S. makes and sells optical tweezers., (Copyright © 2022 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Cellular senescence is a plausible mediator of inflammation-related tissue dysfunction. In the aged brain, senescent cell identities and the mechanisms by which they exert adverse influence are unclear. Here we used high-dimensional molecular profiling, coupled with mechanistic experiments, to study the properties of senescent cells in the aged mouse brain. We show that senescence and inflammatory expression profiles increase with age and are brain region- and sex-specific. p16-positive myeloid cells exhibiting senescent and disease-associated activation signatures, including upregulation of chemoattractant factors, accumulate in the aged mouse brain. Senescent brain myeloid cells promote peripheral immune cell chemotaxis in vitro. Activated resident and infiltrating immune cells increase in the aged brain and are partially restored to youthful levels through p16-positive senescent cell clearance in female p16-InkAttac mice, which is associated with preservation of cognitive function. Our study reveals dynamic remodeling of the brain immune cell landscape in aging and suggests senescent cell targeting as a strategy to counter inflammatory changes and cognitive decline., (© 2022. The Author(s).)

Carbapenems are commonly used to treat infections caused by multidrug-resistant (MDR) bacteria. Unfortunately, carbapenem resistance is increasingly reported in many gram-negative bacteria, especially Acinetobacter baumannii . Diazabicyclooctane (DBO) β-lactamase inhibitors, such as avibactam (AVI), when combined with sulbactam successfully restore sulbactam susceptibility against certain carbapenem-resistant A. baumannii (CRAB) isolates. In the present study, we tested zidebactam, a novel DBO with an additional mechanism of action, in combination with sulbactam against CRAB isolates, including strains that exhibited resistance against sulbactam/avibactam combination. A panel of 43 geographically and genetically distinct CRAB isolates recovered from different hospitals and containing different mechanisms of resistance were included in the present study. We also tested three reference strains (AB0057, AB5075, and AYE). Minimum inhibitory concentrations (MICs) for sulbactam (range 0.12-512 mg/l) and sulbactam plus 4 mg/l zidebactam were performed using microdilution according to CLSI Standards. A decrease ≥2 dilutions in sulbactam MICs was observed in 84% of the isolates when tested in combination with zidebactam. The sulbactam/zidebactam combination was able to restore sulbactam susceptibility in 91% of the isolates, including isolates that were resistant to sulbactam/avibactam combination. These data encouraged us to further explore sulbactam/zidebactam in other experimental models especially against CRAB isolates resistant to other DBOs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Cedano, Baez, Pasteran, Montaña, Ra, Fua, Corso, Tolmasky, Bonomo and Ramírez.)

Abstract: This article describes an evidence-based project that aimed to identify nursing staff perceptions on the use of remote video monitoring (RVM), collect fall data following the implementation of RVM, and determine the benefits of RVM in reducing falls., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Karyotypes are characterized by traits such as chromosome number, which can change through whole-genome duplication and dysploidy. In the parasitic plant genus Cuscuta (Convolvulaceae), chromosome numbers vary more than 18-fold. In addition, species of this group show the highest diversity in terms of genome size among angiosperms, as well as a wide variation in the number and distribution of 5S and 35S ribosomal DNA (rDNA) sites. To understand its karyotypic evolution, ancestral character state reconstructions were performed for chromosome number, genome size, and position of 5S and 35S rDNA sites. Previous cytogenetic data were reviewed and complemented with original chromosome counts, genome size estimates, and rDNA distribution assessed via fluorescence in situ hybridization (FISH), for two, seven, and 10 species, respectively. Starting from an ancestral chromosome number of x  = 15, duplications were inferred as the prevalent evolutionary process. However, in holocentric clade (subgenus Cuscuta ), dysploidy was identified as the main evolutionary mechanism, typical of holocentric karyotypes. The ancestral genome size of Cuscuta was inferred as approximately 1C = 12 Gbp, with an average genome size of 1C = 2.8 Gbp. This indicates an expansion of the genome size relative to other Convolvulaceae, which may be linked to the parasitic lifestyle of Cuscuta . Finally, the position of rDNA sites varied mostly in species with multiple sites in the same karyotype. This feature may be related to the amplification of rDNA sites in association to other repeats present in the heterochromatin. The data suggest that different mechanisms acted in different subgenera, generating the exceptional diversity of karyotypes in Cuscuta ., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Ibiapino, García, Amorim, Baez, Costea, Stefanović and Pedrosa-Harand.)

Background: Carbohydrate-active enzymes (CAZymes) form the most widespread and structurally diverse set of enzymes involved in the breakdown, biosynthesis, or modification of lignocellulose that can be found in living organisms. However, the structural diversity of CAZymes has rendered the targeted discovery of novel enzymes extremely challenging, as these proteins catalyze many different chemical reactions and are sourced by a vast array of microbes. Consequently, many uncharacterized members of CAZyme families of interest have been overlooked by current methodologies (e.g., metagenomic screening) used to discover lignocellulolytic enzymes., Results: In the present study, we combined phenotype-based selective pressure on the rumen microbiota with targeted functional profiling to guide the discovery of unknown CAZymes. In this study, we found 61 families of glycoside hydrolases (GH) (out of 182 CAZymes) from protein sequences deposited in the CAZy database-currently associated with more than 20,324 microbial genomes. Phenotype-based selective pressure on the rumen microbiome showed that lignocellulolytic bacteria (e.g., Fibrobacter succinogenes, Butyrivibrio proteoclasticus) and three GH families (e.g., GH11, GH13, GH45) exhibited an increased relative abundance in the rumen of feed efficient cattle when compared to their inefficient counterparts. These results paved the way for the application of targeted functional profiling to screen members of the GH11 and GH45 families against a de novo protein reference database comprised of 1184 uncharacterized enzymes, which led to the identification of 18 putative xylanases (GH11) and three putative endoglucanases (GH45). The biochemical proof of the xylanolytic activity of the newly discovered enzyme validated the computational simulations and demonstrated the stability of the most abundant xylanase., Conclusions: These findings contribute to the discovery of novel enzymes for the breakdown, biosynthesis, or modification of lignocellulose and demonstrate that the rumen microbiome is a source of promising enzyme candidates for the biotechnology industry. The combined approaches conceptualized in this study can be adapted to any microbial environment, provided that the targeted microbiome is easy to manipulate and facilitates enrichment for the microbes of interest. Video Abstract., (© 2021. The Author(s).)

Domain swapping is a mechanism of protein oligomerization by which two or more subunits exchange structural elements to generate an intertwined complex. Numerous studies support a diversity of swapping mechanisms in which structural elements can be exchanged at different stages of the folding pathway of a subunit. Here, we used single-molecule optical tweezers technique to analyze the swapping mechanism of the forkhead DNA-binding domain of human transcription factor FoxP1. FoxP1 populates folded monomers in equilibrium with a swapped dimer. We generated a fusion protein linking two FoxP1 domains in tandem to obtain repetitive mechanical folding and unfolding trajectories. Thus, by stretching the same molecule several times, we detected either the independent folding of each domain or the elusive swapping step between domains. We found that a swapped dimer can be formed directly from fully or mostly folded monomer. In this situation, the interaction between the monomers in route to the domain-swapped dimer is the rate-limiting step. This approach is a useful strategy to test the different proposed domain swapping mechanisms for proteins with relevant physiological functions., (Copyright © 2021 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Gold nanoparticles (GNP) are tunable nanomaterials that can be used to develop rational therapeutic inhibitors against the formation of pathological aggregates of proteins. In the case of the pathological aggregation of the amyloid-β protein (Aβ), the shape of the GNP can slow down or accelerate its aggregation kinetics. However, there is a lack of elementary knowledge about how the curvature of GNP alters the interaction with the Aβ peptide and how this interaction modifies key molecular steps of fibril formation. In this study, we analysed the effect of flat gold nanoprisms (GNPr) and curved gold nanospheres (GNS) on in vitro Aβ42 fibril formation kinetics by using the thioflavin-based kinetic assay and global fitting analysis, with several models of aggregation. Whereas GNPr accelerate the aggregation process and maintain the molecular mechanism of aggregation, GNS slow down this process and modify the molecular mechanism to one of fragmentation/secondary nucleation, with respect to controls. These results can be explained by a differential interaction between the Aβ peptide and GNP observed by Raman spectroscopy. While flat GNPr expose key hydrophobic residues involved in the Aβ peptide aggregation, curved GNS hide these residues from the solvent. Thus, this study provides mechanistic insights to improve the rational design of GNP nanomaterials for biomedical applications in the field of amyloid-related aggregation., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Resistance to third-generation cephalosporins (3GC) in Escherichia coli has been reported worldwide from humans and animals, but the situation in Cuba is still poorly understood. This study aimed to gain new insights into the phenotypic and genotypic characteristics of third-generation cephalosporin-resistant (3GC-R) E. coli isolated from pigs in Cuba. Rectal swabs from 215 healthy pigs were taken from different municipalities in the western region of Cuba and spread on MacConkey agar supplemented with cefotaxime and ceftazidime. Ninety-six isolates were identified as 3GC-R E. coli and 87.5% of them were resistant to at least three antibiotic classes as determined by the measurement of the minimum inhibitory concentration (MIC) of 14 antibiotics. Twenty-seven different isolates were selected for Illumina next-generation sequencing, and subsequent in silico analysis was performed for the detection of antibiotic resistance and virulence genes, plasmid incompatibility (Inc) groups, multilocus sequence typing (MLST), and core genome MLST (cgMLST). The sequenced isolates contained extended-spectrum β-lactamase genes bla CTX-M-32 ( n  = 17), bla CTX-M-15 ( n  = 5), and bla CTX-M-55 ( n  = 4) as well as with pAmpC gene bla CMY-2 ( n  = 2). They also harbored genes for resistance to other clinically important classes of antibiotics, as well as several diverse virulence factors. The 3GC-R E. coli were genetically highly diverse, belonging to 16 different sequence types. IncX1 was the most frequent Inc group. The presence of 3GC-R E. coli in pigs from Cuba containing several different antibiotic resistance mechanisms emphasizes the need for surveillance programs and the establishment of strategies for the prudent use of antibiotics in food-producing animals.

An increasing number of untreatable infections are recorded every year. Many studies have focused their efforts on developing new β-lactamase inhibitors to treat multi-drug resistant (MDR) isolates. In the present study, sulbactam/avibactam and sulbactam/relebactam combination were tested against 187 multi-drug resistant (MDR) Acinetobacter clinical isolates; both sulbactam/avibactam and sulbactam/relebactam restored sulbactam activity. A decrease ≥2 dilutions in sulbactam MICs was observed in 89% of the isolates when tested in combination with avibactam. Sulbactam/relebactam was able to restore sulbactam susceptibility in 40% of the isolates. In addition, the susceptibility testing using twenty-three A. baumannii AB5075 knockout strains revealed potential sulbactam and/or sulbactam/avibactam target genes. We observed that diazabicyclooctanes (DBOs) β-lactamase inhibitors combined with sulbactam restore sulbactam susceptibility against carbapenem-resistant Acinetobacter clinical isolates. However, relebactam was not as effective as avibactam when combined with sulbactam. Exploring novel combinations may offer new options to treat Acinetobacter spp. infections, especially for widespread oxacillinases and metallo-β-lactamases (MBLs) producers.

Rye (Secale cereale L.) is an exceptionally climate-resilient cereal crop, used extensively to produce improved wheat varieties via introgressive hybridization and possessing the entire repertoire of genes necessary to enable hybrid breeding. Rye is allogamous and only recently domesticated, thus giving cultivated ryes access to a diverse and exploitable wild gene pool. To further enhance the agronomic potential of rye, we produced a chromosome-scale annotated assembly of the 7.9-gigabase rye genome and extensively validated its quality by using a suite of molecular genetic resources. We demonstrate applications of this resource with a broad range of investigations. We present findings on cultivated rye's incomplete genetic isolation from wild relatives, mechanisms of genome structural evolution, pathogen resistance, low-temperature tolerance, fertility control systems for hybrid breeding and the yield benefits of rye-wheat introgressions.

A total of 434 poultry cloacal samples were collected from seven different farms in different years (2013-2015) in the Cuban province of Mayabeque and analyzed for the presence of third-generation cephalosporin-resistant Escherichia coli (3GC-R- Ec ). Sixty-two 3GC-R- Ec isolates were recovered in total from the farms, with detection rates of 2.9% in 2013, 10.3% in 2014, and 28.7% in 2015. Characterization of 32 3GC-R- Ec isolates revealed the presence of the extended-spectrum β-lactamase (ESBL) genes bla CTX-M-1 ( n = 27), bla CTX-M-15 ( n = 4), and bla CTX-M-1 together with bla LAP-2 ( n = 1). The isolates also contained different proportions of genes conferring decreased susceptibility to sulfonamides ( sul1 , sul2 , sul3 ), trimethoprim ( dfrA1 , dfrA 7, dfrA 12, dfrA 14, dfrA 17), tetracyclines ( tet (A), tet (B)), aminoglycosides ( aac(6')-Ib-cr , strA , strB ), chloramphenicol ( cmlA1 , floR ), macrolides ( mph (A), mph (D)), and quinolones ( qnrS , qnrB , aac(6')-Ib-cr ) as well as mutations in the fluoroquinolone-resistance determining regions of GyrA (S83L, D87N, D87Y) and ParC (S80I, E84G). The isolates belonged to 23 different sequence types and to phylogroups A ( n = 25), B1 ( n = 5), and D ( n = 2), and they contained plasmid-associated incompatibility groups FII, X1, HI1, HI2, N, FIA, and FIB. These findings reveal a genetically diverse population of multiresistant ESBL-producing E. coli in poultry farms in Cuba, which suggests multiple sources of contamination and the acquisition of antibiotic resistance genes.

Introduction: Estrogen are hormones linked to different stages of reproductive female system. Also are involved in detailed effects on the vestibular system. A review was performed about changes related to estrogens on the vestibular system, according to each phase of the female reproductive system. Materials: Results: Since menarch to menopause, several vestibular symptoms have been found as vertigo, tinnitus, fullness and hearing loss. Conclusion:, Materials: Search on medical databases which included PubMed, Cochrane, Scielo, Google Scholar and Bvsalud of articles published between 1964 to 2020 with the following keywords ‘estrogen and dizziness’, ‘estrogen and vertigo’, ‘estrogen and vestibular disorders., Results: 207 articles were found, 29 in english with highlighted information about estrogen related changes on the vestibular system., Conclusion: According to each female reproductive stage, changes related to estrogen have been reported on the vestibular system., (Universidad Nacional de Córdoba.)

For a long time, the Cyperid clade (Thurniceae-Juncaceae-Cyperaceae) was considered a group of species possessing holocentromeres exclusively. The basal phylogenetic position of Prionium serratum (Thunb.) Drège (Thurniceae) within Cyperids makes this species an important specimen to understand the centromere evolution within this clade. In contrast to the expectation, the chromosomal distribution of the centromere-specific histone H3 (CENH3), alpha-tubulin and different centromere-associated post-translational histone modifications (H3S10ph, H3S28ph and H2AT120ph) demonstrate a monocentromeric organisation of P. serratum chromosomes. Analysis of the high-copy repeat composition resulted in the identification of two centromere-localised satellite repeats. Hence, monocentricity was the ancestral condition for the Juncaceae-Cyperaceae-Thurniaceae Cyperid clade, and holocentricity in this clade has independently arisen at least twice after differentiation of the three families, once in Juncaceae and the other one in Cyperaceae. In this context, methods suitable for the identification of holocentromeres are discussed.

Background: This review studies technology-supported interventions to help older adults, living in situations of reduced mobility, overcome loneliness, and social isolation. The focus is on long-distance interactions, investigating the (i) challenges addressed and strategies applied; (ii) technology used in interventions; and (iii) social interactions enabled., Methods: We conducted a search on Elsevier's Scopus database for related work published until January 2020, focusing on (i) intervention studies supported mainly by technology-mediated communication, (ii) aiming at supported virtual social interactions between people, and (iii) evaluating the impact of loneliness or social isolation., Results: Of the 1178 papers screened, 25 met the inclusion criteria. Computer and Internet training was the dominant strategy, allowing access to communication technologies, while in recent years, we see more studies aiming to provide simple, easy-to-use technology. The technology used was mostly off-the-shelf, with fewer solutions tailored to older adults. Social interactions targeted mainly friends and family, and most interventions focused on more than one group of people. Discussion . All interventions reported positive results, suggesting feasibility. However, more research is needed on the topic (especially randomized controlled trials), as evidenced by the low number of interventions found. We recommend more rigorous methods, addressing human factors and reporting technology usage in future research., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this paper., (Copyright © 2020 Francisco Ibarra et al.)

Knots are remarkable topological features in nature. The presence of knots in crystallographic structures of proteins have stimulated considerable research to determine the kinetic and thermodynamic consequences of threading a polypeptide chain. By mechanically manipulating MJ0366, a small single domain protein harboring a shallow trefoil knot, we allow the protein to refold from either the knotted or the unknotted denatured state to characterize the free energy profile associated to both folding pathways. By comparing the stability of the native state with reference to the knotted and unknotted denatured state we find that knotting the polypeptide chain of MJ0366 increase the folding energy barrier in a magnitude close to the energy cost of forming a knot randomly in the denatured state. These results support that a protein knot can be formed during a single cooperative step of folding but occurs at the expenses of a large increment on the free energy barrier.

Background: Intergenerational relationships are beneficial for both grandparents and grandchildren. A positive grandparent-grandchild relationship can improve the psychological well-being of older adults and be a source of social support, family history, and identity development. Maintaining meaningful interactions can be, however, a challenging endeavor, especially as life events lead to relocating geographically. Grandparents and grandchildren can have different preferences in terms of communication mediums and different assumptions about the real conversational needs of the other., Objective: In this study, we will investigate the feasibility and effect of sharing memories of older adults with their grandchildren in social media. This intervention focuses on bringing snippets of the lives of the grandparents into the grandchildren's social media feed and analyzing the potential effect on relational quality, relational investment, and conversational resources from the perspective of the grandchildren., Methods: A randomized controlled trial will be used to measure the effectiveness of sharing family memories through social media on intergenerational relationships from the perspective of the grandchildren. The study will be implemented in Mongolia among 60 grandparent-grandchild pairs who will be assigned to either a control or intervention group. Pictures and stories will be collected during reminiscence sessions between the researchers and the grandparents before the intervention. During an intervention period of 2 months, grandchildren in the intervention group will receive pictures and stories of their grandparents on their social media account. Pre- and postintervention questionnaires will measure relationship quality, relationship investment, and conversational resources and will be used to assess the effectiveness of the intervention., Results: We conducted a pretest pilot from January to April 2018 among 6 pairs of participants (6 grandparents and 6 grandchildren). The validation of the protocol was focused on the process, instruments, and technological setup. We continued the study after the validation, and 59 pairs of participants (59 grandparents and 59 grandchildren) have been recruited. The data collection was completed in November 2019., Conclusions: The results of this study will contribute to strategies to stimulate social interactions in intergenerational pairs. A validation of the study process is also presented to provide further operational recommendations. The lessons learned during the validation of the protocol are discussed with recommendations and implications for the recruitment, reminiscence sessions, technological setup, and administration of instruments., International Registered Report Identifier (irrid): DERR1-10.2196/16315., (©Zoljargalan Gantumur, Marcos Baez, Nomin-Erdene Ulamnemekh, Francisco Ibarra, Sugarmaa Myagmarjav, Fabio Casati. Originally published in JMIR Research Protocols (http://www.researchprotocols.org), 30.04.2020.)