Phosphorylation at the disordered N-end makes HuR accumulate and dimerize in the cytoplasm.

Human antigen R (HuR) is an RNA binding protein mainly involved in maintaining the stability and controlling the translation of mRNAs, critical for immune response, cell survival, proliferation and apoptosis. Although HuR is a nuclear protein, its mRNA translational-related function occurs at the cytoplasm, where the oligomeric form of HuR is more abundant. However, the regulation of nucleo-cytoplasmic transport of HuR and its connection with protein oligomerization remain unclear. In this work, we describe the phosphorylation of Tyr5 as a new hallmark for HuR activation. Our biophysical, structural and computational assays using phosphorylated and phosphomimetic HuR proteins demonstrate that phosphorylation of Tyr5 at the disordered N-end stretch induces global changes on HuR dynamics and conformation, modifying the solvent accessible surface of the HuR nucleo-cytoplasmic shuttling (HNS) sequence and releasing regions implicated in HuR dimerization. These findings explain the preferential cytoplasmic accumulation of phosphorylated HuR in HeLa cells, aiding to comprehend the mechanisms underlying HuR nucleus-cytoplasm shuttling and its later dimerization, both of which are relevant in HuR-related pathogenesis.

Acidochromic Free-Standing Multilayered Chitosan-Pyranoflavylium/Alginate Membranes toward Food Smart Packaging Applications.

Food smart packaging has emerged as a promising technology to address consumer concerns regarding food conservation and food safety. In this context, we report the rational design of azide-containing pyranoflavylium-based pH-sensitive dye for subsequent click chemistry conjugation toward a chitosan-modified alkyne. The chitosan-pyranoflavylium conjugate was characterized by infrared (ATR-FTIR), ultraviolet-visible (UV-vis), nuclear magnetic resonance (NMR) spectroscopies, and dynamic light scattering (DLS), as well as its thermodynamic parameters related to their pH-dependent chromatic features. The fabrication of thin-films through electrostatic-driven layer-by-layer (LbL) assembly technology was first screened by quartz crystal microbalance with dissipation monitoring (QCM-D) onto gold substrates, and then free-standing (FS) multilayered membranes from polypropylene substrate were obtained using a homemade automatic dipping robot. The membranes' characterization included morphology analysis and thickness evaluation, assessed by scanning electron microscopy (SEM), pH-responsive color change performance tests using buffer solutions at different pH levels, and biogenic amines-enriched model solutions, demonstrating the feasibility and effectiveness of the chitosan-pyranoflavylium/alginate biomembranes for food spoilage monitoring. This work provides insights toward the development of innovative pH-responsive smart biomaterials for advanced and sustainable technological packaging solutions, which could significantly contribute to ensuring food safety and quality, while reducing food waste.

Chitosan/Polyvinyl Alcohol-Based Biofilms Using Ternary Deep Eutectic Solvents towards Innovative Color-Stabilizing Systems for Anthocyanins.

Anthocyanins are amazing plant-derived colorants with highly valuable properties; however, their chemical and color instability issues limit their wide application in different food industry-related products such as active and intelligent packaging. In a previous study, it was demonstrated that anthocyanins could be stabilized into green plasticizers namely deep eutectic solvents (DESs). In this work, the fabrication of edible films by integrating anthocyanins along with DESs into biocompatible chitosan (CHT)-based formulations enriched with polyvinyl alcohol (PVA) and PVA nanoparticles was investigated. CHT/PVA-DES films' physical properties were characterized by scanning electron microscopy, water vapor permeability, swelling index, moisture sorption isotherm, and thermogravimetry analysis. Innovative red-to-blue formulation films were achieved for CHT/PVA nanoparticles (for 5 min of sonication) at a molar ratio 1:1, and with 10% of ternary DES (TDES)-containing malvidin-3-glucoside (0.1%) where the physical properties of films were enhanced. After immersion in solutions at different pH values, films submitted to pHs 5-8 were revealed to be more color stable and resistant with time than at acidic pH values.

Field Collection and Laboratory Maintenance of Canopy-Forming Giant Kelp to Facilitate Restoration.

Canopy-forming kelps are essential foundation species, supporting biodiversity and providing ecosystem services valued at more than USD$500 billion annually. The global decline of giant kelp forests due to climate-driven ecological stressors underscores the need for innovative restoration strategies. An emerging restoration technique known as 'green gravel' aims to seed young kelps over large areas without extensive underwater labor and represents a promising restoration tool due to cost-effectiveness and scalability. This video article illustrates a protocol and tools for culturing giant kelp, Macrocystis pyrifera. It also provides a resource for further studies to address the successes and limitations of this method in field settings. We outline field and laboratory-based methods for collecting reproductive tissue, sporulating, inoculating, rearing, maintaining, and monitoring substrates seeded with early life stages using the 'green gravel' technique. The protocol simplifies and centralizes current restoration practices in this field to support researchers, managers, and stakeholders in meeting kelp conservation objectives.

+Ciencia: a training program to increase evidence-based science communication and literacy for Hispanic high school and undergraduate students.

Science misinformation represents a significant challenge for the scientific community. Hispanic communities are particularly vulnerable due to language barriers and the lack of accessible information in Spanish. We identified that a key step toward enhancing the accessibility of information for non-native English-speaking communities involves imparting science communication education and training to Hispanic youth. Our goal was to provide them with the skills to become science ambassadors who can effectively engage with their communities and bridge communication gaps. To address this, we developed the first science communication training program in Spanish for Hispanic high school and undergraduate students in Puerto Rico. The program called +Ciencia aims to provide training and education on science communication for Hispanic minorities through experiential and collaborative learning. In the short term, our multifaceted approach works to counter misinformation and promote science literacy within the broader community. Over the long term, our grassroots efforts with students will evolve into a generation of professionals equipped with strong engagement skills and comprehensive training in science communication with a specific focus on Hispanic audiences. Herein, we describe the components of this educational program and provide open access to educational materials and articles developed by three cohorts.

Relationship between psychoactive substance, alcohol and cigarette use in nursing students. A cross-sectional study.

BACKGROUND: The increase in the use of psychoactive substances, alcohol and cigarettes in young people has become a public health problem. The identification of factors that increase or reduce the risk of exposure to these substances and the possible relationship between them is essential for planning strategies with a risk approach; hence the reason for this study. The objective was to establish the profile of use of psychoactive substances, alcohol and cigarettes and the factors associated with such use in nursing students of a higher education institution. METHODS: Quantitative, observational, analytical cross-sectional study. RESULTS: We included 310 students from 1 st to 9th semester of a Nursing programme from a private higher education institution in Bogotá. The prevalence of psychoactive substance use in the last year was 2.96% (95%CI, 1.36-5.54), with marijuana being the substance most used (55.55%). The prevalence of alcohol and cigarette use during the last 12 months was estimated at 86.64% (95%CI, 83.24-91.0) and 12.16% (95%CI, 8.43-15.88) respectively. A statistically significant association was found between the use of these substances: alcohol use was associated with cigarette use (OR = 3.22; P = 0.006) and smoking was associated with psychoactive substance use (OR = 15.4; P < 0.001). CONCLUSIONS: Alcohol use increases the likelihood of smoking cigarettes, and this in turn increases the likelihood of psychoactive substance use, in this university population.

Biochemical and structural impact of two novel missense mutations in cystathionine ß-synthase gene associated with homocystinuria.

Homocystinuria is a rare disease caused by mutations in the CBS gene that results in a deficiency of cystathionine ß-synthase (CBS). CBS is an essential pyridoxal 5'-phosphate (PLP)-dependent enzyme in the transsulfuration pathway, responsible for combining serine with homocysteine to produce cystathionine, whose activity is enhanced by the allosteric regulator S-adenosylmethionine (SAM). CBS also plays a role in generating hydrogen sulfide (H2S), a gaseous signaling molecule with diverse regulatory functions within the vascular, nervous, and immune systems. In this study, we present the clinical and biochemical characterization of two novel CBS missense mutations that do not respond to pyridoxine treatment, namely c.689T > A (L230Q) and 215A > T (K72I), identified in a Chinese patient. We observed that the disease-associated K72I genetic variant had no apparent effects on the spectroscopic and catalytic properties of the full-length enzyme. In contrast, the L230Q variant expressed in Escherichia coli did not fully retain heme and when compared with the wild-type enzyme, it exhibited more significant impairments in both the canonical cystathionine-synthesis and the alternative H2S-producing reactions. This reduced activity is consistent with both in vitro and in silico evidence, which indicates that the L230Q mutation significantly decreases the overall protein's stability, which in turn, may represent the underlying cause of its pathogenicity.

Catalytic specificity and crystal structure of cystathionine γ-lyase from Pseudomonas aeruginosa.

The escalating drug resistance among microorganisms underscores the urgent need for innovative therapeutic strategies and a comprehensive understanding of bacteria's defense mechanisms against oxidative stress and antibiotics. Among the recently discovered barriers, the endogenous production of hydrogen sulfide (H2S) via the reverse transsulfuration pathway, emerges as a noteworthy factor. In this study, we have explored the catalytic capabilities and crystal structure of cystathionine γ-lyase from Pseudomonas aeruginosa (PaCGL), a multidrug-opportunistic pathogen chiefly responsible for nosocomial infections. In addition to a canonical L-cystathionine hydrolysis, PaCGL efficiently catalyzes the production of H2S using L-cysteine and/or L-homocysteine as alternative substrates. Comparative analysis with the human enzyme and counterparts from other pathogens revealed distinct structural features within the primary enzyme cavities. Specifically, a distinctly folded entrance loop could potentially modulate the access of substrates and/or inhibitors to the catalytic site. Our findings offer significant insights into the structural evolution of CGL enzymes across different pathogens and provide novel opportunities for developing specific inhibitors targeting PaCGL.

Hypomagnesaemia with varying degrees of extrarenal symptoms as a consequence of heterozygous CNNM2 variants.

Variants in the CNNM2 gene are causative for hypomagnesaemia, seizures and intellectual disability, although the phenotypes can be variable. This study aims to understand the genotype-phenotype relationship in affected individuals with CNNM2 variants by phenotypic, functional and structural analysis of new as well as previously reported variants. This results in the identification of seven variants that significantly affect CNNM2-mediated Mg2+ transport. Pathogenicity of these variants is further supported by structural modelling, which predicts CNNM2 structure to be affected by all of them. Strikingly, seizures and intellectual disability are absent in 4 out of 7 cases, indicating these phenotypes are caused either by specific CNNM2 variant only or by additional risk factors. Moreover, in line with sporadic observations from previous reports, CNNM2 variants might be associated with disturbances in parathyroid hormone and Ca2+ homeostasis.

Application of Doxorubicin-loaded PLGA nanoparticles targeting both tumor cells and cancer-associated fibroblasts on 3D human skin equivalents mimicking melanoma and cutaneous squamous cell carcinoma.

Nanoparticle (NP) use in cancer therapy is extensively studied in skin cancers. Cancer-associated fibroblasts (CAFs), a major tumor microenvironment (TME) component, promote cancer progression, making dual targeting of cancer cells and CAFs an effective therapy. However, dual NP-based targeting therapy on both tumor cells and CAFs is poorly investigated in skin cancers. Herein, we prepared and characterized doxorubicin-loaded PLGA NPs (DOX@PLGA NPs) and studied their anti-tumor effects on cutaneous melanoma (SKCM)(AN, M14) and cutaneous squamous cell carcinoma (cSCC) (MET1, MET2) cell lines in monolayer, as well as their impact on CAF deactivation. Then, we established 3D full thickness models (FTM) models of SKCM and cSCC using AN or MET2 cells on dermis matrix populated with CAFs respectively, and assessed the NPs' tumor penetration, tumor-killing ability, and CAF phenotype regulation through both topical administration and intradermal injection. The results show that, in monolayer, DOX@PLGA NPs inhibited cancer cell growth and induced apoptosis in a dose- and time-dependent manner, with a weaker effect on CAFs. DOX@PLGA NPs reduced CAF-marker expression and had successful anti-tumor effects in 3D skin cancer FTMs, with decreased tumor-load and invasion. DOX@PLGA NPs also showed great delivery potential in the FTMs and could be used as a platform for future functional study of NPs in skin cancers using human-derived skin equivalents. This study provides promising evidence for the potential of DOX@PLGA NPs in dual targeting therapy for SKCM and cSCC.

SUMOylation controls Hu antigen R posttranscriptional activity in liver cancer.

The posttranslational modification of proteins critically influences many biological processes and is a key mechanism that regulates the function of the RNA-binding protein Hu antigen R (HuR), a hub in liver cancer. Here, we show that HuR is SUMOylated in the tumor sections of patients with hepatocellular carcinoma in contrast to the surrounding tissue, as well as in human cell line and mouse models of the disease. SUMOylation of HuR promotes major cancer hallmarks, namely proliferation and invasion, whereas the absence of HuR SUMOylation results in a senescent phenotype with dysfunctional mitochondria and endoplasmic reticulum. Mechanistically, SUMOylation induces a structural rearrangement of the RNA recognition motifs that modulates HuR binding affinity to its target RNAs, further modifying the transcriptomic profile toward hepatic tumor progression. Overall, SUMOylation constitutes a mechanism of HuR regulation that could be potentially exploited as a therapeutic strategy for liver cancer.

Importance of Selected Nutrients and Additives in the Feed of Pregnant Sows for the Survival of Newborn Piglets.

This systematic review analyzed the effect of selected nutrients and additives in the feed of pregnant sows on the survival of newborn piglets. We analyzed 720 peer-reviewed publications in English in PubMed® and Web of Science®, dated July 2023 to January 2024, related to the effect of dietary supplementation with fatty acids and various percentages of protein, amino acids, and/or sources of dietary fiber on the offspring of gestating sows. While several papers evaluated the effect of nutrition on gestating sows, only a few delved into the distinct feeding strategies required at each stage of gestation to meet the NRC's nutritional requirements for maternal tissue gain and postnatal neonatal survival and growth. This body of research suggests that as gestation progresses the sow's nutritional requirements increase, as the NRC established, to satisfy their own metabolic needs and those of their fetuses. Additional research is needed to determine an optimal feeding strategy.

Host-Guest Chemosensor Ensembles based on Water-Soluble Sulfonated Calix[n]arenes and a Pyranoflavylium Dye for the Optical Detection of Biogenic Amines.

Biogenic amines (BAs) are biologically active nitrogen-containing compounds formed during the food spoilage process and are often related as key markers of food quality, safety, and freshness. Because their presence in foods at high levels can cause significant health problems, researchers have been focused on developing novel strategies and methods for early detection and capture of these analytes. Herein, water-soluble sulfonated calix[n]arene macrocycles (SC4, SC6, and SC8) and a pH-sensitive dye (4'-hydroxy-10-methylpyranoflavylium) were investigated as host-guest systems for BA sensing. The hosts were able to bind the flavylium cation of the dye with association constants of 103 to 104 M-1. The dye complexation also allowed tuning its pKa from 6.72 (free) toward high values: 7.68 (SC4), 7.79 (SC6), and 8.45 (SC8). These data were crucial to optimize the host-guest complexes as optical sensing systems for putrescine/tyramine (pH 7.2-7.6), yielding a colorimetric redshift from yellow to red. The BA sensing was also demonstrated by fluorescence quenching for the calix[n]arene/dye complexes and fluorescence recovery after the addition of BAs. 1H NMR spectroscopy was used to demonstrate the interaction mode, confirming an encapsulation-driven mechanism. Overall, these host-guest systems demonstrated great potential for the detection of BAs, one of the main key markers of food spoilage.

Short-term effects of community-based marine reserves on green abalone, as revealed by population studies.

Marine reserves (MRs) are implemented worldwide to protect, restore, and manage marine ecosystems and species. However, it is important to document the positive effects those marine reserves have on slow-growth, temperate invertebrates such as abalone. Abalone, Haliotis spp., are marine gastropods of high economic value extracted worldwide for decades, which has led to fisheries-driven population decreases. In this work, we focused on a case study and assessed the short-term (1-2 years) effects of marine reserves established and managed by a local fishing cooperative at Guadalupe Island, Mexico. We evaluated the population status of green abalone, H. fulgens, by conducting (1) an assessment of the green abalone population around Guadalupe Island through subtidal monitoring and (2) an evaluation of the effect of two recently established marine reserves on population parameters such as the increase in density (individuals·m2), biomass, number of aggregated abalone, egg production, and proportion of individuals bigger than 150 mm (minimum harvest size) compared to fished areas. To assess the population around Guadalupe Island, we surveyed 11,160 m2 during 2020 and 2021. We recorded 2327 green abalones with a mean ± SE shell length of 135.978 ± 0.83 mm and a mean density of 0.21 ± 0.02 individuals·m2. All variables were statistically higher at the MRs except for shell length in 2021. In this work, we report for the first time the green abalone population status at Guadalupe Island and a positive short-term biological response to community-based marine reserves. This study suggests that a network of MRs combined with good management could help abalone populations in the short term in Guadalupe Island, potentially leading to more sustainable fishing practices and social-ecological resilience.

Anti-miR-873-5p improves alcohol-related liver disease by enhancing hepatic deacetylation via SIRT1.

Background & Aims: Current therapies for the treatment of alcohol-related liver disease (ALD) have proven largely ineffective. Patients relapse and the disease progresses even after liver transplantation. Altered epigenetic mechanisms are characteristic of alcohol metabolism given excessive acetate and NAD depletion and play an important role in liver injury. In this regard, novel therapeutic approaches based on epigenetic modulators are increasingly proposed. MicroRNAs, epigenetic modulators acting at the post-transcriptional level, appear to be promising new targets for the treatment of ALD. Methods: MiR-873-5p levels were measured in 23 liver tissue from Patients with ALD, and GNMT levels during ALD were confirmed using expression databases (transcriptome n = 62, proteome n = 68). High-resolution proteomics and metabolomics in mice following the Gao-binge model were used to investigate miR-873-5p expression in ALD. Hepatocytes exposed to 50 mM alcohol for 12 h were used to study toxicity. The effect of anti-miR-873-5p in the treatment outcomes of ALD was investigated. Results: The analysis of human and preclinical ALD samples revealed increased expression of miR-873-5p in the liver. Interestingly, there was an inverse correlation with NNMT, suggesting a novel mechanism for NAD depletion and aberrant acetylation during ALD progression. High-resolution proteomics and metabolomics identified miR-873-5p as a key regulator of NAD metabolism and SIRT1 deacetylase activity. Anti-miR-873-5p reduced NNMT activity, fuelled the NAD salvage pathway, restored the acetylome, and modulated the levels of NF-κB and FXR, two known SIRT1 substrates, thereby protecting the liver from apoptotic and inflammatory processes, and improving bile acid homeostasis. Conclusions: These data indicate that targeting miR-873-5p, a repressor of GNMT previously associated with NAFLD and acetaminophen-induced liver failure. is a novel and attractive approach to treating alcohol-induced hepatoxicity. Impact and implications: The role of miR-873-5p has not been explicitly examined in the progression of ALD, a pathology with no therapeutic options. In this study, inhibiting miR-873-5p exerted hepatoprotective effects against ALD through rescued SIRT1 activity and consequently restored bile acid homeostasis and attenuated the inflammatory response. Targeting hepatic miR-873-5p may represent a novel therapeutic approach for the treatment of ALD.

Correction: Yu et al. Achieving Effective Multimodal Imaging with Rare-Earth Ion-Doped CaF2 Nanoparticles. Pharmaceutics 2022, 14, 840.

There was an error in the original publication [...].

Identification and structural characterization of a novel (+)-catechin-caffeic acid adduct present in wines.

A new compound with a molecular ion mass of m/z 467 in the negative ion mode was found to occur in a white wine aged 30 months in bottle. In this latter, fragment ions compatible with the loss of a carboxylic acid (-44 a.m.u.), a caffeic acid unit (-178 a.m.u.), and a Retro-Diels Alder (-152 a.m.u.) were observed. The present work reports the synthesis of a (+)-catechin-caffeic acid adduct resulting from the condensation reaction between caffeic acid and (+)-catechin. The structural characterization by NMR showed that this adduct is formed by the linkage between carbon 8 at ring A from (+)-catechin and carbon 9 from caffeic acid. In addition, the similarity in the HPLC retention time and UV-Visible spectra of the synthesized compound with the one detected in white wine and the bottling experiments, confirms the presence of this novel (+)-catechin-derived compound in those matrices.

Stapling Amantadine to Melanostatin Neuropeptide: Discovery of Potent Positive Allosteric Modulators of the D2 Receptors.

This work describes the synthesis and pharmacological and toxicological evaluation of melanostatin (MIF-1) bioconjugates with amantadine (Am) via a peptide linkage. The data from the functional assays at human dopamine D2 receptors (hD2R) showed that bioconjugates 1 (EC50 = 26.39 ± 3.37 nM) and 2 (EC50 = 17.82 ± 4.24 nM) promote a 3.3- and 4.9-fold increase of dopamine potency, respectively, at 0.01 nM, with no effect on the efficacy (Emax = 100%). In this assay, MIF-1 was only active at the highest concentration tested (EC50 = 23.64 ± 6.73 nM, at 1 nM). Cytotoxicity assays in differentiated SH-SY5Y cells showed that both MIF-1 (94.09 ± 5.75%, p < 0.05) and carbamate derivative 2 (89.73 ± 4.95%, p < 0.0001) exhibited mild but statistical significant toxicity (assessed through the MTT reduction assay) at 200 µM, while conjugate 1 was found nontoxic at this concentration.

Correction: Yu et al. Rare-Earth-Metal (Nd3+, Ce3+ and Gd3+)-Doped CaF2: Nanoparticles for Multimodal Imaging in Biomedical Applications. Pharmaceutics 2022, 14, 2796.

In the original publication [...].

Effects of different load weights on the work performance and physiological and hematobiochemical responses in working water buffalo.

Background and Aim: Working animals are important in agriculture because they play a role in various agricultural activities, including milk and meat production. Thus, they contribute to the development of rural communities. In this study, we aimed to evaluate the effects of different load weights on the work performance and the physiological and hematological responses of working water buffalo (Bubalus bubalis). Materials and Methods: The work performances of 12 buffaloes (average weight 782.16 ± 21.62 kg) transporting 200, 350, and 500 kg of African palm fruits in metal baskets placed on their backs were evaluated. Work performance variables evaluated immediately after work were as follows: total number of trips (n), total weight (kg), distance traveled (km/day), working period (h), stopped time (h), and average speed (km/h). In addition, we evaluated physiological, biochemical, and hematological variables at three different times: before the start of work, immediately after work, and on the rest day. Results: Among the load weights, 500 kg (total load carried = 4,138.88 kg) improved work efficiency compared to 200 kg loads (total load = 3,322.22 kg) (p = 0.0281). However, 500 kg loads resulted in slower average speed (2.4 km, p = 0.0164), shorter working period (2.39 h, p < 0.0001) and distance traveled (7.29 km, p < 0.0001), and less total number of trips (8.27 trips, p < 0.0001) compared to 350 and 200 kg load weights (3.45 and 3.52 km/h, 2.55 and 2.79 h, 8.71 and 9.75 km, 10.94 and 16.61 trips, respectively); and the heaviest loads resulted in significantly higher (p < 0.005) respiratory rate, pulse, heart rate, rectal temperature, glucose, lactate dehydrogenase, creatine kinase, total protein, white blood cell count, neutrophils, lymphocytes, monocytes, eosinophils, and basophils. In contrast, was associated with lower levels of red blood cells, hemoglobin, and hematocrit compared to lower loads. All differences were more pronounced in 500 kg (p < 0.005) compared to 200 kg loads. Conclusion: Working buffaloes responded to work related to the transport of African palm fruits through various physiological, biochemical, and hematological changes. However, some variables remained close to the reference values reported in the literature for water buffaloes, and in general, all variables were reestablished during the rest day, indicating that these animals have adapted to working conditions.