AntimiR treatment corrects myotonic dystrophy primary cell defects across several CTG repeat expansions with a dual mechanism of action.

This study evaluated therapeutic antimiRs in primary myoblasts from patients with myotonic dystrophy type 1 (DM1). DM1 results from unstable CTG repeat expansions in the DMPK gene, leading to variable clinical manifestations by depleting muscleblind-like splicing regulator protein MBNL1. AntimiRs targeting natural repressors miR-23b and miR-218 boost MBNL1 expression but must be optimized for a better pharmacological profile in humans. In untreated cells, miR-23b and miR-218 were up-regulated, which correlated with CTG repeat size, supporting that active MBNL1 protein repression synergizes with the sequestration by CUG expansions in DMPK. AntimiR treatment improved RNA toxicity readouts and corrected regulated exon inclusions and myoblast defects such as fusion index and myotube area across CTG expansions. Unexpectedly, the treatment also reduced DMPK transcripts and ribonuclear foci. A leading antimiR reversed 68% of dysregulated genes. This study highlights the potential of antimiRs to treat various DM1 forms across a range of repeat sizes and genetic backgrounds by mitigating MBNL1 sequestration and enhancing protein synthesis.

Hierarchical exsolution in vertically aligned heterostructures.

Metal nanoparticle exsolution from metal oxide hosts has recently garnered great attention to improve the performance of energy conversion and storage devices. In this study, the nickel exsolution mechanisms in a vertically aligned nanostructure (VAN) thin film of heteroepitaxial (Sr0.9Pr0.1)0.9Ti0.9Ni0.1O3-δ-Ce0.9Gd0.1O1.95 with a columnar architecture was investigated for the first time. Experimental results and Density Functional Theory (DFT) calculations reveal that the multiple vertical interphases in a VAN with a hierarchical arrangement provide faster and more selective Ni diffusion pathways to the surface than traditional bulk diffusion in epitaxial films. Kinetic studies conducted at different temperatures and times indicate that the nucleation process of the exsolved metal nanoparticles primarily takes place at the surface through the phase boundaries of the columns. The vertical strain is crucial in preserving the film's microstructure, yielding a robust heteroepitaxial architecture after reduction. This innovative heteromaterial opens up new possibilities for designing efficient devices through advanced structural engineering to achieve controlled nanoparticle formation.

Population screening for colorectal cancer in Spain - A public health success with remaining challenges.

The development of organized colorectal cancer screening programs in developed countries has led to a reduction in the incidence and mortality of this cancer. The implementation of the national screening program in Spain represents a major achievement in public health, aligning us with other European countries, but it still faces ongoing challenges. One of the main issues is the low participation rate of the eligible population, averaging 45%, with significant variation between autonomous communities. This requires allocating specific resources to overcome barriers that hinder participation. Another key aspect to improving the screening program is extending the screening age up to 74 years, in line with the recommendations of the European Union, as many member states are already doing.

Table of life for beneficiaries of the Institute of Security and Social Services of State Workers (ISSSTE), Mexico 2021.

OBJECTIVE: Determine the life expectancy in the covered population of the Institute of Security and Social Services of State Workers in México for 2021. METHOD: We used the abrogated method from Reed-Merrel, for calculate the life expectancy in age groups. RESULTS: By 2021, life expectancy general was 79.51 years; 81.40 years and 78.91 years for woman and men, respectively. CONCLUSIONS: The calculated life expectancy not show a reduction in the population of federal and State employees in Mexico.

OBJETIVO: Determinar la esperanza de vida en la población amparada del Instituto de Seguridad y Servicios Sociales de los Trabajadores del Estado en México para 2021. MÉTODO: Se utilizó el método abreviado de Reed-Merrel para calcular la esperanza de vida en grupos quinquenales de edad. RESULTADOS: Para 2021, la esperanza de vida general fue de 79.51 años; de 81.40 años y 78.91 años para mujeres y para hombres, respectivamente. CONCLUSIONES: La esperanza de vida calculada no presentó reducción después de la pandemia de COVID-19 para la población de empleados federales de México en 2021.

Deletion of exons 45 to 55 in the DMD gene: from the therapeutic perspective to the in vitro model.

BACKGROUND: Gene editing therapies in development for correcting out-of-frame DMD mutations in Duchenne muscular dystrophy aim to replicate benign spontaneous deletions. Deletion of 45-55 DMD exons (del45-55) was described in asymptomatic subjects, but recently serious skeletal and cardiac complications have been reported. Uncovering why a single mutation like del45-55 is able to induce diverse phenotypes and grades of severity may impact the strategies of emerging therapies. Cellular models are essential for this purpose, but their availability is compromised by scarce muscle biopsies. METHODS: We introduced, as a proof-of-concept, using CRISPR-Cas9 edition, a del45-55 mimicking the intronic breakpoints harboured by a subset of patients of this form of dystrophinopathy (designing specific gRNAs), into a Duchenne patient's cell line. The edited cell line was characterized evaluating the dystrophin expression and the myogenic status. RESULTS: Dystrophin expression was restored, and the myogenic defects were ameliorated in the edited myoblasts harbouring a specific del45-55. Besides confirming the potential of CRISPR-Cas9 to create tailored mutations (despite the low cleavage efficiency of our gRNAs) as a useful approach to generate in vitro models, we also generated an immortalized myoblast line derived from a patient with a specific del45-55. CONCLUSIONS: Overall, we provide helpful resources to deepen into unknown factors responsible for DMD-pathophysiology.

Level of Compliance with Spanish Guideline Recommendations in the Management of Asthma.

Objective: To describe the clinical management of asthmatic patients and the level of compliance with the Guía Española para el Manejo del Asma (GEMA, Spanish Guidelines for Asthma Management) in the influential al area of Virgen de la Victoria University Hospital, Málaga (Spain). Materials and Methods: Descriptive, cross-sectional study in asthma patients from the Virgen de la Victoria University Hospital influential area, Málaga (Spain). Asthma Control Test (ACT) results, demographic and clinical characteristics, and GEMA indicators of quality asthma care were obtained from each participating patient. Results: Two hundred and eighty-eight patients (64.2% woman, mean age 48.6 years) were included in the study. The evaluation of the level of compliance with the GEMA quality indicators showed that in 20.8% of the patients the diagnosis of asthma was confirmed by spirometry and bronchodilator test, and 52.5% of the patients with suspicion of allergic asthma performed a sensitization test. Among the smoker patients, 76.3% were recommended to stop smoking; 14.2% of the patients received a paper-based educational plan. Regarding asthma treatment, 92.3% of patients with persistent asthma received inhaled glucocorticoids (IGC). IGC and LABA were the most common treatments for asthma, used in combination (ICG + LABA) in 139 (48.3%) patients; 32.6% discontinued SABA treatment during the study period. Asthma was well controlled in 54.9% of the patients (ACT score ≥20), and 13.9% of the patients registered exacerbations during the previous year with a mean (SD) of 1.38 (0.74) events. Conclusion: Most of the GEMA quality indicators of asthma care are not followed in real-world clinical practice. Specialists select the best option among the available treatments but there is an unmet need for training plans in other relevant aspects, such as diagnosis and/or disease monitoring.

Tunable Ferroionic Properties in CeO2/BaTiO3 Heterostructures.

Ferroionic materials combine ferroelectric properties and spontaneous polarization with ionic phenomena of fast charge recombination and electrodic functionalities. In this paper, we propose the concept of tunable polarization in CeO2-δ (ceria) thin (5 nm) films induced by built-in remnant polarization of a BaTiO3 (BTO) ferroelectric thin film interface, which is buried under the ceria layer. Upward and downward fixed polarizations at the BTO thin film (10 nm) are achieved by the lattice termination engineering of the SrO or TiO2 terminated Nb:SrTiO3 (NSTO or STN) substrate. We find that the ceria layer punctually replicates the polarization of the BTO interface via a dynamic reconfiguration of its intrinsic defects, i.e., oxygen vacancies and small polarons. Tunable oxidative or reducing properties (redox) also arise at the surface from the built-in polarization. Opposite polarities at the ceria termination tune the chemo-physical dynamics toward water molecule adsorbates. The inversion of the surface potential leads to a modulation of the water adsorption-desorption equilibrium and water ionization (splitting) redox overpotentials within ±400 mV at room temperature, depending on the ceria termination's charges. Such tunability opens up the perspectives of using ferroionics for wireless electrochemically enhanced catalysis.

Putting computational models of immunity to the test - an invited challenge to predict B. pertussis vaccination outcomes.

Systems vaccinology studies have been used to build computational models that predict individual vaccine responses and identify the factors contributing to differences in outcome. Comparing such models is challenging due to variability in study designs. To address this, we established a community resource to compare models predicting B. pertussis booster responses and generate experimental data for the explicit purpose of model evaluation. We here describe our second computational prediction challenge using this resource, where we benchmarked 49 algorithms from 53 scientists. We found that the most successful models stood out in their handling of nonlinearities, reducing large feature sets to representative subsets, and advanced data preprocessing. In contrast, we found that models adopted from literature that were developed to predict vaccine antibody responses in other settings performed poorly, reinforcing the need for purpose-built models. Overall, this demonstrates the value of purpose-generated datasets for rigorous and open model evaluations to identify features that improve the reliability and applicability of computational models in vaccine response prediction.

Epigenetic patient stratification via contrastive machine learning refines hallmark biomarkers in minoritized children with asthma.

Identifying and refining clinically significant patient stratification is a critical step toward realizing the promise of precision medicine in asthma. Several peripheral blood hallmarks, including total peripheral blood eosinophil count (BEC) and immunoglobulin E (IgE) levels, are routinely used in asthma clinical practice for endotype classification and predicting response to state-of-the-art targeted biologic drugs. However, these biomarkers appear ineffective in predicting treatment outcomes in some patients, and they differ in distribution between racially and ethnically diverse populations, potentially compromising medical care and hindering health equity due to biases in drug eligibility. Here, we propose constructing an unbiased patient stratification score based on DNA methylation (DNAm) and utilizing it to refine the efficacy of hallmark biomarkers for predicting drug response. We developed Phenotype Aware Component Analysis (PACA), a novel contrastive machine-learning method for learning combinations of DNAm sites reflecting biomedically meaningful patient stratifications. Leveraging whole-blood DNAm from Latino (discovery; n=1,016) and African American (replication; n=756) pediatric asthma case-control cohorts, we applied PACA to refine the prediction of bronchodilator response (BDR) to the short-acting ß2-agonist albuterol, the most used drug to treat acute bronchospasm worldwide. While BEC and IgE correlate with BDR in the general patient population, our PACA-derived DNAm score renders these biomarkers predictive of drug response only in patients with high DNAm scores. BEC correlates with BDR in patients with upper-quartile DNAm scores (OR 1.12; 95% CI [1.04, 1.22]; P=7.9 e-4) but not in patients with lower-quartile scores (OR 1.05; 95% CI [0.95, 1.17]; P=0.21); and IgE correlates with BDR in above-median (OR for response 1.42; 95% CI [1.24, 1.63]; P=3.9e-7) but not in below-median patients (OR 1.05; 95% CI [0.92, 1.2]; P=0.57). These results hold within the commonly recognized type 2 (T2)-high asthma endotype but not in T2-low patients, suggesting that our DNAm score primarily represents an unknown variation of T2 asthma. Among T2-high patients with high DNAm scores, elevated BEC or IgE also corresponds to baseline clinical presentation that is known to benefit more from biologic treatment, including higher exacerbation scores, higher allergen sensitization, lower BMI, more recent oral corticosteroids prescription, and lower lung function. Our findings suggest that BEC and IgE, the traditional asthma biomarkers of T2-high asthma, are poor biomarkers for millions worldwide. Revisiting existing drug eligibility criteria relying on these biomarkers in asthma medical care may enhance precision and equity in treatment.

Microbial community response to temperature reduction during anaerobic treatment of long chain fatty acids-containing wastewater.

Acclimating mesophilic biomass to low temperatures have been used to start-up psychrophilic anaerobic reactors, but limited microbial information is available during the acclimation. To investigate microbial responses to temperature reductions, duplicate lab-scale anaerobic digestion (AD) reactors were operated for 166 days, with the temperature being reduced from 37°C to 15°C, using synthetic long chain fatty acid (LCFA)-containing wastewater as the feedstock. The acclimated biomass at 15°C exhibited efficient removal of organic matter (total COD>75%, soluble COD>88%, and LCFA>99%). Temperature reductions lead to significant reductions in microbiome diversity. Fermentative bacteria were highly dynamic and functional redundant during temperature reductions. Smithella was the dominant syntrophic bacteria involved in LCFA degradation coupled with Methanothrix and Methanocorpusculum at 15°C. Membrane modifications and compatible cellular solutes production were triggered by temperature reductions as microbial response to cold stress. This study provided molecular insights in microbial acclimation to low temperatures for psychrophilic AD.

Effects of monensin and a blend of magnesium oxide on performance, feeding behavior, and rumen morphometrics of Zebu beef cattle fed high-starch diets.

This study aimed to evaluate the effects of a blend of different sources of magnesium oxide associated or not with monensin, on productive, ruminal, and nutritional parameters of steers. Eighty-four Nellore steers with an initial body weight (BW) of 367.3 ±â€…37.9 kg were allocated to one of 28 pens, with three steers per pen. Each pen was considered an experimental unit. Using a completely randomized design with a 2 × 2 factorial arrangement, the following treatments were assigned to each pen: 1) Control (CON)-a basal diet without additive inclusion; 2) Magnesium oxide blend (MG)-basal diet plus a magnesium-based product (pHix-up, Timab Magnesium, Dinard, France) provided at 0.50% of dry matter (DM); 3) Monensin (MON)-basal diet plus 25 mg/ kg of DM of sodium monensin (Rumensin, Elanco Animal Health, Greenfield, IN); and 4) MG association with MON-basal diet plus MG + MON, at the same doses of the individual treatments. The experimental period lasted 100 d. Blood samples were collected on days 0, 13, and 70 to determine d-lactate levels. Daily feed intake was recorded, and animal ingestive behavior was visually observed on days 66 and 67. On day 70, skeletal muscle tissue samples were obtained through biopsy for gene expression analysis. At the end of the experimental period, carcass ultrasonography was conducted. Subsequently, the steers were slaughtered, and rumen epithelium samples were collected for morphometric analysis. The use of monensin, of magnesium oxide blend, and their interactions, were treated as fixed effects, while the pens were considered as a random effect. Statistical differences were considered when P < 0.05. Steers-fed MG-containing diets consumed approximately 0.6 kg more DM per day than those fed diets without this additive (P = 0.01; 11.3 vs. 11.9 kg/d). The inclusion of MG in the diet increased (P = 0.02) the average daily gain. There was a greater Longissimus muscle area (LMA) and LMA per 100 kg of BW (P ≤ 0.03) for steers-fed diets with MG. Steers-fed MON exhibited reduced mRNA expression of the Atrogin-1 and mTOR compared to steers-fed MG + MON diets (MON × MG: P ≤ 0.04). Steers-fed MON had 6.9% greater feed efficiency (P = 0.02). Papillae width was lesser for CON than other treatments (MON × MG: P = 0.02). In conclusion, the magnesium oxide blend improved performance and carcass traits in high-energy feedlot diets, while monensin enhanced feed efficiency, suggesting potential for their use as alternatives or complements in beef cattle nutrition.

Generation of arbitrarily patterned polarizers using 2-photon polymerization.

Patterned polarizers are prepared using liquid crystals (LC) doped with a black dichroic dye and in combination with a linear polarizer. The pattern is achieved with a nanostructured LC alignment surface, that is generated using a two-photon polymerization direct laser write (2PP-DLW). This technique creates a pattern of high-resolution grooves in the photoresist at any arbitrary angle. The angle governs the LC orientation at any substrate surface point, determining the transmitted light linear polarization angle. This paper presents the first use of a 2PP-DLW cured positive tone photoresist for dichroic dye-doped LC alignment. Two complementary photoresists have been employed: conventional negative tone SU-8 photoresist and, in this context novel, positive tone S1805 photoresist. The alignment quality of the polarizers has been assessed by analyzing the transmission using an additional polarizer. For SU-8, the resulting grayscale pattern and a contrast ratio (CR) of 14 has measured. The uniformity of the alignment has been measured to be 65% using normalized Shannon entropy (H). For S1805, a CR of 37 was measured, and a uniformity of 63% was obtained. 2PP-DLW allows for shaping complex patterns in submicron dimensions and for the fabrication of arbitrarily patterned polarizers and other LC devices.

Health-Related Quality of Life and Satisfaction in Atrial Fibrillation Patients on Anticoagulant Therapy: Differences between Vitamin K Antagonists and Direct Oral Anticoagulants; Results from the Multicentre REGUEIFA Registry.

Background: Oral anticoagulation (OAC) is pivotal in the clinical management of atrial fibrillation (AF) patients. Vitamin K antagonists (VKAs) and direct oral anticoagulants (DOACs) prevent thromboembolic events, but information about the quality of life (QoL) and patient satisfaction in relation with the anticoagulant treatment is limited. Methods: REGUEIFA is a prospective, observational, and multicentre study that included patients with AF treated by cardiologists. We included patients treated with VKAs or DOACs. The EuroQol-5D (EQ-5D) questionnaire evaluated QoL, and the Anti-Clot Treatment Scale (ACTS) questionnaire investigated patient satisfaction with OAC. Results: A total of 904 patients were included (532 on VKA and 372 on DOACs). A total of 846 patients completed the EQ-5D questionnaire, with results significantly worse in patients on VKAs than on DOACs: more mobility limitations (37.6% vs. 24.2%, p < 0.001), more restriction in usual activities (24.7% vs. 18.3%, p = 0.026), more pain/discomfort (31.8% vs. 24.2%, p = 0.015), a lower visual analogue scale (VAS) score (66.4 ± 16.21 vs. 70.8 ± 15.6), and a lower EQ-D5 index (0.79 ± 0.21 vs. 0.85 ± 0.2, p < 0.001). After adjusting for baseline characteristics, VKA treatment was not an independent factor towards worse EQ-5D results. Also, 738 patients completed the ACTS questionnaire, and burden and profit scores were lower in patients on VKAs than for DOACs (52.1 ± 8.4 vs. 55.5 ± 6.8, p < 0.001 and 11.1 ± 2.4 vs. 11.8 ± 2.6, p < 0.001, respectively). The negative impact score was higher for VKAs than for DOACs (1.8 ± 1.02 vs. 1.6 ± 0.99, p < 0.001), with a general positive impact score lower for VKAs than for DOACs (3.6 ± 0.96 vs. 3.8 ± 1.02, p < 0.001). Conclusions: Patients on VKA have more comorbidity and worse EQ-5D and VAS scores than those on DOACs. VKA has a greater burden and higher negative impact on the patient's life than DOACs.

Early life movements and mortality of Egyptian vultures: Implications for transcontinental conservation.

Understanding the movements and mortality of individuals across different life stages is crucial for the effective conservation of wild populations. We used data from 32 Egyptian vultures (Neophron percnopterus) tagged with GPS transmitters as nestlings in three Iberian breeding areas to study their dependence period, migration routes, movements in Africa, and mortality at each stage. Our results show no significant differences in the timing of nest departure or the duration of the dependence period between individuals of different sexes or breeding nuclei. Most juveniles migrated to sub-Saharan Africa in their first year, but some (3 of 32, 9.4%) remained in the Iberian Peninsula. Individuals that migrated to Africa did so annually, while those remaining in Iberia never migrated to the Sahel, indicating distinct migratory and non-migratory strategies. Non-migratory individuals consistently moved northward during the breeding season to their natal territories. Siblings did not coordinate their migration strategy or timing. All juveniles showed extensive overlap in the vast areas used in Africa, where females arrived before males, and in the Iberian Peninsula. Our study also revealed that no juveniles died immediately after fledging, but that none of the tagged individuals lived more than 7 years or were recruited as breeders. Although most casualties occurred during the longer stay in the Sahel, the mortality rate was highest during the few days of the first migration. Our results show that despite small variations in movement patterns between breeding nuclei and sexes, Egyptian vultures face similar challenges during the years before recruitment as breeders, mostly determined by their migratory strategy. These findings are relevant for designing conservation strategies, both in breeding areas and, more importantly, in wintering areas and along migration pathways. Such strategies will significantly impact the entire Iberian population of this endangered species.

Different Physiochemical Properties of Novel Fibre Sources in the Diet of Weaned Pigs Influence Animal Performance, Nutrient Digestibility, and Caecal Fermentation.

The effect of including micronised fibre sources (FS) differing in fermentability and hydration capacity (HC) on growth performance, faecal digestibility, and caecal fermentation was investigated in piglets. There were four dietary treatments: a control diet (CON) and three treatments differing in the HC and fermentability of FS added at 1.5% to prestarter (28-42 d) and starter (42-61 d) diets. These were: LHC (low-HC by-product-based insoluble fibre (IF) with a prebiotic fraction (PF) from chicory root); MHC (medium-HC by-product-based IF with a PF); and HHC (high-HC non-fermentable wood-based IF with no PF). There were eight replicates per treatment. Over the entire period, LHC and MHC piglets showed a 10% increase in daily growth and feed intake (p ≤ 0.019) and tended to have a reduced feed conversion ratio (p = 0.087) compared to HHC piglets. At 42 d, faecal protein digestibility increased by 5% in the LHC and MHC groups compared with the HHC group (p = 0.035) and did not differ from the CON group. Both LHC and MHC fibres were more fermented in vitro with caecal inocula from 61 d old piglets than HHC fibre (p ≤ 0.003). These results suggest that balanced soluble and insoluble fibre concentrates can improve piglet performance.

Cardiac-Sympathetic Contractility and Neural Alpha-Band Power: Cross-Modal Collaboration during Approach-Avoidance Conflict.

As evidence mounts that the cardiac-sympathetic nervous system reacts to challenging cognitive settings, we ask if these responses are epiphenomenal companions or if there is evidence suggesting a more intertwined role of this system with cognitive function. Healthy male and female human participants performed an approach-avoidance paradigm, trading off monetary reward for painful electric shock, while we recorded simultaneous electroencephalographic and cardiac-sympathetic signals. Participants were reward sensitive but also experienced approach-avoidance "conflict" when the subjective appeal of the reward was near equivalent to the revulsion of the cost. Drift-diffusion model parameters suggested that participants managed conflict in part by integrating larger volumes of evidence into choices (wider decision boundaries). Late alpha-band (neural) dynamics were consistent with widening decision boundaries serving to combat reward sensitivity and spread attention more fairly to all dimensions of available information. Independently, wider boundaries were also associated with cardiac "contractility" (an index of sympathetically mediated positive inotropy). We also saw evidence of conflict-specific "collaboration" between the neural and cardiac-sympathetic signals. In states of high conflict, the alignment (i.e., product) of alpha dynamics and contractility were associated with a further widening of the boundary, independent of either signal's singular association. Cross-trial coherence analyses provided additional evidence that the autonomic systems controlling cardiac-sympathetics might influence the assessment of information streams during conflict by disrupting or overriding reward processing. We conclude that cardiac-sympathetic control might play a critical role, in collaboration with cognitive processes, during the approach-avoidance conflict in humans.

DNA Origami - Lipid Membrane Interactions Controlled by Nanoscale Sterics.

DNA nanostructures designed to interact with bilayer membranes are of fundamental interest as they mimic biological cytoskeletons and other membrane-associated proteins for applications in synthetic biology, biosensing, and biological research. Yet, there is limited insight into how the binary interactions are influenced by steric effects produced by 3D geometries of DNA structures and membranes. This work uses a 3D DNA nanostructure with membrane anchors in four different steric environments to elucidate the interaction with membrane vesicles of varying sizes and different local bilayer morphology. It is found that interactions are significantly affected by the steric environments of the anchors -often against predicted accessibility- as well as local nanoscale morphology of bilayers rather than on the usually considered global vesicle size. Furthermore, anchor-mediated bilayer interactions are co-controlled by weak contacts with non-lipidated DNA regions, as showcased by pioneering size discrimination between 50 and 200 nm vesicles. This study extends DNA nanotechnology to controlled bilayer interactions and can facilitate the design of nanodevices for vesicle-based diagnostics, biosensing, and protocells.

Full-scale study on high-rate low-temperature anaerobic digestion of agro-food wastewater: process performances and microbial community.

The fast-growing global population has led to a substantial increase in food production, which generates large volumes of wastewater during the process. Despite most industrial wastewater being discharged at lower ambient temperatures (<20 °C), majority of the high-rate anaerobic reactors are operated at mesophilic temperatures (>30 °C). High-rate low-temperature anaerobic digestion (LtAD) has proven successful in treating industrial wastewater both at laboratory and pilot scales, boasting efficient organic removal and biogas production. In this study, we demonstrated the feasibility of two full-scale high-rate LtAD bioreactors treating meat processing and dairy wastewater, and the microbial communities in both reactors were examined. Both reactors exhibited rapid start-up, achieving considerable chemical oxygen demand (COD) removal efficiencies (total COD removal >80%) and generating high-quality biogas (CH4% in biogas >75%). Long-term operations (6-12 months) underscored the robustness of LtAD bioreactors even during winter periods (average temperature <12 °C), as evidenced by sustained high COD removal rates (total COD removal >80%). The stable performance was underpinned by a resilient microbial community comprising active acetoclastic methanogens, hydrolytic, and fermentative bacteria. These findings underscore the feasibility of high-rate low-temperature anaerobic wastewater treatment, offering promising solutions to the zero-emission wastewater treatment challenge.