Biology and Life Sciences

Abstract: Background: Effective influenza molecular epidemiology depends on curated, temporally representative sequence datasets. The transition from raw GISAID or NCBI/GenBank batch downloads to analysis-ready FASTA often requires command-line pipeline configuration or custom scripting, which may be inaccessible to some field and clinical virologists. To our knowledge, a targeted, non-systematic comparison of widely used viral-genome tools did not identify a single browser-delivered workflow combining supported GISAID-style and NCBI/GenBank-compatible header parsing, interactive human-in-the-loop temporal subsampling, exact-sequence identity-group analysis, and six selectable language catalogues.Methods: We developed VirSift (Viral Sequence Intelligence and Filtering Toolkit), a seven-page Streamlit web application implementing: (1) a custom parser handling four supported GISAID-style and NCBI/GenBank-compatible FASTA header variants with host inference and species normalization; (2) vectorized quality filtering and MD5-based identification of exact-sequence redundancy; (3) five documented human-in-the-loop (HITL) temporal sampling strategies with an operational date-span recommendation; (4) ten Plotly visualization types, including a configurable N-level Sankey flow diagram and an exact-sequence persistence matrix; and (5) structured export to FASTA, CSV, and segment-organized ZIP bundles. The interface exposes six structurally aligned 817-key language catalogues, all of which contain complete native‑language values with no English fallback. This tool-description paper documents implemented functions and includes a limited H3N2 demonstration and multi-file round-trip check; comprehensive analytical, usability, performance, and multilingual validation is reserved for separate work.Results: Applied to 59 H3N2 haemagglutinin sequences from Novosibirsk, Russia, spanning October 2021 to April 2025, VirSift identified 12 records redundant by exact nucleotide identity (20.3% of records), retaining 47 unique sequences across 9 calendar months. The observed 1,279-day collection-date span was assigned to the software-defined long-span category (labelled "Endemic" in the version 1.0 interface), for which Custom Checkpoints was recommended. The Molecular Timeline reported 47 exact-sequence identity groups. Clade 3C.2a1b.2a.2a.3a.1 accounted for 69.5% of records. Multi-file ingestion and metadata-based split export were verified by reconciliation of record counts across five source files. All described operations were completed without programming.Conclusions: VirSift provides a reproducible and auditable browser-delivered workflow for pre-phylogenetic curation of influenza sequence datasets. Its six selectable language catalogues, supported parsing rules, HITL sampling options, visualization modules, and structured exports are described here as implemented capabilities. The H3N2 case study and count-reconciliation exercise demonstrate workflow operation but do not constitute comprehensive software validation. VirSift may support institutions seeking an accessible interface for documented sequence-data curation before downstream phylogenetic analysis.

Abstract: Osteoporosis (OP) is a systemic skeletal disease characterized by low bone mass and micro-architectural deterioration of bone tissue, but no effective clinical therapies exist. To address this unmet need, we employed a computational pipeline drug repositioning method based on single-cell data and Mendelian randomization analysis to screen potential candidate drugs for the treatment of osteoporosis. Quinidine was identified as a potential therapeutic agent for osteoporosis. Mendelian Randomization analysis indicated a causal relationship between Quinidine's drug target SCN5A and osteoporosis. Larval zebrafish experiments confirmed that Quinidine can ameliorate the Dexamethasone-induced osteoporosis model and promote cranial bone mineralization. qPCR showed that Quinidine can promote osteoblast-related gene expression and inhibit osteoclast-related genes expression. This study systematically revealed Quinidine as a potential drug in the prevention and treatment of osteoporosis through an integrated strategy of single-cell drug repositioning, genetic causal inference, and animal model validation.

Abstract: The endocannabinoid system (ECS) has been extensively mapped at the level of receptors, ligands, enzymes, and signaling pathways, forming a detailed component inventory of a major homeostatic network. However, prevailing ECS models largely omit the nutritional substrates required to sustain ligand synthesis, membrane composition, signaling capacity, and regenerative function, leaving the system operationally incomplete from a systems-biology perspective. This Hypothesis identifies this gap by integrating evidence from nutritional biochemistry, lipid metabolism, and regenerative physiology, and argues that inclusion of dietary inputs is necessary to advance toward a nutritionally complete model of the ECS. By reframing the ECS as a metabolically sustained regulatory network rather than a purely signaling system, this framework has implications for understanding resilience, regeneration, and system failure under chronic stress, nutritional insufficiency, and environmental disruption. This synthesis is intended as a hypothesis-generating foundation to guide future experimental and clinical investigation.

Abstract: Acidic cannabinoids (e.g., THCA, CBDA) are the dominant phytoconstituents in Cannabis sativa L. and serve as precursors to neutral forms (THC, CBD) via decarboxylation. This is the third work in an integrated series exploring how dietary cannabis inputs interact with Endocannabinoid System (ECS) pathways. This paper examines the role of physiological environments — stomach acidity, blood pH, and hepatic metabolism — in determining the fate, bioavailability, and independent pharmacological activity of ingested acidic cannabinoids. Integrating organic chemistry and pharmacokinetics, the study finds that gastric decarboxylation of acidic cannabinoids is negligible: the reaction's activation energy barrier is largely insurmountable at physiological temperature, and gastric acidity plays no direct catalytic role in overcoming it. Upon absorption, systemic blood pH (7.35–7.45) further stabilizes acidic cannabinoids, which exist almost entirely (>99%) as non-reactive carboxylate anions. Hepatic first-pass metabolism preserves this pattern: acidic cannabinoids are predominantly conjugated intact via UGT1A9, in contrast to neutral THC, which undergoes CYP-mediated oxidation to its own active and inactive metabolites; CBD's position between these two pathways remains unresolved in the literature. The gut microbiome acts as a secondary modulator via β-glucuronidase-mediated deconjugation, potentially enabling enterohepatic recirculation and extending systemic exposure, though this mechanism remains an inference by analogy for cannabinoids specifically rather than a directly demonstrated finding. Beyond their role as precursors, THCA and CBDA act directly on distinct molecular targets independent of any conversion to their neutral forms: CBDA through selective COX-2 inhibition and 5-HT1A receptor potentiation, and THCA through potent PPARγ agonism and weak partial TRPA1 activation. Direct detection of THCA in oral fluid following cannabis use corroborates delivery of the intact acidic form to peripheral tissues. Taken together, these findings indicate that ingested acidic cannabinoids reach systemic circulation and target tissues largely unconverted. Their therapeutic relevance is therefore shaped primarily by their own direct pharmacological activity and by metabolic and microbial processing, rather than by thermal decarboxylation to THC or CBD — a transformation the body's physiological conditions do not provide.

Abstract: While it remains highly challenging to create oral medicines against so-called “undruggable” targets, progress is accelerating. Using multiple approaches to analyze druggability, we estimate that 17–32% of the genome may now plausibly be considered druggable. This range was derived by extrapolating evidence of tractability from targets with high-quality ligands to related genes within several distinct curated classification schemes. Four distinct scientific advances underlie this progress: more aggressive exploration of chemical space; an improved ability to deliver pharmacologically relevant doses of these medicines; a vastly expanded range of mechanistic drug modalities; and dramatically improved biochemical and cellular screening tools to identify and characterize promising ligands. Underlying these scientific advances is a more optimistic mindset that supports the pursuit of challenging targets of high medical interest. We review recent examples, document progress, and identify remaining bottlenecks. While many challenges undeniably remain, the rapid progress suggests that we need to reconsider the term “undruggable.”

Abstract: Classical lipoproteins are classified by density, size, lipid composition, apolipoprotein content, metabolic origin, and receptor routing. These variables explain much of lipid transport but do not establish whether supramolecular organization of the lipid interface adds causal information about particle behavior. We hypothesize that classical lipoproteins are apolipoprotein-scaffolded lipid-state particles in which a measurable, perturbable, and persistent lipid-interface state cooperates with a biogenetic scaffold to influence stability, extracellular remodeling, routing, and recipient-cell responses. Source-state continuity may be material, as during ATP-binding cassette transporter A1-mediated transfer of cellular phospholipids and cholesterol to apolipoprotein A-I, or configurational, as when hepatic or intestinal physiology biases microsomal triglyceride transfer protein-dependent assembly of apolipoprotein B-containing particles. Apolipoprotein B provides a non-exchangeable scaffold with high continuity, whereas apolipoprotein A-I forms a more adaptable scaffold that is initiated by ATP-binding cassette transporter A1-dependent lipidation and subsequently remodeled by lecithin–cholesterol acyltransferase and other plasma factors. After biogenesis, exchangeable apolipoproteins, enzymes, and other associated proteins form an editable identity layer. Structural studies of apolipoprotein B, high-density lipoprotein biogenesis and heterogeneity, and inflammatory remodeling of high-density lipoprotein provide mechanistic precedents, but do not establish the proposed incremental causal layer. The hypothesis predicts that controlled differences in interfacial packing, accessibility, topology, oxidation, or electrostatics will alter protein acquisition, routing, or function after particle number, size, bulk composition, scaffold abundance, and established receptor pathways are controlled. It is weakened if these variables add no reproducible causal or predictive information beyond conventional lipoprotein biology.

Abstract: Background: Adolescence is a critical developmental stage during which preventive mental healthcare interventions may strengthen psychological resources and reduce vulnerability. School-based mental health promotion programmes delivered by mental health nurses have emerged as promising strategies to improve adolescents' well-being. This pilot study evaluated the effectiveness of the Brilho da Mente Program in promoting Positive Mental Health (PMH), hope, reducing psychological vulnerability, and decreasing mental health stigma among adolescents. Methods: A quasi-experimental pilot study with a pre-test/post-test design was conducted with a convenience sample of 91 Portuguese adolescents enrolled in the 8th and 9th grades. Participants were allocated by classroom clusters to an intervention group (n = 71) or a comparison group (n = 20). The intervention consisted of an eight-session school-based PMH programme delivered by specialist mental health nurses during Citizenship classes. Outcomes were assessed using the Positive Mental Health Questionnaire (PMHQ), the Psychological Vulnerability Scale (PVS), the Attribution Questionnaire (AQ-8-C), and the Hope Thermometer. Paired-samples t-tests and Pearson's correlation analyses were performed. Results: Participants presented high baseline levels of Positive Mental Health. Following the intervention, hope significantly increased (p = 0.03), whereas psychological vulnerability significantly decreased (p < 0.05). Although improvements in overall PMH and reductions in mental health stigma were observed, these changes did not reach statistical significance. Positive Mental Health was moderately and negatively correlated with psychological vulnerability (r = −0.629, p < 0.001). Conclusions: The Brilho da Mente Program showed promising effects in strengthening adolescents' psychological resources, particularly hope and psychological vulnerability. These findings support the potential contribution of school-based mental health nursing interventions to improving preventive healthcare services for adolescents, although larger controlled studies with long-term follow-up are required.

Abstract: Pain is a key symptom in hidradenitis suppurativa (HS) and up to one-third of patients use cannabis. Despite this clinical signal, the endocannabinoid system (ECS) and molecular nociceptive networks remain essentially uncharacterized. Transcriptomic comparisons between HS patients and healthy controls were conducted utilizing bulk RNA-seq (blood and skin) alongside cutaneous scRNA-seq. A curated 134-gene panel mapped the peripheral nociceptive network, encompassing neurotrophic and immune signaling, extracellular matrix remodeling, and bioactive lipid mediators. Compared with healthy controls, bulk transcriptomic profiling of HS patients revealed nine ECS-related genes differentially expressed in lesional skin and eight in the blood. Additionally, seven distinct bioactive lipid mediator genes were dysregulated within HS lesions, suggesting a lesion-level shift in prostaglandin, thromboxane, and leukotriene biosynthesis. At single-cell resolution, of the five major cell populations analyzed, keratinocytes and T cells played a relatively minor role in the broader pain signature. In contrast, mononuclear phagocytes (MNPs) and fibroblasts emerged as central cellular amplifiers of peripheral nociception. MNPs significantly dysregulated 48 panel genes, including the biosynthetic machinery for nociceptive lipid mediators across the endocannabinoid, prostaglandin, leukotriene, and thromboxane axes. Moreover, MNPs amplify pain networks by inducing pronociceptive chemokines, cytokines, alarmins, neuropeptides, mechanotransducers, and matrix-remodeling proteases, a shift potentiated by substantial hypoxic and oxidative adaptations. Fibroblasts exhibited a broad transcriptomic shift altering 46 pain-network genes, dysregulating matrix-remodeling, inflammatory, and lipid mediator pathways. Critically, the alteration of neurotrophic factors, plasminogen activators, and axon guidance molecules suggest a permissive environment for disorganized hyperinnervation. Furthermore, the sensory ion channel TRPA1 emerged as a significantly upregulated stromal sensor, with this entire nociceptive signature correlating with CXCL13-positive immunofibroblasts. Additionally, leveraging the high B cell density in HS lesions enabled a novel intratissue trajectory analysis, revealing constitutive lipid mediator expression without significant transcriptomic shifts across memory, plasmablast, and plasma cell maturation stages. Finally, in silico intersection of this pathogenic profile with the druggable proteome identified precision therapeutic candidates, encompassing targetable GPCRs, FDA-approved monoclonal antibodies against inflammatory nodes, and experimental sphingolipid and leukotriene cascade inhibitors. Collectively, this study establishes the first systematic transcriptomic map of the peripheral nociceptive network and endocannabinoidome in hidradenitis suppurativa, providing both a mechanistic framework for disease-associated nociception and a biological rationale for targeted therapeutic intervention.

Abstract: Radiological emergencies necessitate biomarkers that not only estimate absorbed ionizing radiation (IR) dose but also guide timely interventions to prevent or delay multi-organ injury. Conventional LC–MS-based metabolomics of bulk plasma is constrained by matrix effects that mask low-abundance species. Extracellular vesicles (EVs) constitute a metabolically enriched, underexplored compartment that can provide complementary insight into systemic metabolic and redox responses to IR. Female WAG/RijCmcr rats were exposed to 13.0 Gy leg-out partial-body X-rays and treated with one of three activated protein C (APC) variants-rat wild-type (WT), rat 3K3A-APC, or human WT APC-administered 24- and 48-hours post-irradiation. Longitudinal plasma collections (days 1, 14, 30, and 90) were subjected to metabolomic and lipidomic profiling of whole plasma and matched EV-enriched fractions to define signatures of acute radiation syndrome (ARS) and delayed effects of acute radiation exposure (DEARE), and their modulation by APC. ARS was marked by early dyslipidemia and widespread metabolic disruption, evolving into DEARE with persistent alterations in energy metabolism, protein homeostasis, and nucleotide biosynthesis, consistent with sustained oxidative and inflammatory stress. EV profiles showed matrix-specific, time-dependent trajectories distinct from plasma, with prominent lipid dysregulation and enrichment of fatty acid β-oxidation, sphingolipid, and cholesterol pathway metabolites at day 90. Rat 3K3A-APC promoted early EV metabolic normalization, whereas rat WT APC more effectively mitigated late DEARE-associated changes. Elevated sphingomyelins in plasma EVs at day 90 suggested a compensatory or anti-inflammatory lipid response. These data establish plasma-derived EVs as a sensitive matrix for radiation biomarker discovery and for elucidating APC-mediated modulation of IR-induced metabolic and redox disturbances.

Abstract: Fungi recovered from human fecal samples represent an understudied component of the gastrointestinal microbiome, yet fungi can play important roles in health and disease, particularly in hospitalized and immunocompromised patients. Here, we report the culture-based characterization of fungi isolated from fecal samples of 39 hospitalized patients with nosocomial diarrhea in Mexico City. Fungal growth was observed in approximately 40% of samples, yielding 26 isolates: 20 yeasts and 6 filamentous fungi. The predominant genera were Candida (mainly C. albicans) and Rhodotorula, with less common species including Nakaseomyces glabratus, Lodderomyces elongisporus, and yeasts previously explored for probiotic applications, Meyerozyma and Metschnikowia spp. Among filamentous fungi, Paecilomyces variotii and multiple Penicillium species were identified. Phenotypic assays demonstrated that several yeast isolates tolerated gut-associated stress conditions, including elevated temperature, bile salts, and oxidative stress, while thermotolerant filamentous fungi remained viable at temperatures up to 42°C. Cell-free supernatants and organic extracts of selected thermotolerant filamentous fungi showed moderate cytotoxic activity against non-differentiated Caco-2 cells.

Abstract: In this paper, we present our discovery of physicochemical purine-pyrimidine symmetries in DNA genomes and the Supersymmetry genetic code (SSyGC) table but now extend both systems to symmetries of the hydrogen-bonds of bases and the free energy symmetry of trinucleotides/codons which are important for the energy stability of all species. For the first time, we show mirror symmetry in some quadruplets of single-strand RNA coronaviruses and DNA viruses such as acellular infectious agents and obligate intracellular parasites. However, complete quadruplet symmetries are present in all DNA species from protist and prokaryote Cyanobacteria (Blue-green Algae) and eukaryotes. We discovered that DNA quadruplet symmetries and free energy symmetries present a fundamental difference between free-living organisms which have autonomous replication, and non-free-living organisms which are parasites and need a host for replication: only free-living organisms have DNA and free energy symmetries while non-free-living organisms do not. On the other hand, the SSyGC table, with its physicochemical purine-pyrimidine symmetries, is common for all RNA and DNA species as non-free-living and free-living organisms, according to their important symmetry role in the translation process without misreading. After the discovery of the same physicochemical DNA and genetic code symmetries, and the Natural law of DNA creation and conservation as dominant concepts in biological systems responsible for the origin of life and evolution, it is necessary to extend theory of evolution to three balanced, basic core principles of evolution - symmetries, mutations and natural selection.

Abstract: Physiology has long explained organismal stability through homeostasis, allostasis, feedback regulation, and related systems concepts. These frameworks remain essential, but each is most precise at a particular scale or mode of response. This manuscript proposes stratodynamics as a layered theory of biological response in vertebrate physiology. The framework holds that biological response is organized through seven functional layers: molecular, subcellular, cellular, tissue, organ, systemic, and organismal. Each layer reduces its own form of mismatch between actual and required state through a scale-appropriate architecture: molecular-state transition, intracellular integration, cellular-state transition, tissue program, organ output, systemic feedback, or organismal anticipation. The layers are coupled upward, downward, and laterally, allowing local events to become systemic responses and allowing organismal state to shape lower-layer readiness. Stratodynamics therefore locates rather than replaces previous theories: homeostasis is preserved as the canonical systemic architecture, allostasis as organismal predictive regulation, and tissue repair, resolution biology, bioelectricity, and thermodynamic self-organization as complementary layer-specific contributors. The framework is presented as a theoretical synthesis and research program for veterinary and medical sciences.

Abstract: Stress urinary incontinence (SUI) is a highly prevalent condition associated with pelvic floor damage, fibroblast dysfunction, and impaired extracellular matrix (ECM) remodeling. This study aimed to investigate the regenerative potential and underlying molecular mechanisms of decidua-derived mesenchymal stromal cells (DMSCs) in a rat model of vaginal distension (VD) and in human suburethral fibroblasts from SUI patients. Adult female rats were subjected to VD and treated with periurethral DMSC injections, followed by functional and transcriptomic analyses, one and six weeks after VD. In parallel, an in vitro co-culture system was used to evaluate the paracrine effects of DMSCs on SUI fibroblasts. DMSC treatment demonstrated a substantial improvement in continence in rats, as measured by leak point pressure (LPP) and accelerated recovery, while modulating key pathways associated with tissue repair. Gene expression analyses revealed transient increases in proliferative and inflammatory markers, followed by earlier normalization compared to untreated animals. In vitro, DMSCs reduced p16 expression, increased p21 and Klotho levels, and rebalanced eECM remodeling by decreasing MMP-1 and increasing MMP-2. These findings indicate that DMSCs promote a regenerative microenvironment by modulating senescence and ECM dynamics. Overall, DMSCs may represent a promising disease-modifying strategy for SUI by enhancing tissue repair and functional recovery.

Abstract: Hepatic ischemia-reperfusion injury (IRI) is one of the main clinical challenges in liver surgery and transplantation, contributing to postoperative complications and graft dys-function. The pathogenesis of hepatic IRI is complex and multifactorial, involving meta-bolic consequences induced by ischemia, oxidative stress, inflammatory responses, endo-thelial dysfunction, and the activation of immune pathways upon reperfusion. Despite extensive research efforts, the translation of preclinical findings into effective clinical in-terventions has been limited. This review provides a critical overview of the most im-portant models employed to investigate hepatic IRI. Conventional two-dimensional (2D) in vitro systems, including monocultures and co-culture models, offer controlled envi-ronments for mechanistic studies and high-throughput screening but fail to fully repro-duce the structural and cellular complexity of the liver microenvironment. Animal mod-els, particularly those based on mice, rats, and pigs, remain essential for studying the sys-temic and multicellular aspects of hepatic IRI. However, species-specific physiological differences, ethical concerns, high costs, and limited translational predictability represent significant limitations. In this context, three-dimensional (3D) liver models have emerged as promising alternatives able to bridge the gap between in vitro systems and animal ex-perimentation. By better recapitulating tissue architecture, cell-cell interactions, and func-tional heterogeneity, 3D platforms offer improved physiological relevance and transla-tional potential. In detail, we discuss the strengths and limitations of each experimental approach and highlight the role of advanced 3D models as complementary tools that may facilitate more accurate investigations of hepatic IRI and accelerate the development of therapeutic strategies.

Abstract: Background: This scoping review aims to provide healthcare professionals with comprehensive scientific evidence regarding the health complications of navel piercing in women. Currently, navel piercings are attributed to a social identity apart from their aesthetic purpose, particularly among young women. Objectives: To identify and summarize existing literature on the complications of navel piercing in women. This review also aims to propose evidence-based counseling strategies for healthcare professionals, women who wear piercings and intend to become pregnant, and professional piercers. Methods: The databases PubMed, Scopus, Medline, Cinhal, and Sport Discus were consulted using the terms Body Piercing, Cicatrix, Pregnancy, and Abdomen. Selected articles included women with navel piercings and explored the associated implications and complications. Results: The scoping review identified a range of literature addressing the complications associated with navel piercing. Physiological complications included infections, migration of jewelry, and trauma to the piercing site, while psychological implications encompassed body image concerns and self-perception. Obstetric risks highlighted potential difficulties during childbirth, such as tearing and the need for special precautions. Counseling strategies to mitigate these risks focused on education, preventive measures, and culturally sensitive support. Conclusion: This scoping review highlights the importance of providing evidence-based information about navel piercings, focusing on the health risks and the need for clear guidelines. It promotes open communication between health professionals and patients and highlights the importance of future research into the long-term effects and psychological and social impacts of these piercings on women's health, especially during pregnancy and postpartum.

Abstract: HMGB-1 is an alarmin representative of DAMP playing a central role in the immuno-genic cell death (ICD), a necessary condition in the dialogue stablished between dying tumor cells and immune system during some anticancer therapies. Therefore, early screening for ICD inducers represents a major priority in drug development today. In this work, we investigated the effect elicited by the clinical-grade CIGB-300 peptide, which impairs the Protein Kinase CK2-mediated phosphorylation and other CK2 signaling connected kinases. Here, the HMGB-1 extracellular release was investigated in an 18-cell line panel from blood malignancies, uterine-cervical cancer and NSCLC treated with CIGB-300 at equipotent doses (IC50) during 24 hours. Interestingly, CIGB-300 treatment upregulated the HMGB-1 protein levels at the culture supernatant in most of the cell lines (p=0.01) and fold-change increase ≥2 trended to be associated with intrinsic cell line sen-sitivity towards CIGB-300 cytotoxic effect. However, the HMGB-1 release by CIGB-300 was context-specific with clear induction on blood and uterine-cervical cancer cells and a diffused response pattern in NSCLC. Importantly, CIGB-300 treatment of blood cancer patients enrolled in a Phase I study induced plasma HMGB-1 alarmin in 4 out of 7 subject who received the entire treatment plan. Altogether, our data reveal for the first time that CIGB-300 treatment is able to induce extracellular HMGB-1 release in vitro and in vivo which could be indicative of ICD induction in some kind of tumors, also the induction of extracellular HMGB-1 alarmin as putative CIGB-300 response biomarker merits further investigation.

Abstract: Pre-exercise internal cooling through cold water-induced hyperhydration may attenuate increases in core body temperature (T_C) and improve endurance performance. Quantifying the extent of reduction in T_C induced by hyperhydration is important for optimizing its timing before exercise. We compared T_C values obtained from a wearable sensor, the CORE, with those of a gastrointestinal temperature telemetric sensor (G_TS) during hyperhydration. Eleven participants (2 women; age: 24 ± 4 yrs) completed a 120 min seated period where they consumed, over the first 60 min, four boluses of 4 °C water (7.5 mL · kg fat-free mass [FFM]⁻¹), each containing 0.35 g · kg FFM⁻¹ of glycerol. Measures of T_C were taken every 20 min with both sensors. According to the G_TS, hyperhydration induced a peak T_C decline of − 0.76 ± 0.31 °C at min 60; at this time, the change in T_C from baseline estimated by the CORE was − 0.09 °C ± 0.20 °C. The greatest decline in T_C detected by the CORE was − 0.11 ± 0.19 °C. Bland and Altman analyses revealed that average T_C declines of − 0.2, − 0.3, − 0.4, − 0.5 and − 0.6 °C from baseline were associated with T_C values estimated by the CORE that were respectively + 0.41, + 0.57, + 0.65, + 0.77 and + 0.89 °C higher than the G_TS. An intraclass correlation coefficient of 0.12 suggested poor agreement between instruments. These results raise concerns about the ability of the CORE to detect changes in T_C induced by a hyperhydration protocol generating a substantial heat sink.

Abstract:

Antimicrobial resistance (AMR) is a growing public health concern that requires surveillance approaches capable of capturing resistance circulation at the community level. Wastewater-based monitoring provides an opportunity to assess antibiotic-resistant bacteria and antimicrobial resistance genes (ARGs) beyond clinical settings. This study investigated selected bacterial indicators and ARGs in untreated urban wastewater from the municipal wastewater treatment plant of Patras, Greece. Influent wastewater samples were analysed for Escherichia coli, Pseudomonas aeruginosa and Enterococcus spp. using culture-based methods. Isolates were tested for antimicrobial susceptibility by disk diffusion and Etest, and results were interpreted according to EUCAST epidemiological cut-off values. Molecular detection of ARGs was performed by real-time PCR targeting intl1, sul1, qnrS1, blaTEM, blaVIM, vanA and ermB. A total of 16 E. coli, 13 P. aeruginosa and 17 Enterococcus spp. isolates were included in the phenotypic analysis. Among E. coli, non-wild-type profiles were detected in 14/16 isolates for meropenem and 15/16 isolates for ciprofloxacin, while only 1/16 isolates showed a non-wild-type profile for ampicillin. In contrast, all P. aeruginosa isolates were classified as wild type for meropenem but non-wild type for ciprofloxacin, while all Enterococcus spp. isolates were classified as wild type for vancomycin and ampicillin. Molecular screening showed that blaTEM was the most frequently detected gene in E. coli isolates, followed by lower detection frequencies of qnrS1, intl1 and sul1. In P. aeruginosa, intl1 and sul1 were detected in a subset of isolates, whereas blaVIM was not detected. None of the targeted ARGs were detected in Enterococcus spp. These findings highlight the potential of untreated urban wastewater as a complementary matrix for community-level AMR surveillance and support the combined use of phenotypic and molecular approaches to better characterize resistance patterns in environmental settings.

Abstract: The restoration of tropical dry forest (TDF) faces low community integration and limited cultural relevance, especially in the territories of Afro-descendant communities. This study proposes a quantitative framework to systematize the biocultural memory contained in oral tradition as an input for the biocultural restoration of the TDF. A corpus of 401 works from the Afro-descendant community of the Patía Valley (Cauca, Colombia) was analyzed, coded in a multidimensional database of 10 categories. Three indices are proposed: the Biocultural Density Index (IDBC), the Biocultural Vulnerability Index (IVB), and an adaptation of Winter's framework for flora (IVBw), reported in a total version and a version restricted to wild/native species. The results identify the works of greatest biocultural density (IDBC max. = 84) and distinguish between cultivated/introduced species of high cultural value (Limón, Yuca, Caña de azúcar), unsuitable for ecological restoration of the TDF, and wild/native species (e.g., Caña brava, Cañafístula, Guayacán, Ceiba) that constitute the restorable core. Cultural practices are the hubs of the system, and traditional medicine is the most at risk of loss; food security concentrates 43% of the flora records. These findings provide replicable tools to integrate Afro-descendant biocultural knowledge into TDF restoration protocols.

Abstract: Background/Objectives: DNA methylation dynamics are closely linked to ageing. While epigenetic clocks predict biological age from average methylation levels, they may miss structural rearrangements. The "epigenetic rejuvenation" hypothesis posits that embryonic biological age decreases toward a minimum ("ground zero") at gastrulation. This study tested whether DNA methylation entropy decreases from stage E4.5 to E6.5. Methods: Publicly available single-cell nucleosome, methylation and transcription sequencing (scNMT-seq) data (GSE121690; 364 mouse cells) were analysed. Five entropy measures were calculated: Shannon, Renyi, Tsallis, Lempel-Ziv (LZ) complexity, and local gradient entropy. Persistent entropy (PE) and topological analysis (Rips complex, persistence diagrams H₀ and H₁) were computed. A support vector machine (SVM) classifier was trained to distinguish developmental stages. Results: Mean Shannon entropy decreased from 0.841 (E4.5) to 0.805 (E6.5; p < 0.01). All five entropy measures and PE decreased significantly. LZ complexity showed the largest reduction (−28.4% binary, −11.4% ternary; p < 0.001). Ternary LZ correlated with Shannon entropy (r = 0.71). PE decreased from 15.91 to 14.89 (p = 0.01). Topological analysis revealed reorganisation: E6.5 showed stable H₁-cycles (9 intervals), absent at E4.5 (2 intervals), reflecting lineage-specific diversification. Regional entropy and disorder decreased (RE: −25.5%, RD: −27.4%, p < 10⁻¹³). The SVM achieved 93.4% accuracy (AUC = 0.981). Conclusions: The decrease in DNA methylation entropy from E4.5 to E6.5 supports epigenetic rejuvenation as an approach to "ground zero." Topological analysis revealed a qualitative reorganisation: E6.5 showed stable H₁-cycles (9 intervals) that were largely absent at E4.5 (2 intervals). Comprehensive characterisation with balanced subsampling revealed a significant decrease in max persistence (0.446 → 0.218, p = 0.03) and total persistence (2.170 → 1.070, p < 0.001), indicating a transition from a single dominant cycle to multiple smaller, evenly distributed cycles.