Biology and Life Sciences

Abstract: Extracellular vesicles (EVs) have emerged as promising therapeutic carriers owing to their bio-compatibility, low immunogenicity, and ability to transport diverse biological cargos. However, successful EV-based therapy depends not only on carrier engineering but also on selecting an ad-ministration route appropriate for the target disease. This review examines major administration routes for therapeutic EVs, with lipid nanoparticles (LNPs) as a comparative platform, and dis-cusses how anatomical barriers, biodistribution, clearance, inflammatory microenvironments, and target-cell localization influence delivery efficiency. Systemic administration provides broad accessibility but often results in substantial off-target distribution, whereas localized or barri-er-bypassing routes can enhance exposure at specific pathological sites. Importantly, anatomical proximity does not necessarily guarantee efficient delivery, highlighting the need for quantitative assessment of target-tissue bioavailability. We propose a shift from a carrier-centered toward a disease- and route-centered paradigm integrating disease pathology, anatomical accessibility, biodistribution, and carrier engineering.

Abstract: Marine invertebrates and their microbial partners provide distinctive chemical scaffolds, but computational prioritization alone can overstate translational progress when experimental support and material supply remain limited. This critical narrative review proposes a linked-record framework for evaluating exact entity-indication pairs on two independent axes: therapeutic-evaluation stage (E1, in silico, to E4, human intervention) and production readiness (S1, discovery access, to S4, GMP or commercial manufacture). E records the highest stage documented in the reviewed sources, not evidence strength or therapeutic success. Computational contribution, evidence direction, study quality, provenance, and scenario-specific supply adequacy remain separate attributes; E and S are not combined into a composite score. Contrasting lineages involving halichondrin B and eribulin, (+)-aeroplysinin-1, a sea-star steroid, acanthomanzamine C, cynthichlorine, gukulenin A, and meridianin mixtures illustrate how uncertain chronology, test-article mismatch, incomplete validation, and poorly characterized production routes affect interpretation. A worked mass balance distinguishes physical access from the amount required for a defined experiment. Supply-by-design therefore integrates identity, route feasibility, demand, ecological constraints, and access obligations at nomination. The framework is preliminary and requires independent reliability and prospective utility testing.

Abstract: Background/Objectives: Single-cell RNA sequencing enables reconstruction of developmental trajectories, but identifying regions of major transcriptional state reorganization remains challenging, particularly when no explicit dynamical model is available. We developed GDIS-Bio, a framework that applies the Generalized Dynamical Instability Score (GDIS) to pseudotemporally ordered single-cell state spaces to localize transition-associated instability. Methods: GDIS-Bio was evaluated in pancreatic endocrine differentiation (GSE114412) and externally validated in human induced pluripotent stem cell-derived cardiac differentiation (GSE175634). Biological transition landmarks and primary analytical settings were defined independently of GDIS. Robustness was assessed across alternative PCA dimensions and sliding-window configurations, and localization was benchmarked against total variance, Gaussian differential entropy, pseudotemporal step distance, and lag-1 pseudotemporal autocorrelation using structure-preserving null models and individual-level paired tests. Results: In GSE114412, GDIS peaks localized reproducibly near the NEUROG3-early transition across both endocrine lineages and differentiation replicates, with significant localization under circular-shift null testing. In GSE175634, significant localization was retained for MES→CMES, PROG→CM, and PROG→CF transitions without dataset-specific retuning. GDIS was not universally superior to conventional metrics; variance, entropy, and step distance were competitive or better in several settings, whereas GDIS showed its clearest comparative advantage over lag-1 pseudotemporal autocorrelation in the terminal cardiac transitions. Conclusions: GDIS-Bio provides a transferable, model-independent framework for localizing transition-associated dynamical instability in single-cell trajectories and is best interpreted as a complementary integrative measure rather than a universally superior early-warning statistic.

Abstract:

Three-dimensional (3D)-printed clear aligners represent a new alternative for treating orthodontic malocclusions. However, their biocompatibility remains a critical factor requiring careful evaluation. In the current study, the in vitro cytotoxicity of three innovative 3D-printed resins (Tera Harz TC-85 DAC, TA-28, and TR-07, Graphy, Seoul, Korea) and two conventional thermoformed materials (Smart Track, Invisalign, Align Technology, USA, and TruGEN, Spark, Ormco, USA) was assessed by measuring the cell viability of human primary gingival fibroblasts (HGFs) exposed to specimens via direct contact. The MTT assay revealed that, at 7 days, 3D-printed resins exhibited a statistically significant reduction in HGF metabolic activity compared to both the control group and the thermoformed groups (p < 0.0001). This was categorized as "slight cytotoxicity" according to the Ahrari et al. classification. A similar overall biological trend was observed at 14 days, with no progressive impairment in HGF viability, indicating that the materials maintain a stable in vitro biocompatibility profile over time. Notably, all tested materials maintained cell viability levels above the 70% threshold, in compliance with the International Organization for Standardization (ISO) 10993-5 standards. While thermoformed materials are currently considered the conventional reference in orthodontics, the tested 3D-printed resins revealed no relevant in vitro cytotoxic risk, aligning with their ongoing integration into clinical orthodontic practice.

Abstract: Colorectal cancer (CRC) is associated with high mortality rates and therapeutic limitations due to drug toxicity and resistance. Molecular hybridization has emerged as an innovative strategy, highlighting Mel-Fur (6f), a novel hybrid of melatonin and furanochalcone with antitumor potential. However, characterizing its pharmacokinetic profile is essential to support its preclinical applicability. This study aimed to develop and partially validate a high-performance liquid chromatography method with diode array detection (HPLC-DAD) for the quantification of Mel-Fur in serum and murine organs, and to apply it to an in vivo distribution analysis. Chromatographic separation was performed on an Agilent 1200 using a C30 column and acetonitrile:water (85:15) mobile phase at 0.8 mL/min and detection at 342 nm. The method met ICH M10 and FDA validation guidelines, demonstrating high selectivity, linearity (R²>0.999), precision, accuracy, recovery (>89.9%), and autosampler stability for up to 8 days. Following single oral administration of Mel-Fur (1000" mg/kg" ) in BALB/c mice (n=3 per sampling time point), the hybrid displayed rapid absorption and elimination, with measurable systemic exposure, preferential accumulation in the lungs and liver, and sustained presence in the colon. The partially validated HPLC-DAD method provides a robust analytical tool for preliminary pharmacokinetic and tissue distribution studies of Mel-Fur.

Abstract:

Fungal cell factories have played a central role in industrial biotechnology, with Saccharomyces cerevisiae and filamentous fungi such as Aspergillus niger, Penicillium chrysogenum and Acremonium chrysogenum serving as established platforms for the production of fuels, organic acids and pharmaceuticals. This review examines the development of these fungal production systems from conventional fermentation and empirical strain improvement towards metabolic engineering, genome-scale analysis and targeted genome modification. Particular emphasis is placed on ethanol and advanced biofuel production by S. cerevisiae, citric acid production by A. niger, and β-lactam antibiotic production by P. chrysogenum and A. chrysogenum. Recent advances in pathway engineering, transporter manipulation, stress tolerance, comparative genomics and genome editing have substantially expanded the capacity to optimise fungal cell factories. However, limitations including substrate utilisation, product toxicity, metabolic burden, genetic stability and strain-specific regulatory requirements continue to constrain industrial implementation. The review also considers the regulatory implications of using genetically modified fungal organisms, with particular attention to the European Food Safety Authority (EFSA) framework for the safety assessment of microorganisms used in the food chain. Overall, the transition from classical strain improvement to genome-informed engineering is transforming fungal cell factories from empirically optimised production strains into increasingly rationally designed microbial platforms.

Abstract: High clean-water throughput does not necessarily translate into rapid fermentation-broth filtration. We hypothesized that a macroporous upstream layer combined with an asymmetric polyethersulfone (PES) membrane could unite rapid filtration at low broth concentration with improved performance under a greater fouling challenge. Development progressed from symmetric and asymmetric manufacturing platforms through layered proof-of-concept tests to a PES composite. Preliminary asymmetric-PES/nitrocellulose assemblies increased average broth flux 4.81-fold at 100-fold dilution and 3.74-fold at 50-fold dilution relative to their respective bare-base controls. A separately cast PES upstream layer was then developed using mixed polyethylene-glycol additives and vapor/non-solvent-induced phase separation. The selected 40-µm layer had an open particulate morphology and a clean-water flux of 146,813 L m⁻² h⁻¹. Its composite filtered 100 mL of 50-fold diluted Escherichia coli broth in 52.35 s versus 185 s for an asymmetric-base reference. Subsequent upstream incorporation of 0.5 wt% acid-form sulfonated PES improved composite broth flux 1.77–2.25-fold despite lower isolated-layer water flux; both tested coupons exceeded two commercial references in the same session. The final PES and PES/sPES-H composites were compared at 50-fold dilution only. These results support the architectural hypothesis and establish a third-generation PES composite through coordinated development of upstream morphology, composition and interlayer contact.

Abstract: Macroalgae host complex microbial communities that influence surface colonization, development, nutrient transformation, carbohydrate turnover, environmental responses, and the production of potentially valuable metabolites. Advances in shotgun metagenomics and genome-resolved analysis have greatly expanded access to this functional diversity, but they have also increased the risk of conflating genomic potential with biological activity or host benefit. This narrative review evaluates the bacterial component of macroalgal microbiomes — the component for which mechanistic and experimentally validated evidence is most extensive — through a framework that separates what has been proven from what is merely predicted, distinguishing functional prediction from expression, biochemical activity, metabolite exchange, host response, and experimental causality. Particular attention is given to bacteria-dependent morphogenesis, metabolic complementarity, environmental acclimation, carbohydrate-active enzymes and polysaccharide-utilization loci, biosynthetic gene clusters, and the translational potential of seaweed-associated microorganisms. Comparative perspectives from microalgal and cyanobacterial systems are used to identify principles that generalize across algal microbiomes while emphasizing the distinctive spatial and chemical complexity of macroalgal surfaces. Invasive and bloom-forming macroalgae provide an additional ecological context in which microbial functional redundancy, recruitment flexibility, and dominant populations may shape whole-community profiles. An illustrative, hypothesis-generating taxon-attribution case study, based on single metagenomic datasets from Rugulopteryx okamurae and Sargassum sp. holobionts in the Azores, which confounds host identity with sampling location, time, and thallus condition, shows that the disproportionate genomic contribution of the genus Cobetia measurably alters the interpretation of a specific subset of community-level KEGG pathway contrasts, while the majority of pathway-level and taxon-specific signals discussed remain robust to this taxon's exclusion. Together, these analyses argue that macroalgal microbiome research should move beyond inventories of predicted functions toward explicit taxonomic attribution, experimental validation, and ecological context. Such integration will be essential for establishing which microbial functions are ecologically consequential and which can be translated reproducibly into biotechnology.

Abstract: EraGene is a platform that compiles experimentally validated engineering targets and production data from engineered Escherichia coli strains for metabolite overproduction. By integrating published metabolic engineering studies, it includes recurrently modified reactions, enabling identification of successful engineering strategies for biochemically related metabolites. We demonstrate its utility through two applications: identifying conserved engineering targets for pathway reinforcement and improving constraint-based metabolic model predictions using experimentally validated modifications. Leave-one-out analysis showed that 75.52% of a metabolite’s engineered reactions were, on average, already annotated for other metabolites, indicating substantial conservation of central metabolism and opportunities for knowledge transfer. Incorporating reactions with conserved modulation direction into QPAML simulations reduced the average search space from 180.21 to 43.51 candidate reactions per metabolite while recovering 81.88% of experimentally validated modifications in the prediction set, corresponding to a 3.41-fold enrichment. At publication, EraGene contained 3,833 production measurements from 2,611 strains across 360 studies, comprising 342 native reactions involving 921 genes. The analytical dataset included 300 native reactions associated with 88 metabolites producible from glucose as the sole carbon source. EraGene is publicly available at https://eragene.com for visualization and data download.

Abstract: Injected botulinum neurotoxins (BoNT) reduce dynamic wrinkles by cleaving SNARE proteins at the neuromuscular junction; however, their large size mostly limits their use to injectable procedures. Certain polyphenols, including myricetin, have been shown in neuronal systems to reduce acetylcholine (ACh)-induced muscle contraction via inhibition of SNARE complex zippering, suggesting potential for topical use as modulators of skin cholinergic activity. This study aimed to identify a natural extract blend similarly capable of attenuating ACh release followed by characterization of the activity of this extract on human skin explants, alongside reference molecules BoNT/A, α-bungarotoxin, and myricetin. The natural extract blend (“Blend 1”) inhibited vesicle fusion and attenuated norepinephrine release in a cell culture system, inhibited muscle cell contraction within nerve/muscle co-culture, and attenuated ACh release consistent with myricetin and reference neurotoxins within human skin explants. Bulk transcriptomics of treated ex vivo skin explants revealed that BoNT/A, myricetin and Blend 1 all shared a similar downregulated inflammatory and anti-viral gene profile separate from α-bungarotoxin, but that myricetin and Blend 1 shared a large degree of overlap for upregulated genes associated with skin homeostasis and repair that was not observed for either toxin. These data support Blend 1 as a candidate for topical cosmetic use through modulation of BoNT/A-like cholinergic pathways, alongside activation of additional pathways associated with skin homeostasis and repair.

Abstract: Background: Ribonucleic acid (RNA)-loaded lipid nanoparticles (LNPs) are established platforms for the delivery of messenger RNA (mRNA), small interfering RNA (siRNA), and other functional RNA molecules. Their biological performance is determined by interconnected factors extending from manufacturing and particle assembly to physicochemical characteristics, interactions with biological fluids, cellular trafficking, and in-vivo function. This systematic review aimed to integrate the available evidence on manufacturing parameters, physicochemical characteristics, protein-corona interactions, and biological performance of RNA-loaded LNPs. Methods: This systematic review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) guidelines. PubMed, Scopus, and Web of Science were searched from inception to 12 July 2026 without language or publication-date restrictions. The first 50 pages of Google Scholar and the reference lists of relevant articles were additionally screened. Eligible experimental studies investigated RNA-loaded LNPs and contributed evidence to at least one prespecified domain: manufacturing/process-related physicochemical characteristics, primary in-vitro biological delivery, downstream or in-vivo functional outcomes, or protein-corona and serum/protein interactions. Methodological quality was assessed using the quantitative Standard Quality Assessment Criteria for Evaluating Primary Research Papers from a Variety of Fields (QualSyst). Owing to substantial methodological and experimental heterogeneity, the evidence was synthesized systematically without meta-analysis. Results: A total of 45 unique parent studies were included. Thirty studies evaluated manufacturing/process-related physicochemical outcomes, 26 investigated primary in-vitro biological delivery, 37 assessed downstream or in-vivo functional outcomes, and 17 examined protein-corona or serum/protein interactions, with substantial overlap among evidence domains. Manufacturing variables, including mixer architecture, total flow rate, flow-rate ratio, scale-up conditions, dilution, and post-processing, were associated with changes in particle size, polydispersity, encapsulation efficiency, surface characteristics, and particle structure; however, the direction and magnitude of these effects varied among formulations and platforms. Similar conventional physicochemical characteristics did not consistently correspond to equivalent biological performance, and greater cellular association or uptake was not invariably accompanied by greater productive RNA expression or functional activity. Protein-corona studies demonstrated formulation- and biofluid-dependent interactions that were associated with changes in cellular uptake, intracellular trafficking, biodistribution, and functional RNA delivery. Apolipoprotein E was a frequently investigated mediator, particularly in hepatic delivery, although multiple other protein classes were also implicated. Conclusions: The available evidence indicates that RNA-LNP performance should be considered as the product of an interconnected relationship among manufacturing conditions, physicochemical and structural characteristics, biological-fluid interactions, and subsequent cellular and in-vivo function. Manufacturing effects are highly dependent on formulation and platform context, while conventional physicochemical attributes alone are insufficient to predict biological performance. Protein-corona formation represents an additional context-dependent component of LNP biological identity. Greater methodological standardization and integrated process–physicochemical–biological studies are required to enable more reliable cross-study comparisons and future quantitative synthesis.

Abstract: This study aimed to evaluate the ability of microbes from the rumen contents of cattle as antagonists of pathogenic fungi and as liquid organic fertilizer (LOF) containing P-solubilizing bacteria in rice plants. The mixture consisting of 5 kg rumen contents of cattle, 2 kg of rice bran, 1 kg of molasses, and 10 L of tap water was fermented for 14 days. After fermentation, the mixture was filtered to separate the liquid from the solid components. The solid portion was collected for the isolation and screening of contained bacteria. Subsequent tests included biochemical tests, antagonistic activity tests, DNA extraction, PCR amplification, phosphate solubilization ability testing, inhibitory ability study, and application of LOF in a screen house. One bacterial isolate (PTK2TB code) exhibited the highest antagonistic activity against three tested pathogenic fungi (inhibits more than 50%). Subsequent 16SrDNA sequence analysis showed that this isolate belongs to the same group as the type strain and reference strain of Bacillus amyloliquefaciens. Furthermore, out of the 90 bacterial isolates obtained, 20 demonstrated a high capacity for phosphate solubilization. Additionally, LOF containing phosphate-solubilizing bacterial isolates positively impacted rice growth and yield.

Abstract: Long-duration human exploration beyond low-Earth orbit exposes crews to a convergent hazard field that includes galactic cosmic radiation, solar particle events, hypogravity, hypokinesia, immune dysregulation, microbiome disturbance, circadian disruption, psychological stress, and constrained habitat conditions. Current radioprotection strategies (exposure limitation, active dosimetry, space-weather forecasting, physical shielding, storm-shelter logic, ALARA-style dose management, and biomedical countermeasures) remain essential, but they will not fully solve the biological problem of cumulative radiation injury during Mars-class missions. This article proposes a hypoxia-centred integrated radioprotection framework in which habitat atmosphere composition and oxygen partial pressure are treated as active design variables rather than passive life-support parameters. Controlled normobaric hypoxic engineering may reduce oxygen-dependent fixation of radiation injury, lower oxidative pressure, support mitochondrial resilience, and improve fire safety in enclosed vehicles and habitats. This approach is not a replacement for shielding, monitoring, or pharmacological countermeasures. It is a missing environmental layer designed to integrate with them. The central operational concept is a two-level architecture: long-term adaptation to moderate habitat hypoxia, provisionally corresponding to approximately 16-12% O2 under carefully controlled normobaric conditions, combined with short, monitored, deeper hypoxic excursions to approximately 9-10% O2 during solar particle event sheltering, when crew are already in structurally protected niches with minimal physical activity and reduced oxygen consumption. These values are proposed research windows, not operational recommendations, and require staged validation before any mission use. The proposed framework integrates mechanistic rationale, operational architecture, safety objections, fire-safety co-benefits, adjunctive countermeasures, and a four-prototype validation pathway for controlled oxygen modulation. Controlled hypoxic habitat engineering emerges as a mission-critical countermeasure candidate for deep-space exploration and as a possible translational bridge to terrestrial geromedicine through mitochondrial resilience, hypoxic conditioning, and healthspan infrastructure.

Abstract: Microbial source tracking (MST) estimates the contributions of candidate source communities to an observed sink community. However, taxon-aligned approaches can degrade when biological turnover or technical differences introduce "drift" between source and sink profiles, because abundance on related but nonmatching taxa is treated as separate features. We present MARSH, a phylogeny-aware MST framework that models source attribution as constrained optimal transport on the microbial tree using the tree-Wasserstein distance. By leveraging evolutionary structure, MARSH allows abundance to be matched across closely related taxa rather than enforcing strict feature identity. The formulation limits the loss to local tree structure so that broad clade-level differences do not dominate the fit, and it estimates unobserved contribution from the residual left by the named-source mixture. Across synthetic and real 16S rRNA benchmarks, MARSH attains the lowest mean attribution error in every evaluated drift condition and remains competitive in the no-drift settings. These results show that integrating tree geometry into community deconvolution can improve source tracking when ecological or experimental drift preserves local phylogenetic structure.

Abstract: Hyaluronan (HA) has traditionally been viewed as a structural component of the extracellular matrix, valued primarily for its viscoelastic, lubricating, and hydrating properties. However, accumulating evidence suggests that ultra-high molecular weight hyaluronan (UHMW-HA) functions far beyond passive tissue support, acting as a dynamic regulator of inflammation, mechanotransduction, immune homeostasis, cellular senescence, and regeneration. In this narrative review, we examine HA and specifically focus upon UHMW-HA operating as a “living hydrogel”, an adaptive regenerative macromolecule that continuously interacts with its microenvironment to shape tissue function and resilience. We examine evidence spanning evolutionary biology, glycobiology, biomaterials science, and regenerative medicine to examine how polymer size encodes biological activity and how UHMW-HA influences stem cell behaviour, extracellular matrix remodelling, and responses to injury and ageing. Attention is given to unique biological models, including the naked mole-rat, whose exceptional longevity and cancer resistance have been linked to abundant UHMW-HA, offering insights into the relationship between extracellular matrix architecture and organismal health. We further discuss how advances in synthetic biology and biomaterials engineering are enabling the development of next-generation HA-based hydrogels that mimic these adaptive properties for therapeutic applications in tissue engineering, in addition to potential roles in dynamically optimising wound repair, and musculoskeletal rejuvenation, through precision medicine approaches. Finally, we argue that reclassifying UHMW-HA as a living regenerative material rather than an inert scaffold provides a new concept for designing bioinspired therapeutics and understanding extracellular matrix function. This review demonstrates HA-adaptive macromolecules as critical entities in future regenerative strategies and highlights the specific and unique potential of UHMW-HA to combine material science developments with enhancement of preferable biological outcome.

Abstract: Triple-negative breast cancer (TNBC) is the most aggressive form of breast cancer and poses significant treatment challenges due to fewer available options. Creating highly specific and precise anticancer therapies continues to be a significant challenge for TNBC. Previous studies showed that combining drugs or adding chemotherapy to other therapies has significantly improved patient prognosis and proved to be an effective strategy for treating TNBC compared to using chemotherapeutic drugs alone targeting TNBC pathways. One such combination is a PARP inhibitor (PARPi) with an immune checkpoint inhibitor (ICI). In this study, we explored the transcriptomic profiles of the combination of Olaparib (a PARP inhibitor) and atezolizumab (a monoclonal antibody targeting PD-L1) in cells derived from African American (AA) and White populations. Our results demonstrated that combining Olaparib (Ola) and Atezolizumab (Atz) significantly reduced cell viability in both AA and White cell lines, compared to either agent alone. Our transcriptional profiling results showed that in both up- and down-regulated genes, there was widespread suppression of proliferation and biosynthetic gene networks, and selective activation of immune and stress-related pathways. Our transcriptional profiling revealed that both up- and down-regulated genes exhibited widespread suppression of proliferation and biosynthesis pathways, along with selective activation of immune and stress-related pathways. Our functional enrichment analysis revealed significant changes in DNA damage response, chromatin organization, RNA regulatory processes, immune signaling, and inflammatory responses. In conclusion, our study’s results provide valuable insights into key features of combination therapy, which may help guide the more targeted development of these regimens.

Abstract: This review synthesizes regulatory and scientific evidence accumulated over the past decade supporting the safety of newly expressed proteins (NEPs) in genetically modified (GM) crops. Using five case studies representing diverse protein classes, including dicamba monooxygenase (DMO), protoporphyrinogen IX oxidase (PPO), Cry1B.2/Cry1B.3, Mpp75Aa1.1, and Vip3Cb1, it illustrates how science-based, case-by-case, weight-of-evidence approaches are applied in food and feed safety assessment and ecological risk assessment (ERA) for regulatory submissions. These examples demonstrate how hazard identification, hazard characterization, and exposure assessment are integrated across proteins with diverse biological functions, structural characteristics, and levels of familiarity. It further examines the safety relevance of protein processing and sequence variation arising from protein engineering or biological processing and discusses the scientific basis supporting bridging and read-across approaches, including phylogenetic relationships, sequence similarity, structural conservation, domain architecture, and functional equivalence. The increasing application of safe-by-design principles and bridging strategies is highlighted as an important advancement enabling efficient, scientifically robust evaluations while reducing unnecessary testing. The review also discusses the convergence of food/feed safety assessment and ERA and future directions. Overall, accumulated evidence demonstrates that NEPs in commercialized GM crops have not introduced safety concerns and supports the continued evolution and modernization of protein safety assessment frameworks.

Abstract: Somatic genome editing is emerging as a potential strategy for achieving durable therapeutic effects in cardiovascular disease. This review examines current genome-editing platforms, therapeutic targets, delivery systems, and translational hurdles, contrasting liver-directed and myocardial applications. CRISPR-Cas nucleases, base editors, and prime editors offer complementary capabilities, enabling durable gene disruption or precise nucleotide correction. However, key translational challenges include unintended alterations, immune responses, irreversibility, dose optimization, long-term surveillance, regulatory requirements, and equitable access. Clinical translation has advanced for liver-directed applications, with single-administration editing of hepatocyte targets including PCSK9, ANGPTL3, and TTR producing decreases in circulating proteins and atherogenic lipids in early-phase studies, although evidence of cardiovascular event reduction and lifelong safety remains unavailable. In contrast, direct myocardial editing remains predominantly preclinical, with studies targeting MYH7 and MYBPC3 pathogenic variants demonstrating proof of concept and disease rescue in selected models, while revealing challenges in cardiomyocyte delivery, spatial coverage, allele selectivity, and tissue-level safety. Future progress will require integrating target biology, editor selection, organ-specific delivery, and rigorous molecular and clinical outcome assessment. Cardiovascular genome editing is therefore entering clinical translation in selected liver-directed applications, whereas direct myocardial editing remains at an earlier stage, with delivery, safety, and myocardial coverage as major barriers.

Abstract: Background: Long-term biospecimen preservation requires sustained institutional investment in storage equipment, energy or cryogen, monitoring, maintenance, and inventory management. Although biobank economics and biospecimen utilization have been examined independently, the recurring direct cost associated with maintaining existing inventories in relation to storage capacity and research utilization remains less well characterized. We introduce Annual Carrying Cost (ACC) as an operational metric for quantifying this continuing institutional investment. Methods: A retrospective observational analysis was conducted within a large academic biobank using long-term storage infrastructure and retrieval data from 2023–2025. ACC incorporated annualized equipment and monitoring-probe costs, energy and cryogen consumption, maintenance and repair, environmental monitoring, and inventory management. Storage capacity was characterized according to inventory-bearing, online contingency, and offline/staged reserve capacity. ACC was normalized to available and occupied vial positions and evaluated in relation to annual researchdirected vial-equivalent retrievals. Results: Sixteen storage units provided 832,120 vial positions, of which 272,476 were occupied (32.7%). Eight inventory-bearing units were 56.1% occupied, while 41.6% of total capacity was intentionally maintained as online contingency or offline/staged reserve infrastructure. Estimated ACC was $131,252 annually, comprising annualized equipment and monitoring probes (40.5%), energy and cryogen (30.8%), environmental monitoring (14.4%), maintenance and repair (8.4%), and inventory management (6.0%). ACC corresponded to $0.16 per available vial position and $0.48 per occupied vial position annually. Research-directed retrieval volume varied more than 13-fold across the study period, from 217,348 vial equivalents in 2023 to 16,414 in 2025, resulting in retrieval-normalized ACC ranging from $0.60 to $8.00 per vial-equivalent retrieved. Conclusions: Long-term biospecimen retention creates a measurable recurring institutional obligation that persists despite variation in annual research utilization. ACC provides a practical framework for characterizing this investment and distinguishing routinely occupied storage from infrastructure maintained for resilience and future capacity. When considered alongside scientific value, utilization, uniqueness, and anticipated future demand, ACC may support more informed capacity planning, lifecycle management, resource allocation, and sustainable long-term biospecimen stewardship.

Abstract: Background: Cannabidiol (CBD), isolated from hemp (Cannabis sativa or Cannabis indica), is receiving increasing attention for its potential against different pathologies, including viral infections. CBD has been reported to alter lateral membrane diffusion and cholesterol accessibility. Therefore, it remains unclear which signaling pathways are affected by CBD treatment. Here, we sought to determine the effect of CBD treatment on human cell lines and primary human cells. Methods: We performed bioinformatics analysis to identify differentially expressed genes (DEGs) in published RNA-seq datasets of CBD-treated cells, including A549, THP-1, normal human epidermal keratinocytes (NHEK), monocyte-derived dendritic cells (DC), and monocyte-derived macrophages (MDM). We then performed RT-qPCR to validate A549, THP-1, and MDM sequencing data. Results: Comparative transcriptional analysis across five CBD-treated cell types identified 21 genes shared across all cell types, 142 among four, and 694 among three, enriched in ER stress, lipid metabolism, and cholesterol metabolism pathways. Transcription factor activity analysis identified ATF4, ATF6, XBP1, SREBF1, and SREBF2, which encode key regulators of the ER stress response and cholesterol metabolism. Conclusions: Results highlight CBD’s pivotal role in regulating transcription factor expression, which is crucial for controlling gene expression involved in ER stress, lipid, and cholesterol metabolism. Consequently, CBD treatment modulates gene expression programs in a cell-dependent manner and promotes lipotoxicity by dysregulation ER stress signaling pathways.