Biology and Life Sciences

Abstract: Multicellular individuality depends upon constraints that limit the establishment and evolutionary success of genetically foreign cellular lineages. Naturally transmissible cancers expose the permeability of these constraints because malignant cells can propagate between unrelated organisms. Although ecological barriers to transmission, allorecognition and host–parasite genotype-by-genotype interactions are established evolutionary concepts, their integration into a stage-resolved account of somatically derived cellular parasitism remains incomplete. This review critically synthesises evidence from transmissible cancers in dogs, Tasmanian devils, bivalves and freshwater catfish alongside genetically regulated cellular recognition in colonial invertebrates. Historical experimental transmission in soft-shell clams, recent environmental-survival studies, comparative tumour genomics and contemporary host population-genetic responses provide complementary evidence. A Genetic Firewall Framework is proposed as a restricted application of established host–parasite compatibility theory to clonally transmissible malignant cells. It distinguishes exposure, viable-cell entry, recognition-mediated elimination, post-entry niche compatibility and onward transmission. An explicit recipient-genotype-by-clone-genotype establishment model is linked to host fitness and malignant-lineage frequency dynamics. The primary empirical prediction concerns reproducible, stage-specific compatibility effects, with susceptibility-rank reversal representing a particularly discriminating but non-essential interaction pattern. A falsification-oriented programme centred on Mya arenaria separates inherited resistance from tumour-intrinsic attenuation, environmental confounding and phylogenetic pseudoreplication. The framework neither assumes homologous recognition systems across taxa nor claims an established universal antitumour mechanism. Its value lies in determining when genetic compatibility explains cellular parasitism and when simpler ecological or physiological explanations prevail.

Abstract: Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive human cancers and is characterized by a high frequency of activating KRAS mutations, most commonly KRASG12D. Yet oncogenic KRAS alone is insufficient for malignant progression, indicating that additional cellular and environmental factors shape disease evolution. Obesity is a major modifiable risk factor for PDAC and creates a chronic metabolic environment marked by excess nutrient and lipid availability, inflammation, and altered redox homeostasis. In acinar cells, these conditions can increase metabolic and mitochondrial stress, promoting reactive oxygen species (ROS) generation and molecular and organellar damage. Cellular responses to such damage, however, do not necessarily lead to either complete recovery or cell death. Antioxidant defenses, autophagy, mitochondrial quality control, proteostasis, and DNA repair can contain or process damage, allowing cells to remain viable despite persistent or incompletely resolved injury. Thus, cellular fitness under chronic stress may depend not only on the amount of damage generated, but also on the capacity to tolerate damage while maintaining sufficient function. Here, we conceptualize the relationship between damage burden and cellular fitness as a dynamic redox damage tolerance threshold. Transient metabolic and redox stress may allow acinar cells undergoing acinar-to-ductal metaplasia (ADM) to return toward homeostasis, whereas sustained metabolic stress, as occurs in obesity, in the presence of oncogenic KRAS may enable ADM cells to remain functional despite residual damage. Such persistence may increase the opportunity for additional alterations and progression toward pancreatic intraepithelial neoplasia (PanIN). This framework places cellular damage tolerance, rather than damage generation alone, at the center of early PDAC development and highlights the mechanisms that determine this threshold as potential therapeutic vulnerabilities.

Abstract: Cisplatin-induced hearing loss (CIHL) is a debilitating side effect in cancer patients undergoing chemotherapy. So far, epigenetic studies in the inner ear have largely focused on developmental biology to regenerate hair cells. In post-mitotic hair cells, on the other hand, DNA methylation regulates transcriptomic response to external stimuli. Adult mammalian hair cells are locked in the G0 phase, and passive demethylation via DNA replication does not occur under normal physiological conditions. However, external stimuli such as cisplatin can aberrantly upregulate DNMT methyltransferases (DNMTs), resulting in promoter methylation and silencing protective genes. From this angle, DNMTs modulation is an attractive therapeutic strategy to protect hearing during chemotherapy. Recent studies suggest that non-nucleoside inhibitors such as RG108 and natural polyphenolic compounds downregulate DNMTs, and at the same time, protect auditory cells. These compounds function via the inhibition of the DNMT catalytic site but can also act through downregulating DNMT1 and DNMT3A/B at mRNA and protein levels. Polyphenols are also antioxidants, decreasing reactive oxygen species, thereby increasing cell survival. Furthermore, it is suggested that upregulating TET-mediated active demethylation can be another angle to modify DNA methylation in cancer and CIHL models. However, functional DNA methylation should be distinguished from non-functional, passenger DNA methylation to elucidate the compounds’ otoprotective effects. In addition, the candidate compounds should not interfere with cisplatin’s antitumor effect. In this review, we investigate the relationship between DNA methylation and CIHL in the post-mitotic inner ear and evaluate the therapeutic potential of candidate DNMT inhibitors.

Abstract: Bluetongue is an economically important arboviral disease affecting domestic and wild ruminants health and well-being, caused by bluetongue virus (BTV) and transmitted by Culicoides midges. More than 30 BTV serotypes may co-circulate regionally, while current vaccines face limitations related to safety and incomplete cross-protection. Here, reverse vaccinology and immunoinformatics were used to design four serotype-specific and one multiserotype multi-epitope vaccine candidates targeting BTV-1, BTV-3, BTV-4 and BTV-8. Cytotoxic T-lymphocyte (CTL) and helper T-lymphocyte (HTL) epitopes were predicted from VP2, VP5, VP7 and NS1, and linear B-cell epitopes from VP2 and VP5. Twenty-five CTL, 24 HTL and 15 B-cell epitopes met the computational antigenicity, non-allergenicity and non-toxicity screening criteria; HTL epitopes were also predicted to induce interferon-γ. Five constructs comprising 13-25 epitopes (252-471 amino acids) were fused to bovine β-defensin 2 as a putative adjuvant. Molecular docking with bovine Toll-like receptor 4 ectodomain yielded predicted binding energies ranging from -19.6 to -10.7 kcal/mol. Molecular dynamics simulations (100 ns), immune simulation, codon optimization and in silico cloning further characterized the candidates. These computational findings support the prioritization of these constructs for experimental evaluation as potential serotype-specific and multiserotype BTV vaccine candidates.

Abstract: This cross-sectional study evaluated the validity of peak oxygen uptake (V̇O₂peak) elicited during a 200-m front crawl test (T200) for predicting maximal oxygen uptake (VO_2max) in master swimmers. Twenty-one master swimmers (age = 46.1 ± 13.7 years; World Aquatics Performance = 345 ± 28 points) completed two protocols: a 200-m all-out test (T200) and an incremental intermittent swimming test (ISST; n x 200-m) as a reference. Respiratory variables (VO₂, V̇CO₂, VE) were measured directly using a portable gas analyzer, heart rate was monitored, and stroke kinematics were evaluated via video analysis. Mean performance times were 186.7 ± 33.4 s (T200) and 193.5 ± 34.0 s (final repetition of the ISST). Cardiorespiratory fitness values reached 45.9 ± 8.5 mL·kg⁻¹·min⁻¹ (V̇O₂peak) and 44.2 ± 8.2 mL·kg⁻¹·min⁻¹ (VO_2max), with no significant differences observed between protocols for VO₂, V̇CO₂, or VE (p > 0.05). Bland-Altman analysis demonstrated strong agreement between measurements, and a significant intraclass correlation was found (VO₂ ICC = 0.74; p = 0.002). The T200 is a valid, practical, and time-efficient field-based method for assessing cardiorespiratory fitness. Its strong agreement with the ISST supports its use for estimating VO_2max, thereby facilitating accurate training prescriptions and health assessments for master swimmers.

Abstract: Due to oncoming climate changes, drought is recognized as one of the most important factors of crop yield losses. Water stress differentially affects plant longevity and productivity depending on the ontogenesis step. Specifically, drought occurring at the reproductive stage can reduce crop productivity and the quality of harvested seeds. Therefore, breeding new cultivars preserving both productivity and seed quality under drought conditions is urgently mandatory. However, no straightforward and reliable markers of drought tolerance are established for this ontogenesis step so far. It is generally unclear whether genetic differences between genotypes known as drought-tolerant and drought-sensitive can be traced in performance of the progeny mature seeds and their metabolic responses after rewatering. Therefore, here we address physiological, reproductive and seed metabolic responses in 29 pea genotypes varying in the degree of documented drought tolerance. The plants were subjected to transient irrigation withdrawal during pod formation. Based on the physiological data, metabolomes of nine most contrasting genotypes were profiled by GC-MS. The results revealed strong genotype-dependency of the stress-induced metabolic shifts, which only partly matched the physiological data. Thus, only complementation of physiological plant and seed traits with metabolic data might give access to interpretation of drought tolerance in grain legumes.

Abstract: The exposure to electric amenities, both fields and current, affects microbial growth and its systematic study in species, substrates and specific settings defines the Electroculturom-ics. In this context, the effect of microcurrents on facultative microbial pathogens remains controversial, and thus restricts the wide clinical use of such amenities. This study pre-sents preliminary observations in a low-replicate in vitro system. Solid cultures in Sabouraud Glucose Agar of the ubiquitous mold Aspergillus fumigatus were repeatedly treated with a commercially available device performing wireless conductive microelec-trostimulation by 3.5 μA microcurrent routed by spraying negatively charged air particles onto solid cultures in modified petri dishes. The treated cultures displayed increased growth compared to standard ones, apperceived as visually apparent differences in myce-lial texture; the radial growth rate of the mycelium remained unaltered. The increased growth was positively related to the duration of the treatment. At the same time, second-ary development (new mycelial loci within the dish) was greatly upheld due to treatment, possibly due to the spraying creating microairstreams dislocating the fungal spores. These results imply complex kinetics of mycelial growth both with and without treatment, since the folding of the mycelial mat is observed regularly. Both the fungus' response to the elec-trostimulation and the possible revision of growth kinetics might translate to prospects for biotechnological and bioremediation applications but may also imply biomedical com-plications, regarding infection dynamics of mycelial fungi and their in situ resistance to immune responses and therapeutic treatment if subjected to such ES in vivo.

Abstract: Hit-to-lead optimization must balance binding-related performance with drug-likeness, synthetic accessibility, and early safety considerations. We developed an iterative large language model agent that integrates target and starting-hit context, literature and computational tools, multi-property scoring, and feedback from previously scored analogs. A matched benchmark covered six kinase targets, five comparison strategies, a 40-candidate budget, and six independent campaigns per method. The agent achieved the highest mean aggregate score on four targets under an objective combining docking, quantitative estimate of drug-likeness (QED), synthetic accessibility, and a rule-based property panel. Re-evaluating the same candidate sets using drug-likeness-gated docking and docking-drug-likeness hypervolume produced different target-level winners, showing that metric choice changes the comparative interpretation of optimization campaigns. Candidate distributions showed higher-QED regions alongside improved docking, and ablation identified scored-candidate feedback as the most consistent contributor. In an epidermal growth factor receptor campaign, an erlotinib analog retained the 4-anilinoquinazoline core while improving docking from -6.80 to approximately -8.6 kcal mol⁻¹ and QED from 0.418 to approximately 0.73. These computational results support target-specific agent evaluation and candidate prioritization for subsequent biochemical, cellular, and ADMET characterization.

Abstract: Inhalation represents a primary entry route for airborne pollutants, aerosols, pathogens, and therapeutic agents. Their pathological impacts depend on a complex interplay of physical transport, deposition patterns, cellular crosstalk, mechanical forces, and localized microenvironmental responses. However, traditional 2D cell cultures and animal models fall short in mimicking these human-specific microenvironments, particularly due to the lack of air-exposed barriers, dynamic fluid flow, physiological motion, and precise aerosol dosage control. Respiratory organ-on-a-chip technologies address these limits by integrating air–liquid interfaces, microfluidic perfusion, and tissue-specific mechanics. In this review, we examine the principles of inhaled exposure, disease mechanisms across the respiratory tract, and recent advances in chip design and exposure delivery. We highlight key applications in modeling respiratory infections, chronic inflammatory and fibrotic diseases, airway injury, and carcinogenesis. Finally, we discuss critical requirements for translational impact, emphasizing that advanced model complexity must be supported by accurate dosimetry, robust tissue validation, and defined contexts of use.

Abstract: Background: Snakebite envenomation is a common and potentially life-threatening emergency in dogs, particularly in regions where elapid and viperid snakes are endemic. Secreted phospholipase A2 is a major toxic component of snake venom responsible for myotoxicity, neurotoxicity, hemolysis, and coagulopathy. Currently, no point-of-care tool exists to objectively quantify circulating venom burden at presentation, so clinicians rely on subjective assessment of clinical signs to guide antivenom dosing, often resulting in under- or over-treatment. Methods: We developed and analytically validated a lateral flow immunoassay strip for rapid, quantitative detection of serum snake venom (sv)PLA2 following envenomation by Micrurus fulvius (eastern coral snake). The assay employs a sandwich immunodetection strategy in which svPLA2 is captured between a FITC-labeled and a biotinylated anti-PLA2 polyclonal antibody, forming a gold nanoparticle-labeled complex that produces a visible test line at a streptavidin capture zone and a control line at an anti-FITC line. A standard curve was generated from serial dilutions of svPLA2 and fit to a log-linear regression model. The assay was then applied to serum samples from eight clinical canine patients. Results: The standard curve followed the equation y = 277,524·ln(x) − 571,808 (R² = 0.973), yielding a limit of detection of approximately 2.4 µg/mL and a limit of quantification of approximately 14.3 µg/mL. Among the eight patient samples, three showed no detectable svPLA2 (true negatives), while five showed clearly elevated signal consistent with envenomation, with intensity varying widely across positive samples, reflecting known variability in venom yield. One sample fell between the LOD and LOQ and was interpreted qualitatively as positive without quantitative estimation. Conclusions: This lateral flow strip assay reliably and rapidly distinguishes envenomated from non-envenomated dogs and stratifies relative svPLA2 burden. This platform offers a practical, equipment-light diagnostic tool to guide antivenom dosing decisions and may serve as a companion diagnostic for emerging PLA2-targeted therapies.

Abstract: Electronic cigarettes are widely promoted as a less harmful alternative to conventional cigarettes, but their cellular effects remain incompletely defined and may vary by flavouring composition. This study examined the effects of four flavoured e-cigarette aerosols, cigarette smoke, and nicotine on human THP-1 monocytic cells, focusing on cytotoxicity, oxidative stress, inflammatory activation, adhesion, and phagocytic function. Cells were exposed for 24 hours to conditioned media generated using a standardised puffing protocol and then analysed by microscopy, trypan blue exclusion, flow cytometry, and dual-fluorescence scatterplot analysis. Cigarette smoke-conditioned medium produced the greatest acute toxicity, significantly increasing cell death, membrane damage, and DNA fragmentation. In contrast, the tested e-cigarette aerosol-conditioned media and nicotine alone produced little overt cytotoxicity under the same experimental conditions. However, e-cigarette exposures increased cell adhesion and were associated with formulation- and cell subpopulation–dependent changes in CD11a, CD11b, and CD54 expression. E-cigarettes increased intracellular DCF-associated oxidative fluorescence and TNF expression, with the watermelon and strawberry formulations producing the strongest responses. Nicotine alone did not reproduce these pro inflammatory/pro oxidant effects to the same extent as the full aerosol. Instead, in non adherent cells, nicotine behaved more like an antioxidant/anti inflammatory agent, while in adherent cells, it showed a tendency toward pro oxidant/pro inflammatory behaviour, though generally weaker than cigarette smoke and e cigarette aerosols. These effects indicate that other aerosol components substantially contribute to biological activity, and the effect may be bimodal. Phagocytosis was reduced in selected groups, suggesting that some exposures may impair this cellular function. Bivariate fluorescence revealed exposure-dependent differences in the relationship between oxidative stress and TNF expression, with cigarette smoke producing the most pronounced disruption. Overall, these findings support the potential utility of bivariate fluorescence analysis as a complementary in vitro approach for detecting and comparing oxidative and inflammatory cellular responses to different e-cigarette aerosols.

Abstract: Plant growth-promoting rhizobacteria (PGPR) offer a sustainable alternative to chemical fertilizers in rice production, yet the mechanisms underlying multifunctional growth promotion by individual strains remain insufficiently characterized. In this study, strain ST2023006 was isolated from sugarcane rhizosphere and systematically characterized through phenotypic assays and whole-genome sequencing. The strain was identified as Pseudomonas chlororaphis based on morphological, physiological, biochemical, and multilocus sequence analyses. It exhibited tolerance to temperature (25-37 ℃), pH (5.0-9.0), and salinity (≤2% NaCl). ¹⁵N isotope dilution analysis revealed nitrogen fixation contribution rates of 13.4% and 9.72% to rice roots and shoots, respectively. Pot experiments demonstrated that inoculation significantly promoted growth in two rice varieties, increasing plant height by up to 86.6%, while enhancing root surface area, root volume, and biomass accumulation. In vitro antagonism against Rhizoctonia solani and quantitative IAA analysis (59.44 µg·mL⁻¹) provided experimental support for genome-predicted biocontrol and IAA production traits. Whole-genome sequencing revealed genes potentially associated with nitrogen fixation (nifU, fixI), nutrient mobilization (potassium/zinc solubilization, siderophores), phytohormone synthesis (IAA), stress tolerance (heavy metal resistance, heat/cold shock proteins), and complete biosynthetic gene clusters for the antifungal compounds pyrrolnitrin, phenazine, and hydrogen cyanide. These findings indicate that P. chlororaphis ST2023006 is a promising multifunctional PGPR candidate with demonstrated plant growth-promoting effects and experimentally supported genomic potential for biocontrol.

Abstract: Understanding human cardiac development, disease, and therapeutic response requires experimental models that reproduce defined aspects of myocardial organization and function. However, no single in vitro or ex vivo preparation captures the complete structural, electrophysiological, contractile, metabolic, vascular, stromal, and immune context of the human heart. This review critically compares five partly overlapping model families: cardiac spheroids and aggregate-based microtissues, developmentally self-organizing cardiac organoids and assembly-guided constructs, engineered heart tissues, heart-on-a-chip systems, and living myocardial slices. The comparison distinguishes how tissue organization is generated or preserved, which variables can be controlled, how function is measured, and which conclusions are supported by the resulting evidence. Across model families, interpretation is shaped by species and cell source, maturation state, cellular composition, conditioning, transport, measurement design, and donor variables. Representative studies illustrate applications in cardiac development, functional perturbation, fibrosis, immune -cardiac interactions, genotype attribution, and retained adult tissue physiology. Major limitations include domain-specific and incomplete maturation, restricted oxygen delivery, variable cellular persistence, technical reproducibility, limited access to human tissue, and insufficient consideration of donor sex and hormonal context. Rather than identifying a universally superior platform, this review proposes a claim-validation alignment in which model selection is guided by the biological question, the required tissue features, the relevant endpoint, and the level of inference supported by validation.

Abstract: Background and Objective: Maximal strength is a critical indicator of neuromuscular function essential for assessing physical performance and prescribing resistance training. Nevertheless, indirect estimation of one-repetition maximum (1RM) depends on both the predictive model and joint positioning during task execution. The aim of this study was to evaluate the influence of different glenohumeral joint positions on the estimated 1RM of the triceps brachii muscle in trained and untrained individuals, using the Matveev, Adams, and Epley equations. Methods: Utilizing a descriptive, cross-sectional, and quantitative design, this study examined 20 men separated into trained (n = 10) and untrained (n = 10) groups. Unilateral elbow extension was executed at three glenohumeral positions: 10 degrees flexion, 45 degrees extension, and 130 degrees flexion. The 1RM was estimated via the Matveev, Adams, and Epley equations, with data assessed using analysis of variance and a subsequent Tukey test (p <0.01). Results: Trained individuals exhibited higher estimated 1RM values in all evaluated positions. Glenohumeral joint position influenced estimated 1RM, with higher values at 10° of flexion and lower values at 130° of flexion. No significant differences were observed between the Matveev, Adams, and Epley equations. Conclusion: Training level and glenohumeral joint positioning influence the estimated 1RM of the triceps brachii, whereas the three equations show similar performance when applied using a standardized protocol.

Abstract: Background/Objectives: Simple, reproducible in vitro liver models are increasingly important for early screening strategies aligned with the 3Rs principles. However, three-dimensional culture effects depend strongly on spatial organization, and greater structural complexity does not nec-essarily improve functional performance. This study compared five AML12 culture configura-tions to evaluate their effects on hepatocyte-associated characteristics. Methods: AML12 cells were cultured for 7 days under matched conditions using conventional tissue-culture plastic (2D-TC), two-dimensional collagen-supported culture (2D-Coll), three-dimensional culture on a collagen gel (3D-Coll), collagen sandwich culture (3D-SW), and collagen-supported spheroid cul-ture (3D-Sph). Cellular organization, cell abundance and size, metabolic and biochemical param-eters, and transcriptional markers were evaluated using four biological replicates for gene-expression analyses and six for biochemical measurements. Multivariate analyses were used to characterize transcriptional patterns. Results: Cell number remained comparable across con-figurations, whereas cell size, metabolic activity, biochemical endpoints, and transcriptional pro-files differed. The collagen sandwich produced the most consistent hepatocyte-associated profile, with increased Alb, Hnf4a, Ttr, and Trf expression and the highest urea production. Spheroids showed the highest cell-normalized resazurin activity and multicellular aggregation, while 2D collagen exhibited a stronger stress-associated profile. PERMANOVA confirmed an overall effect of culture configuration on transcriptional profiles (R² = 0.442, p < 0.001). Conclusions: AML12 responses depended on culture configuration rather than following a simple 2D-versus-3D pattern, highlighting the importance of spatial organization when selecting in vitro liver models.

Abstract: Urbanization and macroclimatic geographic gradients exert distinct selective pressures on the morphology of widespread insect species. However, the combined effects of these factors on sexual dimorphism remain under-explored. This study investigated variations in body size and shape, and sexual dimorphism, in the ground beetle Pterostichus melanarius (Illiger, 1798) across a three-level urbanization gradient (rural, suburban, and urban). Linear morphological traits were extracted and analyzed for 4,616 specimens collected across nine regions spanning Europe and Russia. Morphological trajectories were established using a Random Forest machine learning approach for sex prediction, alongside Principal Component Analysis to separate general size from size-independent shape. We used a Random Forest machine learning approach to establish morphological trajectories for sex prediction, alongside Principal Component Analysis to separate general size from size-independent shape. We observed persistent female-biased dimorphism in both size and shape, with scaling occurring isometrically between sexes across populations. Overall body size increased significantly as populations transitioned from rural to urban habitats. However, this size increase was severely dampened in males within the most urbanized areas, resulting in wider sexual size dimorphism. Furthermore, human-altered habitats caused a widespread shift in population shape towards wider, larger-headed phenotypes, a transformation driven disproportionately by females. These statistically significant differences suggest that, while broad geographic clines establish baseline morphology, localized urbanization gradients drive rapid, sex-specific phenotypic adaptations. Ultimately, females exhibit greater morphological plasticity in response to urbanization pressures.

Abstract: Consumer distrust of synthetic additives has reshaped ingredient strategy across the global bakery sector, pushing formulators toward “clean-label” products defined by short, recognizable ingredient lists and minimal processing. This review synthesizes the peer-reviewed literature on clean-label reformulation in industrial baked goods, covering sodium reduction, sugar reduction, saturated-fat replacement, additive-free shelf-life extension, natural colorant and whole-grain/fiber fortification, and the replacement of synthetic dough conditioners with enzymes, sourdough fermentation, and natural hydrocolloids. Across these domains, a consistent pattern emerges: ingredients removed for clean-label reasons rarely perform a single function, so their replacement typically requires a combination of technologies rather than a one-for-one swap. Sourdough fermentation surfaces repeatedly as a multifunctional tool addressing sodium perception, sugar replacement, and antifungal shelf-life extension simultaneously, while enzymes and oleogel-based fat-structuring systems offer more targeted, single-function solutions. The review considers where these technologies sit on the path from laboratory demonstration to commercial deployment, and closes with a discussion of unresolved scientific and definitional gaps. A subset of the numeric claims made throughout this review were checked directly against open-access full texts rather than relying on abstracts alone; a verification appendix at the end states, claim by claim, which figures carry that higher confidence level and which remain single-source or abstract-derived.

Abstract: Background/Objectives: This study evaluated potential temporal associations between international histopathological classifications, staging updates, and the long-term divergence observed between papillary thyroid cancer (PTC) and follicular thyroid cancer (FTC) incidence trends. Applying an advanced statistical protocol comprising the recursive Supremum Chow Test, Piecewise Segmented Regression with Regime Switching (PSR-RS), and a non-linear Markov-Switching Intercept and Slope (MSIA) model, we analyzed population-level annual incidence rates (AIR) in the United States from 1975 to 2023. Methods: We implemented a two-pronged approach to identify and validate structural trend changes in incidence trends. Approach 1 tracked endogenous, data-driven incidence shifts, while Approach 2 tested the fixed calendar milestones corresponding to the official publication dates of major classifications. Crucially, this multistage analytical protocol was applied directly to the individual PTC and FTC series, as well as to the scale-independent log ratio, ln ( PTC /FTC ) and aggregate diagnostic gap timelines. Results: The results show that the incidence gap between the two cancers widened until 2014 and stabilized thereafter. AG quadrupled from 2.9 per 100,000 population in 1975 (PTC AIR = 3.6 minus FTC AIR = 0.7) to 11.6 in 2014 (PTC AIR = 12.8 minus FTC AIR = 1.2) with no further increase thereafter (AG = 11.6 in 2022 and 2023). Under Approach 1, the natural breakpoints partitioned the timelines into highly diverse behavioral segments. Specifically, the data identified five evolutionary regimes for PTC, three independent regimes for FTC, and four distinct regimes for AG. The breakpoint identified in 1987 coincided with the staging updates introduced that year and marked a clear change in the incidence trends of both histotypes. Between 1987 and 1991, PTC incidence increased, while FTC incidence declined toward its historical minimum. Despite these changes, the aggregate diagnostic gap continued along its existing trend, with no structural break detected. The 2004 WHO third edition corresponded to a systematic upward level shift (β2 = +4.1500) associated with the formal codification of microcarcinomas. Finally, both analytical approaches converged on 2014 as a significant historical turning point in the slowing of incidence growth. This phase is characterized by a significant negative slope change (Δβ = - 0.35) consistent with conservative classification updates and subsequent stabilization at a structural plateau. Conclusions: The progressively widening AIR gap between PTC and FTC does not appear to reflect a purely biological phenomenon. Instead, these population-level changes are highly consistent with systematic reporting shifts and diagnostic thresholds introduced in successive international staging and histopathological classifications.

Abstract: Strawberry (Fragaria × ananassa) is an economically important berry crop in protected horticulture, mainly produced through soil‑based and solid‑substrate soilless systems. Soil cultivation possesses strong physicochemical buffering and abundant indigenous microbial resources, but suffers from continuous cropping obstacles including soil degradation and soil‑borne pathogen accumulation. Solid‑substrate cultivation can isolate soil‑borne pathogens and enables precise rhizosphere manipulation; however, it is constrained by low buffering capacity, unstable microbial assembly, peat‑related ecological costs, and waste‑substrate disposal risks. This review builds a unified analytical framework centered on the cascade "medium physicochemical properties – microbial community – extracellular enzymes – root physiology – yield‑quality performance" and clarifies the conceptual boundaries among soil cultivation, solid‑substrate cultivation, and hydroponics. We analyze the sources of heterogeneity across published results, including cultivar genotype, medium properties, water‑fertilizer management, growth stage, and analytical methodology. Most existing evidence is correlational, whereas rigorous causal manipulation experiments remain scarce, which explains why no universal consensus has been reached on the relative superiority of the two production modes. Key research gaps include fragmented rhizospheric mechanism research, lack of cross‑study standardized experimental conditions, insufficient normalization of enzyme and microbiome datasets, and a shortage of long‑term multi‑objective field assessments. We highlight that microbial–functional decoupling frequently occurs in both soil and solid‑substrate systems, indicating that microbial community composition alone cannot represent real rhizosphere biogeochemical processes. Based on evidence synthesis, differentiated optimization strategies are proposed: soil systems should prioritize soil‑health restoration, whereas solid‑substrate systems need peat‑substitute development, microbial–habitat compatible inoculation, and substrate circular regeneration. Shared future directions cover in‑situ rhizosphere sensing, causal validation experiments, and multi‑objective evaluation balancing yield, fruit quality, resource cost, and environmental footprints. This review provides theoretical references for sustainable protected strawberry production.

Abstract: Can a stellar configuration identify a particular DNA helical shape? We test a map from two Cygnus ecliptic angles to helical repeat and rise-to-radius ratio that matches canonical B-DNA within 0.92% relative RMS error. The original route uniquely wins all five retained B-DNA structures and none of the A/Z controls in the historical panel, with continuous dominance across the B-DNA region. Selecting the five brightest stars of all 88 constellations preserves the Northern Cross and the unchanged map’s first rank at the benchmark. Cygnus alone has a minimum spanning tree with four arms meeting at one star; brightness and opposition recover its axis before molecular scoring. Repeating this retrospective selection on new quintets gives 0.17–0.21% molecular-score exceedance overall and 1.08–1.46% among admissible panels under two source models. Expanded formulas and target substitution remove canonical first rank; the exploratory molecular graph extension identifies Cygnus under H2 but not H1. Historical exceedance rises from 2.7% to 55.9% when coordinate roles and formulas are released. The geometric “stellar DNA” hypothesis links an astronomically recoverable configuration to a measured B-DNA preference, with role asymmetry defining its specificity and explicit predictions for independent tests.