Appl. Biosci., Volume 5, Issue 1 (March 2026) – 22 articles

Musculoskeletal (MSK) diseases present major health and economic challenges globally. Advancing age, diseases like osteoarthritis (OA), osteoporosis (OP), fracture and other conditions significantly reduce the quality of life (QOL) of these patients. Current pharmaceutical approaches are able to manage symptoms for some of these; however, they do not provide long-term solutions. Surgeries which are usually the final resort, present an added layer of challenges with the risk of post-surgical complications. The last couple of decades have observed an increase in the use of tissue engineering and regenerative medicine (TERM) for bone tissue engineering (BTE) applications. With the advent of artificial intelligence (AI), there will inevitably be an intersection of AI with TERM for MSK conditions. As of 2025, AI is already in use for small-scale applications in BTE including data extraction, image analysis, scaffold design and fabrication using three-dimensional (3D) printing techniques. This review outlines the convergence of these three fields and discusses the potential of their intersection. The author describes the need for this convergence, a brief update of TERM in MSK in the last decade, followed by the potential of AI in MSK-TERM. The review concludes on the challenges and future directions of the emerging field and hopes to encourage bold and ambitious collaborations between industry, academia, hospitals and health-care start-ups to realize the potential of this unique intersection. Full article

This work presents a compact microwave sensor for noninvasive blood glucose monitoring based on a substrate-integrated waveguide loaded with a complementary split-ring resonator on RO4350. The sensing principle uses shifts in resonance frequency and changes in S-parameters to track the dielectric dispersion of glucose-containing tissue. The resonator is constructed using Substrate-Integrated Waveguide (SIW) technology, which mimics the propagation characteristics of a conventional rectangular waveguide. To validate its versatility, the sensor implements three practical sample delivery modes: direct liquid contact with the sensing surface, a glass tube holder mounted over the active region, and a non-invasive fingertip interface. Electromagnetic simulations and benchtop measurements confirm clear glucose-dependent frequency shifts with stable matching and insertion levels. Across the physiological range of 20 to 200 mg·dL⁻¹, the sensor exhibits clear glucose-dependent resonance shifts in all configurations. In direct contact mode, the resonance frequency shifts from 10.83 GHz to 10.45 GHz with sensitivities up to 2.47 MHz per mg·dL⁻¹. The tube configuration shows a shift from 10.49 GHz to 10.38 GHz with sensitivity up to 0.80 MHz per mg·dL⁻¹, while reducing contamination. In the non-invasive fingertip mode, the resonance shifts from 2.56 GHz to 2.52 GHz with sensitivities up to 0.25 MHz per mg·dL⁻¹. These results confirm the sensor’s compactness, reliability, and suitability for portable, low-cost glucose monitoring. The results indicate that the proposed sensor can support practical continuous or spot monitoring and offers a clear path toward portable and low-cost glucose assessment. Full article

Cutaneous melanoma is one of the most aggressive skin cancers, and early diagnosis remains essential to reduce mortality. Reflectance Confocal Microscopy (RCM) provides non-invasive, quasi-histological images of the epidermis, dermoepidermal junction (DEJ), and dermis, enabling real-time assessment of melanocytic lesions. However, interpretation still relies on expert visual evaluation, which is time-consuming and subjective. In this context, Artificial Intelligence (AI) and Computer-Assisted Detection (CAD) systems are emerging as valuable tools to improve diagnostic accuracy and reproducibility. This review summarizes research on AI applications in RCM imaging for melanoma, focusing on three major areas: delineation of skin strata, segmentation of tissues and morphological patterns, and classification of benign versus malignant lesions. Early approaches included Bayesian classifiers, wavelet-based decision trees, and logistic regression, while recent studies have employed support vector machines, random forests, and increasingly deep learning architectures such as convolutional and recurrent neural networks. The results demonstrate encouraging accuracy in DEJ localization, the segmentation of diagnostically relevant patterns, and the discrimination of melanoma from benign nevi. We distinguish the maturity of dermoscopy-based AI (AUC (ROC) > 0.80 on large multicenter cohorts) from the still-exploratory evidence for RCM-based AI. Nonetheless, current studies are often limited by small datasets, heterogeneous protocols, and a lack of multicenter validation. Overall, progress in AI applied to RCM supports the development of CAD systems that could assist clinicians during acquisition and diagnosis, reducing unnecessary biopsies and improving early melanoma detection. Future work should address standardization, dataset expansion, and the integration of advanced AI methods to move closer to clinical implementation. Full article

Zoonotic pathogens could persist in their environment and be introduced into the food-chain. With careful screening and selection, lactic acid bacteria (LAB) could be used as direct-fed microbials (DFMs) to control these pathogens in food animals. Previously isolated LAB (n = 250) were evaluated for inhibition against Shiga-toxigenic Escherichia coli (STEC) and Salmonella enterica, using agar spot test. Tests revealed that LAB were more effective against Salmonella than STEC, with 67% showing excellent (>15 mm) inhibition. LAB (n = 65) exhibiting significant pathogen inhibition (zones > 10 mm) were tested for acid (pH: 2, 4, 5, 7) and bile (0, 0.1, 0.3, 0.5%) tolerance, and biofilm-forming capabilities. About half of the tested LAB exhibited excellent to very good tolerance. All LAB formed biofilms, with 33% forming strong biofilms. LAB (n = 59) were also examined for susceptibility to commonly used antibiotics due to their intrinsic or acquired antibiotic resistance (AR), transferrable to pathogens. Only S. thermophilus S-2 showed susceptibility to all the antibiotics. The majority were susceptible to erythromycin (88%), followed by ampicillin (85%), clindamycin (64%), tetracycline (58%), vancomycin (44%), streptomycin (15%), and gentamicin (9%). Overall, LAB showed strong inhibition against pathogens, along with survival capabilities for environmental stress conditions, and could be considered for potential DFM applications. Full article

Growing interest in natural therapies has increased the demand for essential oils; however, the complex interactions within their mixtures that dictate their final efficacy remain poorly understood. This study aimed to optimize a blend of ginger, cinnamon, tea tree, and geranium essential oils to develop an active ingredient, with synergistic multifunctional bioactivities, that was relevant to cutaneous healing. Initially, the composition and cytotoxicity for individual oils were determined; subsequently, a D-optimal mixture design was employed to evaluate three biological responses related to skin recovery: ultraviolet B radiation absorption, red blood cell lysis inhibition, and catalase enzyme activity. GC-FID analysis revealed the following major components (% w/w): cinnamon (cinnamaldehyde, 77.56%), ginger (α-zingiberene, 33.77%), geranium (citronellol, 33.6%), and tea tree (terpinen-4-ol, 38.38%). Dose–response data from essential oils tested against Detroit ATCC 551 skin fibroblasts revealed a clear cytotoxic hierarchy (IC₅₀ µg/mL): cinnamon (21.03) > ginger (25.3) > tea tree (41.67) > geranium (92.51). Cinnamaldehyde content was the primary contributor to photoprotective capacity, with a maximum sun protection factor (SPF) of 4.5. Inhibition against erythrocyte membrane lysis was not attributable to a single component; maximum protection (98.4%) was achieved through synergy between oxygenated monoterpenoids (geranium and tea tree), sesquiterpenes (ginger), and aromatic aldehydes (cinnamon). Highest catalase activity (160.86 kU/g Hb) was reached in mixtures with high cinnamaldehyde and eugenol contents, whereas an antagonistic effect was observed between tea tree and geranium oils. Finally, an optimal formulation (desirability = 0.927) was identified (% w/w): 31.7% ginger, 39.1% cinnamon, 14.5% tea tree, and 14.7% geranium. Experimental validation confirmed no significant difference compared with developed predictive models. This optimized mixture constitutes a bioactive natural component with potential for use in products aimed at promoting skin health, warranting further investigation into direct models of skin healing. Full article

Hyperlipidemia is a major global risk factor for cardiovascular disease, underscoring the need for safe, multi-target preventive strategies. In this study, two novel dietary supplements were developed by blending freeze-dried aqueous extracts of chamomile (CDS) or thyme (TDS) with linseed oil (1:1, w/w) and evaluated for their phytochemical composition, antioxidant capacity, and hypolipidemic efficacy. Total phenolics, total flavonoids, fatty acid composition, volatile constituents, and individual phenolic profiles were characterized, while antioxidant activity was assessed using DPPH· radical-scavenging and FRAP assays. Hypolipidemic activity was investigated in a Triton X-100-induced hyperlipidemia rat model through an assessment of plasma lipid parameters, oxidative stress and inflammatory markers, and liver and kidney function indices, supported by hepatic histopathology. Molecular docking was performed to explore the interactions of major bioactive compounds with AMP-activated protein kinase (AMPK) and HMG-CoA reductase. Both CDS and TDS exhibited strong antioxidant activity and high polyphenol content, with kaempferol and chlorogenic acid identified as the predominant phenolics in CDS and TDS, respectively. β-Farnesene and carvacrol were the main volatile constituents. In vivo, both formulations significantly reduced total cholesterol, triglycerides, LDL-C, lipid peroxidation markers, and TNF-α, while increasing HDL-C and improving cardiac risk indices, with more pronounced effects observed for TDS. Histopathological analyses confirmed marked hepatoprotection, particularly in the TDS-treated group. Docking analyses identified ellagic acid as the strongest dual binder to both AMPK and HMG-CoA reductase. Overall, these findings demonstrate that chamomile-linseed and thyme-linseed formulations exert synergistic, multi-target antioxidant and hypolipidemic effects, supporting their potential as nutraceutical strategies for the early prevention and management of hyperlipidemia and cardiometabolic risk. Full article

Nanotechnology has emerged as a promising option in combating the worsening situation with antibiotic resistance. We studied the antimicrobial effectiveness of four types of green synthesized zinc oxide nanoparticles (ZnO-NPs), obtained via Pluronic-assisted co-precipitation by lavender and thyme essential oils and their praseodymium-doped variants. Resazurin Microtiter Assay was applied to a panel of Gram-positive and Gram-negative bacteria from Risk 1 and 2 groups and the ESKAPE group. In relation to the pro-oxidative features of the ZnO-NPs, the production of superoxide dismutase (SOD) and catalase (CAT) in the tested microorganisms was also investigated, as these enzymes are important participants in the antioxidant defense of the bacterial cell and are considered virulence factors. We hypothesized that the sensitivity of microorganisms to the action of ZnO-NPs is related to their innate levels of antioxidant enzyme activity. The results showed that all types of studied ZnO-NPs had an antibacterial effect against the entire panel of tested strains, but with different potencies. The strongest effect was found for Arthrobacter nicotianae, Oerskovia paurometabola, Bacillus subtilis, and Escherichia coli. Less inhibition was observed for bacteria from Risk group 2 maybe due to their better antioxidant protection, especially for Pseudomonas aeruginosa. Praseodymium doping contributed to enhancing the bactericidal effect. A correlation between susceptibility of bacteria to ZnO-NPs and their antioxidant enzyme activity was observed. Full article

Seafood products are highly perishable and particularly susceptible to contamination by pathogenic and spoilage microorganisms, including Listeria monocytogenes, Vibrio spp., Salmonella spp., and Escherichia coli. Conventional control strategies in seafood processing and storage largely rely on chemical preservatives and thermal treatments, which may negatively affect sensory quality and increasingly conflict with consumer demand for minimally processed, “clean-label” foods. In this context, bacteriophages, viruses that specifically infect and lyse bacterial hosts, have emerged as natural, targeted, and environmentally sustainable biocontrol agents for food safety applications. This review provides a comprehensive assessment of bacteriophage applications in seafood processing and storage, with particular emphasis on their mechanisms of action, host specificity, and ability to selectively reduce pathogenic bacteria without compromising nutritional or sensory attributes. Recent advances in phage-based technologies, including phage cocktails, immobilized phage systems, and genetically engineered phages, are discussed in relation to their efficacy against major seafood-associated pathogens under both laboratory and industrial conditions. Key challenges limiting large-scale implementation such as phage resistance development, regulatory considerations, stability during processing and storage, and consumer perception are critically evaluated. In addition, the review highlights emerging evidence on the synergistic use of bacteriophages with complementary preservation strategies, including natural antimicrobials and innovative packaging systems. Overall, this review underscores the potential of bacteriophage-based interventions as practical and sustainable tools to enhance seafood safety, extend shelf life, and support modern seafood processing practices aligned with evolving regulatory and consumer expectations. Full article

Complex transcriptional and epigenetic regulation, including variation in promoter-level cis-regulatory architecture, influences infertility. In this study, we performed a purely in silico analysis of the −1000 to −1 bp promoter regions (relative to the annotated TSS) of 13 human fertility-related genes using an integrated motif-discovery and annotation workflow (NNPP, MEME/STREME, Tomtom, FIMO/CentriMo, GOMo, and MethPrimer). Motif discovery identified multiple statistically supported de novo promoter motifs, and motif scanning mapped their occurrences across the analyzed promoters. Similarity searches against curated PWM databases did not yield significant matches under stringent criteria, consistent with divergent or under-represented motif patterns. Functional association analysis and CpG island profiling further highlighted promoter segments that merit prioritization for follow-up testing. As the analysis is purely in silico and restricted to a fixed promoter window, the identified motifs should be interpreted as candidate regulatory elements pending experimental validation. Full article

The present study aims to evaluate the interaction of gliclazide with proteins related to inflammation—{inhibitor of nuclear factor kappa-B kinase subunit beta (IKKα) and NF-kappa-B-inducing kinase (NIK)}; oxidative stress—{kelch domain of Kelch-like ECH-associated protein 1 (KKeap1)} and ER stress—{inositol-requiring enzyme-1alpha (IRE1α)}. X-ray crystal structure of IKKα, (PDB ID: 5EBZ), KKeap1 (PDB ID: 4L7B), NIK (PDB ID: 8YHW) and IRE1α (PDB ID: 4YZ9) were obtained from Protein Data Bank and Open Babel 3.1.1 was used to prepare the ligands. Prior to docking, protein structures were prepared by removing water molecules, adding hydrogen atoms, and optimizing side chain conformations using Maestro (Schrödinger Suite, version 2024-2) along with the OPLS4 force field. Ligand docking was performed using the Glide application. Molecular dynamics simulation was performed with Desmond (Schrödinger Suite) within the Maestro interface for 100 ns for the NPT ensemble at 300 K and 1 atm pressure. Physicochemical and pharmacokinetics properties were analyzed using ADMETlab 3.0 and SwissADME. The binding energies of gliclazide with IKKα, NIK, KKeap1 and IRE1α were −8.3, −7.9, −8.4 and −8.8, respectively. Root mean square displacement (RMSD), root mean square fluctuation (RMSF) and radius of gyration analyses predicted relatively strong and stable interactions between gliclazide and the proteins, with favourable pharmacokinetic properties. It was also observed that CYP3A4 metabolizes gliclazide, in addition to CYP2C9 and CYP2C19. The activity of gliclazide against inflammation, oxidative stress and endoplasmic reticulum stress might be via interaction with these proteins. Full article

Despite significant advances in neurosurgical and critical care, traumatic brain injury (TBI) remains a major cause of morbidity and mortality. Surgical treatment of intracranial hemorrhagic lesions can only target the primary mechanical injuries and their immediate consequences but fails to address the biochemical pathological cascade that unfolds during the second injury. This review synthesizes current knowledge regarding the use of several biomarkers in diagnosis and prognosis assessment. A structured literature search was conducted by querying the PubMed database. Articles evaluating diagnostic and prognostic biomarkers in adult TBI were screened according to Prisma guidelines, and data regarding biomarkers type, cut-off values, and correlations with the outcome were extracted and summarized. Among Central Nervous System (CNS)-Specific markers, S100 calcium-binding protein (S100B) emerged as a remarkably strong negative predictor for Computed Tomography (CT)-visible intracranial lesions (NPV = 97.3–100%), whereas glial fibrillary acidic protein (GFAP) yielded both high NPV and brain specificity. Coagulation parameters such as the international normalized ratio (INR) and fibrinogen were independently correlated with mortality and unfavorable outcomes. Fibrinogen displayed a bidirectional relationship with increased mortality risk at both low (<2 g/L) and high (>4.5 g/L) values. In conclusion, biomarkers quantify the otherwise invisible progression of secondary traumatic brain injury that persists even after successful surgery. Full article

Background: Caveolin-1 (Cav-1) is a protein found in various forms and locations within cells and tissues throughout the body. Studying its structure and function provides valuable insights into key cellular processes such as growth, death, and cell signaling. This review synthesizes evidence from human studies and animal models to elucidate the complex role of Caveolin-1 (Cav-1) in regulating nitric oxide (NO) synthesis within the vasculature and perivascular adipose tissue (PVAT) during atherosclerosis. Cav-1 is a master regulator of endothelial NO synthase (eNOS), a relationship well-defined in rodent endothelial cells and cell lines. In humans, loss-of-function CAV1 mutations are linked to pulmonary arterial hypertension, suggesting a protective vascular role. Paradoxically, Cav-1 is upregulated in atherosclerotic plaques. Whether this represents a pathological process reducing NO bioavailability or a compensatory response remains unclear. Furthermore, the direct translation of the Cav-1/eNOS axis to PVAT—a metabolically active tissue expressing Cav-1—is not fully established outside of preclinical models. PVAT influences vascular tone and inflammation, potentially contributing to the paradoxical, stage-specific roles of Cav-1 in disease. Resolving these questions requires integrating human observational data with mechanistic insights from animal models to evaluate Cav-1 as a therapeutic target in vascular disease. Full article

Metabolic profiling enables comprehensive characterisation of the small molecules that are part of the biochemical composition of biological fluids. The most widely profiled biofluids include serum and plasma. Additionally synovial fluid provides a direct reflection of the metabolomic environment of joints and holds promise for biomarker discovery in arthropathies. However, the reproducibility of metabolomics data is highly sensitive to pre-analytical variation, and at the present time, standardised protocols for synovial fluid remain underdeveloped. This review aims to identify and evaluate the existing literature on effects of biofluid pre-analytical handling treatments on metabolic profiles. This review was conducted and reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines. A search was carried out to identify studies employing LC-MS, GC-MS, and NMR spectroscopy for the investigation of factors including sample collection variables, pre-centrifugation conditions, centrifugation parameters, post-centrifugations conditions, sample storage conditions, and freeze/thaw cycling. Best practice recommendations emerging from this review include the use of additive free serum and heparin plasma tubes, the centrifugation of samples within two hours of collection, immediate storage of samples at −80 °C, and avoidance of repeated freeze/thaw cycling. However, while pre-analytical influences have been extensively characterised for plasma and serum, evidence for synovial fluid remains limited. Overall, the findings highlight the existing recommendations for plasma and serum and demonstrate the need for standardised pre-analytical protocols and validation of quality control markers to advance synovial fluid metabolomics. Full article

Background: The Allium cepa test is a widely used, cost-effective, and versatile model for assessing cytogenotoxicity. Cytotoxicity is determined through changes in root growth and the mitotic index, while genotoxicity is identified through chromosomal aberrations such as breaks, bridges, and micronuclei. Objective: To synthesize the methodological principles, applications, and interpretation of the assay’s endpoints, with emphasis on environmental monitoring, nanotoxicology, and the evaluation of emerging materials. Methods: An exploratory analytical approach was applied to identify and compare studies employing the Allium cepa assay across different contexts. The literature, selected from scientific databases, was organized to highlight methodological diversity and biomarker performance. Conclusions: Compared with other models, Allium cepa stands out for its simplicity, the availability of multiple cytogenotoxic markers, and its minimal ethical constraints, making it especially suitable for research in low-infrastructure settings. Future studies should work toward the international standardization of methodologies, the integration of this model with molecular and omics-based approaches, and its incorporation into predictive frameworks for environmental and human health risk assessment. In an increasingly complex toxicological landscape, Allium cepa emerges as a pivotal tool for enhancing toxicological surveillance and safeguarding biological systems. Full article

Escherichia coli is a major pathogen responsible for urinary tract infections, septicemia, and other clinically relevant conditions, with increasing multidrug resistance (MDR) limiting available treatment options. In this context, bacteriophages represent a valuable resource for exploring novel antimicrobial and biotechnological tools. Here, we report the isolation and genomic characterization of BorMax, a novel lytic phage infecting multiple MDR E. coli. Transmission electron microscopy revealed a tailed morphology consistent with Dhillonvirus. Whole genome sequencing and de novo assembly showed a linear double-stranded DNA genome of 45,502 bp, encoding 70 predicted coding sequences (CDSs) and lacking tRNAs. Bioinformatic analyses confirmed the absence of lysogeny-associated genes, as well as virulence and antimicrobial resistance determinants. Comparative genomics using classified BorMax within the genus Dhillonvirus as a new species, sharing <77% intergenomic similarity with known members. Notably, predictions using PaCRISPR and AcRanker identified four CDSs with strong anti-CRISPR (Acr) potential, representing previously undescribed Acr candidates in this group. These genomic features highlight the novelty, safety, and potential biotechnological relevance of BorMax and contribute to the expanding genomic and functional diversity of Dhillonvirus and E. coli-infecting phages. Full article

The heat shock protein 60 (Hsp60) is a highly conserved molecular chaperonin belonging to the chaperone system, a complex network that maintains proteostasis and regulates numerous cellular processes beyond protein folding. Initially described as a mitochondrial protein essential for the folding of newly imported polypeptides, Hsp60 is now recognized as a multifunctional molecule. Its expression, localization, and post-translational modifications dynamically influence cell fate and tissue homeostasis. Alterations in Hsp60 quantity, structure, or distribution underlie a heterogeneous group of disorders known as chaperonopathies, which may occur “by defect,” “by excess,” or “by mistake” (also called “by collaborationism”). Genetic Hsp60’s chaperonopathies are associated with rare neurodegenerative and cardiovascular diseases, whereas acquired forms contribute to widespread conditions, including autoimmune, inflammatory, and malignant pathologies. This review provides a comprehensive overview of Hsp60 biology across human systems, emphasizing its structural plasticity, context-dependent functions, and dual role in health as both a biomarker and a therapeutic target. The emerging paradigm of chaperonotherapy, encompassing positive strategies to restore protective chaperones and negative strategies to inhibit pathogenic ones, highlights the translational potential of targeting Hsp60. Understanding the molecular mechanisms governing its activity will be essential for developing precision medicine approaches aimed at modulating the chaperone system in human disease. Full article

Probiotics, whether consisting of a single strain or multiple strains, are attracting growing interest in the management of type 2 diabetes mellitus (T2DM). However, their efficacy remains a matter of controversy and requires careful consideration. Accordingly, this meta-analysis aimed to compare the efficacy of single-strain to that of multi-strain probiotics supplementation on glycated haemoglobin (HbA1c) and fasting blood glucose (FBG) levels in adults with T2DM. Nineteen articles published between 2017 and 2024 obtained from 4 databases (Cochrane, Web of Science, Scopus, and PubMed) were included. These interventions, conducted in a total of 1159 participants, lasted from 6 to 24 weeks and were based on clearly identified probiotic formulations, with assessments of HbA1c and FBG. The results showed that, overall, probiotic supplementation had no significant effect on HbA1c (−0.24%; 95% CI [−0.76; 0.27]; p = 0.36), although a trend towards reduction was observed for single-strain formulations (−0.57%; p = 0.05). Regarding FBG, only the multi-strain group showed a significant reduction (−0.76; 95% CI [−1.18; −0.34]; p < 0.001), while the effect of the single-strain formulation was not significant. The comparison between the two formulations (Wald test) showed that there was no significant difference (p ≤ 0.05). However, high heterogeneity (I² > 75%) and variable strains/doses limit confidence in these findings. Full article

This study aimed to develop a robust morphometric-based framework for classifying Apis cerana populations using deep learning and machine learning approaches. Previous studies on Apis cerana population differentiation have primarily relied on manual morphometrics or genetic markers, which are labor-intensive and often lack scalability for large image-based datasets. Forewing landmarks were automatically detected through a deep learning model employing a heatmap regression and Hourglass Network architecture. The extracted coordinates were processed by Principal Component Analysis (PCA) for dimensionality reduction, and shape alignment was further refined through Procrustes ANOVA to minimize non-biological variation. Nine machine learning algorithms were trained and compared under identical preprocessing and validation settings. Among them, the Extra Trees classifier achieved the highest accuracy (99.7%) in distinguishing the three populations—A. cerana cerana from China and A. cerana indica from Thailand, the northern and southern populations. After applying error-based data filtering and retraining, classification accuracy improved further, with almost perfect population separation. The Procrustes ANOVA confirmed that individual variation significantly exceeded residual error (Pillai’s trace = 1.13, p < 0.0001), validating the biological basis of shape differences. Mahalanobis distance and permutation tests (10,000 rounds) revealed significant morphological divergence among populations (p < 0.0001). The integration of geometric alignment and ensemble learning demonstrated a highly reliable strategy for population identification, supporting morphometric and evolutionary studies in Apis cerana. Full article

Mtx1 is a mosquitocidal protein that exhibits high toxicity toward Culex species. It is produced during the vegetative phase of Lysinibacillus sphaericus but at very low levels and is rapidly degraded. The low expression appears to result from a weak promoter and a potential regulatory stem-loop structure in the 5′ untranslated region. To investigate this regulation, promoter variants of mtx1 were constructed to disrupt stem-loop formation, and promoter activity was assessed using green fluorescent protein (GFP) as a reporter. Disruption of the inverted repeat resulted in approximately twofold higher fluorescence compared with the wild-type promoter in L. sphaericus 2297, indicating partial derepression of translation. To improve protein stability, Bacillus subtilis WB800N, a protease-deficient host, was employed for heterologous expression. Truncated Mtx1 (tMtx1) was secreted into the culture medium, and no obvious degradation products were detected by Western blot analysis under the conditions tested. Although the overall yield was low and not quantitatively determined, the secreted protein retained biological activity. Larvicidal assays showed elevated mortality in tMtx1-containing culture supernatants, with an estimated LC₅₀ at approximately a 1:83 dilution and detectable activity up to a 1:512 dilution relative to control cultures. These results demonstrate that the upstream inverted repeat contributes to partial repression of mtx1 expression in L. sphaericus and that protease-deficient B. subtilis can be used as a host for producing biologically active tMtx1, although further optimization will be required to improve yield. Full article

One of the global challenges is the deficit of food. Food production is highly dependent on the productivity of agricultural plants used by humans and livestock. Various chemical and natural compounds are used to stimulate plant growth and increase their resistance to stress. The aim of our study was to analyze the chemical composition of extracts of the most common Ural tinder fungi and their effect on the early stages of wheat growth. Water–alcohol extracts from five wood-destroying fungi contained biologically active compounds (BACs), such as phenolics, free amino acids and reducing sugars. F. pinicola was characterized by the smallest amount of extracted substances. F. fomentarius has the largest amount of phenolic compounds and sugars, and I. obliquus had the highest concentration of free amino acids. Qualitative analysis revealed alkaloids in P. betulinus, and anthraquinones in F. fomentarius. Saponins were found in all tested species, except F. fomentarius. The extracts stimulated the early stages of wheat development at concentrations of 1.0–0.2 g of fungal biomass per liter. Seed germination rate was comparable to the control samples or exceeded it, and the length of roots and shoots increased. Thus, extracts from fruiting bodies of studied fungi can be recommended for priming wheat seeds, and for biotechnological cultivation. Full article

Fruit softening in Carica papaya L. is a significant postharvest limitation, primarily driven by the dynamic remodeling of cell wall polysaccharides. In this study, we conducted a genome-wide identification and in silico characterization of gene families involved in cell wall assembly and disassembly in papaya. A total of 181 genes were identified and classified into metabolic pathways: hemicellulose (58), pectin (69), extensin (24), expansin (13), and cellulose (17). These genes encode 176 predicted proteins, ranging in size from 100 to 1093 amino acids, featuring family-specific catalytic domains, including glycosyl hydrolases, transferases, and serine/threonine kinases. Phylogenetic analyses revealed strong conservation within the expansin-A and pectin polygalacturonase subfamilies, while hemicellulose-related XTH genes exhibited significant diversification. Experimental validation of nine XTH members confirmed this diversification, with amplicons ranging from 322 to 1370 bp, consistent with computational predictions. Notably, CpXTH1 and CpXTH32 produced bands of approximately 1200 and 1400 bp, respectively. These findings underscore the complexity of papaya cell wall gene families and provide a molecular framework for understanding fruit softening. Given that postharvest losses of papaya in Mexico exceed 34.7% of production (approximately 150,000 tons annually), our results offer valuable genomic resources for biotechnological strategies aimed at extending shelf life and reducing economic losses. Full article

The search for effective anti-cancer therapies is one of the most significant goals of modern medicine. The combination of oncolytic viruses (OV) and mRNA immunoadjuvants can significantly improve the outcome or even substitute traditional immunotherapy. In addition to the direct OV-mediated cytotoxic elimination of tumor cells, both OV and mRNA immunoadjuvants can significantly alter the immunosuppressive tumor microenvironment (TME) supporting cancer cells and unleash the immune response against the malignant cells. The present study is aimed at assessing the therapeutic effects of recombinant vesicular stomatitis virus (rVSV) and lipid nanoparticles (LNP) delivering mRNA coding for murine interleukin-12 (mIL12) and granulocyte-macrophage colony-stimulating factor (mGMCSF) (mRNA-LNP) in colorectal carcinoma CT26-induced tumors both as independent therapies and in combination with each other. The results of the in vivo experiment on BALB/c mice demonstrated that rVSV monotherapy did not have a significant effect, with the tumor growth inhibition index (TGII) ranging from 13.7 to 29.8% on days 6–10 after the therapy start. While monotherapy with mRNA-LNP was more effective (TGII of 48.6–53.7%), it was the therapy combining the two approaches (rVSV and mRNA-LNP) that resulted in the highest TGII of 66.7% on day 10. While these results can be further improved by optimizing the experimental design, they show the great potential of combination immunotherapy for the treatment of oncological diseases. Full article