Terephthalic acid (TPA) from poly(ethylene terephthalate) (PET) depolymerization is a promising substrate for microbial upcycling, yet complete degradation at high concentrations has not been reported above 120mM. Here, we report seven Rhodococcus strains isolated from compost enrichment cultures using TPA as the sole carbon source. Three isolates completely consumed 240mM TPA in minimal medium under high osmolarity conditions, surpassing the previously reported threshold. Strain TA18 produced β-linked exopolysaccharides and a polyhydroxyalkanoate composed mainly of 3-hydroxyvalerate from TPA as the sole carbon source, neither of which has been previously described from this substrate in Rhodococcus, alongside C16 and C18 fatty acids. These strains expand the set of robust TPA-assimilating bacteria available for PET upcycling and provide a foundation for developing routes that convert plastic waste streams into multiple microbial products.

Exosomes, a specialized group of extracellular vesicles (EVs) ranging from 30 to 150nm in size, have emerged as important mediators of intercellular communication across both mammalian and plant kingdoms. By facilitating the transfer of bioactive cargo, these vesicles orchestrate complex signalling networks essential for physiological homeostasis and pathological progression. Extensive research into both plant and mammalian exosomes has established their inherent biocompatibility and potential as cell-free systems for the targeted delivery of therapeutic drugs in the management of various diseases, including cancer. Moreover, exosomes have unique molecular signatures that enable the identification of their cell of origin and serve as high-fidelity biomarkers for precise disease diagnosis. This review provides a comprehensive overview of exosomal biology, beginning with the critical importance of standardized isolation and enrichment protocols to ensure sample purity and yield. This review also evaluates contemporary detection and characterization methodologies, ranging from nanoparticle tracking analysis to super-resolution microscopy, which are essential for defining exosomal identity. Moreover, the dual role of exosomes in cancer and disease progression, emphasizing their ability to modulate the tumour microenvironment and serve as non-invasive biomarkers for liquid biopsies, has also been explored. Bioengineering strategies such as genetic manipulation of the host cell or chemical modification of the exosome surface, can enhance their targeting precision and therapeutic loading capacity. The review also discusses the capacity of these vesicles to be loaded with exogenous drugs, including small molecules and RNA-based therapeutics, transforming them into potent, site-specific delivery vehicles. Despite their immense potential as biocompatible, cell-free drug delivery vehicles, several translational hurdles, including large-scale manufacturing consistency and regulatory complexities, remain as significant challenges, which if addressed, can lead to a shift toward personalized exosome-based diagnosis and therapy that could revolutionize the management of cancer and other complex systemic disorders.

Basal stem rot (BSR), a devastating disease caused by Ganoderma boninense (synonym G. orbiforme) significantly affects the oil palm industry, particularly in Indonesia and Malaysia. Current control measures for BSR are limited, prompting the need for environmentally sustainable biocontrol alternatives. In this study, the antagonistic potential of medicinal mushrooms against G. orbiforme to mitigate BSR in oil palm were evaluated. Among the tested species, G. lucidum exhibited the highest percentage inhibition of radial growth (PIRG) at 60.96%, followed by G. neojaponicum (50.37%) and Lignosus rhinoceros (13.50%). The biocontrol efficacy of G. lucidum was further assessed using a modified air-lift bioreactor (MALB) system, demonstrating that G. lucidum is non-pathogenic to oil palm and enhanced plant growth by 41.67% in terms of dry weight. In contrast, G. orbiforme inoculation resulted in typical symptoms such as leaf necrosis, yellowing and browning, Cercospora leaf spots and extensive hyphal mass growth, thereby confirming the pathogenicity of this species in oil palm. Infected plants experienced a significant reduction in dry weight by 115.31%. Furthermore, inoculating G. lucidum into plants either before or after inoculation with G. orbiforme positively impacted plant growth, with more pronounced effects when G. lucidum was introduced first, followed by the pathogenic G. orbiforme as tested in MALB system. These findings suggest that G. lucidum could be effectively utilized as a biocontrol agent to control BSR.

The increasing reliance on fossil fuels for global energy production has intensified greenhouse gas emissions, highlighting the need for sustainable energy alternatives. Hydrogen is considered a promising green fuel due to its high energy density and conversion efficiency. Among various production pathways, microalgae-based biohydrogen generation via biophotolysis is particularly attractive owing to its high biomass productivity, adaptability to diverse water sources, flue gas mitigation potential, and low land requirements.This study investigates the effects of key microalgal growth parameters on biohydrogen production by Chlorella sp. through biophotolysis. The impacts of nitrogen purging during the transition from aerobic to anaerobic conditions, different photoperiod regimes (continuous illumination, continuous darkness, and a light-dark cycle), glucose supplementation (5, 10, and 15 g L⁻1), and temperature (25, 30, and 35 °C) were systematically evaluated. Microalgal cell density was monitored during hydrogen production to elucidate its relationship with hydrogen yield. Initial experiments were conducted in 10 mL test tubes to identify optimal conditions, which were subsequently applied to scale-up experiments in a 1000 mL jacketed reactor. Nitrogen purging significantly enhanced hydrogen production by removing oxygen and activating hydrogenase, resulting in a peak hydrogen concentration of 11 ppm. Continuous illumination yielded higher hydrogen levels than darkness and light-dark cycling. Glucose addition substantially increased hydrogen production, with the highest yield observed at 15 g L⁻1 (30 ppm). An optimal temperature of 30 °C also maximized hydrogen production. Under these conditions, hydrogen production increased as cell density decreased due to metabolic shifts. Scale-up experiments achieved a 405-fold increase in hydrogen yield, demonstrating the scalability potential of the process. These findings emphasize the importance of optimizing algal growth conditions to balance microalgal growth and biohydrogen production for future industrial applications.

Fecal calprotectin (FCP) is a crucial non-invasive biomarker for diagnosing and monitoring inflammatory bowel disease (IBD). However, the standard enzyme-linked immunosorbent assay (ELISA) is time-consuming and labor-intensive. This study aimed to develop a novel, independent latex particle-enhanced turbidimetric immunoassay (PETIA) suitable for open-channel automated biochemical analyzers, offering a flexible and cost-effective alternative to existing commercial PETIA kits. Recombinant human S100A8 and S100A9 proteins (the subunits of calprotectin) were expressed inE. coliand purified. Polyclonal antibodies were generated in rabbits by immunization with these proteins. The antibodies were then covalently coupled to carboxylated latex particles (188 nm) to create the PETIA reagent. The assay was validated on an LC-400 specific protein analyzer. Analytical performance-including repeatability, within-lab precision, and interference-was assessed. The method was evaluated using clinical fecal samples (n = 104) and compared with a commercial ELISA kit (BÜHLMANN fCAL). The developed PETIA demonstrated a measuring range of 0-1500 µg/g based on a six-point calibration curve (R2 = 0.99). A strong correlation was observed between the new PETIA and the reference ELISA (r = 0.98). The regression equation was y = 1.04x + 8.82. Bland-Altman analysis confirmed good agreement between the two methods, with 95.2% of data points within the 95% limits of agreement. The PETIA showed strong correlation with the established ELISA for FCP quantification, offering a rapid, automated alternative suitable for high-throughput laboratories. This study, encompassing development from antibody to reagent, supports its translational potential.

Reproductive disorders remain a major cause of subfertility and infertility in human and animals, encompassing ovarian dysfunction, endometritis, impaired endometrial receptivity, and treatment-induced gonadotoxicity. These conditions substantially reduce reproductive efficiency and quality of life. In recent years, advances in translational medicine have shifted focus toward regenerative strategies, particularly stem cell-based therapies, as alternatives to conventional pharmacological or surgical interventions that often yield limited or transient benefits. Among these, mesenchymal stem/stromal cells (MSCs) have emerged as promising candidates for infertility management due to their self-renewal ability, multilineage differentiation potential, low immunogenicity, and robust paracrine activity. Growing preclinical and early clinical evidence demonstrates that MSC-based therapies can restore ovarian function, enhance folliculogenesis, promote endometrial regeneration, and modulate inflammatory and fibrotic microenvironments associated with female infertility. Notably, these effects are mediated primarily through the secretion of bioactive factors, stimulation of angiogenesis, regulation of apoptosis, and immunomodulation rather than direct cell replacement alone. Despite their therapeutic promise, challenges related to targeted delivery, therapeutic precision, and safety remain. In this context, emerging targeting strategies, particularly aptamer-based approaches, represent a rational advancement to enhance MSC efficacy. Aptamers are short single-stranded DNA or RNA molecules with high affinity and specificity for target proteins, offering advantages over antibodies, including improved tissue penetration, minimal immunogenicity, and ease of synthesis and modification. This review aimes to summarizes current progress in MSC-based therapies for female infertility and highlights aptamer-guided targeting strategies to improve therapeutic precision, efficacy, and safety.

Millettia pinnata (M. pinnata) is a medicinally important plant containing many phytoconstituents capable of curing various diseases. Flavonol synthase (FLS, EC 1.14.20.6) has a significant role in flavonoid biosynthesis like kaempferol having various pharmacological properties. The present study aimed to characterize the FLS gene and its encoded protein (MpFLS) in M. pinnata comprehensively. FLS gene was characterized by isolating RNA from M. pinnata leaves, cDNA synthesis, PCR-based amplification of the desired FLS gene, sequencing of the PCR amplified product and sequence analysis followed by detailed protein-protein interaction and protein flexibility analysis. PCR product sequencing and analysis revealed that the full-length cDNA of M. pinnata FLS contains a 952-base pair long open reading frame (ORF) encoding 234 amino acids that has been submitted to NCBI (OM469017). The NCBI blastp analysis of 234 amino acid sequences of FLS shows 81.9% similarity to Vigna unguilata, whereas blastn analysis exhibited 83.31% identity to Clitoria ternatea L. that belongs to the 2OG-FeII Oxy superfamily. In silico characterization of primary, secondary, and tertiary structure, docking of MpFLS with dihydrokaempferol showed a strong binding (-10.0kcalmol⁻1) stabilized by hydrogen bonds (Gly54, Glu256) and hydrophobic/π-π interactions, highlighting a specific and stable enzyme-substrate complex. STRING based protein-protein interaction analysis and detailed ProtScale and MEDUSA based protein flexibility analysis of FLS protein revealed that FLS it is highly stable and hydrophilic. M. pinnata FLS gene have 952-base pair long ORF while encoding FLS protein of 234 amino acids long. FLS belong to 2-oxoglutarate/Fe(II)-dependent oxygenase superfamily that utilize molecular oxygen and 2-oxoglutarate to oxidize various substrates. Molecular docking results indicate that MpFLS binds dihydrokaempferol with high specificity and stability, supporting its key role in kaempferol biosynthesis in Pongamia pinnata. Present study provides systematic and comprehensive analysis on FLS gene and its encoded FLS protein in M. pinnata revealing that FLS protein is highly stable and hydrophilic in nature.

Glycoprotein hormones, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), and thyroid-stimulating hormone (TSH), play essential roles in the reproductive endocrinology of teleosts. However, research on these hormones in non-model species such as the Japanese eel (Anguilla japonica) has been constrained by the lack of specific immunological detection tools. Recombinant A. japonica FSH, LH, and TSH were expressed using a Bombyx mori-based silkworm-baculovirus system. Polyclonal antibodies were then generated using two distinct immunogen strategies: (i) chimeric constructs fusing β-subunits with a murine α-subunit, and (ii) synthetic peptides representing unique, non-conserved β-subunit epitopes. Both approaches induced antibody responses, but only peptide-based immunogens yielded polyclonal antisera with high specificity and sensitivity. These antibodies distinguished homologous glycoprotein hormones without cross-reactivity and retained target selectivity in mixtures simulating physiological serum conditions. Peptide-derived antibodies provide a robust and precise tool for the immunological discrimination of FSH, LH, and TSH in A. japonica. This strategy establishes a scalable framework for species-specific hormone assay development and offers translational potential for endocrine monitoring, reproductive management, and diagnostic applications in aquaculture biotechnology.

The isolate A85 was identified as Streptomyces enissocaesilis, and it might serve as a novel actinobacteria source for uricase production. The uricase was successfully produced and purified from this novel strain, demonstrating that it could be an efficient source for future applications. Characterization studies of the purified enzyme showed maximal activity at pH 9 and 37°C. From the differential scanning calorimetry, a melting point of 55.6°C was observed, which ensured that the purified uricase is thermostable. Overall, 32 fold purification with a 48% yield of uricase was recovered at the end of purification steps. Inhibitor studies revealed that the purified uricase is metalloprotease. Vmaxand Kmvalues were found to be 30.39 U/mL and 0.0212mM, respectively. The molecular mass of the purified uricase was found to be around 32 KDa. Further MALDI-TOF MS analysis identified four peptides as uricase, with a significant score. The purified enzyme was classified as an uricase based on a comprehensive analysis that integrated its amino acid sequence and conserved regions. The secondary structure of the purified uricase using PDBsum revealed 1 beta sheet, 1 beta hairpin, 1 beta bulge, 2 strands, and 2 beta turns. The 3D structure of the purified uricase was predicted through homology modeling, and the structure was validated using Ramachandran plot analysis. Docking results revealed significant binding affinity of -4.3kcal/mol, which indicates strong and stable interaction of uricase and uric acid substrate.

Bacillus subtilis strain AF2 was obtained from the Mediterranean Sea and its identification was determined by its phylogenetic analysis of 16S rRNA sequences, as well as their morphological, physiological, and biochemical characteristics. The isolate yielded EPSS4 at a concentration of 7.4g/l. EPSS4 constituted a significant proportion of the EPS recovered through purification using DEAE-Cellulose. The sample consisted of sulfate (24.71%) and uronic acid (18.55%). The fraction's viscosity was measured to be 1.1mm2/sec, and the total hexose amine content was measured to be 9.06%. The monosaccharide composition of this fraction consists of glucose, fructose, xylose, and glucuronic acid in a molar ratio of 1.0:0.5:1.0:1.0, respectively. The toxicity studies on EPSS4 demonstrated its safety at 5g/kg doses. EPSS4 underwent evaluation for its antioxidant, anticancer, and anti-inflammatory properties. The results indicated that the antioxidant activity percentages were as follows: 85.17 ± 0.77% for DPPH, 79.63 ± 1.05% for ABTS, 52.20 ± 0.67% for Fe2+ ion chelation ability, 75.80 ± 0.28% for Lipid peroxidation Inhibition capacity, 84.66 ± 1.21% for O2- radicals scavenging capacity, and 55.09 ± 0.34% for NO scavenging capacity. The IC50 value of EPSS4 for HepG-2 is 238.05 µg/ml. A-549 has determined the IC50 values to be 102.71µg/ml. The concentration of HCT-116 was measured to be 328.01µg/ml, while the IC50 value for the MCF-7 cell line was determined to be 204.77µg/ml. The IC50 values for the HEP-2 and PC-3 cell lines were 118.95 and 128.94µg/ml, respectively. The anti-inflammatory activity of EPSS4 was assessed using various methods, including measuring its inhibitory effects on lipoxygenase (LOX) and cyclooxygenase (COX2). At a dosage of 100µg/ml, EPSS4 had a LOX inhibitory activity of 80.54 ± 1.09% and a COX2 inhibitory activity of 84.70 ± 1.05%. Due to its antioxidant capabilities, the EPS produced from Bacillus subtilis strain AF2 is a promising option for application as a potent antioxidant agent in the healthcare system. So, from observed activitiesdemonstrate that it has the potential to be employed as an anticancer, antioxidant and non-steroidal anti-inflammatory substance via improving adaptive immune responses.