Search Results (6,065)

The proteolytic processing of the amyloid precursor protein (APP) is a core pathological event in Alzheimer’s disease (AD) pathogenesis, yet the global genetic regulatory networks modulating this process have not been fully characterized. To systematically identify novel regulators of APP cleavage, we performed a genome-wide CRISPR/Cas9 knockout screen utilizing an optimized UAS-GAL4-based cellular reporter, and identified genetic modulators governing amyloidogenic and non-amyloidogenic processing. The screen uncovered distinct functional gene clusters regulating the APP, prominently involving cellular metabolism, protein modification, and vesicular trafficking. Specifically, LDHB, PIAS2, CCDC53, and TRIM61 emerged as novel functional modulators. Biochemical validation confirmed that ablating these genes significantly alters the metabolic balance between sAPPα and amyloid-β (Aβ) production. Finally, integration with human AD transcriptomic datasets demonstrated that these identified modulators undergo significant dysregulation in clinics. Together, these findings establish a reporter-based functional screening framework for APP processing and identify candidate regulatory nodes linked to metabolism, protein modification, and vesicular trafficking. These candidates provide a resource for future mechanistic investigation and validation in more disease-relevant AD models. Full article

Background: Arginase 1 deficiency (ARG1-D) is an ultra-rare urea cycle disorder characterized by hyperargininemia and progressive neurological impairment, including spasticity, loss of motor function, and reduced quality of life. Conventional management based on dietary protein restriction and ammonia scavengers rarely achieves adequate metabolic control or prevents neurological deterioration. Pegzilarginase, a recombinant human arginase 1 enzyme, is the first disease-modifying therapy for ARG1-D. Methods: We report the first Italian real-world experience with pegzilarginase in three pediatric patients with genetically confirmed ARG1-D enrolled in the phase 3 PEACE trial. Clinical, biochemical, functional, nutritional and quality-of-life data were retrospectively collected over a long-term follow-up (2003–2025). Outcomes were evaluated across three phases: treatment initiation (Start), a 13-month treatment interruption due to trial closure (Stop), and therapy re-initiation through an early access program (Restart). Results: Pegzilarginase rapidly normalized plasma arginine levels and was associated with improvements in motor function, spasticity, walking endurance, dietary protein tolerance, bone mineral density, and quality of life. During treatment interruption, all patients experienced biochemical worsening and clinical deterioration, including increased spasticity, reduced mobility, and emotional distress. Re-initiation of pegzilarginase restored metabolic control and led to progressive neurological and functional recovery, including partial reversal of long-standing motor deficits. Conclusions: This real-world experience supports pegzilarginase as a disease-modifying therapy for ARG1-D. Sustained normalization of plasma arginine, rather than subthreshold biochemical control, correlates with functional and neurological improvement and may partially reverse non-lesional metabolic brain injury. Early initiation of pegzilarginase, including in newborn-screened patients, may further modify the natural history of ARG1-D. Full article

Growing municipal solid wastes, environmental deterioration, and the world’s increasing energy demand highlight the urgent need for effective, sustainable energy recovery solutions. Uncontrolled municipal solid wastes contribute explicitly to the global crises of climate change, pollution, and biodiversity loss. Food and organic waste are converted into value-added products using biochemical and thermochemical techniques. Anaerobic digestion (AD) is a versatile, multi-phase waste-to-energy technology that transforms organic waste into renewable energy in an oxygen-free environment. AD uses microorganisms to break down waste, yielding biogas (mostly methane and carbon dioxide) and digestate, a nutrient-fortified by-product. Compared with traditional Single-Stage Anaerobic Digesters (SSAD), Two-Stage Anaerobic Digesters (TSAD) offer notable benefits by separating hydrolysis–acidogenesis from acetogenesis–methanogenesis. These include increased methane yield, improved process control, increased microbial stability, and resistance to inhibitory substances. According to the literature, TSAD systems have been shown to increase methane yield by about 10–30% compared to SSAD. This article covers the dynamics of the microbial population at various stages, the impact of operational factors (HRT, OLR, pH, and temperature), and novel reactor designs with modular and multi-state functions. In line with Oman’s Vision 2040, this study discusses the continuous operation of a two-phase AD co-digestion process and the in-depth techno-economic feasibility of decentralized waste management through optimized biogas production. Optimizing the carbon-to-nitrogen (C/N) ratio within the range of 20–30 in co-digestion systems significantly enhances microbial activity and methane production. The potential of recent developments, such as microbial immobilization, biogas generation techniques, and hybrid integration with photobioreactors or electrochemical systems, to enhance the scalability and efficiency of bioconversion is addressed in a TSAD system. In addition to encouraging circular economy principles through efficient organic waste valorization, this review identifies TSAD as a promising approach to achieving the SDGs related to sustainable cities, clean energy, and responsible consumption. Full article

As global demand grows for natural health-promoting food ingredients, the agri-food industry’s organic wastes are emerging as promising alternatives thanks to attributes such as biocompatibility, nutritional value and nutraceutical effect. During saffron (Crocus sativus L.) production, approximately 53 kg of petals are obtained as a by-product for every 1 kg of saffron spice. The use of saffron petals and petal extracts in bakery products improves products’ shelf life due to the petals’ high content of nutraceuticals and minerals acting as natural preservatives. Petal-enriched bakery products contain high levels of fiber, minerals and antioxidants. Addition of saffron petals into bread dough reduces gluten network strength, increases crumb hardness, enhances acidity, improves water retention, alters color profiles and increases the duration of the shelf life. The formulation for incorporating saffron petals or petal extracts into food products must address three primary criteria: the maximum concentration of bioactive compounds per 100 g of the finished matrix, the thermal stability of these compounds during the baking process, and their bioavailability (in the food matrix) within the human gastrointestinal tract. Nutraceuticals with pharmacological potential are also influenced by the compositional profile: the proximate composition, minerals, phenolic content, flavonols, and antioxidant capacity of saffron petals and bakery products containing saffron petals. Saffron petals exhibit diverse therapeutic potentials, acting as antidepressants, anxiolytics, anticonvulsants, and neuroprotective agents. They also offer metabolic, cardiovascular, hepatoprotective, and renoprotective benefits, along with anti-inflammatory, antimicrobial, and antitumor activities. This article proposes a roadmap for developing bakery products enriched with saffron petals or petal extracts, targeting both pharmacological applications and consumer goods focused on disease prevention and general wellness. This study investigates the biochemical composition of saffron petals and their integration into bakery products. It evaluates the influence of petal-derived additives on rheological properties, shelf stability, and organoleptic characteristics, alongside an assessment of their bioactivity and toxicological profiles. Furthermore, the analytical methodologies employed for ingredient and biological sample characterization are discussed, emphasizing their role in verifying the authenticity, safety, and nutritional functionality of both raw materials and finished formulations. Full article

Cisplatin is a potent chemotherapeutic agent whose clinical application is frequently limited by severe nephrotoxicity. N-acetylserotonin (NAS), a precursor of melatonin and a selective agonist of the TrkB receptor, has demonstrated significant antioxidant and neuroprotective properties. This study aimed to evaluate the potential renoprotective effects of NAS against cisplatin-induced acute kidney injury (AKI) in a rat model. Thirty-five Wistar Albino rats were divided into five groups: Control, Sham, NAS (5 mg/kg), Cisplatin (CP; 7.5 mg/kg), and CP + NAS. NAS was administered daily for seven days, while cisplatin was given as a single dose on the fourth day. Renal function was assessed via serum urea and creatinine. Oxidative stress markers, including Malondialdehyde (MDA), Superoxide Dismutase (SOD), Total Antioxidant Status (TAS), and Total Oxidant Status (TOS), were measured in kidney tissue. Comprehensive histopathological evaluations were performed to assess tubular and glomerular damage. Cisplatin administration significantly increased serum creatinine levels and induced severe histopathological damage (p < 0.05). While cisplatin reduced SOD and TAS levels, NAS treatment showed a trend toward biochemical recovery without reaching statistical significance in oxidative markers. Notably, NAS administration significantly ameliorated cisplatin-induced histopathological lesions, specifically reducing tubular epithelial loss, glomerular degeneration, interstitial inflammation, and vacuolization (p < 0.05). Our findings indicate that NAS exerts a profound structural protective effect against cisplatin-induced renal injury. The preservation of renal parenchyma, despite modest systemic biochemical shifts, suggests that NAS-mediated protection may involve localized TrkB-dependent pro-survival signaling and stabilization of mitochondrial integrity. NAS represents a promising therapeutic candidate for mitigating chemotherapy-induced nephrotoxicity. Full article

5-Fluorouracil (5-FU) is a cornerstone chemotherapeutic agent that is extensively utilized in the management of malignancies; however, its clinical utility is constrained by its narrow therapeutic index and dose-limiting toxicities. The study aimed to study the hepato-nephroprotective effects of epigallocatechin gallate (EGCG) and EGCG mediated selenium nanoparticles and their effect in mitigating the toxicity induced by 5-FU. EGCG-functionalized selenium nanoparticles (EGCG-SeNPs) were produced by mixing sodium selenite, with EGCG acting as both the reducing and stabilizing agent. Nanoparticles were characterized using UV-vis spectroscopy, FT-IR, dynamic light scattering, zeta potential analysis, and transmission electron microscopy. 35 adult rats were randomly assigned to control, 5-FU, 5-FU + Na₂SeO₃, 5-FU + EGCG, and 5-FU + EGCG-SeNPs groups. Hepatorenal toxicity was induced by intraperitoneal 5-FU administration during the final five days of the experiment. Serum biochemical markers, tissue oxidative stress, antioxidant enzyme, inflammatory cytokine levels, and apoptosis-related gene expression were evaluated. Immunohistochemical analysis of Nrf2 and Keap1 and histopathological examination of tissues were performed. 5-FU induced severe hepatorenal toxicity, evidenced by marked elevations in liver and kidney function biomarkers, excessive oxidative stress, inflammatory cytokine overproduction, NF-κB activation, and apoptotic signaling. Treatment with EGCG-SeNPs markedly ameliorated 5-FU-induced hepatic and renal dysfunction, restoring liver enzyme and kidney biomarker levels to near-normal levels more effectively than EGCG or sodium selenite alone. EGCG-SeNPs significantly suppressed lipid peroxidation, NGAL, and inflammatory mediators while robustly enhancing antioxidant defenses and activating the Nrf2/HO-1 pathway with concomitant Keap-1 downregulation, strongly inhibited NF-κB signaling, normalized cytokine balance, reduced poly (ADP-ribose) (PAR) activation, and attenuated apoptosis. EGCG–SeNPs confer superior protection against 5-FU–induced hepatorenal toxicity compared to EGCG or inorganic selenium alone. The potent protective effects of EGCG–SeNPs are mediated through coordinated antioxidant, anti-inflammatory, and anti-apoptotic mechanisms, primarily via activation of the Nrf2/HO-1 axis and suppression of NF-κB signaling. Full article

Neutrophils are the most abundant circulating leukocytes and are characterized by a proteome in which granule-associated proteins synthesized during granulopoiesis constitute a major fraction of total cellular protein, reflecting their preloaded effector nature in innate immune defense. A striking feature of neutrophil biology is the unusual abundance of the calcium-binding proteins S100A8 and S100A9, which together form the heterodimeric complex known as calprotectin. Early biochemical studies estimated that S100A8/A9 constitutes a substantial fraction of the soluble cytosolic proteome in neutrophils, with later studies often describing it as one of the most abundant protein complexes in these cells. Despite extensive studies on the antimicrobial and inflammatory activities of calprotectin, the biological rationale for this unusual abundance remains incompletely understood. In this review, we examine the structural, biochemical, and regulatory features of S100A8/A9 and explore the potential explanations for its high abundance in the neutrophil cytosol. We first discuss the unique organization of the neutrophil proteome and the transcriptional programs governing granulopoiesis that lead to large-scale production of neutrophil effector proteins. We then review the structural and biochemical properties of S100A8/A9, including its calcium-dependent conformational dynamics and high-affinity transition metal binding, which contribute to antimicrobial defense through nutritional immunity. Several functional hypotheses are considered to explain calprotectin abundance, including roles as an antimicrobial reservoir, a metal-sequestering molecule, a regulator of oxidative stress, and a source of damage-associated molecular patterns. Finally, we discuss the evolutionary logic of neutrophil protein preloading and the implications of calprotectin biology in inflammatory diseases and the tumor microenvironment. Resolving the abundance paradox of S100A8/A9 may reveal fundamental principles governing the organization of innate immune cell proteomes and provide new insights into the strategies used by neutrophils to achieve rapid and effective host defense. Full article

GDSL esterase/lipase (GELP) proteins constitute an evolutionarily conserved yet functionally diversified hydrolase family in land plants. They participate in cuticle and secondary cell wall biosynthesis, seed lipid remobilization, reproductive development, and hormone-mediated responses to biotic and abiotic stresses. Despite extensive genome-wide and comparative genomic studies that have categorized large GELPs across numerous crops and model species, only a fraction of members have been functionally characterized in plants, and their catalytic mechanisms and regulatory architectures remain poorly understood. Recent population genomics and cross-species orthogroup analyses in 46 angiosperms have uncovered substantial natural variation within GELP coding sequences and regulatory regions, providing a powerful framework to link allelic diversity to evolutionary trajectories and physiological functions. This review synthesizes current knowledge on GELP evolution, biochemical properties, and roles in development and stress adaptation, and critically evaluates how these insights can be translated into biotechnology and molecular breeding strategies. It highlights emerging resources and concepts from orthogroup-based classification and multi-species datasets that enable systematic discovery of GELP alleles affecting agronomic traits. It further outlines research exploiting GELPs in crop improvement, emphasizing the integration of reverse and forward genetics with multi-omics profiling, biochemical and structural characterization, and gene regulatory network reconstruction. Systematic assessment of the phenotypic impacts of single and combinatorial GELP perturbations on yield, quality, and stress resilience is proposed as a key step toward translating basic insights into breeding and engineering strategies. Full article

Background/Objectives: Chronic glomerulonephritis (CGN) is a progressive chronic kidney disease that can ultimately advance to end-stage renal disease (ESRD). Zhuangyang Bushen Pill (ZYBSW) is a traditional Chinese herbal formulation derived from the Yi ethnic medicine of Yunnan Province, and it has been widely employed in folk practice for the amelioration of chronic nephritis and renal dysfunction. This study was designed to evaluate the therapeutic efficacy of ZYBSW against CGN and to provide preliminary insights into its underlying mechanisms of action. Methods: The nephropathy model was induced in mice by tail vein injection of ADR (10 mg/kg). Renal function was evaluated by measuring relevant biochemical parameters, and renal histopathological alterations were examined using HE staining. Chemical constituents of ZYBSW were analyzed by LC-MS/MS. Its mechanisms of action were investigated using network pharmacology, WGCNA, molecular docking, multiplex immunofluorescence, and Western blotting. Results: ZYBSW significantly reduced ACR by 88.9%, SCr by 56.4%, and BUN by 30.4%, increased ALB by 32.4%, and alleviated renal histopathological damage (all p < 0.01). LC-MS/MS analysis identified 419 chemical constituents in ZYBSW. Network pharmacology, WGCNA, and molecular docking experiments identified EGFR and DUSP1 as potential targets, and indicated the MAPK pathway as a key pathway. Mechanistic studies revealed that ZYBSW significantly inhibited EGFR expression in renal tissue, enhanced DUSP1 expression, and reduced the phosphorylation levels of ERK, JNK, and p38. Conclusions: This study reveals ZYBSW can effectively alleviate CGN, with EGFR and DUSP1 as likely therapeutic targets, and its mechanism of action primarily involves regulating the MAPK signaling pathway. Full article

This study explored the differences in slaughter performance, blood biochemical indices, and ruminal and colonic microbiota between 6-month-old male and female Small-tailed Han sheep, a typical meat-wool dual-purpose breed in China. Twenty weaned lambs (10 males and 10 females) with uniform body condition were reared under unified feeding management until 6 months of age, followed by slaughter sampling and microbial sequencing detection. Results showed that male lambs had significantly higher pre-slaughter live weight, carcass weight and serum ALP content than females (p < 0.05), with lower BUN and β-BHBA levels (p < 0.05). High-throughput sequencing of the 16S rDNA gene in rumen fluid and colon contents revealed that microbial alpha diversity in the rumen was extremely significantly higher than that in the colon (p < 0.01), and their microbial community structures were distinctly separated (p = 0.001). Sex had no significant effect on overall microbial diversity, but altered specific flora and functional pathways: male rumen had higher Actinobacteriota abundance, while female colon had enriched galactose metabolism and male colon had enhanced folate-mediated one-carbon pool pathway. These findings clarify the tissue specificity of gastrointestinal microbiota and sex-related phenotypic differences, providing a theoretical basis for sex-specific feeding of Small-tailed Han sheep. Full article

Fermented dairy products with probiotic and functional properties are a promising matrix for modulation of the human microbiome. The functionality of such products will depend not only on the technological properties of the lactic acid bacteria included in the starter culture but also on the combined effects of metabolites, enzymatic activity, stress tolerance, and strain-specific adaptation mechanisms. The aim of this work was to conduct a comprehensive analysis of Lactobacillus strains to facilitate the design of microbial consortia for the development of fermented products with diverse functional properties. Twenty Lactobacillus strains from different species were investigated using microbiological, physicochemical, and biochemical methods to evaluate antagonistic activity against opportunistic microorganisms and to assess changes in amino acid and organic acid profiles, vitamin content, fatty acid composition, and enzymatic activity. Additionally, proteomic analysis was performed to create a matrix of functional complementarity of the studied strains, representing proteins associated with antimicrobial activity, bacteriocin transport, resistance to oxidative stress, surface structure formation, and adhesion. It was shown that the studied strains exhibit pronounced functional heterogeneity, demonstrating the feasibility of scientifically based selection of strains to create next-generation fermented dairy products with predictable properties. Full article

Automated multiplex immunohistochemistry (IHC) and in situ hybridization (ISH) require staining platforms that combine stable reagent exchange, low-volume operation, process observability, and protocol flexibility. Existing autostainers are often rigid and costly, whereas microfluidic and sensing solutions remain largely component-specific rather than system-oriented. This study proposes and partially validates a layered cyber-physical architecture for multiplex histological staining. The architecture integrates five functional layers—biochemical workflow, fluidic processing, capacitive sensing, protocol-driven control, and software-based process representation—within a unified formal framework and is supported at the subsystem level by experimental characterization of its fluidic and sensing layers. Fluidic experiments on a slot-type microfluidic chamber identified a practical operating window in which upper-feed operation, moderate calibrated flow conditions, and low chamber angles between 10° and 40° provide stable filling and acceptable drainage. The differential slot-line capacitive sensing subsystem detected liquid volumes as low as 0.5 µL, with stable threshold-based interpretation at a practical detection threshold of approximately 5 fF after digital filtering. The control and software layers are specified at the architectural and formal model level; their hardware implementation and closed-loop validation remain subjects of future work. Together, the reported results demonstrate that controlled reagent transport and sensing-based process observability are jointly feasible within the proposed modular framework, establishing a conceptual and experimental foundation for scalable, flexible, and resource-efficient multiplex IHC/ISH systems. Full article

Smooth muscle (SM) dysfunction contributes to several pathological conditions, including atherosclerosis; current treatment strategies often fail to restore functional contractility. Adipose-derived stem cells (ADSCs) offer a promising cell source for regenerative medicine due to their accessibility and multipotency. Their differentiation into smooth muscle cells (SMC) is commonly driven by biochemical cues such as retinoic acid and transforming growth factor β; however, supporting this process with additional, non-invasive stimuli may enhance outcomes. Photobiomodulation (PBM) has emerged as a potential modulator of cellular metabolism, mitochondrial function and lineage commitment; however, its role in ADSCs to SMC differentiation remains insufficiently defined. ADSCs were irradiated with green (525 nm), near-infrared (825 nm) or dual wavelengths at 5 J/cm² and 10 J/cm² alongside the growth factors. Proliferation, cytotoxicity, mitochondrial membrane potential, collagen production, migration and smooth muscle marker expression were assessed. PBM induced a fluence-dependent biphasic response. 5 J/cm² fluences enhanced proliferation, mitochondrial activity, collagen deposition and organized SMC marker expression, whereas 10 J/cm² fluences lowered proliferation and membrane potential, reduced collagen and increased migration. PBM at 5 J/cm², especially greenlight, most effectively promoted ADSCs’ progression towards a SMC-like phenotype, with features consistent with a more contractile-like state. Full article

Oxidative stress appears to be implicated in both the initiation and progression of autoimmune thyroiditis. Selenomethionine, which exhibits antioxidant properties, has been shown to reduce thyroid antibody titers in patients with autoimmune thyroiditis. Recent evidence suggests that vitamin E, a fat-soluble antioxidant, may protect against the development of autoimmune thyroiditis, and that its supplementation has been associated with improvements in female sexual function. The objective of the present pilot study was to determine whether vitamin E intake modulates the effects of selenomethionine on female sexual function and depressive symptoms in individuals with thyroid autoimmunity. The study enrolled three groups of reproductive-age women with euthyroid autoimmune thyroiditis, with 26 participants in each group. The groups were matched for age, thyroid peroxidase antibody titers, and TSH levels and differed according to vitamin E intake: adequate intake (group A), low intake (group B), and high intake (group C). All participants received selenomethionine supplementation (200 µg/day) for six months. Antibody titers and hormone levels were measured, and participants completed questionnaires assessing female sexual function (FSFI) and depressive symptoms (BDI-II). At baseline, no differences in biochemical outcomes were observed between the groups, except for testosterone levels. The study groups differed in sexual desire and arousal domain scores, which were higher in group A than in the other two groups. Total FSFI scores, the remaining FSFI domain scores, and BDI-II scores did not differ between groups at baseline. Across all groups, selenomethionine reduced thyroid peroxidase and thyroglobulin antibody titers and increased SPINA-GD and the ratio of free triiodothyronine to free thyroxine; however, the effects on antibody titers were most pronounced in group A. An increase in SPINA-GT and testosterone levels following selenomethionine supplementation was observed only in group A. In this group, selenomethionine also led to significant improvements in total FSFI scores and all individual domain scores. In contrast, in the remaining groups, the effects of supplementation were limited to increases in domain scores for lubrication, sexual satisfaction, and pain. A treatment-related reduction in total BDI-II scores was observed exclusively in women with adequate vitamin E intake. These findings suggest, for the first time, that dietary intake of a natural antioxidant may influence the effects of exogenous selenomethionine on sexual function and depressive symptoms in reproductive-age women with euthyroid autoimmune thyroiditis. Full article

Subterranean termites, particularly Odontotermes obesus, cause severe damage to forest nurseries and plantations in arid and semi-arid ecosystems. This study demonstrates the dual functional role of endophytic entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, in termite suppression and induction of plant defense responses. Laboratory bioassays revealed significantly higher virulence of M. anisopliae, with a lower LT₅₀ (lethal time required to cause 50% mortality) of 33.1 h compared to B. bassiana (46.7 h), a steeper probit slope (5.4 ± 0.3), and strong model fit (R² = 0.95), indicating rapid and synchronized mortality. Endophytic colonization varied across host species and application methods, with soil incorporation consistently outperforming foliar inoculation. Maximum colonization (82.5%) was recorded in Tecomella undulata and exceeded 80% in Azadirachta indica under M. anisopliae. Biochemical analyses revealed significant increases in protein (up to 3.5 mg g⁻¹), phenols (3.7 mg g⁻¹), and tannins (2.7 mg g⁻¹). Activity of defense enzymes was significantly enhanced, with catalase reaching 263.5 U mL⁻¹, while Phenylalanine ammonia-lyase and Tyrosine ammonia-lyase exceeded 170 and 198 U mL⁻¹, respectively, indicating activation of antioxidant and phenylpropanoid pathways. Molecular docking analysis further revealed strong interactions between fungal metabolites and termite cellulase, with Bassianin (−8.4 kcal mol⁻¹) and Tenellin (−8.1 kcal mol⁻¹) showing the highest binding affinities. These findings highlight the combined biochemical and molecular mechanisms underlying fungal-mediated termite suppression and plant defense induction, and future research should prioritize transcriptomic validation, rhizosphere microbiome interactions, formulation optimization, and long-term multi-location field evaluation to support sustainable termite management strategies. Full article