Search Results (16,699)

Bioanalytical methods to quantitate conjugated payloads are essential for assessing antibody-drug conjugate (ADC) stability and pharmacokinetics (PK). Dual-payload ADCs present analytical challenges; different linker chemistries can require complex digestion conditions to perform the cleavage. Developing separate methods for each linker combination can be time and resource demanding. Rat tritosomes—purified lysosomal fractions from Triton-treated rat liver—provide a comprehensive enzymatic mixture that mimics the lysosomal environment. The presented bioanalytical method combines immunoaffinity purification with tritosome-mediated digestion for simultaneous quantitation of dual-conjugated payloads. The method was applied to a model dual-payload ADC containing two different cytotoxic payloads, conjugated using different enzymatically cleavable linkers, with an unrelated DAR (drug-to-antibody ratio). Method validation in mouse plasma demonstrated excellent accuracy (bias ± 20%, LLOQ and ULOQ ± 25%) and precision (coefficient of variation CV% ≤ 20%, LLOQ and ULOQ ± 25%) across all concentration levels (lower to upper limit of quantitation, LLOQ to ULOQ) for both payloads, with 100% of quality control samples (QCs) meeting acceptance criteria for hybrid LC-MS/MS quantitation methods. This tritosome-based approach provides a unified, efficient platform for multi-payload ADC bioanalysis, eliminates linker-specific method optimization, and enables robust support for preclinical studies. The method has been tested for accuracy and precision on 4 different model ADCs and employed to quantify the conjugated payloads in in vivo samples from a homozygous hFcRn transgenic mouse model (Tg32) PK study, resulting in reliable data in accordance with total antibody measurements. Full article

The complex genus Achillea L. comprises more than 140 species distributed widely throughout the Northern Hemisphere. Several species are widely used in traditional medicine for their therapeutic properties, yet few studies have correlated their biological properties with the plant’s phytochemical composition. Among these, Achillea setacea Waldst. & Kit. is a perennial species traditionally used to treat digestive and inflammatory disorders. In this study, the essential oil of A. setacea, collected wild in North Macedonia, was analyzed spectrometrically and spectroscopically by GC-MS and NMR, respectively. A total of nineteen compounds were identified, with camphor (31.3%), 4-terpineol (11.3%), and eucalyptol (10.6%) being the main constituents. Furthermore, the biological activities of pure oil were evaluated, showing notable antioxidant properties, as well as antimicrobial effects against a panel of clinically relevant microorganisms, including Gram-positive and Gram-negative bacteria. Furthermore, its impact on human intestinal epithelial (Caco-2) cells was assessed, highlighting its potential relevance for gastrointestinal applications, in agreement with the traditional use of Achillea species for digestive disorders. Full article

Pomegranate (Punica granatum), a shrub belonging to the Lythraceae family, has long been recognized for its diverse pharmacological benefits, including potential roles in managing inflammation and diabetes. The present study explored the insulin-secretory and β-cell proliferative properties of the ethanol extract of P. granatum fruit peel (EEPG) and assessed its influence on glucose regulation in high-fat-fed diet-induced obese mice (HFDi-OM) through in vivo and in silico studies. In vitro, EEPG was found to activate cAMP-dependent pathways and regulate K_ATP channels, thereby enhancing glucose-stimulated insulin secretion from BRIN-BD11 β-cells, with partial reliance on extracellular calcium. EEPG promoted β-cell proliferation, as indicated by an increase in Ki-67 positive cells, and displayed inhibitory effects on glucose diffusion and starch hydrolysis, suggesting a capacity to delay carbohydrate digestion and absorption. Furthermore, EEPG demonstrated antioxidant activity by neutralizing free radicals. In an acute test, EEPG (at doses of 150 and 250 mg/5 mL/kg) improved oral glucose tolerance and elevated plasma insulin levels. Long-term oral treatment for 21 days to HFDi-OM led to a significant reduction in fasting blood glucose, body weight, and food and fluid intake. It also enhanced gastrointestinal motility and improved lipid profiles by increasing HDL and lowering total cholesterol, LDL, and triglycerides. The therapeutic properties of EEPG are likely attributed to its rich bioactive components, including flavonoids (quercetin, kaempferol, catechin, and epicatechin) and phenolic acids (ellagic acid), which exhibited strong multi-target binding affinities in in silico molecular docking studies toward SUR1, PDE4, PI3K, and α-amylase, thereby supporting enhanced insulin secretion, β-cell function and glucose homeostasis. Full article

Aurelia coerulea is a bloom-forming scyphozoan that recurs in Masan Bay, a semi-enclosed embayment on the southern coast of Korea. To quantify its population-level predation impact on mesozooplankton, medusae and zooplankton were sampled monthly at six stations from May to September 2013, except for August. Individual ingestion rates were estimated from gut-content analysis combined with temperature-dependent digestion time, and population ingestion rates were evaluated relative to mesozooplankton biomass and production. Medusa abundance peaked in May (2.11 medusae m⁻³) and declined sharply thereafter, whereas the individual body size and the total population biomass increased. Oithona similis was the dominant prey, indicating positive selectivity from June to September despite its low biomass in the ambient zooplankton community. Mesozooplankton abundance, biomass, and production ranged from 4300 to 26,900 ind. m⁻³, 6.9 to 20.6 mg C m⁻³, and 2.26 to 5.82 mg C m⁻³ d⁻¹, respectively. Individual ingestion rates of the jellyfish on the mesozooplankton ranged from 0.31 to 5.31 mg C medusa⁻¹ d⁻¹. Population ingestion rates ranged from 0.06 to 0.66 mg C m⁻³ d⁻¹, equivalent to 0.3–6.6% and 1.1–11.8% of the biomass and production of the mesozooplankton, respectively. Predation impact of A. coerulea on mesozooplankton in Masan Bay varied seasonally and depended primarily on medusa abundance, because population ingestion rates decreased after May despite increasing individual ingestion rates associated with medusa growth. A modified calculation of the population ingestion rate, considering jellyfish distribution only in the upper layer under water-stratification conditions in summer, was also discussed. Full article

Anti-nutritional factors (ANFs) in terrestrial plant feeds constrain efficient herbivore production, an issue intensified by rising feed costs and growing demand for animal products. Unlike previous reviews that focus on single ANFs or feed types, this review provides an integrated, cross-species framework linking ANF chemistry, rumen microbial interactions, and mitigation strategies. It examines major ANF classes—tannins, phytates, saponins, oxalates, protease inhibitors, lectins, glucosinolates, and gossypol—and their distribution and biochemical modes of action. Mechanistic pathways are grouped into digestive effects (reduced palatability and enzyme inhibition), microbial effects (altered rumen microbiota and fermentation), metabolic effects (impaired absorption), and mineral interactions (nutrient complexation and chelation). Species-specific responses are evaluated, emphasizing the partial detoxification capacity of the rumen microbiome and the dose-dependent nature of ANF effects. Mitigation strategies—physical, chemical, microbial, enzymatic, probiotic, and genetic—are critically assessed for efficacy, scalability, and sustainability. Emerging metabolomic and metagenomic evidence shows that certain ANFs confer functional benefits at controlled doses; for example, tannins improve nitrogen retention, saponins reduce methane, and phytic acid scavenges free radicals. This synthesis supports strategic management rather than complete elimination, informing safe and sustainable use of terrestrial feeds under evolving food-security and environmental challenges. Full article

The quality and productivity of tomato (Solanum lycopersicum L.) crops largely depend on the quality of the seedlings used for cultivation. Several factors, including cultivation strategy, fertilization practices, and abiotic and biotic stressors during early plant development, influence seedling quality. Recently, anaerobic digestate has attracted attention as a potential organic fertilizer and substrate component; however, information about its effects on tomato seedling quality remains limited, particularly when comparing different seedling establishment methods such as direct sowing and transplanting. Therefore, this study aimed to evaluate the effects of different concentrations of solid grain waste digestate (further digestate) in the peat substrate on the growth and physiological characteristics of tomato seedlings grown by means of direct sowing and transplanting. The experiment was conducted at the Institute of Horticulture of the Lithuanian Research Centre for Agriculture and Forestry in unheated greenhouses covered with double polymer film. Two cultivation strategies were applied (factor A): transplanting and direct sowing into pots. To evaluate the influence of digestate (factor B), different substrate compositions were used: peat (control) and peat mixed with 10%, 20%, 30%, 40%, and 50% digestate. The strong decline in growth parameters with increasing digestate concentration indicates that higher proportions of digestate created unfavorable conditions for seedling development in both cultivation stategies. A 10% digestate addition improved certain plant characteristics, while 20% improved some physiological indices but was associated with reduced growth. However, higher digestate concentrations (≥30%) negatively affected plant growth and physiological activity. Seedlings grown in substrates with higher digestate levels exhibited reduced transpiration rates and lower gas exchange indices, suggesting impaired water relations and stomatal regulation. These effects were more pronounced in transplanted plants compared with direct-sown seedlings, indicating greater sensitivity to changes in substrate composition after transplanting. Overall, the results demonstrate that digestate can be used as a substrate component for tomato seedling production. Still, its concentration must be carefully optimized to avoid negative effects on plant growth and physiological performance. Full article

Repeated heat–moisture treatment (RHMT) is an efficient approach for modifying starch. However, the role of treatment temperature, a critical parameter, remains poorly understood. Therefore, this study investigated the effects of RHMT temperatures (80, 100, 120 °C) and cycles (1, 3, 5, 7) on the multi-scale structure and in vitro digestibility of hard proso millet starch, using native starch as a control. Compared with the severe 120 °C treatment, processing at 100 °C better preserved double-helical organization (supported by moderately retained enthalpy, ΔH) and short-range order, while maintaining granule integrity. These structural retentions restricted swelling, improved pasting stability, and reinforced the macroscopic gel network. Furthermore, multivariate analysis suggested that the rigidified internal granular architecture delayed initial enzymatic hydrolysis, maximizing slowly digestible starch (SDS) formation (47.44% in 100-RHMT-5). Conversely, 120 °C caused severe granular collapse and a drastic drop in ΔH, diminishing gel elasticity and triggering a surge in rapidly digestible starch (RDS, 59%). Overall, 100 °C RHMT yields an SDS-enriched starch, which may be a promising ingredient for the development of starch-based foods with slower in vitro digestibility. Full article

The second stomach compartment (C2) of the dromedary camel (Camelus dromedarius) plays an important role in digestion. However, detailed morphological and histochemical data remain limited. This study aimed to investigate the gross anatomy, histological organization, histometric features, and histochemical distribution of muco-substances in C2. The study was conducted on twenty dromedary camels, including fetuses and adults. Gross anatomical observations were performed on eight fresh and fixed specimens, while histological, histometric, and histochemical analyses were carried out on samples from twelve adult camels using routine and special staining techniques to identify neutral and acidic mucins. C2 was the smallest gastric compartment, located on the right side of the abdominal cavity and partially continuous with C1. Its mucosa formed chambered zones supported by prominent longitudinal muscular bands. Histologically, C2 comprised glandular and non-glandular regions. The glandular mucosa contained gastric pits and branched tubular glands with mucous, chief, and parietal cells, whereas the non-glandular region was lined by keratinized stratified squamous epithelium. Submucosal lymphoid aggregations were observed near the C2–C3 junction. Histometric analysis revealed a markedly developed tunica muscularis. Strong PAS and Alcian blue reactions indicated abundant neutral and acidic mucins. These findings demonstrate that C2 is a structurally specialized compartment supporting digestion, mucosal immune defense, and adaptation to arid environments, clearly distinguishing it from the reticulum of true ruminants. Full article

Soybean seeds have long been regarded as “storehouses of high-quality proteins”. The breakdown of dietary proteins by digestive proteases is essential for achieving adequate protein digestibility in animals and humans. However, plants have evolved a diverse array of protease inhibitors that regulate or restrict protease activity. In soybean, these inhibitors are concentrated primarily within the 2S protein fraction. Trypsin inhibitors (TIs) of Kunitz trypsin inhibitor (KTI) and Bowman–Birk inhibitor (BBI) are the most impactful due to their strong anti-tryptic activity, which interferes with digestive proteases in humans and animals. Elevated TI levels render raw soybeans unsuitable for direct food or feed use unless thermal or processing inactivation treatments are applied. Elimination or reduction in KTI and BBI using classical and biotechnology-based breeding efforts is a promising strategy. Soybean germplasm harboring BBI null alleles has not been reported. Breeding only for low or null KTI content in soybean would not be sufficient for practical applications. Hybridizing IT105782 × PI 547656 and using the reported Kompetitive Allele-Specific PCR (KASP) markers represents an effective classical breeding strategy. Simultaneous CRISPR/Cas9-mediated knockout of key KTI and BBI genes is expected to enable the development of soybean lines with substantially reduced TI levels, an outcome that cannot be readily achieved through classical introgression of null alleles, as naturally occurring null BBI alleles have not yet been identified. Moreover, this approach avoids the linkage drag associated with donor-derived null KTI alleles. However, this approach remains challenging due to functional redundancy and compensatory effects among KTI and BBI family members, extensive sequence homology among KTI and BBI genes that complicates the minimization of off-target effects, and the genotype dependency of Agrobacterium-mediated soybean transformation. Microtiter plate AACCI/AOCS could be one practical option for measuring TIA in breeding programs in terms of precision. Potential trade-offs associated with reduced trypsin inhibitor levels, including possible effects on plant defense and stress resistance, should be investigated in future studies, as these aspects have received little attention in previous research. Full article

This study investigated possibilities to increase the efficiency of anaerobic digestion of sewage sludge using biochar produced by pyrolysis of digested sludge (sludgechar). Experiments were conducted in continuous laboratory bioreactors operated at the same loading rate, gradually increased from 3.2 to 4.5 g/(L·d) (COD) under mesophilic conditions (40 °C). Sludgechar (SCH) was dosed into the experimental bioreactor at a rate of 0.4–1.3 g/(L·d), corresponding to 12–28% of the added COD. Biogas production in the experimental bioreactor increased by 6.9–33% compared with the control bioreactor, while the CH₄ concentration remained comparable, averaging 62.8%. The COD removal efficiency remained high in the sludgechar-supplemented bioreactor as the loading rate increased, whereas it decreased in the control bioreactor, corresponding to lower biogas production. The adsorption capacity, alkalinity, and mineral buffering properties of sludgechar prevented pH decline and the accumulation of volatile fatty acids (VFAs) at higher substrate loading. The pH values were less affected by increasing organic loading in the experimental than in the control bioreactor and remained within 6.9–7.0. Continuous experiments confirmed that sludgechar can facilitate stable operation at loading rates that would otherwise cause process failure. However, the low carbon-to-inorganic ratio of sludgechar is its significant disadvantage. Full article

Host-adapted probiotics offer a promising strategy for improving stingless bee nutrition and colony sustainability. In this study, gut-derived lactic acid bacteria (LAB) isolated from Heterotrigona itama were evaluated for probiotic potential and used to develop fermented bee pollen. Of 37 presumptive LAB isolates, three strains (BP-2, BP-3, and BPW-B1) exhibited strong tolerance to simulated gastrointestinal conditions, favorable adhesion-related properties, and acceptable safety profiles. Phylogenetic and biochemical analyses identified the selected isolates as Apilactobacillus kunkeei. The LAB strains were co-cultured with the osmophilic yeasts Zygosaccharomyces bailii TSU_YK2 and Starmerella meliponinorum TSU_YP10 to establish a host-associated LAB–yeast co-fermentation model that mimics stingless bee pollen fermentation. Co-fermentation significantly improved protein digestibility, organic acid production, antioxidant activity, and microbial viability relative to spontaneous fermentation controls (p < 0.05). Feeding experiments demonstrated that probiotic-fermented pollen increased feed intake, body weight, abdominal lipid reserves, hypopharyngeal gland development, and survival among H. itama workers. In addition, probiotic supplementation was associated with shifts in the dominant gut-associated bacterial taxa, including Lactobacillus, Bifidobacterium, and Snodgrassella. This study demonstrates the potential of combining gut-derived A. kunkeei with osmophilic yeasts as a functional fermentation starter culture to develop biologically relevant probiotic feed supplements for stingless bees. Full article

Heavy metal-contaminated post-mining soils remain persistent sources of ecological degradation and contaminant dispersion. This study provides a quantitative field-based assessment of sorghum (Sorghum bicolor), maize (Zea mays) and soybean (Glycine max) cultivated on heavy metal-affected mining soil from the Jiu Valley, Romania, within a Living Lab platform. Soil properties, pseudo-total metal concentrations, multi-year biomass production, growth indicators, vegetation cover and aboveground plant metal concentrations were evaluated. The soils showed slightly acidic to near-neutral pH, low organic matter and multi-metal contamination, with Cr, Cu, Ni, Zn and Pb ranging from 82 to 146, 51 to 92, 41 to 79, 156 to 287 and 64 to 121 mg kg⁻¹, respectively. Total fresh biomass increased from 85 kg in the first cultivation year to 487 kg in the third cultivation year, with sorghum showing the highest final production (230 kg) and vegetation cover (55–86%). Aboveground Cr, Cu, Ni, Zn and Pb concentrations measured at species-specific levels of 6.21–8.06, 8.77–10.64, 7.48–12.92, 38.01–47.11 and 4.32–6.46 mg kg⁻¹ dry weight, respectively. Sorghum showed the highest preliminary phytomanagement suitability, mainly through stronger vegetation cover formation, higher fresh biomass production and lower visible stress under the investigated field conditions. Maize showed intermediate feasibility, whereas soybean appeared more sensitive to the degraded substrate. Biomass reuse should be considered only under controlled non-food pathways, such as pyrolysis or anaerobic digestion, and should only be considered after a dedicated assessment of dry biomass, conversion residues and metal fate. Full article

Heat and Cd²⁺ stress are major environmental challenges for marine benthic invertebrates. This study examined their combined effects on growth, physiology, and transcriptomic responses in the peanut worm (Sipunculus nudus). After 30 days, Cd²⁺ reduced survival at 26 °C without significantly affecting growth, whereas at 32 °C, both survival and growth declined with increasing Cd²⁺ concentration, indicating that heat stress exacerbates Cd²⁺ toxicity. Cd accumulation increased with exposure concentration but was not affected by temperature. Heat stress increased immune (AKP) and antioxidant (SOD, CAT) enzyme activities, although significant increases in SOD and CAT were observed only under Cd²⁺ exposure. AKP activity rose at low Cd²⁺ concentrations and fell at high Cd²⁺ concentrations at 26 °C, whereas no significant difference occurred at 32 °C between 0 and 0.25 mg/L Cd²⁺. At the same temperature, SOD and CAT activities were significantly higher under high Cd²⁺ exposure than under low Cd²⁺ exposure. Transcriptome analysis showed that Cd²⁺ exposure activated longevity-related pathways, protein processing, and translation initiation. Heat stress activated Jak-STAT signaling and endoplasmic reticulum protein processing while inhibiting the ribosome pathway. Under combined stress, pathways related to xenobiotic metabolism, nutrient digestion and absorption, and amino acid derivative metabolism were broadly suppressed. These results highlight that heat stress exacerbates Cd²⁺ toxicity, affecting growth, enzyme activity, and transcriptomic responses, and provide insights into the adaptive strategies of marine benthic organisms under the combined pressures of climate change and heavy metal pollution. Full article

Early weaning can disrupt the intestinal function and microbial community balance of piglets, and trigger inflammation and oxidative stress, thereby affecting their production performance. In recent years, butyric acid has gained considerable interest as a functional feed additive. However, practical limitations such as its pungent odor and low absorption efficiency in the digestive tract have led to the development of more stable forms, including sodium butyrate, coated butyrate, and butyrate glycerides, etc. Research has shown that butyric acid and its derivatives can serve as effective feed additives by enhancing pigs’ resistance to pathogenic colonization, stabilizing the intestinal microbiota, and alleviating oxidative stress to mitigate challenges such as weaning stress and pathogenic infections. This review systematically highlights the role of butyric acid and its derivatives as dietary supplements for weaned piglets. Importantly, it underscores the potential of butyric acid and its derivatives may contribute to antibiotic-reduction strategies in weaned piglet nutrition, while also highlighting the need for optimized supplementation strategies and further investigation into synergistic effects with other feed additives. This review aims to offer both theoretical and practical insights for the application of butyric acid in weaned piglet nutrition. Full article

Nuts are an important dietary source of phenolic compounds, which are associated with potential health benefits. In the present study, the bioaccessibility of phenolic compounds from almonds, peanuts, pistachios and their corresponding butters was investigated after in vitro gastro-intestinal digestion. High-resolution mass spectrometry analysis provided a comprehensive picture of the phenolic profiles across samples, with a total of 55–95 compounds identified per matrix. Pistachios showed the highest amount of phenolic compounds (60.603 ± 1.170 mg/100 g), followed by peanuts and almonds. Processing into butter affected phenolic concentration in a matrix-specific manner. Almond and peanut butters showed higher phenolic content compared to whole nuts, whereas pistachio butter exhibited lower levels than the whole nut. After digestion, differences in bioaccessibility were observed. Peanuts showed the highest phenolic bioaccessibility (84.31%), while almonds and pistachios exhibited lower values (<15%). Overall, phenolic bioaccessibility was mainly influenced by nut type and phenolic compound structure, rather than by their initial concentration or processing. These findings are based on an in vitro model representing an estimate of bioaccessibility rather than in vivo absorption. However, nut type and processing appear to influence the release of phenolic compounds, which is relevant for the nutritional evaluation of nuts and nut-derived products. Full article