The induction of triploidy, a strategy to mitigate unwanted pre-harvest sexual maturation and a genetic containment measure for escaped farmed Atlantic salmon (Salmo salar), may give rise to challenges because of the distinct environmental and dietary requirements of sterile triploid fish. Smoltification is a critical phase in the life cycle of Atlantic salmon, so knowledge about parr–smolt transformation in triploids is important for the salmon farming industry. This study covered an investigation of hepatic expression of growth hormone receptor (GHrec) and insulin-like growth factor-I (IGF-I) genes, both of which are intimately involved in the regulation of osmoregulation and growth. Additionally, hepatic presence and location of IGF-I mRNA were examined using RNAscope®, an advanced in situ hybridization technique. Triplicate groups of juvenile diploid and triploid salmon were reared at low temperature (10 °C) and fed either a standard diet or one enriched with hydrolyzed fish proteins from the start of feeding onwards. Liver samples were collected from three fish per tank each month from October to December (2454–3044 degree-days post-start feeding), the period encompassing smoltification, and hepatic expression of IGF-I and GHrec genes was quantified by real-time PCR. The results indicated that neither ploidy nor diet significantly influenced IGF-I or GHrec gene expression, suggesting that, under our conditions, triploidy and diet did not adversely affect this molecular pathway linked to growth and osmoregulation. IGF-I gene expression exhibited significant temporal variation, correlating with the progression of smoltification, while GHrec gene expression showed a similar, albeit non-significant, trend. Triploids exhibited IGF-I and GHrec gene expression patterns comparable to diploids, and both the temporal changes and lack of difference between triploids and diploids were mirrored in the quantification of IGF-I mRNA within the liver cells. The potential applicability to a commercial aquaculture setting requires further investigation.

Background: Chickpea (Cicer arietinum L.) is an important grain legume valued for its high protein content and adaptability to diverse environments. Understanding the relationship between morphological, phenological and genetic variability is essential for improving breeding strategies and cultivar performance under contrasting agroecological conditions. Methods: Sixteen chickpea genotypes of diverse seed shape and size were evaluated under uniform field conditions for morphological and phenological traits. Genetic diversity was assessed using iPBS markers. Based on preliminary results, five genotypes were selected for multi-location trials under contrasting agronomic management systems to assess yield components. Data were analyzed using analysis of variance, correlation analysis and cluster analysis. Result: Significant variation was observed in plant height, seed size, pod number and seed yield, with seed shape emerging as the most consistent phenotypic marker for genotype differentiation. Molecular analysis supported phenotypic grouping, though with low bootstrap values, likely due to partial cross-pollination and seed admixture. Location had the largest effect on yield components, with higher rainfall contributing to increased productivity. Small-seeded genotypes demonstrated greater yield stability across environments, whereas large-seeded types performed best under favorable conditions.

Abstract Background Cognitive frailty, defined as the presence of physical frailty and cognitive impairment in the absence of dementia, is a common finding among older adults. The causative factors for cognitive frailty are not well understood. It is known that vascular factors such as arterial stiffness are associated with ageing and frailty. In the Frailty and arterial stiffness-role of oxidative stress and inflammation (FRAXI) study, the correlation between cognitive frailty (assessed by the mini-mental state examination (MMSE)), clinical frailty score (CFS) and arterial stiffness was explored. Methods The longitudinal FRAXI study included fifty community dwelling adults ≥70 years (mean age ± standard deviation: 79 ± 5 years, 46% male), with CFS ≤ 6 and no active malignancy, who were followed up for six months. Measures of arterial stiffness included pulse wave velocity (PWV, Complior®) and cardio-ankle vascular index, measured at baseline. Other study measurements: MMSE, timed up and go test), sarcopenia, geriatric depression scale, interleukin-6 and high sensitivity C-reactive protein biomarkers were measured at baseline and 6 months. Results All fifty participants were assessed for cognition using MMSE, with mean CFS at baseline of 3.5 (±SD 1.4) and at follow up, 4.0 (± SD 1.5). At baseline, MMSE strongly correlated with both functional and phenotypic frailty as assessed by Charlson’s Comorbidity Index (r = −0.3; p < 0.05) and CFS (r = −0.5; p < 0.001). Similarly, MMSE strongly correlated with measures of arterial stiffness; PWV-carotid femoral (r = −0.4; p = 0.01) and PWV-carotid radial (r = −0.4; p < 0.005). At follow up, MMSE remained strongly correlated with CFS (r = −0.3; p < 0.01). Conclusion Cognitive frailty correlates strongly with measures of vascular ageing. Arterial stiffness can be used as a vascular measure to identify older adults at risk of cognitive impairment.

Background The poliovirus receptor (PVR) is an emerging therapeutic target currently under clinical investigation in combination with checkpoint inhibitors. Although it is broadly expressed across many malignancies, its prognostic and predictive value in renal and bladder cancers remains underexplored. Methods Quantitative immunohistochemical analysis of PVR expression was performed in tumor tissue from patients with bladder cancer or renal cell carcinoma, including clear cell and papillary subtypes. Matched serum samples were analyzed using a custom enzyme-linked immunosorbent assay for total circulating PVR. To distinguish isoforms, we generated an antibody specific to the secreted isoform and developed an isoform-selective assay. Results Serum PVR levels were elevated in both bladder and renal cancer patients but did not correlate with tumor tissue expression, whether total or isoform-specific. In bladder cancer, tissue levels correlated with tumor grade and lymphovascular invasion. In renal cancer, expression was subtype-dependent, with only a subset of clear cell carcinoma samples showing tissue positivity. Importantly, reduced overall survival in clear cell carcinoma was linked to PVR expression, and rare cases with high levels consistently progressed to metastatic disease. Conclusion In renal tumors, a preliminary subtype-specific pattern emerged: PVR expression was generally low in clear cell carcinoma but enriched in papillary carcinoma. Notably, in the expanded clear cell cohort all cases with high expression were associated with metastatic progression. These findings highlight PVR in tumor biopsies as a potential prognostic marker in clear cell carcinoma and support its further investigation as a biomarker and therapeutic target in renal cancers.

Identification of N-tert-butyloxycarbonyl protected amino acids as novel hydrophobic tags to induce targeted protein degradation.

The emergence of insecticide resistance in pests affecting cotton and other key crops presents an escalating challenge to global agriculture, resulting in considerable declines in both yield and quality. Resistance mechanisms, including target site mutations and improved detoxification processes, have diminished the effectiveness of traditional insecticides. This study identified NPA001937 (di-O-demethylspirilloxanthin), a bacterial carotenoid obtained from the Natural Products Atlas, as a promising natural insecticidal compound. Employing in silico methods, we focused on three essential insect targets: Ryanodine Receptor (RyR), Arginine Kinase (ArgK), and Serine/Threonine Protein Phosphatase (STPP), which are known for their conserved sequences and structures across various pest species. Structural models were produced using AlphaFold2 and SwissModel, followed by validation through SAVES. The virtual screening of 30,052 natural products conducted with AutoDock Vina revealed NPA001937 as a leading binder to all three targets. Subsequent 500 ns molecular dynamics simulations (GROMACS) validated the stability and advantageous interaction profiles of the ligand- protein complexes. This compound presents several advantages over chemical insecticides, including its natural origin, ability to target multiple pathways, and a lower chance of resistance development. The results emphasize the effectiveness of computational methods in speeding up the discovery of agrochemicals which upon further experimental validation may serve as ecofriendly options for sustainable pest management.

Pulses are essential components of agricultural production systems, post-harvest losses caused by storage insect pests remain a major challenge. Callosobruchus maculatus (F.) is a widespread pest of stored legumes, and its host preference is influenced by seed type and associated physico-biochemical traits. This study examined the determinants of C. maculatus preference among nine legume species using controlled laboratory assays and seed trait analyses. Under no-choice conditions, chickpea ( Cicer arietinum L), pigeon pea (Cajanus cajan L.), and moth bean (Vigna aconitifolia (Jacq.) Maréchal) supported high oviposition, larval development, and adult emergence, suggesting their overall biological suitability as hosts. In contrast, the common bean (Phaseolus vulgaris L.) was the most preferred oviposition substrate under free-choice conditions but exhibited poor hatching and emergence, indicating its role as an oviposition trap. Black gram (Vigna mungo L.), soybean (Glycine max (L.) Merr), and moth bean, though less preferred, supported high post-embryonic success, revealing latent susceptibility. Green gram (Vigna radiata L.), black gram, red gram, cowpea (Vigna unguiculata L.), and moth bean exhibited high intrinsic suitability, reflected by positive growth indices, shorter developmental durations (notably in black gram and green gram), and balanced oviposition with adult longevity. Principal component analysis identified two major axes of host discrimination: (i) physical traits governing oviposition preference and (ii) biochemical composition determining larval development. Seeds with greater surface area, thinner seed coats, and moderate sphericity were favoured for egg deposition, whereas high protein and carbohydrate content enhanced larval growth. In contrast, anti-nutritional factors such as tannins and phytic acid negatively affected larval performance. These findings underscore the significance of integrating physico-biochemical resistance traits into legume breeding and storage management strategies to reduce bruchid-related post-harvest losses.

Background: Fish crackers are traditional starch-based snack foods widely consumed in Southeast Asia and are increasingly recognized as convenient protein-enriched products. Surimi technology has been widely recognized as an effective method for converting mechanically recovered fish meat into functional protein suitable for value-added products. Aims: The present study develops surimi-based fish crackers from snapper filleted frame meat and to evaluate their physicochemical, nutritional, microbiological, sensory, and storage stability characteristics to assess the feasibility of value addition and sustainable utilisation of fish processing waste. Study Design: An experimental laboratory study involving recovery of snapper frame meat, optimisation of surimi through water-washing, development of surimi-based fish crackers, and evaluation of physicochemical, nutritional, microbiological, sensory, and storage stability under ambient and refrigerated conditions. Place and Duration of Study: The experimental work was carried out during the period July 2019 to March 2020 at the Department of Fish Processing Technology, College of Fisheries, Mangaluru, Karnataka Veterinary, Animal and Fisheries Sciences University, India. The laboratory is located in the coastal region of Karnataka at approximately 12.87° N latitude and 74.88° E longitude. Methodology: Snapper filleted frame meat recovered by mechanical mincing was evaluated for quality and processed into surimi using standardised chilled water-washing cycles. Fish crackers were developed by blending surimi with tapioca flour, millet flour, and soya chunk powder in different ratios, followed by cooking, drying, and frying. The optimal formulation was selected through sensory evaluation. Dried and fried crackers were analysed for nutritional, biochemical, microbiological, and sensory quality, and storage stability was assessed under ambient and refrigerated conditions. Results: Snapper filleted frames yielded substantial recoverable meat (n = 10), with a mean total length and weight of 660.9 ± 28.4 mm and 2550 ± 506 g, respectively, and an average fillet yield of 1369 ± 264 g. Progressive water-washing significantly increased moisture while reducing lipid content, TVB-N, TMA-N, free fatty acids, TBARS, and microbial load (p < 0.05), improving surimi quality. Among four formulations evaluated by 25 panellists, crackers containing surimi (50%), tapioca flour (25%), millet flour (25%), and soya chunk powder (5%) showed the highest overall acceptability (8.61 ± 0.04; p < 0.01). Dried crackers had higher protein (20.41 ± 0.58%) and lower lipid content (0.95 ± 0.02%) than fried crackers (p < 0.001). Initial microbial counts were low (≤1.84 × 10² cfu/g), with no mould or yeast detected. During 60-day storage, biochemical spoilage indices and microbial counts increased significantly (p < 0.05), with slower deterioration and better sensory retention under refrigerated conditions. Conclusion: The study confirms the feasibility of converting snapper filleted frame waste into nutritionally acceptable, microbiologically safe, and sensory-appealing surimi-based fish crackers, with improved functional quality through controlled washing and superior storage stability under refrigerated conditions, supporting sustainable fish processing and value addition.

Prion diseases originate from the pathological misfolding of the cellular sialoglycoprotein prion protein (PrPC), universally found across mammalian species, into an aberrant conformation termed PrPSc, which exhibits high aggregation propensity and neurotoxicity. Distinct conformations of the misfolded and aggregated PrPSc, termed prion strains, can cause different disease phenotypes and transmission characteristics. Different prion strains exhibit well-defined and distinct glycoform preferences arising from two sialylated, N-linked glycans. Glycosylation, and in particular sialylation, have been demonstrated to modulate the replication rate of PrPSc, with profound implications for the propagation of prion diseases. In this work, we leverage high-resolution cryo-EM structural data and all-atom molecular dynamics simulations to elucidate the molecular basis of the glycoform preferences in mouse strains RML and ME7. We show that these preferences are determined by differential engagement of the major basic patch and palindromic region of PrP, shedding light on a long elusive, fundamental aspect of prion biology.

Abstract The aim of this study is to identify and establish biochemical parameters that can predict shunt blockage or malfunction in patients undergoing ventriculoperitoneal (VP) shunt surgery for hydrocephalus by analyzing cerebrospinal fluid (CSF) obtained during surgery. This retrospective study was conducted among patients with hydrocephalus who underwent VP shunt surgery in the Departments of Neurosurgery at two medical institutions during the periods 2011 to 2014 and 2021 to 2024, respectively. CSF samples were collected during shunt surgery and analyzed. Individual biochemical parameters were studied to identify potential predictive factors for shunt blockage. Each patient was followed-up for a minimum of 6 months to assess shunt malfunction or blockage. Patients with shunt malfunction due to infection or malposition were excluded. A high CSF protein value (>100 mg/dL) was found to be statistically significant in predicting the possibility of shunt blockage during follow-up in patients undergoing VP shunt surgery. The CSF protein level was also inversely proportional to the symptom-free interval following VP shunt placement. The ability to predict shunt malfunction using CSF parameters prior to surgery may help a group of patients avoid unnecessary VP shunt-related complications and help surgeons in appropriate surgical planning and in explaining prognosis.

Super-resolution microscopy in combination with genetic labeling methods allows imaging of single proteins in cells. However, visualizing endogenous proteins on primary cells remains challenging due to the use of sterically demanding antibodies for labeling. Here, we demonstrate how immunolabeling conditions and antibody cross-linking influence the quantification and identification of membrane receptor stoichiometry on cells using single-molecule localization microscopy. We developed an optimized immunolabeling and analysis protocol and demonstrate the performance of the approach by resolving the molecular organization of endogenous CD45, CD69, and CD38 on Jurkat T cells. To demonstrate the usefulness of the method for immunotherapy applications, we investigated the interaction of primary multiple myeloma cells with the therapeutic monoclonal antibodies daratumumab and isatuximab and a polyclonal anti-CD38 antibody. Our approach might lay the foundation for improved personalized diagnostics and treatment with therapeutic antibodies.

White maize (Zea mays L.) is an important cereal for human consumption in temperate regions, yet its genetic improvement remains limited due to inadequate evaluation of adapted germplasm and insufficient integration of agronomic and nutritional traits. The present study aimed to assess the extent of agro-morphological variability, yield potential, and nutritional quality among 100 white maize inbred lines, along with three locally adapted checks, under temperate agro-climatic conditions of Kashmir. Field evaluation was conducted during two consecutive kharif seasons (2022 and 2023) using an Augmented Block Design. Thirteen agro-morphological and yield-related traits were recorded to quantify phenotypic diversity and genotypic performance. Descriptive statistics and analysis of variance revealed highly significant differences among inbred lines for flowering, maturity, plant height, ear characteristics, kernel traits, and grain yield across both years, indicating a broad and stable genetic base. Several inbred lines consistently outperformed the checks for yield and maturity traits, demonstrating their suitability for temperate environments with short growing seasons. The consistency of trait expression across years further confirmed the reliability of the evaluated germplasm for selection and breeding. Biochemical profiling of selected inbred lines showed substantial variation in glycemic index (GI), protein content, and resistant starch (RS). Notably, inbred lines G37, G41, G72, and G63 exhibited low GI, high protein content, and elevated RS, identifying them as nutritionally superior and suitable for health-oriented maize improvement. An inverse association between GI and RS/protein content highlighted the scope for simultaneous improvement of yield and nutritional quality. Overall, the study identified genetically diverse, agronomically superior, and nutritionally enriched white maize inbred lines that can serve as valuable parental resources for developing early-maturing, high-yielding, and nutritionally enhanced hybrids adapted to temperate agro-climatic conditions.

Mechanisms underlying differential utilization of carbohydrates from diverse structures and sources in Nile tilapia (Oreochromis niloticus): Insights from glycolipid metabolism, protein deposition and liver health.

Antimicrobial resistance is a silent pandemic responsible for 1.14 million deaths worldwide in 2021, with a major contribution from Klebsiella pneumoniae. β-lactams are the most commonly used antibiotics in humans, and there is an urgent need to characterize the resistance mechanisms to these antibiotics in Enterobacterales other than Escherichia coli. Mecillinam is a narrow-spectrum β-lactam targeting a single penicillin-binding protein, PBP2. Pivmecillinam, its oral prodrug, is used as a first-line treatment for uncomplicated urinary tract infections. It has been used for decades in Europe but was only authorized by the US Food and Drug Administration in 2024. Here, we decipher mecillinam resistance mechanisms in K. pneumoniae by characterizing its resistome in a pan-susceptible strain. The chromosomally encoded SHV β-lactamase led to spontaneous mecillinam-resistant mutants appearing at a higher rate, growing faster and at higher mecillinam concentrations in K. pneumoniae than in E. coli. The most frequent genetic event was an unstable duplication leading to heteroresistance. The selected mutations leading to resistance affected a wide range of functions, with resistance being dependent or independent of the RelA (p)ppGpp synthetase. Through an in-depth characterization of six mutant strains, we showed that, in the presence of mecillinam, they all experienced different growth defects despite high minimal inhibitory concentrations. Overall, our results in K. pneumoniae suggest different mechanisms to escape the complex mode of action of β-lactams in synergy with the β-lactamase SHV.

Different neuron types show distinct susceptibility to age-dependent degeneration, yet the underlying mechanisms are poorly understood. Here we applied aging clocks to single neuron types in Caenorhabditis elegans and found that distinct neurons differ in their biological age. Ciliated sensory neurons with high neuropeptide and protein biosynthesis gene expression show accelerated aging and degeneration, correlating with loss of function, which could be prevented by pharmacological inhibition of translation. We show that the C. elegans neuronal aging transcriptomes correlate with human brain aging patterns and anticorrelate with geroprotective interventions. We performed an in silico drug screen to identify potentially neuroprotective small molecules. We show that the natural occurring plant metabolite syringic acid and the piperazine derivative vanoxerine delay neuronal degeneration, and propose these compounds as neuroprotective interventions. Furthermore, we identify neurotoxins that accelerate neurodegeneration, indicating that distinguishing aging trajectories between neuron types can inform on protective interventions as well as risk factors.

Fish protein hydrolysates hold great promise as nutraceuticals, yet their application as food ingredients or nutraceuticals is currently limited by their fish-like odor. This odor is mainly due to the presence of trimethylamine (TMA), a volatile biogenic amine resulting from the breakdown of naturally occurring trimethylamine-N-oxide (TMAO) in marine fish. The bacterial trimethylamine monooxygenase mFMO can oxidize TMA into TMAO using molecular oxygen and the cofactor nicotinamide adenine dinucleotide phosphate (NADPH). We have established an enzyme cascade that takes advantage of glucose dehydrogenase to recycle NADPH from NADP+, significantly decreasing the cost of the reaction and paving the way for using the enzyme system in fish protein hydrolysates targeted for human consumption. We demonstrate that the dual enzyme system works in an industrially relevant substrate. Salmon protein hydrolysate treated with an mFMO/glucose dehydrogenase cocktail showed a 75% reduction in TMA. A trained sensory panel perceived an improved odor across several parameters, including a reduction in the characteristic TMA smell.IMPORTANCEMarine by-products are a valuable source of high-quality peptide ingredients; however, their application in the food market is limited by the unpleasant fishy odor caused by trimethylamine (TMA). An enzyme that oxidizes TMA to the odor-free trimethylamine-N-oxide (TMAO) in salmon protein hydrolysates is known, but it requires excessive amounts of NADPH, an expensive cofactor. Here, we describe a cofactor regeneration system that allows using less cofactor in the enzyme-driven TMA removal process. This dual enzyme system removed 75% of TMA from a salmon protein hydrolysate, resulting in a significantly reduced fishy odor as confirmed by a trained sensory panel compared to the untreated control. This enzyme cascade is an important step toward making targeted TMA removal economically feasible for marine biomass valorization.

High Mobility Group Protein B1 and its downstream elements, syntaxins, contribute to temporal tight junction assembly in a human keratinocyte cell line.

Ribonucleases are associated with processing and degradation of diverse RNA substrates. These enzymes act on the substrate with high specificity, often in association with their interacting partners. Functionally redundant exoribonucleases are indispensable for maintaining the physiological homeostasis under normal and challenging conditions for growth. In this review article, we will report the comprehensive role of exoribonucleases in RNA metabolism especially the processing, maturation and degradation. Structural aspects of exonucleases, tendency to oligomerize, and their association with other cellular proteins that facilitate their interaction with RNA substrate and modulate their activity have also been highlighted. Transcription and post-transcriptional regulatory mechanisms of gene expression that are crucial for maintaining levels of exoribonuclease expression has been briefly discussed.

CRISPR-Cas9 is natively present in the adaptive immune systems of a multitude of bacteria and has been adapted as an effective genome engineering tool. The Cas9 effector enzyme, which is composed of a single Cas9 protein and a single-guide RNA (sgRNA), identifies and cleaves double-stranded DNA targets through a series of conformational changes that require DNA distortion and unwinding. While most studies of Cas9 specificity have focused on the DNA sequence, the role of intrinsic DNA physical properties ("DNA shape") in modulating Cas9 activity remains insufficiently defined. We previously showed that with a 16-nucleotide (-nt) truncated guide, the intrinsic DNA duplex dissociation energy at the PAM+(17-20) segment beyond the RNA-DNA hybrid tunes Cas9 cleavage rates of linear substrates. Here, we examined the impact of DNA supercoiling on Cas9 cleavage with the 16-nt truncated guide. Enzyme kinetic analysis revealed that PAM+(17-20) DNA sequences beyond the RNA/DNA hybrid preserve their effects on Cas9 cleavage in the supercoiled state. Furthermore, combining a novel asymmetric hairpin construct with a parallel-sequential kinetics model, rates for first-step nicking and second-step cleavage by Cas9 were obtained for both supercoiled and linear substrates. With both topologies, it was found that first-step nicking is clearly impacted by PAM+(17-20) DNA sequences, and the effects can be correlated with DNA unwinding, which dictates R-loop dynamics. This work expands our understanding of DNA target recognition by Cas9, and the methods developed, in particular those for analyzing the progression of Cas9-induced nicks, will aid in further in-depth mechanistic investigation.

ABSTRACTDate palm leaves (DPL), a widely available lignocellulosic by‐product, are used as ruminant feed but are limited by high lignin and low protein content. This study evaluated the enhancement of DPL's nutritional value using a lignocellulose‐degrading bacterial consortium (Staphylococcus sp., Brevibacterium sp., and Enterobacter sp.) isolated from the leopard moth (Zeuzera pyrina L.) gut, supplemented with microbial growth stimulators. Six treatments were applied: untreated DPL (control), DPL with M9 medium (T1), DPL with bacterial inoculum (T2), T2 + 0.5% glucose (T3), T2 + 0.5% urea (T4), and T2 + 0.5% glucose + 0.5% urea (T5). Parameters assessed included chemical composition, lignin peroxidase (LiP) activity, in vitro gas production (IVGP), fermentation characteristics, nutrient digestibility, and ruminal enzyme activities. LiP activity was highest in T5 (0.328 U/mL/min), representing a significant increase over controls. Inoculated treatments significantly reduced acid detergent lignin (ADL) and increased crude protein (CP), with T5 showing the greatest improvement: ADL decreased from 97.8 to 83.8 g/kg DM, and CP increased from 52.2 to 70.3 g/kg DM. T5 also exhibited the highest dry matter (DM) loss (61.1 g/kg DM), IVGP (61.2 mL), metabolizable energy (5.61 MJ/kg DM), short‐chain fatty acid concentration (2.43 mmol/g DM), microbial protein synthesis (405 mg/g DM), and ammonia‐N (10.2 mg/dL). Activities of carboxymethyl cellulase, microcrystalline cellulase, and filter paper‐degrading enzymes were significantly elevated in all inoculated treatments, with T5 consistently yielding the highest values. These results demonstrate that co‐application of the leopard moth gut‐derived bacterial consortium with glucose and urea effectively delignifies DPL, substantially enhances its fermentability and nutritive value, and offers a sustainable strategy for valorizing agricultural residues in ruminant nutrition.