Search Results (100)

Bamboo shoot dietary fiber (BSDF) is dominated by an insoluble fraction, which severely restricts its physicochemical performance and food application. In this study, the soluble dietary fiber (SDF) content of bamboo shoots was enhanced using three enzyme fermentation sequences: enzymatic hydrolysis followed by fermentation (EH), fermentation followed by enzymatic hydrolysis (F-EH), and integrated enzymatic hydrolysis and fermentation (IEHF). EH-F treatment resulted in the highest SDF content (17.27%). Variations in pH, biomass, enzyme activity, and short-chain fatty acids were assessed to understand the differences in the modification efficiency among the treatment sequences. Fourier transform infrared (FTIR) spectroscopy indicated that sequential enzymatic and fermentation treatments altered the chemical structure of bamboo shoot powder, consistent with the conversion of insoluble to soluble fractions. SDF from the EH-F treatment exhibited superior water-holding capacity (6.71 g/g), oil affinity (6.42 g/g), and DPPH radical scavenging rate (65.69% at 1.2 mg/mL). Moreover, SDF from the EH-F treatment achieved an 87.82% carbohydrate residue retention rate during simulated gastrointestinal digestion. These enhanced properties were associated with improved hydration properties resulting from the sequential tailoring process. This preliminary study explored the effects of different enzyme fermentation sequences on BSDF modification, providing a reference for the utilization of BSDF. Full article

This review examines sorghum digestibility from molecular structure to clinical implications, focusing on compositional factors, processing methods, and health outcomes. We evaluate how sorghum’s unique protein–starch interactions influence digestibility and explore emerging technologies that can modulate these properties for targeted nutritional benefits. Cooked sorghum generally has lower digestibility than raw sorghum and other cereals due to heat-induced protein–starch cross-linking and the formation of disulfide bonds by sorghum proteins (kafirins), which restrict enzymatic access. Enzyme inhibitors in sorghum further reduce starch hydrolysis. This reduced digestibility may negatively impact malnourished individuals and those relying on sorghum as a dietary staple. However, it can be advantageous to individuals with diabetes by lowering postprandial blood glucose levels. Sorghum consumption may also beneficially influence the gut microbiome. Certain processing methods have been shown to significantly enhance digestibility while preserving beneficial bioactive compounds. Improving digestibility through these strategies may enhance sorghum’s value for vulnerable populations while maintaining its metabolic advantages. Balancing increased nutrient bioavailability with preservation of beneficial functional properties is critical for optimizing sorghum as a health-promoting grain across diverse populations. Full article

Corn dust is an abundant agro-industrial by-product with potential as an alternative energy source. Its use in animal feeding, however, is restricted by high fiber content and low digestibility. This study evaluated the effects of non-starch polysaccharide (NSP) enzymes and yeast (Candida tropicalis KKU20) on the chemical composition, fermentation characteristics, and microbial populations of fermented corn dust. The experiment was conducted using a completely randomized design with a 3 × 2 factorial arrangement plus an additional control treatment. Factor A consisted of three levels of enzyme supplementation (0.02%, 0.04%, and 0.06% of dry matter), and Factor B consisted of yeast supplementation (without yeast or with C. tropicalis KKU20, approximately 1 × 10¹³ cells/g of inoculum). The control treatment consisted of fermented corn dust without enzyme or yeast supplementation. Samples were fermented for 15 days prior to analysis. Yeast inoculation increased crude protein and non-fiber carbohydrate contents while reducing neutral detergent fiber, acid detergent fiber, and acid detergent lignin (p < 0.05). Significant enzyme × yeast interactions were observed for several components, particularly fiber fractions (p < 0.05). The reduction in fiber was more pronounced when enzymes were combined with yeast. Predicted energy values, including metabolizable and digestible energy, were increased following yeast supplementation (p < 0.05). Fermentation characteristics were mainly affected by yeast. Yeast-treated samples exhibited higher pH and ammonia–nitrogen concentrations, indicating increased nitrogen turnover during fermentation. In contrast, lactic and propionic acid concentrations were higher in treatments without yeast, while yeast inoculation was associated with lower acetic acid and slightly higher butyric acid levels. Microbial analysis indicated interactions between treatments for lactic acid bacteria populations, reflecting competition for available substrates. No coliform bacteria were detected, indicating acceptable hygienic quality. Overall, yeast inoculation modified the chemical composition of corn dust, particularly by increasing crude protein and reducing fiber fractions, while NSP enzymes contributed to fiber degradation, especially when combined with yeast. However, these changes reflect compositional modification rather than confirmed feeding value, and further evaluation under rumen or in vivo conditions is required. Full article

DNA cloning traditionally relies on two approaches: restriction endonuclease digestion-ligation, and homologous recombination involving exonucleases, polymerases, and other enzymes. Here, we present a novel cloning method that requires only restriction endonucleases, eliminating the need for exonucleases or polymerases. The linearized cloning vector and the foreign DNA fragment (FDF) containing overlapping sequences were mixed and incubated at the melting temperature of the overlapping DNA sequences for 5 min, then cooled slowly to 0 °C. The mixture was transformed into E. coli and positive transformants were obtained. This cloning method was named DNA ‘breathing’ recombination (DBR) cloning. The overlapping sequence between the linearized vector and the FDF is preferably from 12 to 16 base pairs. Even when the ends of the linearized vector contain mismatches of up to 20 base pairs with the ends of the FDF, the DBR cloning method can still proceed efficiently, enabling truly seamless assembly. Meanwhile, the DBR method supports one-step assembly of multiple fragments. Therefore, the DBR cloning method simplifies experimental operations and reduces experimental costs while maintaining high cloning efficiency. Full article

Tissue decellularization aims to obtain bioscaffolds for regenerative applications by removing all cellular components while preserving the extracellular matrix (ECM) architecture. Although decellularization removes the majority of linear nuclear DNA (nDNA), residual amounts remain detectable. However, the fate of circular mitochondrial DNA (mtDNA) after decellularization has not yet been reported. Cell death or injury can cause the release of mtDNA, which is resistant to breakdown by exonucleases. Extracellular mtDNA acts as a damage-associated molecular pattern (DAMP) that can trigger immune responses. The aim of this study is to assess the presence of residual mtDNA in the liver, bile duct, and vascular scaffolds after decellularization and whether this causes inflammatory responses in macrophages. Decellularized tissues showed a marked reduction in total DNA content well below the threshold of 50 ng/mg tissue. However, in liver and vascular scaffolds, a relative increase in the mtDNA:nDNA ratio was detected in the remnant DNA fraction. Residual mtDNA in bioscaffolds acted as DAMPs causing macrophage activation, as shown by increased cell proliferation and cytokine production. Strategies to further reduce remnant mtDNA were tested. We found that treatment with the endonuclease enzyme HpaII was effective in degrading residual mtDNA. Importantly, mtDNA removal resulted in a significantly reduced macrophage activation. In conclusion, our study shows that mtDNA is relatively resistant to the decellularization procedure and can act as a DAMP in bioscaffolds. This underscores the importance of removing mtDNA from decellularized bioscaffolds to improve the immunocompatibility for biomedical applications. Full article

The rapid expansion of anaerobic digestion (AD) as a key technology for producing renewable energy has led to a substantial increase in digestate generation. This has intensified the need for sustainable management strategies that align with circular economy principles. While the solid fraction of digestate (SD) is traditionally applied to land or composted, its heterogeneous composition, regulatory constraints, and handling challenges restrict its wider use. This review aims to clarify the current state of SD treatment and highlight emerging opportunities to convert this underexploited resource into value-added bioproducts. A systematic bibliographic analysis of the past decade was conducted to identify consolidated and emerging SD valorisation technologies, supported by an evaluation of EU-level regulatory frameworks and the role of mechanical solid–liquid separation in enabling downstream valorisation. In addition, a comprehensive comparative table compiling physicochemical characterisation data of SD from various feedstocks and separation methods is presented, emphasising the significant variability in composition and its implications for valorisation pathways. The results show that, while composting and thermochemical routes, particularly pyrolysis, remain predominant, novel approaches such as advanced drying, pelletisation, vermicomposting, insect bioconversion, and fermentation-based pathways (including submerged and solid-state fermentation) are rapidly gaining interest. These emerging technologies enable the production of high-value products such as biochar, pellets, enzymes, microbial biopesticides, protein sources, and fungal biomass. However, their adoption is currently limited by feedstock heterogeneity, process complexity, scalability constraints, and economic considerations. Overall, SD is a versatile feedstock whose valorisation is expanding beyond agricultural applications. However, regulatory harmonisation, quality assurance, and process optimisation are still needed to encourage industrial uptake and to fully integrate SD into circular bioeconomy frameworks. Full article

The modified DNA base 2,6 aminopurine (2-aminoadenine, (d)Z base) was originally found in phages to counteract host-encoded restriction systems. However, only a limited number of restriction endonucleases (REases) have been tested on dZ-modified DNA. Here, we report the activity results of 147 REases on dZ-modified PCR DNA. Among the enzymes tested, 53% are resistant or partially resistant, and 47% are sensitive when their restriction sites contain one to six modified bases. Sites with four to six dZ substitutions are most likely to resist Type II restriction. Our results support the notion that dZ-modified phage genomes evolved to combat host-encoded restriction systems. dZ-modified DNA can also reduce phage T5 exonuclease degradation, but has no effect on RecBCD digestion. When two genes for dZ biosynthesis and one gene for dATP hydrolysis from Salmonella phage PMBT28 (purZ (adenylosuccinate synthetase), datZ (dATP triphosphohydrolase), and mazZ ((d)GTP-specific diphosphohydrolase) were cloned into an E. coli plasmid, the level of dZ incorporation reached 19–20% of adenosine positions. dZ levels further increased to 29–44% with co-expression of a DNA polymerase gene from the same phage. High levels of dZ incorporation in recombinant plasmid are possible by co-expression of purZ, mazZ, datZ and phage DNA helicase, dpoZ (DNA polymerase) and ssb (single-stranded DNA binding protein SSB). This work expands our understanding of the dZ modification of DNA and opens new avenues for engineering restriction systems and therapeutic applications. Full article

This study investigates whether fecal microbiota transplantation (FMT) can alleviate gut microbiota dysbiosis induced by a high-fat diet (HFD) through modulation of fatty acid metabolism, competition for nutrients, production of short-chain fatty acids (SCFAs), and restoration of mucus layer integrity. To elucidate the mechanisms by which FMT regulates colonic microbial function and host metabolic responses, 80 male Bal b/c mice were randomly assigned to four experimental groups (n = 20 per group): Normal Diet Group (NDG), High-Fat Diet Group (HDG), Restrictive Diet Group (RDG), and HDG recipients of NDG-derived fecal microbiota (FMT group). The intervention lasted for 12 weeks, during which body weight was monitored biweekly. At the end of the experiment, tissue and fecal samples were collected to assess digestive enzyme activities, intestinal histomorphology, gene expression related to gut barrier function, and gut microbiota composition via 16S rRNA gene sequencing. Results showed that mice in the HDG exhibited significantly higher final body weight and greater weight gain compared to those in the NDG and RDG (p < 0.05). Notably, FMT treatment markedly attenuated HFD-induced weight gain (p < 0.05), reducing it to levels comparable with the NDG (p > 0.05). While HFD significantly elevated the activities of α-amylase and trypsin (p < 0.05), FMT supplementation effectively suppressed these enzymatic activities (p < 0.05). Moreover, FMT ameliorated HFD-induced intestinal architectural damage, as evidenced by significant increases in villus height and the villus height-to-crypt depth ratio (V/C) (p < 0.05). At the molecular level, FMT significantly downregulated the expression of pro-inflammatory cytokines (IL-1β, IL-1α, TNF-α) and upregulated key tight junction proteins (Occludin, Claudin-1, ZO-1) and mucin-2 (MUC2) relative to the HDG (p < 0.05). 16S rRNA analysis demonstrated that FMT substantially increased the abundance of beneficial genera such as Lactobacillus and Bifidobacterium while reducing opportunistic pathogens including Romboutsia (p < 0.05). Furthermore, alpha diversity indices (Chao1 and ACE) were significantly higher in the FMT group than in all other groups (p < 0.05), indicating enhanced microbial richness and community stability. Functional prediction using PICRUSt2 revealed that FMT-enriched metabolic pathways (particularly those associated with SCFA production) and enhanced gut barrier-related functions. Collectively, this study deepens our understanding of host–microbe interactions under HFD-induced metabolic stress and provides mechanistic insights into how FMT restores gut homeostasis, highlighting its potential as a therapeutic strategy for diet-induced dysbiosis and associated metabolic disorders. Full article

Bovine mastitis remains a globally prevalent disease, with the limitations of antibiotic-based treatments—such as the rise in antimicrobial resistance and the presence of drug residues—highlighting the urgent need for alternative therapeutic approaches. Inflammation is intricately linked to various cytokines and immunomodulatory proteins, among which cyclophilin A (CypA) serves as a pivotal inflammatory mediator, significantly contributing to the initiation and amplification of inflammatory responses under such conditions. The acquisition of high-purity recombinant protein is a fundamental prerequisite for in vitro functional studies of bovine CypA. This study aimed to construct a eukaryotic expression vector for bovine CypA and verify its expression in CHO-K1 cells. Utilizing the bovine CypA gene sequence available in GenBank, the coding region was artificially synthesized and optimized for codon usage, subsequently being inserted into the pPB[Exp] backbone vector via BsrGI and BstEII double digestion. The resulting polycistronic expression vector contained a CAG promoter driving the CypA transcription, an EF1α promoter driving the EGFP reporter gene, a PGK promoter controlling the puromycin resistance gene, and a C-terminal His-tag. Restriction enzyme digestion and bidirectional Sanger sequencing confirmed that the inserted fragment sequence was completely consistent with the optimized design. Robust EGFP fluorescence was observed 24 h post-transfection and remained stable after puromycin selection. qPCR analysis showed that the Ct value of CypA in the experimental group was 16.20 ± 0.04, while no amplification signal was detected in the control group. Additionally, Western blot analysis identified a CypA-specific band at approximately 18 kDa, confirming the correct expression of the exogenous CypA protein in CHO-K1 cells. Collectively, these results demonstrate the successful construction and validation of a bovine CypA eukaryotic expression vector. The established CHO-K1 expression system exhibited stable and efficient expression, thereby providing a robust foundation for future research on the production and application of recombinant CypA protein. Full article

This study investigated how amylopectin (AP) chain-length distribution (CLD) influences the structural and functional properties of ten waxy rice varieties through a multiscale analysis. The fitting CLD parameters revealed that varieties such as YN12 and GMN2 contained the highest proportions of intermediate chains (h_iii~14.5~10.2) and long chains (h_v~0.552~0.477), resulting in higher gelatinization temperatures, including onset temperatures (T_o~60.2~66.4 °C), temperature peak (T_p~72.0~72.2 °C), and the conclusion temperatures (T_c~80.8~79.4 °C). Sample YN12 exhibited the lowest breakdown viscosity (BDV~748 cP), indicating higher thermal stability and stable pasting behavior. All of the samples showed relative crystallinity (Rc%) ranging from 19.3 to 22.8% with lamellar spacing of 8.8~9.4 nm. Sample YN12 exhibited the lowest enzymatic digestion rate (k~1.48 × 10⁻² min⁻¹) and the highest resistant starch (C_res~9.81%), reflecting restricted enzyme accessibility. In contrast, sample SN9714 and ZN106, characterized by a higher proportion of shorter branches in the long-chain region of AP CLD (β_vi~3.75~2.53), exhibited greater BDV (1407 cP) and faster digestion kinetics (k~1.90 × 10⁻² min⁻¹). The Rc% (22.83%) of NGXN was positively correlated with gelatinization enthalpy (ΔH~14.8 J/g). These findings highlight the pivotal role of AP CLDs in determining the structural, thermal, pasting, and digestive properties of waxy rice starch, offering molecular-level guidance for the development of rice-based ingredients with tailored functional characteristics. Full article

Pediculosis produced by the presence of the human head louse (Pediculus humanus capitis DeGeer, 1767) and the body louse (Pediculus humanus humanus L., 1758) remains a neglected tropical disease in Nigeria, where permethrin-based pediculicides are widely used. However, the resistance status of lice populations has not been previously assessed. Knockdown resistance (kdr) to pyrethroids is primarily driven by two mutations—T917I and L920F—in the voltage-sensitive sodium channel (VSSC) gene. This study investigated the presence of these mutations in 85 head and body lice collected from school-age children in two settlements in Nigeria. The T917I mutation was detected in head lice at frequencies ranging from 21% to 76%, and in body lice from 10% to 95%, with significant variation between sites and louse types. Remarkably, all lice examined carried the L920F mutation, regardless of T917I genotype, a pattern not previously reported in body lice. These findings suggest that pyrethroid resistance is well established or under active selection in the study populations. This is the first report of kdr mutations in human lice from Nigeria and highlights the urgent need for resistance monitoring programs. Early genetic surveillance of these mutations can inform treatment strategies and help prevent widespread resistance in lice populations, preserving the efficacy of available pediculicides. Full article

This research assessed the efficacy of glutamate (Glu) supplementation to feed in counteracting growth restriction and intestinal stress-induced injury in juvenile groupers (Epinephelus coioides; initial weight 15.11 ± 0.03 g). The study comprised five isonitrogenous and iso-lipidic diets: a fish-meal-based (FM) diet, a soya-meal-based (SBM) diet, and SBM diets containing varying Glu levels of 1.0% (G-1), 2.0% (G-2), or 3.0% (G-3). The trial employed a randomized design with five treatment groups. Each group was housed in triplicate aquariums and received assigned diets for 56 consecutive days. Supplementation with Glu resulted in dose-dependent enhancements in weight gain, specific growth rate, serum high-density lipoprotein cholesterol, intestinal superoxide dismutase activity, digestive enzyme activity (trypsin, lipase, amylase), amino acid metabolic enzyme activity (glutaminase, GLS; glutamine synthetase), and intestinal mRNA levels of GLS, IL-10, and TGF-β1. Maximal values of the G-3 diet were restored to the levels of the FM diet (p > 0.05). Serum total cholesterol, intestinal total antioxidant capacity, and catalase activity followed a similar increasing trend with Glu level, attaining maxima in diet G-3, yet these values remained markedly lower than those of the FM diet (p < 0.05). Conversely, intestinal malondialdehyde content and mRNA levels of genes IL-8, IL-12, IL-1β, and TNF-α exhibited a significant dose-dependent decrease, reaching minimal levels in diet G-3 that were restored to the levels of the FM diet (p > 0.05). The results above demonstrate that Glu addition enhances nutritional status and intestinal structural integrity by augmenting antioxidant and digestive capacity and mitigating inflammatory responses, consequently enhancing growth performance and intestinal health. Full article

Background: Centromeric alpha satellite DNA is organized into higher-order repeats (HORs), whose precise structure is often difficult to resolve in standard genome assemblies. The recent telomere-to-telomere (T2T) assembly of the human genome enables complete analysis of centromeric regions, including the full structure of HOR arrays. Methods: We applied the novel high-precision GRMhor algorithm to the complete T2T-CHM13 assembly of human chromosome 21. GRMhor integrates global repeat map (GRM) and monomer distance (MD) diagrams to accurately identify, classify, and visualize HORs and their subfragments. Results: The analysis revealed a novel Cascading 11mer HOR array, in which each canonical HOR copy comprises 11 monomers belonging to 10 different monomer types. Subfragments with periodicities of 4, 7, 9, and 20 were identified within the array. A second, complex 23/25mer HOR array of mixed Willard’s/Cascading type was also detected. In contrast to the hg38 assembly, where a dominant 8mer and 33mer HOR were previously annotated, these structures were absent in the T2T-CHM13 assembly, highlighting the limitations of hg38. Notably, we discovered a novel 52mer HOR—the longest alpha satellite HOR unit reported in the human genome to date. Several subfragment repeats correspond to alphoid subfamilies previously identified using restriction enzyme digestion, but are here resolved with higher structural precision. Conclusions: Our findings demonstrate the power of GRMhor in resolving complex and previously undetected alpha satellite architectures, including the longest canonical HOR unit identified in the human genome. The precise delineation of superHORs, Cascading structures, and HOR subfragments provides unprecedented insight into the fine-scale organization of the centromeric region of chromosome 21. These results highlight both the inadequacy of earlier assemblies, such as hg38, and the critical importance of complete telomere-to-telomere assemblies for accurately characterizing centromeric DNA. Full article

High-fat diets are commonly used in fish farming due to their protein-sparing effect, contributing to reduced production costs. However, this practice may have adverse effects such as metabolic impairment and inflammation. These problems can be assessed in two ways: by developing functional diets or using food restriction, which leads to compensatory growth. The present study characterized digestion in gilthead sea bream fed a high-fat diet in the presence (HT) or absence (HF) of an olive oil polyphenol as an additive, hydroxytyrosol, under two different dietary regimes: feeding to satiation (ST) or at a 40% restriction (R). Digestive enzyme activities, specifically trypsin-like activities, were mainly upregulated by dietary treatment (HT). In contrast, restriction effects mainly appeared during digestion in the pyloric caeca, where a significant rise in chymotrypsin-like activities was detected. Moreover, those fish tended to have an increased relative intestinal length compared to those fish fed at a standard ration. Feed restriction enhanced the growth of γ-Proteobacteria in pyloric caeca and proximal intestinal regions, without altering their population in the distal intestine. Overall, it is suggested that hydroxytyrosol inclusion at a standard ration could improve digestion processes in gilthead sea bream fed high-fat diets under healthier conditions than without this additive. Full article

Adrenal cortex carcinoma (ACC) in children is a rare tumor that is probably of multifactorial origin and is mainly associated with genetic and environmental alterations. In the south and part of the southeast of Brazil, as well as in the Paraguayan region bordering the Brazilian State of Paraná, ACC prevalence is higher than in any other country, which is associated with the high prevalence of the TP53 p.R337H variant in Paraná (0.30%), Santa Catarina (0.249%), cities around Campinas-SP (0.21%), and the Paraguayan cities on the border with Paraná (0.05%). Recent research suggests that the co-segregation of XAF1-E134* and TP53-R337H mutations leads to a more aggressive cancer phenotype than TP53-R337H alone. Breast cancer may be mildly influenced by co-segregation with XAF1 p.E134*, and this variant can also confer risk for sarcoma. Objectives: The objectives of this study were to (1) estimate the prevalence of the germline TP53 p.R337H and XAF1 p.E134* variants in Paraguay (excluding cities on the border with Paraná State, Brazil) and (2) estimate whether the ethnic origin of TP53 p.R337H carriers in Paraguay is similar to that of ethnic groups in Paraná (possible Portuguese/Spanish origin). Materials and methods: DNA tests for the identification of TP53 p.R337H were carried out from 2016 to 2019 at the Bio-Materials Laboratory of Facultad Politecnica, UNA, and at the Research Center in Biotechnology and Informatics (CEBIOTEC), Asunción, Paraguay. Polymerase chain reaction followed by restriction enzyme digestion (PCR-RFLP) was used to identify TP53 p.R337H, and real-time PCR (RT-PCR) was employed for XAF1 p.E134*. Peripheral blood samples from 40,000 Paraguayan newborns (NBs) were used for the TP53 p.R337H tests. The XAF1 p.E134* tests (RT-PCR) were performed on samples from 2000 Paraguayan newborns at the Pelé Pequeno Principe Research Institute, Curitiba, PR, Brazil. Results: The TP53 p.R337H variant was not found in any of the 14 Paraguayan departments investigated. A total of 12 of the 2000 Paraguayan NBs were positive for one XAF1 p.E134* allele. Conclusions: The hypothesis of Spanish immigrants carrying p.R337H to Paraguay was disproved. TP53 p.R337H neonatal testing in Paraguay is not recommended, except when there are families with Brazilian ancestry presenting cancer cases. Additional epidemiological studies are required to determine the likelihood of the identified prevalence of the XAF1 p.E134* allele (1/153) in NBs from Paraguay without TP53 p.R337H to present cancer risk. This study complements the first national initiative for the DNA screening of newborns aimed at mapping the TP53 p.R337H and XAF1 p.E134* variants in Paraguay (based on the regions of residence of the newborns). Full article