Search Results (6,445)

While chitosan is widely used in aquaculture feed, chitosan nanoparticles (CNPs) offer potential advantages due to their enhanced absorption. This study investigated the safe use of CNP levels in Nile tilapia feed, evaluating its impact on growth, immunity, and disease resistance. Five experimental diets were formulated and supplemented with zero chitosan (served as a control group), 1g/kg of chitosan (CS), and 1, 3, and 5 g/kg of CNPs. Each diet was randomly assigned to three replicate groups of 45 fish per group (15 fish/tank) with an average weight of (42.10 ± 0.05g, mean ± S.E.) twice daily (09:00 a.m. and 4:00 p.m.) to apparent satiation for two months. At the end of the feeding trial, fish fed 5 g/ kg of CNPs had the highest growth performance. However, no significant variations (p > 0.05) in somatic index were seen between the experimental groups. All chitosan and CNP-enriched groups exhibited improved intestinal morphology compared to the control group, characterized by increased villus length and width, reduced necrosis of intestinal tips, and better overall tissue integrity, with the CNP 3g and 5g groups demonstrating the most favorable intestinal structure. The CNP-treated groups (3, 5 g/kg) had significantly higher blood indices and serum globulin. Malondialdehyde (MDA) levels were lower in the CNP-treated groups compared to the chitosan macromolecule group. There was a substantial rise in glutathione (GSH), total antioxidant capacity (TAC), phagocytic index, and respiratory burst activity in the 5 g/kg CNP-treated group. The dietary addition of 5 g/kg of CNPs raised mRNA expression for TLR-2, MUC-2, and IGF-1, but there was no significant difference in HSP70 expression across treatments. After the experimental challenge with Aeromonas veronii biovar sobria, the groups that received 3 and 5 g/kg of CNPs exhibited the lowest mortality rates. Overall, the results suggest that including 5g/kg of CNPs in fish food is safe and effective for enhancing their health and growth, making it a promising addition to aquaculture feed. Full article

Atherosclerosis is a chronic inflammatory disease characterized by lipid accumulation and foam cell formation in the arterial wall. Promoting macrophage autophagy has emerged as a promising therapeutic strategy against atherosclerosis. Dipsacoside B (DB) is an oleanane-type pentacyclic triterpenoid saponin extracted from Lonicerae Flos with potential anti-atherosclerotic properties. In this study, we investigated the effects of DB on atherosclerosis progression in ApoE^−/− mice fed a high-fat diet and explored the underlying mechanisms in oxidized low-density lipoprotein (ox-LDL)-induced foam cells. DB treatment significantly reduced atherosclerotic lesion size, improved plaque stability, and regulated lipid metabolism without impairing liver and kidney function in ApoE^−/− mice. In vitro studies revealed that DB dose-dependently inhibited ox-LDL internalization and intracellular lipid accumulation in RAW264.7 macrophages. Mechanistically, DB induced autophagy, as evidenced by increased autophagosome formation and upregulated expression of autophagy markers LC3-II and p62 both in vivo and in vitro. Inhibition of autophagy by chloroquine abolished the antiatherosclerotic and pro-autophagic effects of DB. Furthermore, DB treatment increased LC3-II and p62 mRNA levels, suggesting transcriptional regulation of autophagy. Collectively, our findings demonstrate that DB exerts anti-atherosclerotic effects by inhibiting foam cell formation via autophagy induction, providing new insights into the pharmacological actions of DB and its potential as a therapeutic agent against atherosclerosis. Full article

Background and objectives: Sudden cardiac death (SCD) is a natural and unexpected death of cardiac origin that occurs within 1 h from the onset of acute symptoms. The major leading causes of SCD are cardiomyopathies and channelopathies. In this review, we focus on channelopathies, inherited diseases caused by mutations affecting genes encoding membrane ion channels (sodium, potassium or calcium channels) or cellular structures that affect Ca²⁺ availability. The diagnosis of diseases such as long QT, Brugada syndrome, short QT and catecholaminergic polymorphic ventricular tachycardia (CPVT) is still challenging. Currently, genetic testing and next-generation sequencing allow us to identify many rare alterations. However, some non-coding variants, e.g., splice-site variants, are usually difficult to interpret and to classify. Methods: In our review, we searched for splice-site variants of genes involved in channelopathies, focusing on variants of unknown significance (VUSs) registered on ClinVar up to now. Results: The research led to a high number of splice-site VUSs of genes involved in channelopathies, suggesting the performance of deeper studies. Conclusions: In order to interpret the correlation between variants and pathologies, we discuss experimental studies, such as RNA sequencing and functional analysis of proteins. Unfortunately, as these in vitro analyses cannot always be performed, we draw attention to in silico studies as future perspectives in genetics. This review has the aim of discussing the potential methods of detection and interpretation of VUSs, bringing out the need for a future reclassification of variants with currently unknown significance. Full article

Aim: The primary endothelial NADPH oxidase isoform 4 (NOX4) is notably induced during hypoxia, with emerging evidence suggesting its vasoprotective role through H₂O₂ production. Therefore, we aimed to elucidate NOX4′s significance in endothelial function under hypoxia. Methods: Human vessels, in addition to murine vessels from Nox4^-/- mice, were explored. On a functional level, Mulvany myograph experiments were performed. To obtain mechanistical insights, human endothelial cells were cultured under hypoxia with inhibitors of hypoxia-inducible factors. Additionally, endothelial cells were cultured under combined hypoxia and laminar shear stress conditions. Results: In human occluded vessels, NOX4 expression strongly correlated with prostaglandin I2 synthase (PTGIS). Hypoxia significantly elevated NOX4 and PTGIS expression and activity in human endothelial cells. Inhibition of prolyl hydroxylase domain (PHD) enzymes, which stabilize hypoxia-inducible factors (HIFs), increased NOX4 and PTGIS expression even under normoxic conditions. NOX4 mRNA expression was reduced by HIF1a inhibition, while PTGIS mRNA expression was only affected by the inhibition of HIF2a under hypoxia. Endothelial function assessments revealed hypoxia-induced endothelial dysfunction in mesenteric arteries from wild-type mice. Mesenteric arteries from Nox4^-/- mice exhibited an altered endothelial function under hypoxia, most prominent in the presence of cyclooxygenase inhibitor diclofenac to exclude the impact of prostacyclin. Restored protective laminar shear stress, as it might occur after thrombolysis, angioplasty, or stenting, attenuated the hypoxic response in endothelial cells, reducing HIF1a expression and its target NOX4 while enhancing eNOS expression. Conclusion: Hypoxia strongly induces NOX4 and PTGIS, with a close correlation between both factors in occluded, hypoxic human vessels. This relationship ensured endothelium-dependent vasodilation under hypoxic conditions. Protective laminar blood flow restores eNOS expression and mitigates the hypoxic response on NOX4 and PTGIS. Full article

Selecting high-quality varieties with disease resistance by artificial crossbreeding is the most fundamental way to address the damage caused by Calonectria spp. in eucalypt plantations. However, understanding the mechanism of disease-resistant heterosis occurrence in eucalypts is crucial for successful crossbreeding. Two eucalypt hybrids, the susceptible EC333 (H1522 × unknown) and the resistant EC338 (W1767 × P9060), were screened through infection with Calonectria isolates, a pathogen that causes eucalypt leaf blight. RNA-Seq was performed on the susceptible hybrid, the disease-resistant hybrid, and their parents. The gene differential expression analysis showed that there were 3912 differentially expressed genes between EC333 and EC338, with 1631 up-regulated and 2281 down-regulated genes. The expression trends of the differential gene sets in P9060 and EC338 were similar. However, the expression trend of W1767 was opposite that of EC338. The similarity of the expression and the advantage of stress resistance in E. pellita suggested that genes with significant differences in expression likely relate to disease resistance. A GSEA based on GO annotations revealed that the carbohydrate binding pathway genes were differentially expressed between EC338 and EC333. The gene pathways that were differentially expressed between EC338 and EC333 revealed by the GSEA based on KEGG annotations were the sesquiterpenoid and triterpenoid biosynthesis pathways. The alternative splicing analysis demonstrated that an AS event between EC338 and EC333 occurred in LOC104426602. According to our SNP analysis, EC338 had 626 more high-impact mutation loci than the male parent P9060 and 396 more than the female parent W1767; W1767 had 259 more mutation loci in the downstream region than EC338, while P9060 had 3107 fewer mutation loci in the downstream region than EC338. Additionally, EC338 had 9631 more mutation loci in the exon region than EC333. Modules were found via WGCNA that were strongly and oppositely correlated with EC338 and EC333, such as module MEsaddlebrown, likely associated with leaf blight resistance. The present study provides a detailed explanation of the genetic basis of eucalypt leaf blight resistance, providing the foundation for exploring genes related to this phenomenon. Full article

Water deficiency and potential drought periods could be important ecological factors influencing cultivation areas and productivity once different crops are established. The principal supply of vegetable oil for oil crops is oil palm, and new challenges are emerging in the face of climatic changes. This study investigated the photosynthetic performance of 12 genotypes of Elaeis exposed to drought stress under controlled conditions. The assay included genotypes of Elaeis guineensis, Elaeis oleifera, and the interspecific O×G hybrid (E. oleifera × E. guineensis). The principal results showed that the E. guineensis genotype was the most efficient at achieving photosynthesis under drought stress conditions, followed by the hybrid and E. oleifera genotypes. The physiological parameters showed good prospects for vegetal breeding with different O×G hybrids, mainly because of their ability to maintain the equilibrium between CO₂ assimilation and stomatal aperture. We validated 11 genes associated with drought tolerance, but no differences were detected. These results indicate that no allelic variants were represented in the RNA during sampling for the validated genotypes. In conclusion, this study helps to define genotypes that can be used as parental lines for oil palm improvement. The gas exchange data showed that drought stress tolerance could define guidelines to incorporate the available genetic resources in breeding programs across the early selection in nursery stages. Full article

Composite produce leaf samples from commercial production rarely test positive for pathogens, potentially due to low pathogen prevalence or the relatively small number of plants sampled. Aggregative sampling may offer a more representative alternative. This pilot study investigated whether aggregative swab samples performed similarly to produce leaf samples in their ability to recover quality indicators (APCs and coliforms), detect Escherichia coli, and recover representative microbial profiles. Aggregative swabs of the outer leaves of romaine plants (n = 12) and composite samples consisting of various grabs of produce leaves (n = 14) were collected from 60 by 28 ft sections of a one-acre commercial romaine lettuce field. Aerobic plate counts were 9.17 ± 0.43 and 9.21 ± 0.42 log(CFU/g) for produce leaf samples and swabs, respectively. Means and variance were not significantly different (p = 0.38 and p = 0.92, respectively). Coliform recoveries were 3.80 ± 0.76 and 4.19 ± 1.15 log(CFU/g) for produce leaf and swabs, respectively. Means and variances were not significantly different (p = 0.30 and p = 0.16, respectively). Swabs detected generic E. coli in 8 of 12 samples, more often than produce leaf samples (3 of 14 positive, Fisher’s p = 0.045). Full-length 16S rRNA microbial profiling revealed that swab and produce leaf samples did not show significantly different alpha diversities (p = 0.75) and had many of the most prevalent bacterial taxa in common and in similar abundances. These data suggest that aggregative swabs perform similarly to, if not better than, produce leaf samples in recovering indicators of quality (aerobic and coliform bacteria) and food safety (E. coli), justifying further method development and validation. Full article

Background/Objectives: The global emergence of antibiotic-resistant zooanthroponotic Escherichia coli strains, producing extended-spectrum beta-lactamases (ESBL-E) and persisting in the intestines of farm animals, has now led to the development of a pandemic of extra-intestinal infectious diseases in humans. The search for innovative probiotic microorganisms that eliminate ESBL-E from the intestines of humans and animals is relevant. Previously, we received three isolates of bifidobacteria: from milk of a calved cow (BLLT1), feces of a newborn calf (BLLT2) and feces of a three-year-old child who received fresh milk from this calved cow (BLLT3). Our goal was to evaluate the genetic identity of BLLT1, BLLT2, BLLT3 isolates using genomic DNA fingerprinting (GDF), to study the tolerance, adhesion, homeostatic and antibacterial activity of BLLT1 against ESBL-E. Methods: We used a complex of microbiological, molecular biological, and immunological methods, including next generation sequencing (NGS). Results: GDF showed that DNA fragments of BLLT2 and BLLT3 isolates were identical in number and size to DNA fragments of BLLT1. These data show for the first time the possibility of natural horizontal transmission of BLLT1 through with the milk of a calved cow into the intestines of a calf and the intestines of a child. BLLT1 was resistant to gastric and intestinal stresses and exhibited high adhesive activity to calf, pig, chicken, and human enterocytes. This indicates the unique ability of BLLT1 to inhabit the intestines of animals and humans. We are the first to show that BLLT1 has antibacterial activity against ESBL-E strains that persist in humans and animals. BLLT1 produced 145 ± 8 mM of acetic acid, which reduced the pH of the nutrient medium from 6.8 to 5.2. This had an antibacterial effect on ESBL-E. The genome of BLLT1 contains ABC-type carbohydrate transporter gene clusters responsible for the synthesis of acetic acid with its antibacterial activity against ESBL-E. BLLT1 inhibited TLR4 mRNA expression induced by ESBL-E in HT-29 enterocytes, and protected the enterocyte monolayers used in this study as a bio-model of the intestinal barrier. BLLT1 increased intestinal alkaline phosphatase (IAP) as one of the main molecular factors providing intestinal homeostasis. Conclusions: BLLT1 shows promise for the creation of innovative functional nutritional products for humans and feed additives for farm animals that will reduce the spread of ESBL-E strains in the food chain. Full article

Background/Objectives: Iron (Fe) is a co-factor for enzymes of the developing brain necessitating sufficient supply. We investigated the effects of administering ferric derisomaltose/Fe isomaltoside (FDI) subcutaneously to Fe-deficient (ID) pregnant rats on cerebral and hepatic concentrations of essential metals and the expression of iron-relevant genes. Methods: Pregnant rats subjected to ID were injected with FDI on the day of mating (E0), 14 days into pregnancy (E14), or the day of birth (postnatal (P0)). The efficacy was evaluated by determination of cerebral and hepatic Fe, copper (Cu), and zinc (Zn) and gene expression of ferroportin, hepcidin, and ferritin H + L in pups on P0 and as adults on P70. Results: Females fed an ID diet (5.2 mg/kg Fe) had offspring with significantly lower cerebral and hepatic Fe compared to female controls fed a standard diet (158 mg/kg Fe). Cerebral Cu increased irrespective of supplying a standard diet or administering FDI combined with the standard diet. Hepatic hepcidin mRNA was significantly lower following ID. Cerebral hepcidin mRNA was hardly detectable irrespective of iron status. Conclusions: In conclusion, administering FDI subcutaneously to ID pregnant rats on E0 normalizes fetal cerebral and hepatic Fe. When applied at later gestational ages, supplementation with additional Fe to the offspring is needed to normalize cerebral and hepatic Fe. Full article

Wastewater surveillance for SARS-CoV-2 provides a broad assessment of community health since wastewater represents all community members, regardless of health care access and status of health (i.e., symptomatic and asymptomatic). Wastewater surveillance also provides early detection of disease transmission since the virus can be present in human waste before the presentation of clinical symptoms. We analyzed wastewater from Lehigh University (Bethlehem, PA) as well as the greater Bethlehem community for SARS-CoV-2 (N1, N2, and E genes) from August 2020 through May 2021. Total RNA was extracted and analyzed for SARS-CoV-2 and pepper mild mottle virus (PMMoV) by RT-qPCR. Of 73 Bethlehem wastewater samples, the number of positive samples depended on which SARS-CoV-2 gene was screened: 62 (84.9%) for N1, 52 (71.2%) for E, and 28 (38.4%) for N2. Of 67 university wastewater samples, the numbers of positive samples were 59 (88.1%) for N1, 51 (76.1%) for E, and 31 (46.3%) for N2. Temporal trends of SARS-CoV-2 in wastewater mirrored trends in COVID-19 positive cases in the Bethlehem community. Normalizing SARS-CoV-2 RNA concentrations to PMMoV (a human fecal indicator) increased the correlation between both N1 (ρ increased from 0.37 to 0.72) and E (ρ increased from 0.32 to 0.61) concentrations in wastewater with COVID-19 cases in the community. Full article

The main objective of this study is to isolate and identify lactic acid bacteria (LAB) from various Egyptian dairy products, examine their antibacterial and hemolysis potential, and ensure their safety when used as starter cultures in different dairy industries. Egyptian dairy products are often made without the use of commercial starter cultures, using raw milk and artisanal methods. The most popular traditional dairy products are Laban Rayeb and Zabady, as well as the cheese varieties of Ras, Domiati, and Karish. The microbial communities used for fermentation and the diversity of lactic acid bacteria are the most important factors that can affect the quality of these products. In order to investigate the diversity of cocci lactic acid bacteria in Egyptian dairy products, 70 samples of raw or fermented milk and cheeses were collected from traditional cheese-making factories, local markets, and farmhouses located in the Delta area of Egypt. Following this, the LAB were isolated from the samples. One hundred fifty-seven isolates of Gram-positive, catalase-negative, and cocci bacterial species were identified via rep-PCR, and some isolates were confirmed using pheS and 16S rRNA gene sequencing, as follows: Streptococcus infantarius subsp. infantarius (three isolates), Enterococcus hirae (three isolates), Enterococcus faecium (ninety-six isolates), Enterococcus faecalis (forty isolates), Enterococcus durans (six isolates), Lactococcus garvieae (one isolate), Pediococcus acidilactici (seven isolates), and Lactococcus lactis subsp. Lactis (one isolate). These findings validate that five strains have strong antibacterial activity against Escherichia coli, Salmonella typhimurium, and Listeria monocytogenes, and one hundred thirty-four strains were safe for hemolysis. The five strains were selected as protective cultures, including Pediococcus acidilactici, Lactococcus lactis subsp. lactis, E. faecalis, and E. faecium. Full article

The NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER (NRT1/PTR) family (NPF) plays an important role in plant nitrate absorption, distribution, and nitrogen use efficiency. Nevertheless, few reports are available regarding Eucalyptus grandis NPF genes and their expression profiles. This study aims to identify and analyze NPF genes and their expression under various nitrogen (N) conditions. In this study, we successfully screened 64 NPF genes within the E. grandis genome. Subsequently, we conducted an extensive analysis, encompassing investigations into chromosome location, gene structure, phylogenetic relationship, promoter region, conserved motif, and gene expression profile. RNA-seq was conducted to analyze the expression profiles of EgNPF genes under different N conditions. The 64 NPF genes were categorized into eight distinct groups, exhibiting an uneven distribution among the 10 chromosomes of E. grandis, and no member was mapped on chromosome (Chr) 9. The examination of cis-regulatory elements revealed that NPF promoters were closely related to light responsive element, MeJA responsiveness, anaerobic induction, gibberellin responsiveness, low-temperature responsiveness, and auxin responsiveness. We used the comparative transcriptome method to identify the 10 differently expressed EgNPF genes of E. grandis under high-nitrogen (N: 119 mg/L) and low-nitrogen (N: 29.25 mg/L) conditions. Expression pattern analyses revealed that EUGRSUZ_G03119 showed an elevated expression in both leaves and roots under high-nitrogen conditions compared to low-nitrogen conditions, suggesting that EUGRSUZ_G03119 might affect nitrogen transport and redistribution, potentially boosting the stress tolerance of E. grandis in response to nitrogen deficiency. These findings may provide valuable insights into the evolutionary development of the NPF gene family in E. grandis and facilitate the clarification of the molecular mechanism underlying EgNPF-mediated N absorption and distribution in E. grandis. Full article

Although fine dust is linked to numerous health issues, including cardiovascular, neurological, respiratory, and cancerous diseases, research on its effects on oral health remains limited. In this study, we investigated the protective effects of mature hemp stem extract-induced exosomes (MSEIEs) on periodontal cells exposed to fine dust. Using various methods, including microRNA profiling, PCR, flow cytometry, immunocytochemistry, ELISA, and Alizarin O staining, we found that MSE treatment upregulated key microRNAs, such as hsa-miR-122-5p, hsa-miR-1301-3p, and hsa-let-7e-5p, associated with vital biological functions. MSEIEs exhibited three primary protective functions: suppressing inflammatory genes while activating anti-inflammatory ones, promoting the differentiation of periodontal ligament stem cells (PDLSCs) into osteoblasts and other cells, and regulating LL-37 and MCP-1 expression. These findings suggest that MSEIEs have potential as functional biomaterials for applications in pharmaceuticals, cosmetics, and food industries. Full article

Eriochloa villosa (Thunb.) Kunth, the main weed in corn fields, has gradually developed resistance to nicosulfuron due to continuous and extensive application. We identified a biotype showing resistance to ALS inhibitor nicosulfuron with a resistant index 13.83, but without any target spot mutation. Herein, transcriptome sequencing was used to analyze the differences in gene expression at the transcriptional level between nicosulfuron-resistant E. villosa HEK-40 varieties and sensitive E. villosa HEK-15 varieties. The resistant and sensitive varieties comparison revealed 9931 DEGs after nicosulfuron application, of which 5426 and 4505 genes were up-regulated and down-regulated, respectively. Some contigs related to metabolic resistance were identified based on differential expression via RNA-Seq, which includes ABC transporters (ko02010), glucosinolate biosynthesis (ko00966), 2-oxocarboxylic acid metabolism (ko01210), alanine, aspartate, and glutamate metabolism pathways (ko00250). Seven CYP450 genes, four GST genes, ten ABC transporter genes, and two GT genes related to metabolic resistance were identified. The 10 candidate genes screened were validated using q-PCR. This validation indicates that activities associated with P450 enzymes, ABC transporters, and glutathione S-transferases (GST) may play a role in conferring resistance, which is important for reducing the impact of weeds on corn fields and ensuring food security. Full article

Melatonin plays a critical role in regulating embryo attachment in ruminants. While numerous studies have investigated its effects on early embryo development in vitro, the precise mechanisms by which melatonin influences the receptivity of endometrial epithelial cells in dairy cows remain unclear. The prerequisite for embryo implantation is the specific physiological condition of the endometrium that allows the embryo to implant, also known as endometrial receptivity. In addition to this, endometrial cells undergo processes such as proliferation, differentiation, and renewal, which makes the embryo more easily implanted. In this study, bovine endometrial epithelial cells were cultured and treated with melatonin, Silent Information Regulator 1 (SIRT1) inhibitor (EX527), and protein kinase B (AKT) phosphorylation inhibitor (periposine). RT-qPCR, Western blot, and immunofluorescence analysis were performed to investigate the effects of melatonin on the expression of target gene (SIRT1); cell proliferative genes, phosphatidylinositol-4,5-bisphosphate 3-Kinase (PI3K), AKT, cyclinD1, cyclinE1; and receptive genes (Leukemia Inhibitory Factor (LIF), Vascular Endothelial Growth Factor (VEGF), Homeobox Structure Gene 10 (HOXA10)). Additionally, microRNA (miRNA) mimics and inhibitors were used to transfect the cells to study the regulatory relationship between miRNA and receptive genes. Results indicated that melatonin activates the PI3K/AKT signaling pathway, upregulates cyclinD1 and cyclinE1, and promotes the proliferation of bovine endometrial epithelial cells. Melatonin also upregulated the expression of VEGF and HOXA10 and downregulated the expression of bta-miR-497 and bta-miR-27a-3p through SIRT1/PI3K/AKT signaling pathway. Further, bta-miR-497 and bta-miR-27a-3p were found to negatively regulate VEGF and HOXA10, respectively. Therefore, melatonin regulates the expression of VEGF and HOXA10 through the SIRT1/PI3K/AKT pathway and promotes the establishment of receptivity in bovine endometrial epithelial cells. Full article