An experiment was conducted to evaluate the effects of broiler breeder age on the growth performance, nutrient metabolism, and intestinal histomorphometry of the progeny during early life. A total of 180 one-day-old Cobb 500® chicks, with an average initial body weight of 45.82 ± 2.21 g, were assigned to a completely randomized design with three treatments, six replicates, and 10 birds per replicate. The treatments consisted of chicks derived from broiler breeders aged 39, 51, and 69 weeks. Productive performance, gastrointestinal tract histomorphometry, and nutrient metabolism were evaluated. Chicks originating from older breeders (69 weeks) presented superior growth performance, whereas those from younger breeders (39 weeks) presented greater nutrient metabolism. With respect to intestinal morphology, progeny from older breeders presented more developed intestinal structures at 1 and 21 days of age, whereas at 7 days of age, chicks from younger breeders presented more pronounced intestinal morphometric characteristics. Broiler breeder age influences the early performance, intestinal development, and nutrient metabolism of progeny. Older breeders favor early growth and intestinal maturation, whereas younger breeders enhance protein metabolism, highlighting the relevance of breeder age in early broiler management. These findings provide integrated evidence linking breeder age to growth performance, intestinal development, and nutrient metabolism during early life.
Intestinal stem cells (ISCs) play critical roles in the self-renewal and regeneration of the intestinal epithelium under physiological conditions and after injury, respectively. However, the underlying mechanisms are not fully understood. In this study, we investigate the role of the G protein-coupled receptor formyl peptide receptor 2 (FPR2) in intestinal epithelium homeostasis and regeneration. In mice, knocking out Fpr2 in either intestinal epithelial cells (IECs) or ISCs significantly reduces villus height and crypt depth by impairing ISC and transit-amplifying (TA) cell proliferation and differentiation, primarily TA cell differentiation. Mechanistic studies using intestinal organoid culture and bulk and single-cell RNA sequencing revealed that activation of FPR2 promotes proliferation and differentiation of ISCs and TA cells by activating the wingless/integrated (Wnt), Notch, and Hippo signaling pathways via protein kinase C (PKC)-extracellular signal-regulated kinase (ERK). Under physiological conditions, the Wnt and Notch signaling pathways mediate the regulation of ISC proliferation and differentiation by FPR2. Fpr2 deficiency in mouse IECs exacerbates X-ray- and 5-fluorouracil-induced villus and crypt injury, and delays intestinal epithelium regeneration by reducing ISC and TA cell proliferation. Administering an FPR2 agonist to mice significantly increases survival rates and accelerates intestinal epithelium regeneration after irradiation. Taken together, these results demonstrate that intestinal epithelial FPR2 plays a key role in intestinal epithelium homeostasis and regeneration by promoting ISC and TA cell proliferation and differentiation. FPR2 is a potential therapeutic target against chemotherapy- and radiotherapy-induced intestinal injury.
Chronic airway diseases (CAD) including Chronic obstructive pulmonary diseases (COPD), persistent asthma (≥2weekly episodes) and bronchiectasis, exhibit overlapping extrapulmonary manifestations potentially affecting health-related quality-of-life (HRQOL). The study objective was to quantify HRQOL of CAD patients and identify impact of associated pulmonary and extrapulmonary manifestations. Total 101 established CAD patients were recruited cross-sectionally (COPD, n=61, persistent asthma, n=30 and bronchiectasis, n=10). Patients completed spirometry, body composition (Dual-energy X-ray Absorptiometry), muscle function (isometric and isokinetic dynamometry), exercise capacity (6-minute walking distance,6MWD) and patient reported outcomes including physical activity, sleep and Hospital-Anxiety-and-Depression-Scale. HRQOL was assessed using EuroQol quality-of-life questionnaire and utility score (EQ-5D index) was stratified into tertiles to compare patient characteristics. Mean age of participants was 60.9±12.8y, and 25.7% were female. Mean EQ-5D index was 0.658(95%CI: 0.596, 0.721), significantly lower than population norms and other chronic diseases including diabetes, cardiovascular diseases and rheumatoid heart disease (p<0.01). Participants in lowest EQ-5D tertile were older, had higher depression, lower 6MWD, lower handgrip and quadriceps muscle strength than other two tertiles, whereas anxiety was higher in both lowest and intermediate tertile compared to highest tertile(p<0.01). In adjusted regression model, age (AOR:1.187) depression (AOR:1.438), anxiety (AOR:1.266), and body fat (AOR:1.114) showed increased odds for poor EQ-5D index while in intermediate vs highest EQ-5D tertile, only age (AOR: 1.125) and anxiety (AOR:1.403) showed a significant association. HRQOL is significantly lower in CAD compared to other chronic non-communicable diseases and significantly associated with extrapulmonary manifestations. Interventions targeting modifiable risk factors may aid in improving HRQOL in CAD.
With stress being causally linked to the most frequently occurring mental and physical health problems, effective coping with acute stress is essential for promoting overall well-being and building long-term stress resilience. However, most interventions aimed at improving stress regulation require extended periods of training and practice. In a previous study, we demonstrated that the Jena Safety Anchor-a brief, hypnosis-based intervention-effectively reduced subjective stress responses in a controlled laboratory setting using the Trier Social Stress Test (TSST). The present study examined whether this effect extends to everyday life. A total of 80 participants (n = 40 female) were randomly assigned to either a hypnosis group or a control group (n = 40 per group). Over the course of two weeks, all participants collected saliva samples at home to assess their cortisol awakening response (CAR), an indicator of anticipatory stress, reflecting the body's preparation for expected demands upon waking. Additionally, all participants continuously recorded their heart rate via smartwatches during the two data collection weeks. The hypnosis group received a single guided session of the Jena Safety Anchor at the end of the first week, while the control group came to the laboratory but received no intervention. CAR was measured via three saliva samples taken within one hour after waking on specified days. Morning heart rate was measured between 6 and 9 a.m. Results revealed a significant reduction in CAR and morning heart rate in the hypnosis group during the second week, following the intervention. In contrast, no change in CAR and morning heart rate was observed in the control group. These findings show that a single session of the Jena Safety Anchor can effectively reduce physiological stress anticipation in the morning. This points to its potential as a rapid, low-effort intervention for enhancing stress coping and fostering resilience. It offers a promising approach to mental health promotion that is both time-efficient and easily integrated into everyday routines.
Probiotic therapy that targets fat absorption in the small intestine holds considerable potential for regulating lipid metabolism, yet it faces challenges related to spatiotemporal delivery and colonization efficiency. To address these issues, an enzyme-probiotic biohybrid (AKK-COD) has been developed and subsequently encapsulated in thiol-modified alginate hydrogel microspheres. This therapeutic system enables sequential functions: intragastric protection, small intestinal adhesion, and probiotic release and colonization. The Ca2+-crosslinked hydrogel maintains structural integrity under acidic gastric conditions, ensuring protection of probiotics and enzymes while facilitating timely gastric emptying. Upon reaching the small intestine, the microspheres adhere to the small intestinal mucosa via disulfide linkage, enhancing local retention. Concurrently, the gradual dissociation of ionic crosslinks within the hydrogel allows the release of AKK-COD, which participates in the catalytic conversion of cholesterol, modulates lipid absorption pathways, and promotes microbial homeostasis. In vivo experiments demonstrate that this approach significantly alleviates high-fat diet-induced hypercholesterolemia, offering a safe and efficient strategy for lipid-lowering therapies.
Polycystic ovary syndrome (PCOS) is a prevalent reproductive and metabolic disorder frequently accompanied by obesity, creating a dual burden of infertility and metabolic disease for women, yet therapeutic options addressing both pathologies remain limited. Here, using high-fat diet-induced obese and dehydroepiandrosterone-induced PCOS mouse models, we demonstrate that alnustone, a naturally derived compound, exerts coordinated benefits on both metabolic and reproductive dysfunctions in females. Long-term alnustone administration effectively attenuated body weight gain, adipocyte hypertrophy, and insulin resistance. Critically, hyperandrogenism, estrous cycle irregularity, and polycystic ovarian morphology were concurrently reversed. Mechanistically, alnustone enhanced energy expenditure by activating brown adipose tissue (BAT) and promoting white adipose tissue browning through induction of thermogenic programs. Transcriptomic profiling further revealed that alnustone corrected lipid metabolic dysregulation in obese and PCOS mice via activating adipose tissue AMPK signaling. These findings establish alnustone as a dual-action therapeutic agent that restores both metabolic homeostasis and reproductive function through adipose tissue remodeling, highlighting its potential as a natural intervention for integrated management of obesity and PCOS.
To evaluate the effects of substituting soybean oil (SO) with soy lecithin (SL) and specialty lipid (CL) on the growth, body composition, and glucose-lipid metabolism of juvenile largemouth bass (Micropterus salmoides). The CL is a composite lipid source formulated by mixing coconut oil, linseed oil, soy lecithin oil, and palm oil in a ratio of 2:2:3:3, with the addition of antioxidants and emulsifiers as fillers. A total of six experimental groups were established: an iso-nitrogenous and iso-lipidic soybean oil (SO) (control group), a 1% soy lecithin supplementation group (1% SL), a 2% soy lecithin supplementation group (2% SL), a 2.5% specialty lipid supplementation group (2.5% CL), a 5% specialty lipid supplementation group (5% CL), and a 7.5% specialty lipid supplementation group (7.5% CL). After a 10-week feeding trial (initial body weight: 12.64 ± 0.02 g), the results showed that final body weight (FBW), weight gain rate (WGR), and specific growth rate (SGR) in the 2.5% CL and 5% CL groups were significantly higher than those in the SO group. The 5% CL group exhibited the lowest feed conversion ratio (FCR) numerically, though no significant differences were observed among groups. Whole-body crude lipid content was significantly reduced in the 7.5% CL. Compared to other groups, the 5% CL and 7.5% CL showed significantly increased levels of saturated fatty acid (SFA), docosahexaenoic acid (DHA), and n-3/n-6 polyunsaturated fatty acid (PUFA), while PUFA levels markedly decreased. Compared to the control, the 7.5% CL showed significant decreases in plasma serum albumin (ALB), total protein (TP), total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL). Regarding lipid metabolism gene expression, the 5% CL exhibited a significantly lower expression level of the peroxisome proliferator-activated receptor-γ (pparγ) in contrast to SO. The expression levels of peroxisome proliferators-activated receptor-α (pparα) were significantly upregulated in the 5% CL and 7.5% CL compared to other groups. Furthermore, the carnitine palmitoyltransferase (cpt1) expression level in the 5% CL was significantly higher than in all other groups. In conclusion, CL enhanced growth performance, effectively reduced whole-body lipid deposition, improved fatty acid composition, and promoted lipid metabolism in juvenile largemouth bass, without adversely affecting liver function. Based on the comprehensive results, the recommended optimal inclusion level for specialty lipid was 5%.
Nitrogen form and concentration are key environmental regulators that mediate symbiotic nitrogen fixation and root development in legumes. To understand the metabolic and molecular mechanisms underlying the effects of distinct nitrogen sources (nitrate and ammonium) on soybean nodulation and root development, this study evaluated root and nodulation phenotypes, and their corresponding transcriptional and metabolomic responses under different concentrations of NH₄Cl or KNO₃. Results showed that both high concentrations of NH₄Cl and KNO₃ significantly suppressed nodulation and promoted root growth, with nitrate exerting a stronger effect than ammonium. Metabolomic analysis revealed that ammonium treatment enhanced nitrogen assimilation and primary metabolism while suppressing symbiosis-related flavonoids. Nitrate specifically activated chemical defense pathways and inhibited parts of central carbon metabolism. Integrated multi-omics analysis indicated that the nitrogen sources differentially regulated key genes and metabolites involved in nitrogen metabolism, flavonoid/isoflavonoid biosynthesis, and arginine metabolism, leading to distinct metabolic fluxes. Our results demonstrate that soybean perceives different nitrogen forms to orchestrate a metabolic trade-off between autonomous growth, defense, and symbiosis, thereby providing new insights into the mechanistic basis of nitrogen-form adaptation in legumes.
Inborn errors in propionyl-CoA carboxylase cause life-threatening propionic acidemia. To understand the contribution of propionyl-CoA metabolism to cellular and systemic metabolic dysfunction, we generated inducible and tissue-specific Pcca knockout mouse models. The inducible whole-body loss of Pcca results in acute metabolic decompensation like the inborn error. The liver-specific loss of Pcca recapitulates these adverse effects, demonstrating the centrality of the liver to systemic disease. Propionate and pyruvate converge in the TCA cycle as major anaplerotic substrates. Strikingly, the lethality of Pcca knockout (KO) mice is reversed by simultaneously inhibiting pyruvate carboxylase (Pcx). Most metabolites suspected as deleterious in propionic acidemia are exacerbated in liver-specific Pcca;Pcx double KO mice with the exception of methylcitrate, suggesting a role of this metabolite in systemic toxicity. These data clarify relevant toxic biomarkers and suggest that rebalancing hepatic TCA cycle metabolism is critical to mitigate the adverse effects from alternative propionyl-CoA metabolic pathways.
In the management of gestational diabetes mellitus (GDM), the usual medical treatment requires frequent visits for glucose monitoring and insulin dose adjustment, and this imposes significant physical, psychological, and economic burdens on pregnant women. As mobile health platforms become increasingly integrated into diabetes care, telemedicine may help alleviate these burdens; however, evidence evaluating its effectiveness as a replacement for routine in-person GDM care remains limited. This study aims to evaluate the impact of telemedicine on the quality of life and costs for patients with GDM requiring insulin therapy. This single-center, 2-arm, randomized, open-label, parallel-group study included patients with GDM who started insulin injection therapy. Participants were randomized to either the telemedicine or standard face-to-face care groups for 10 (SD 2) weeks. The telemedicine intervention used a smartphone-linked platform that enabled the automatic transfer of glucose data from connected glucose meters and facilitated real-time video consultations. Primary end points included costs and patient satisfaction. Costs were assessed using claims data, transportation calculations, and wage-based productivity losses, while patient satisfaction was evaluated through changes in the Problem Areas in Diabetes Survey and Diabetes Therapy-Related Quality of Life questionnaire scores. Secondary outcomes included glycemic control and perinatal outcomes. In total, 38 participants were included, with 18 assigned to the telemedicine group and 20 to the standard care group. Total costs (32,712, 95% CI 15,412-50,013 vs 59,202, 95% CI 42,603-75,800 Japanese yen; $284, 95% CI 134-435 vs $515, 95% CI 370-659, purchasing power parity [PPP]-adjusted; P=.01), direct non-health care costs (922, 95% CI -240 to 2084 vs 2561, 95% CI 1447-3676 yen; $8, 95% CI -2 to 18 vs $22, 95% CI 13 to 32 PPP-adjusted; P=.02), and indirect costs (8981, 95% CI -7119 to 25,082 vs 32,832, 95% CI 17,384-48,279 yen; $78, 95% CI -62 to 218 vs $285, 95% CI 151-420 PPP-adjusted; P=.01) reduced significantly in the telemedicine group compared with the standard care group. The improvements in the Problem Areas in Diabetes Survey (-7.6, 95% CI -13.7 to -1.4; P=.02) and Diabetes Therapy-Related Quality of Life domain 1 (10.5, 95% CI 0.9-20.1; P=.03) scores from the baseline were significantly greater in the telemedicine group than that in the standard care group. Nonetheless, glycemic control and frequency of perinatal complications were comparable between the 2 groups. Consultation time was similar across groups, suggesting no added workload for clinicians. In this randomized trial, mobile health-enabled telemedicine safely replaced routine in-person visits for patients with GDM requiring insulin therapy. Telemedicine significantly reduced psychological and economic burdens without compromising glycemic or perinatal outcomes, demonstrating its value as a patient-centered and cost-efficient model of care. These findings support the broader implementation of mobile-based telemedicine approaches in GDM management.
Circadian rhythms regulate physiological functions and critical biological processes in almost all living forms. The circadian system enables organisms to anticipate and adapt to environmental fluctuations, thereby optimizing physiological responses. Proper synchronization between the master pacemaker in the brain and peripheral clocks throughout the organism is essential for maintaining homeostasis and promoting health. Conversely, circadian misalignment is increasingly recognized as a contributing factor in the pathogenesis of various diseases. Emerging evidence implicates circadian disruption as a driver of carcinogenesis. Indeed, epidemiological studies have associated night-shift work and chronic jet lag with an increased risk of cancer. At the molecular level, the biological clock comprises a complex network of transcriptional-translational feedback loops that generates approximately 24-h cycles in gene expression across all tissues. Notably, many clock-regulated genes participate in key pathways relevant to tumorigenesis, including cell cycle regulation, DNA damage repair, angiogenesis, and metabolism. Moreover, the circadian clock represents a promising therapeutic target in oncology. Numerous clinical trials have demonstrated that chronomodulation of chemotherapeutic agents can enhance treatment efficacy while mitigating adverse effects, thereby improving the quality of life for cancer patients. Hence, advancing our understanding of the molecular interplay between circadian regulation and cancer development may elucidate the role of circadian rhythms in tumorigenesis, facilitate the identification of novel prognostic markers and therapeutic targets, and ultimately improve outcomes in breast cancer patients.
Sepsis remains a life-threatening complication of severe pneumonia in infants and young children, yet early biomarkers are lacking. The gut microbiota modulates host immunity, but the association between the gut microbiota and pediatric pneumonia-associated sepsis is unclear due to confounding factors. In this prospective, 1:1 matched case-control study, we enrolled 100 infants and young children (28 days-36 months) with severe pneumonia, stratifying them into sepsis (n=50) and non-sepsis (n=50) groups matched for age and antibiotic exposure. Fecal samples collected within 48 hours of PICU admission underwent 16S rRNA gene sequencing. Diversity, taxonomic composition, and differential taxa were analyzed. The sepsis group exhibited significantly reduced alpha diversity (Shannon index: 2.30 ± 1.50 vs. 2.83 ± 1.36, P = 0.027), increased Enterobacteriaceae (18.97% vs. 9.44%, P = 0.046), and decreased Lachnospiraceae (2.01% vs. 8.11%, P = 0.010). LEfSe (Linear discriminant analysis Effect Size) further revealed distinct microbial signatures: the sepsis group exhibited enrichment of Lactobacillaceae and Clostridium butyricum, while the non-sepsis group was characterized by higher abundance of Lachnospiraceae and Segatella. Sepsis in infants and young children with severe pneumonia is associated with a specific gut microbiota signature, independent of major confounders. This dysbiotic profile, involving taxa associated with endotoxin production and short-chain fatty acid metabolism, may serve as an early biomarker for risk stratification and could inform microbiota-targeted interventions in critically ill infants and young children.
Agonistic behaviors are crucial and common among animals due to their importance in securing an individual's fitness, and neural signaling molecules are known to mediate these behaviors. Stenopus, a genus of shrimp-like decapod crustaceans characterized by a pair of enlarged pereiopods, exhibits prominent agonistic behaviors when encountering conspecifics of the same sex owing to its monogamous social structure. These shrimps represent another potentially excellent model organism for investigating the neural signaling basis of agonistic behaviors in crustaceans aside from traditional models. Yet, their underpinning molecular aspects have never been studied. Using S. hispidus and S. cyanoscelis as representatives, the present study is the first that systematically examines the genetics of agonistic behaviors in Stenopus. Three organs, including (1) antennae + antennules, (2) central nervous system, and (3) eyestalk ganglia, were RNA-sequenced to identify the differentially expressed genes (DEGs) and pathways potentially conserved in winners and losers of Stenopus after fighting interactions. Our results suggested that Stenopus agonistic interactions might be systemic activities involving the simultaneous modulation and interplay of multiple signaling cascades, organismal systems, and metabolic pathways. In particular, winners and losers typically exhibited enriched gene ontologies involved in neural signaling, and sensory and behavioral processes. Regarding enriched pathways, while those related to glycan biosynthesis and metabolism were enriched in winners, cholesterol metabolism and one-carbon pool by folate were enriched in losers. These different sets of pathways suggested that while fighting interactions in Stenopus were injurious to both combatants, the damage in losers appeared to be more traumatic. Furthermore, four neural signaling systems, including dopamine, acetylcholine, octopamine, and glutamate, were identified as potentially major mediators of agonistic behaviors and fighting interactions in both Stenopus species, with the first two appearing to be relatively more important. A comparison of the neural signaling systems involved in mediating aggression among pan-crustaceans suggested that Stenopus appeared to stand out by its seemingly major reliance on dopamine and acetylcholine, as opposed to the primarily serotonin-based regulation of aggression observed in most examined pan-crustaceans. The different metabolic responses between winners and losers in Stenopus highlight the profound, asymmetric physiological costs of social conflict at the molecular level. Furthermore, their unique reliance on dopamine and acetylcholine reveals diverse evolutionary trajectories in the neuroendocrine regulation of aggression, providing new insights into the current paradigms of invertebrate social behavior.
Myocardial dysfunction is a major determinant of mortality after cardiac arrest, yet the molecular events driving post-resuscitation injury remain incompletely understood. Nitric oxide (NO) has been proposed as a cardioprotective adjunct during extracorporeal life support (ECLS), but its mechanistic impact on myocardial recovery is unclear. We investigated whether NO supplementation during ECLS modulates oxidative stress, metabolic pathways, and apoptotic signaling in the post-cardiac arrest heart. Male Sprague Dawley rats underwent hypothermic cardiac arrest followed by ECLS resuscitation with or without NO supplementation (20 ppm). Myocardial tissue was analyzed using bulk RNA sequencing, quantitative RT-PCR, oxidative stress assays (MDA, 3-nitrotyrosine, total oxidant/antioxidant status), and TUNEL staining to characterize pathway-level alterations. NO supplementation markedly increased cardiomyocyte apoptosis (46%±3 vs. 27%±2; p < 0.0001). Transcriptomic profiling revealed > 550 differentially expressed genes, highlighting upregulation of inflammatory and apoptotic cascades (MAPK, TNF, NF-κB, proteasome/TLR pathways) and profound suppression of metabolic programs essential for myocardial recovery, including fatty acid oxidation, branched-chain amino acid metabolism, and oxidative phosphorylation. NO induced a striking oxidative-nitrosative imbalance, with elevated MDA, 3-nitrotyrosine, and total oxidant status and reduced total antioxidant capacity, resulting in a threefold increase in the oxidative stress index. NO administration during ECLS drives a coordinated oxidative-inflammatory-apoptotic response and disrupts metabolic pathways necessary for myocardial recovery, suggesting a mechanistic basis for worsened post-arrest myocardial injury. These findings have direct implications for optimizing resuscitation strategies in human ECLS and may inform future therapeutic modulation of NO signaling.
Current screening for diabetic kidney disease (DKD) relies on the estimated glomerular filtration rate (eGFR) and albuminuria, which often fail to detect early tubular dysfunction and non-albuminuric phenotypes. The integration of macroscopic urine physical characteristics with metabolic signatures may offer a novel approach to precision stratification. We conducted a multicenter, prospective-retrospective cohort study involving 364 participants with type 2 diabetes. We developed "FluxPro-DKD fusion model," that integrates "Digital Physicalomics" (computer-vision quantification of urine foam stability and chromaticity) and "Dual-Fluid Metabolomics" (serum-to-urine flux ratios). The model was trained in a discovery cohort (n=282) and tested in an independent external validation cohort (n=82). The primary outcome was the detection of early-stage DKD. We also assessed the model's prognostic utility for major adverse renal events over a simulated 3-year period. Metabolic profiling identified a distinct "serum-to-urine flux mismatch" of protein-bound uremic toxins (e.g., indoxyl sulfate), suggesting tubular secretory failure prior to glomerular damage. Digital physicalomics revealed that urine foam half-life was correlated with albuminuria (r=0.78). In the discovery cohort, the FluxPro-DKD fusion model achieved an area under the receiver operating characteristic curve (AUC) of 0.90 (95% confidence interval [CI], 0.87 to 0.93), significantly outperforming the standard clinical model (AUC, 0.78; P<0.001). The model maintained robust discrimination in the external validation cohort (AUC, 0.85; 95% CI, 0.79 to 0.91). Among patients with normoalbuminuria, those classified as high-risk by the model had a significantly higher projected 3-year event rate than those classified as low-risk (35.3% vs. 2.3%). The integration of digital urine physical phenotypes and metabolic flux ratios effectively reveals early tubular secretory dysfunction and improved risk stratification for diabetic kidney disease compared with standard clinical metrics.
Neutrophils are the most common type of white blood cell in the body, acting as crucial mediators of innate immunity. They initiate immune responses against invading microorganisms and threats like cancerous cells. Neutrophil infiltration is observed across inflammatory and autoimmune diseases and many cancers. Neutrophils orchestrate innate and adaptive immune responses to return the body to homeostasis, including after infections and preventing the development of tumours. While both protective and deleterious functions of neutrophils have been reported, this dual functionality reflects the heterogeneity of neutrophil subsets. Previously, neutrophils short lifespan resulted in their underappreciation. Recent advances in technology and methodology have drastically increased understanding of neutrophil biology, heterogeneity and plasticity, leading to a plethora of discoveries around the therapeutic potential of neutrophils, both through their direct cytotoxic effects to remove deleterious populations and through their ability to modulate other components of immunity. How to achieve this therapeutically with a rapidly-turned over population has proven elusive until now. This review highlights physiological and patho-physiological activities of neutrophil populations and their therapeutic potential and challenges. Recent and ongoing efforts to target or exploit neutrophil-type populations therapeutically have included engager-type antibody-based reagents and 'off-the-shelf' allogeneic cell therapy with and 'Immuno-Modulatory Alpha Neutrophils' (IMANs).
Sepsis is a life-threatening syndrome with dysregulated immune responses and multiple organ dysfunction. However, precise diagnostic biomarkers and effective therapeutic targets for this syndrome are still lacking. Protein ubiquitination modulates inflammatory regulation and immune cell function, but the specific immune cell subsets that drive ubiquitination-associated immune dysregulation in human sepsis have not been clearly identified. An integrated analysis was performed using 315,220 single cells from two single-cell RNA sequencing (scRNA-seq) datasets in conjunction with independent bulk transcriptomic cohorts. We quantified cell-type responsiveness using Augur, inferred intercellular communication via CellChat, and identified ubiquitination-related gene networks through weighted gene co-expression network analysis (WGCNA) and subsequent multi-algorithm feature selection. Functional validation was conducted with lipopolysaccharide (LPS)-stimulated murine dendritic cells (DCs) line - DC2.4 in vitro and a cecal ligation and puncture (CLP) mouse model in vivo. conventional Dendritic cells (cDCs) were identified as the most transcriptionally perturbed immune population in sepsis, with subsequent subclustering revealing that the type-1 conventional dendritic cells (cDC1) subset specifically exhibited pronounced activation of ubiquitination signatures. Cell-cell communication analysis identified TNF signaling as a sepsis-specific pathway, in which cDC1 functions as a critical mediator predominantly via the TNF-TNFRSF1B axis. Four ubiquitination-related genes (CUL1, UBE2F, UBE2N and UBE3A) demonstrated reproducible diagnostic performance across three independent cohorts. Notably, UBE2F showed the strongest upregulation and functional relevance in sepsis models. Both in vitro and in vivo experiments showed that silencing UBE2F markedly suppressed dendritic cell activation, decreased proinflammatory cytokine production and organ injury, and ultimately improved survival in septic mice. Our results reveal cDC1 as a key immune cell subset involved in ubiquitination-mediated immune dysregulation in sepsis and suggests that UBE2F may serve as a potential diagnostic biomarker and therapeutic target.
Edwardsiella piscicida is an intracellular bacterial pathogen that causes intestinal injury and hemorrhagic sepsis in marine and freshwater animals. VgrG (valine-glycine repeat protein G) has been identified as a crucial virulence factor in the type VI secretion system (T6SS) of the bacterium, but its role and mechanism involved in E. piscicida-host interactions remain unclear. In this study, we found that the wild-type E. piscicida strain markedly induced host cell ferritin degradation and elevated intracellular Fe²⁺ levels, but a vgrG deletion mutant (ΔvgrG) strain failed to induce similar effects in fish cells, indicating the role of VgrG in the regulation of iron metabolism. However, in the NCOA4 (nuclear receptor coactivator 4, a selective cargo receptor that binds ferritin)-knockout fish cells, VgrG did not alter ferritin protein expression and E. piscicida-promoted intracellular Fe²⁺ levels, suggesting that VgrG caused iron storage disorders via NCOA4-dependent ferritinophagy. In support of this notion, VgrG overexpression was found to facilitate the co-localization of ferritin with autophagosomes and lysosomes, and also drove the interaction of NCOA4 with lysosomes, strengthening the involvement of this effector in mediating NCOA4-dependent ferritinophagy. Notably, in NCOA4-knockout cells, the inhibitory effect of VgrG on the intracellular growth of E. piscicida was further magnified, particularly highlighting the role of VgrG-induced ferritinophagy in augmenting bacterial intracellular survival. Moreover, zebrafish infected with the ΔvgrG strain had a much higher survival rate than those infected with the wild-type strain. In summary, our data revealed a new function of VgrG in maintaining the persistence of E. piscicida within host cells, which advanced our understanding of how intracellular pathogens target ferritinophagy to evade the host's immune surveillance. Moreover, this work also supported the potential for developing defense strategies against E. piscicida based on the VgrG-induced ferritinophagy response in fish.
With increasing mozzarella consumption in the United States, mozzarella innovations are of interest, particularly in functional attributes. One opportunity in mozzarella is the application of Lacticaseibacillus casei adjunct culture to increase desirable flavors and modulate functional properties such as melting. The objective of this study was to determine the influence of the adjunct culture, Lacti. casei, on the sensory, physical, and functional properties of part-skim, pasta filata mozzarella cheese. Two treatments of mozzarella were produced: a control with S. thermophilus, and a modified treatment with Streptococcus thermophilus and Lacti. casei. Samples were stored at 3.3°C and assessed at 5, 25, 50, and 75 days in cubed cold and melted preparations by a trained sensory panel (n = 10). Meltability and physical properties were assessed through instrumental measures. The inclusion of Lacti. casei adjunct modulated melting behavior in baked applications over storage time. Results showed that Lacti. casei suppressed mozzarella meltability at Day 25 (compared to the control [p ≤ 0.05]). Lacticaseibacillus casei did not influence the sensory or texture attributes of cubed cold or melted mozzarella (p ≥ 0.05). However, storage time strongly influenced yellow color, butter flavor, bitterness, adhesiveness, and cohesiveness (p ≤ 0.05). These findings suggest that Lacti. casei may be a beneficial adjunct culture for improving baked mozzarella performance over its shelf life, though it may not be desired for all cheese producers.
Cardiovascular and metabolic diseases account for an increasing share of morbidity and mortality globally. Folic acid supplementation has been linked to a lowered risk of stroke and some metabolic indicators due to its involvement in homocysteine and one-carbon metabolism and its role in the production of nitric oxide; however, the evidence on these associations is inconclusive. We searched MEDLINE, Embase, CINAHL, the Cochrane Library, and the Database of Abstracts of Reviews of Effects from inception to February 2024 for systematic reviews and meta-analyses investigating the associations of folate (dietary intake, supplementation, or blood concentrations) with any cardiometabolic outcome. We performed screening, data abstraction, and risk of bias assessment in duplicate, and assessed the credibility of the evidence using predefined criteria. We identified 113 unique associations from 49 reviews. The included syntheses mostly had low risk of bias of and provided pooled risk estimates from intervention trials or prospective cohorts. A larger volume of evidence was available for composite cardiovascular outcomes, coronary heart disease, and stroke compared to other outcomes. No association reached a convincing or highly suggestive level of credibility. Six directional associations and five null associations met the criteria for a suggestive level of credibility. Three dose-response relationships, all at suggestive levels of credibility, supported an association between higher dietary folate intake and a reduced risk of coronary heart disease and stroke. The available evidence on the association between folate status and cardiometabolic outcomes primarily focuses on secondary prevention of cardiometabolic diseases and substantially underrepresents low- and middle-income countries. More large-scale studies are warranted to validate a relationship between folate status and cardiometabolic events or indicators. Overall, the evidence landscape around folate and cardiometabolic diseases appears to be limited both in volume and scope. PROSPERO: CRD42021265041.