Spider Silk Protein Forms Amyloid-Like Nanofibrils through a Non-Nucleation-Dependent Polymerization Mechanism.

Amyloid fibrils-nanoscale fibrillar aggregates with high levels of order-are pathogenic in some today incurable human diseases; however, there are also many physiologically functioning amyloids in nature. The process of amyloid formation is typically nucleation-elongation-dependent, as exemplified by the pathogenic amyloid-ß peptide (Aß) that is associated with Alzheimer's disease. Spider silk, one of the toughest biomaterials, shares characteristics with amyloid. In this study, it is shown that forming amyloid-like nanofibrils is an inherent property preserved by various spider silk proteins (spidroins). Both spidroins and Aß capped by spidroin N- and C-terminal domains, can assemble into macroscopic spider silk-like fibers that consist of straight nanofibrils parallel to the fiber axis as observed in native spider silk. While Aß forms amyloid nanofibrils through a nucleation-dependent pathway and exhibits strong cytotoxicity and seeding effects, spidroins spontaneously and rapidly form amyloid-like nanofibrils via a non-nucleation-dependent polymerization pathway that involves lateral packing of fibrils. Spidroin nanofibrils share amyloid-like properties but lack strong cytotoxicity and the ability to self-seed or cross-seed human amyloidogenic peptides. These results suggest that spidroins´ unique primary structures have evolved to allow functional properties of amyloid, and at the same time direct their fibrillization pathways to avoid formation of cytotoxic intermediates.

Evaluation of Fermented Soybean Meal to Replace a Portion Fish Meal on Growth Performance, Antioxidant Capacity, Immunity, and mTOR Signaling Pathway of Coho Salmon (Oncorhynchus kisutch).

In this study, we evaluated the effects of fermented soybean meal (FSBM) or/and unfermented SBM replacing a portion of fish meal (FM) on the growth performance, antioxidant capacity, immunity, and mechanistic target of rapamycin (mTOR) signaling pathway of juvenile coho salmon (Oncorhynchus kisutch). Four groups of juvenile coho salmon (initial weight 152.23 ± 3.21 g) in triplicate were fed for 12 weeks on four different iso-nitrogen and iso-lipid experimental diets: G0 diet (28% FM protein, control group), G1 diet (18% FM protein and 10% SBM protein), G2 diet (18% FM protein, 5% SBM protein, and 5% FSBM protein), and G3 diet (18% FM protein and 10% FSBM protein). The main results were compared with the G0 diet; the weight gain rate, specific growth rate, and condition factor of juveniles in G3 were increased significantly (p < 0.05). The content of muscle crude protein, the total protein, glucose, albumin, total cholesterol in serum, and the total antioxidant capacity in the liver of juveniles in G3 was increased significantly (p < 0.05). The activities of pepsin, trypsin, α-amylase, and lipase in the intestine, the superoxide dismutase, catalase, and alkaline phosphatase in the liver of juveniles in G3 were increased significantly (p < 0.05). The expression levels of phosphatidylinositide 3-kinases, serine/threonine kinase, mTOR, and ribosomal protein S6 kinase 1 genes in the liver of juveniles in G3 were upregulated significantly (p < 0.05). The feed coefficient ratio, viscerosomatic index, the contents of muscle moisture, and malondialdehyde in the liver of juveniles in G3 were decreased significantly (p < 0.05). The expression levels of tumor necrosis factor α, interleukin 1ß, and interleukin 6 genes in the liver of juveniles in G3 were downregulated significantly (p < 0.05). However, there was no significant effect (p > 0.05) on the survival rate, food intake, and muscle crude lipid and ash of juveniles among the experimental groups. In conclusion, FSBM to replace a portion FM had a positive effect on the growth performance, protein deposition, antioxidant enzyme activity, digestive enzyme activity, protein synthesis, and immune-related genes of juvenile coho salmon.

Effects of Dietary Vitamin C on the Growth Performance, Biochemical Parameters, and Antioxidant Activity of Coho Salmon Oncorhynchus kisutch (Walbaum, 1792) Postsmolts.

Vitamin C (VC) plays an essential role in fish physiological function and normal growth. However, its effects and requirement of coho salmon Oncorhynchus kisutch (Walbaum, 1792) are still unknown. Based on the influences on growth, serum biochemical parameters, and antioxidative ability, an assessment of dietary VC requirement for coho salmon postsmolts (183.19 ± 1.91 g) was conducted with a ten-week feeding trial. Seven isonitrogenous (45.66% protein) and isolipidic (10.76% lipid) diets were formulated to include graded VC concentrations of 1.8, 10.9, 50.8, 100.5, 197.3, 293.8, and 586.7 mg/kg, respectively. Results showed that VC markedly improved the growth performance indexes and liver VC concentration, enhanced the hepatic and serum antioxidant activities, and increased the contents of serum alkaline phosphatase (AKP) activity, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and total cholesterol (TC) whereas decreased the serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) activities, and triglyceride (TG) level. Polynomial analysis showed that the optimal VC levels in the diet of coho salmon postsmolts were 188.10, 190.68, 224.68, 132.83, 156.57, 170.12, 171.00, 185.50, 142.77, and 93.08 mg/kg on the basis of specific growth rate (SGR), feed conversion ratio (FCR), liver VC concentration, catalase (CAT), hepatic superoxide dismutase (SOD) activities, malondialdehyde (MDA) content, and serum total antioxidative capacity (T-AOC), AKP, AST, and ALT activities, respectively. The dietary VC requirement was in the range of 93.08-224.68 mg/kg for optimum growth performance, serum enzyme activities, and antioxidant capacity of coho salmon postsmolts.

Vitamin C regulates the production of reactive oxygen species through Wnt10b signaling in the gill of zebrafish.

In this study, the mechanism that vitamin C (VC) regulates the production of reactive oxygen species (ROS) through Wnt10b signaling was investigated in the gill of zebrafish (Danio rerio). The results showed that 0.5 and 1.0 g/kg VC diets induced the gene expression of Wnt10b, ß-catenin, SOD, CAT, and GSH-PX in gill. In addition, VC decreased the levels of H2O2, O2·- and ·OH, whereas the activities of SOD, CAT, and GSH-PX were increased by VC in the gill of zebrafish. To evaluate the role of Wnt10b in regulating oxidative stress, Wnt10b RNA was further interfered and the gene expression and activities of antioxidant enzymes were detected in gill. The result of Wnt10b RNA interference showed that Wnt10b signaling played a key role in regulating the gene expression of SOD, CAT, and GSH-PX. In all, VC may regulate the production of ROS through Wnt10b signaling in the gill of zebrafish (Danio rerio).

One-step heating strategy for efficient solubilization of recombinant spider silk protein from inclusion bodies.

BACKGROUND: Spider silk is a proteinaceous fiber with remarkable mechanical properties spun from spider silk proteins (spidroins). Engineering spidroins have been successfully produced in a variety of heterologous hosts and the most widely used expression system is Escherichia coli (E. coli). So far, recombinantly expressed spidroins often form insoluble inclusion bodies (IBs), which will often be dissolved under extremely harsh conditions in a traditional manner, e.g. either 8 mol/L urea or 6 mol/L guanidine hydrochloride, highly risking to poor recovery of bioactive proteins as well as unexpected precipitations during dialysis process. RESULTS: Here, we present a mild solubilization strategy-one-step heating method to solubilize spidroins from IBs, with combining spidroins' high thermal stability with low concentration of urea. A 430-aa recombinant protein (designated as NM) derived from the minor ampullate spidroin of Araneus ventricosus was expressed in E. coli, and the recombinant proteins were mainly present in insoluble fraction as IBs. The isolated IBs were solubilized parallelly by both traditional urea-denatured method and one-step heating method, respectively. The solubilization efficiency of NM IBs in Tris-HCl pH 8.0 containing 4 mol/L urea by one-step heating method was already comparable to that of 7 mol/L urea with using traditional urea-denatured method. The effects of buffer, pH and temperature conditions on NM IBs solubilization of one-step heating method were evaluated, respectively, based on which the recommended conditions are: heating temperature 70-90 °C for 20 min, pH 7.0-10, urea concentration 2-4 mol/L in normal biological buffers. The recombinant NM generated via the one-step heating method held the potential functions with self-assembling into sphere nanoparticles with smooth morphology. CONCLUSIONS: The one-step heating method introduced here efficiently solubilizes IBs under relatively mild conditions compared to the traditional ones, which might be important for the downstream applications; however, this protocol should be pursued carefully in terms of urea-induced modification sensitive applications. Further, this method can be applied under broad buffer, pH and temperature conditions, conferring the potential to apply to other thermal stable proteins.

Vitamin C inhibits lipid deposition through GSK-3ß/mTOR signaling in the liver of zebrafish.

In this study, the mechanism that VC inhibits lipid deposition through GSK-3ß/mTOR signaling was investigated in the liver of Danio rerio. The results indicated that 0.5- and 1.0-g/kg VC treatments activated mTOR signaling by inhibiting GSK-3ß expression. The mRNA expression of FAS, ACC, and ACL, as well as the content of TG, TC, and NEFA, was decreased by 0.5- and 1.0-g/kg VC treatments. Moreover, to confirm GSK-3ß playing a key role in regulating TSC2 and mTOR, GSK-3ß RNA was interfered and the activity of GSK-3ß was inhibited by 25- and 50-mg/L LiCl treatments, respectively. The results indicated that GSK-3ß inactivation played a significant role in inducing mTOR signaling and inhibiting lipid deposition. VC treatments could induce mTOR signaling by inhibiting GSK-3ß, and mTOR further participated in regulating lipid deposition by controlling lipid profile in the liver of zebrafish.

Met-enkephalin inhibits ROS production through Wnt/ß-catenin signaling in the ZF4 cells of zebrafish.

Opioid neuropeptides are developed early in the course of a long evolutionary process. As the endogenous messengers of immune system, opioid neuropeptides participate in regulating immune response. In this study, the mechanism that Met-enkephalin (M-ENK) inhibits ROS production through Wnt/ß-catenin signaling was investigated in the ZF4 cells of zebrafish. ZF4 cells were exposed to 0, 10, 20, 40, 80, and 160 µM Met-enkephalin (M-ENK) for 24 h, and the cell viability was detected with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The cell viability was significantly increased by 10, 20, 40, 80, and 160 µM M-ENK. After ZF4 cells were exposed to 0, 20, 40, and 80 µM M-ENK for 24 h, the mRNA expression of Wnt10b, ß-catenin, and CCAAT/enhancer binding protein α (C/EBPα) was significantly increased by 40 and 80 µM M-ENK. However, the mRNA and protein expression of GSK-3ß was significantly decreased by 40 and 80 µM M-ENK. The protein expression of ß-catenin was significantly induced by 40 and 80 µM M-ENK, while the protein expression of p-ß-catenin was significantly decreased by 20, 40, and 80 µM M-ENK. In addition, the mRNA expression of CAT, SOD, and GSH-PX was significantly increased by 40 and 80 µM M-ENK. The levels of H2O2, ·OH, and O2·- were significantly decreased, but the activity of CAT, SOD, and GSH-PX was significantly increased by 40 and 80 µM M-ENK. The fluorescence intensity of reactive oxygen species (ROS) was decreased, and that of mitochondrial membrane potential (MMP) was increased with the increase of M-ENK concentration in ZF4 cells. The results showed that M-ENK could induce Wnt/ß-catenin signaling, which further inhibited ROS production through the induction of C/EBPα, MMP, and the activities of antioxidant enzymes.

Clinical relevance of different handgrip strength indexes and cardiovascular disease risk factors: A cross-sectional study in suburb-dwelling elderly Chinese.

BACKGROUND: Reduced muscle strength, as measured by handgrip strength (HS), has been associated with an increased risk of cardiovascular disease (CVD). The aim of this study was to examine the association between different HS indexes and CVD risk factors in elderly Chinese individuals. We also determine optimal cutoffs of HS indexes for predicting CVD risk factors. METHODS: Data were obtained from 603 men and 789 women aged ≥60 years (average age 66.8 ± 6.4 y). These study participants were recruited in the suburb area of Tianjin, China. An individual was considered a patient when they exhibited any one of three CVD risk factors: diabetes mellitus, hypertension and dyslipidemia. All participants were interviewed face-to-face. In addition, serum samples were collected from all participants, and all participants underwent measures of anthropometry and HS. RESULTS: The optimal cutoffs were 0.376 of HS/weight in men and 0.726 of HS/body fat mass in women for predicting diabetes mellitus. The adjusted odds ratios (ORs) of at least one CVD risk factor for those with low muscle strength identified by HS/body fat mass were 2.14 (95% confidence interval [CI]: 1.53, 3.44; p < 0.001) in men and 2.32 (95% CI: 1.60, 3.29; p < 0.001) in women. CONCLUSION: HS/body fat mass appear to be the index best associated with CVD risk factors except diabetes mellitus in men. The optimal cutoffs of HS indexes have the potential to identify elderly adults at risk of CVD.

The inhibition of GSK-3ß promotes the production of reactive oxygen species via ß-catenin/C/EBPα signaling in the spleen of zebrafish (Danio rerio).

In this study, the mechanism that the inhibition of glycogen synthase kinase-3ß (GSK-3ß) promotes the production of reactive oxygen species (ROS) via ß-catenin/CCAAT/enhancer binding protein α (C/EBPα) signaling was investigated in the spleen of zebrafish (Danio rerio). The results demonstrated that the inhibition of GSK-3ß induced the mRNA expression of ß-catenin and C/EBPα by lithium (Li) treatments or GSK-3ß RNA interference. The levels of hydrogen peroxide (H2O2), superoxide anion (O2.-), and hydroxy radical (·OH) as well as the activity of superoxide dismutase (SOD) were increased, while the activities of catalase (CAT) and glutathione peroxidase (GSH-PX) were decreased in the spleen and ZF4 cells of zebrafish by Li+ treatments. In addition, GSK-3ß RNA interference increased ROS levels and decreased the activities of CAT and GSH-PX in the spleen. The fluorescence intensity of ROS was increased but the mitochondrial membrane potential (MMP) was decreased by Li+ treatments in ZF4 cells labeled with 2',7'-dichlorofluorescein diacetate (DCFH-DA) and Rhodamine-123, respectively. The results of present study indicated that the inhibition of GSK-3ß promoted the ROS production via ß-catenin/C/EBPα signaling in the spleen of zebrafish, and the balance between ROS and antioxidants could be destroyed by the GSK-3ß/ß-catenin/C/EBPα signaling. The results may be a valuable contribution to understanding the modulatory mechanism of GSK-3ß/ß-catenin/C/EBPα signaling on the antioxidant system in fish species.

Muscle strength rather than muscle mass is associated with osteoporosis in older Chinese adults.

BACKGROUND: Assessment of the association of muscle strength and muscle mass with osteoporosis (OP) is of special interest as muscles are a potential target for interventions (i.e., strength training). METHODS: A cross-sectional descriptive study encompassing people aged ≥ 60 years (average age: 66.9 ± 6.2 years; men, n = 516; women, n = 652) in the Hangu area of Tianjin, China. The study populations were invited to participate in a comprehensive geriatric assessment. OS was identified by measuring the calcaneal using a quantitative ultrasound and a T score of less than -2.5. Muscle characteristics included grip strength and appendicular skeletal muscle mass (ASM). RESULTS: The prevalence of OS in this study was 61.6% (male 52.1%, female 69.1%). Grip strength was negatively related to OS and after adjusting for all other variables, higher grip strength was found to be associated with a lower OS risk (p = 0.023). ASM/height2 was not associated with OS (p = 0.205). CONCLUSION: Based on our study, muscle strength rather than muscle mass is negatively associated with OS in older people; thus, we should pay more attention to muscle strength training in the early stage of the OS.

Clinical Evidence of Exercise Benefits for Stroke.

Even though stroke is the third, not the first, most common cause of disability-adjusted life years in developed countries, it is one of the most expensive to treat. Part of the expense is due to secondary problems in the post-stroke period including: cognition, memory, attention span, pain, sensation loss, psychological issues, and problems with mobility and balance. Research has identified that exercise has both positive physical and psychosocial effects for post-stroke patients. Therefore, this scientific statement provides an overview on exercise rehabilitation for post-stroke patients.We will use systematic literature reviews, clinical and epidemiology reports, published morbidity and mortality studies, clinical and public health guidelines, patient files, and authoritative statements to support this overview.Evidence clearly supports the use of various kinds of exercise training (e.g., aerobic, strength, flexibility, neuromuscular, and traditional Chinese exercise) for stroke survivors. Aerobic exercise, the main form of cardiac rehabilitation, may play an important role in improving aerobic fitness, cardiovascular fitness, cognitive abilities, walking speed and endurance, balance, quality of life, mobility, and other health outcomes among stroke patients. Strength exercise, included in national stroke guidelines and recommended for general health promotion for stroke survivors, can lead to improvements in functionality, psychosocial aspects, and quality of life for post-stroke patients. Flexibility exercises can relieve muscle spasticity problems, improve motor function, range of motion, and prevent contractures. Stretching exercises can also prevent joint contractures, muscle shortening, decrease spasticity, reduce joint stiffness and improve a post-stroke patient's overall function. Neuromuscular exercises can improve activities of daily living (ADL) through coordination and balance activities. Traditional Chinese exercises are used to improve walking and balance ability as well as increase muscle strength, which is important for post-stroke patients.The present evidence strongly supports the power of exercise for post-stroke patients, which in this study combined aerobic exercises, strength training, flexibility exercises, neuromuscular exercises, and traditional Chinese exercises. This research can encourage post-stroke survivors to consider the importance of exercise in the rehabilitation process.

Dietary supplementation of Bacillus subtilis and fructooligosaccharide enhance the growth, non-specific immunity of juvenile ovate pompano, Trachinotus ovatus and its disease resistance against Vibrio vulnificus.

A feeding trial was conducted to investigate the effects of dietary administration of probiotic Bacillus subtilis and prebiotic fructooligosaccharide (FOS) on growth performance, immune responses and disease resistance of juvenile ovate pompano, Trachinotus ovatus. One thousand six hundred and twenty individuals (initial body weight: 10.32 ± 0.46 g, mean ± S.E) were fed nine practical diets according to a 3 × 3 factorial design: the basal diet as the control diet supplemented with three levels of B. subtilis (0, 1.05 × 10(7) or 5.62 × 10(7) CFU g(-1) diet), crossed with 0, 0.2% or 0.4% FOS. After an 8-week feeding experimental period, six fish per cage were sampled for immunity determination. Then 18 fish of each cage left were challenged by Vibrio vulnificus. The results showed that fish fed with 5.62 × 10(7) CFU B. subtilis g(-1) in combination with 0.2% FOS produced the highest specific growth rate, and were significantly higher than the groups fed with 0 and 0.2% FOS without B. subtilis supplementation (P < 0.05). Feed efficiency ratio significantly increased with the increasing doses of dietary FOS without B. subtilis added (P < 0.05). The immune assay showed that fish fed with the control diet produced the lowest respiratory burst activity and was significantly different from the groups fed the diets containing 0.2% FOS at each B. subtilis level and containing 0.4% FOS single (P < 0.05). Phagocytic activity was significantly decreased with the increasing doses of dietary B. subtilis at 0.4% FOS level (P < 0.05). Alternative complement pathway activity of the fish fed with 0.2% FOS single was significantly lower than those fed with 5.62 × 10(7) CFU B. subtilis g(-1) diet supplemented at each FOS level (P < 0.05). Fish fed with the control diet had the lowest lysozyme activity, and were significantly different from those fed with 0.2 or 0.4% FOS at 1.05 and 5.62 × 10(7) CFU B. subtilis g(-1) diet level. Moreover, fish fed with diets supplemented with 0.2% and 0.4% FOS at each B. subtilis level had notably lower cumulative mortality after 10 days following V. vulnificus infection (P < 0.05). Under the experimental conditions, dietary B. subtilis and FOS had a significant interaction on enhancing the immune responses and disease resistance of juvenile ovate pompano (P < 0.05).

Spider minor ampullate silk protein nanoparticles: an effective protein delivery system capable of enhancing systemic immune responses.

Spider silk proteins (spidroins) are particularly attractive due to their excellent biocompatibility. Spider can produce up to seven different types of spidroins, each with unique properties and functions. Spider minor ampullate silk protein (MiSp) might be particularly interesting for biomedical applications, as the constituent silk is mechanically strong and does not super-contract in water, attributed to its amino acid composition. In this study, we evaluate the potential of recombinant nanoparticles derived from Araneus ventricosus MiSp as a protein delivery carrier. The MiSp-based nanoparticles were able to serve as an effective delivery system, achieving nearly 100% efficiency in loading the model protein lysozyme, and displayed a sustained release profile at physiological pH. These nanoparticles could significantly improve the delivery efficacy of the model proteins through different administration routes. Furthermore, nanoparticles loaded with model protein antigen lysozyme after subcutaneous or intramuscular administration could enhance antigen-specific immune responses in mouse models, through a mechanism involving antigen-depot effects at the injection site, long-term antigen persistence, and efficient uptake by dendritic cells as well as internalization by lymph nodes. These findings highlight the transnational potential of MiSp-based nanoparticle system for protein drug and vaccine delivery.

Effect of Dietary Manganese on the Growth Performance, Lipid Metabolism, and Antioxidant Capacity in the Post-Larval Coho Salmon (Oncorhynchus kisutch).

In this study, post-larval coho salmon Oncorhynchus kisutch (initial weight 0.37 ± 0.03 g) were fed with 6 experimental diets with increasing manganese (Mn) content (2.4, 8.5, 14.8, 19.8, 24.6, and 33.7 mg kg-1) for 12 weeks. Our results indicated that the feed conversion rate (FCR), specific growth rate (SGR), condition factor (CF), crude protein, moisture, crude lipid, ash, whole-body Mn, and vertebral Mn were affected by the elevation of Mn content in the diet. The activities of hepatic GSH-PX, Mn-SOD, and CAT were enhanced with increasing Mn content in the diet and reached the highest value at 19.8 mg kg-1 Mn. However, the level of hydrogen peroxide (H2O2), superoxide anion (O2·-), and malondialdehyde (MDA) was reduced with increasing Mn content in the diet. In addition, the activity of hepatic lipase (HL) and lipoprotein lipase (LPL) was increased with the elevation of dietary Mn content and reached a peak value at 14.8 mg kg-1 Mn. The activity of fatty acid synthetase (FAS) and the content of nonesterified fatty acid (NEFA) were increased following the elevation of Mn content from 2.4 to 19.8 mg kg-1 in the diet. The results indicated that the appropriate dietary Mn supplementation improved the feeding efficiency, lipid metabolism, and antioxidant capacity of coho salmon. The dietary Mn requirement for post-larval coho salmon was 17.35 mg kg-1 and 19.75 mg kg-1 based on the SGR and FCR, respectively. An optimal dietary level of Mn enhances hepatic lipid metabolism, and the signaling pathway of PI3K/AKT/mTOR may be involved in regulating the activity of enzymes related to lipid metabolism.

Sex and Race Differences in Obesity-Related Genetic Susceptibility and Risk of Cardiometabolic Disease in Older US Adults.

Importance: The fat mass and obesity-associated gene (FTO) is associated with obesity phenotypes, but the association is inconsistent across populations. Within-population differences may explain some of the variability observed. Objective: To investigate sex differences in the association between FTO single-nucleotide variants (SNVs) and obesity traits among self-identified non-Hispanic Black and non-Hispanic White US adults, to examine whether the SNVs were associated with cardiometabolic diseases, and to evaluate whether obesity mediated the association between FTO SNVs and cardiometabolic diseases. Design, Setting, and Participants: This cross-sectional study used data from the Reasons for Geographic and Racial Differences in Stroke (REGARDS) study, a US population-based cohort study with available genetic data (assayed in 2018) and phenotypic data at baseline (enrolled 2003-2007). Participants were aged 45 to 98 years at baseline. Data were analyzed from October 2021 to October 2022. Exposures: Eleven SNVs in the FTO gene present among both Black and White participants. Main Outcomes and Measures: Objectively measured obesity indicators (body mass index and waist-to-height ratio), objectively measured and/or self-reported cardiometabolic diseases (hypertension, stroke history, heart disease, and diabetes), and self-reported social-economic and psychosocial status. Results: A total of 10 447 participants (mean [SD] age, 64.4 [9.7] years; 5276 [55.8%] women; 8743 [83.7%] Black and 1704 [16.3%] White) were included. In the White group, 11 FTO SNVs were significantly associated with obesity, hypertension, and diabetes using linear models (eg, body mass index: ß = 0.536; 95% CI, 0.197-0.875), but none of the FTO SNVs were associated with obesity traits in the Black group. White males had a higher risk of obesity while White females had a higher risk of hypertension and diabetes. However, 1 FTO SNV (rs1121980) was associated with a direct increase in the risk of heart disease in Black participants not mediated by obesity (c' = 0.145 [SE, 0.0517]; P = .01). Conclusions and Relevance: In this cross-sectional study of obesity phenotypes and their association with cardiometabolic diseases, the tested FTO SNVs reflected sex differences in White participants. Different patterns of associations were observed among self-identified Black participants. Therefore, these results could inform future work discovering risk alleles or risk scores unique to Black individuals or further investigating genetic risk in all US residents.

M1 linear ubiquitination of LKB1 inhibits vascular endothelial cell injury in atherosclerosis through activation of AMPK.

Endothelial cell injury is confirmed to be the initial step in the atherosclerosis (AS) process. Here, we tried to elucidate the role of liver kinase B1 (LKB1) and adenosine phosphate protein kinase (AMPK) in modulating vascular endothelial cells (VECs) in AS. High-fat feed (HFD)-induced AS rat models were prepared and treated with AMPK activator A-769662 alone or combined with chloroquine. An analysis of VEC injury, inflammation response, and autophagy followed it. The M1 linear ubiquitination of LKB1 was assessed by co-immunoprecipitation. The interaction between LKB1 and AMPK was analyzed. Primary aortic VECs were isolated and induced by LPS to verify the effects of LKB1 and AMPK on VEC injury in AS. Activation of AMPK reduced the VEC injury and inflammatory response of VECs and promoted autophagy caused by AS. LKB1 could regulate the activation of AMPK in AS. M1 linear ubiquitination enhanced LKB1 activity and increased AMPK activation to protect against VEC injury in AS, which was validated by in vitro experiments. Our current study highlighted that M1 linear ubiquitination of LKB1 may induce the activation of LKB1 to activate AMPK, which inhibited VEC injury in AS.

Effects of fibrous microplastics on the accumulation of tris(2,3-dibromopropyl) isocyanurate and behavior of zebrafish via water- and foodborne exposure routes.

Fibrous microplastics are abundant in water, and the additives on fibers could also be transported jointly, which is a combined pollution scenario prevalent in the environment. Organisms ingest microplastics directly from the environment or indirectly through trophic transfer. However, there is a dearth of available information on the uptake and effects of fibers and their additives. This study investigated the uptake and depuration of polyester microplastic fibers (MFs, 3600 items/L) by adult female zebrafish via waterborne and foodborne exposure routes and the effects on the fish behavior. Moreover, we used brominated flame-retardant tris(2,3-dibromopropyl) isocyanurate (TBC, 5 µg/L) as a representative plastic additive compound and explored MFs' effects on the accumulation of TBC in zebrafish. Results substantiated that the highest MF concentrations in zebrafish from waterborne exposure (12.00 ± 4.59 items/tissue) were approximately three times higher than foodborne exposure, suggesting waterborne exposure as the primary ingestion route. In addition, environmentally relevant MF concentrations did not affect TBC bioaccumulation via aqueous exposure. However, MFs could decrease TBC accumulation via foodborne exposure by ingesting contaminated D. magna, which was probably because MF co-exposure decreased the TBC burden in daphnids. MF exposure also considerably increased behavioral hyperactivity in zebrafish. Moved speed, travelled distance, and active swimming duration all increased when exposed to MFs-containing groups. This phenomenon remained apparent in the foodborne exposure experiment with a low MF concentration (0.67-6.33 items/tissue) in zebrafish. This study offers a deeper understanding of MF uptake and excretion in zebrafish and the accumulation of the co-existing pollutant. We also confirmed that waterborne and foodborne exposure may lead to abnormal fish behavior even at low in vivo MF burdens.

Dietary L-Lysine Requirement of Coho Salmon (Oncorhynchus kisutch) Alevins.

The suitable dietary L-lysine concentration for coho salmon (Oncorhynchus kisutch) alevins was assessed by a dose response feeding trial. Six experimental diets were made with graded L-lysine concentrations of 2.29%, 2.81%, 3.32%, 3.80%, 4.27%, and 4.78% of the dry matter, respectively, each of which was fed to triplicate groups of 100 alevins (initial body weight: 0.30 ± 0.01 g) in 18 plastic baskets (water volume 240 L). The alevins were cultured in a flowing freshwater system and fed manually to apparent satiation four times a day for 12 weeks. The survival rate of alevins did not differ significantly among the dietary groups. The specific growth rate (SGR), protein efficiency ratio (PER), and body protein deposition (BPD) increased significantly (p < 0.05) with the increase in dietary lysine concentration up to 3.80% and then reduced as lysine level further increased. The feed conversion ratio (FCR) had an inverse trend to SGR. The whole-body crude protein content of the alevins increased significantly with increasing dietary lysine level, while crude lipid content showed the opposite trend. In comparison, the contents of morphological indices, whole-body moisture, and ash were not affected significantly (p > 0.05) by the different dietary lysine concentrations. The highest contents of lysine, arginine, and total essential amino acids (EAAs) were observed in the group with 4.27% dietary lysine concentration, which did not differ significantly from those in the 3.32%, 3.80%, and 4.78% groups but was significantly higher than those in the 2.29% and 2.81% groups. Similarly, valine had the highest content in the group with 4.78%. The variations in dietary lysine had no significant impacts on other EAA and non-EAA contents except glycine, which increased with increasing dietary lysine level. Second-order polynomial model analyses based on SGR, PER, BPD, and FCR evaluated the optimum L-lysine requirements of coho salmon alevins as 3.74%, 3.73%, 3.91%, and 3.77% of the diet or 6.80%, 6.78%, 7.11%, and 6.85% of dietary proteins, respectively.

Partial Replacement of Fishmeal with Poultry By-Product Meal in Diets for Coho Salmon (Oncorhynchus kisutch) Post-Smolts.

The present study evaluated the effects of partially substituting fish meal (FM) with poultry by-product meal (PBPM) on the growth, muscle composition, and tissue biochemical parameters of coho salmon (Oncorhynchus kisutch) post-smolts. Five isonitrogenous (7.45% nitrogen) and isoenergetic (18.61 MJ/kg gross energy) experimental diets were made by substituting 0%, 10%, 20%, 40%, and 60% FM protein with PBPM protein, which were designated accordingly as PBPM0 (the control), PBPM10, PBPM20, PBPM40, and PBPM60, respectively. Each diet was fed to triplicates of ten post-smolts (initial individual body weight, 180.13 ± 1.32 g) in three floating cages three times daily (6:50, 11:50, and 16:50) to apparent satiation for 84 days. Both specific growth rate (SGR) and protein efficiency ratio did not differ significantly (p > 0.05) among the control, PBPM10, and PBPM20 groups, which were remarkably (p < 0.05) higher than those of the PBPM40 and PBPM60 groups. Feed conversion ratio varied inversely with SGR. The PBPM replacement had no remarkable effects on the morphological indices and proximal muscle components. The control and PBPM10 groups led to significantly higher muscle contents of leucine, lysine, and methionine than groups of higher PBPM inclusion. The groups of PBPM40 and PBPM60 obtained significantly (p < 0.05) higher serum alanine aminotransferase and aspartate aminotransferase activities than the control and low PBPM inclusion groups. The control group had significantly higher albumin and total cholesterol contents than the groups with PBPM inclusion. The control group had significantly higher triglycerides content than the PBPM60 group. The PBPM60 group had significantly lower contents of high-density lipoprotein, low-density lipoprotein, and total protein than the control and PBPM10 groups. The high PBPM replacement level up to 40% and 60% had adverse effects on hepatic malondialdehyde levels. The catalase and superoxide dismutase activities were not affected by low PBPM inclusion, but significantly decreased in high-PBPM-inclusion groups. Based on broken-line regression analysis of SGR and PER, the optimum dietary PBPM replacing level was evaluated to be 16.63-17.50% of FM protein for coho salmon post-smolts.