Dissecting Causal Relationships Between Plasma Metabolites and Osteoporosis:A Bidirectional Mendelian Randomization Study

Objective To investigate the causal relationships between plasma metabolites and osteoporosis via Mendelian randomization(MR) analysis.Methods Bidirectional MR was used to analyze pooled data from different genome-wide association studies(GWAS). The causal effect of plasma metabolites on osteoporosis was estimated using the inverse variance weighted method, intersections of statistically significant metabolites obtained from different sources of osteoporosis-related GWAS aggregated data was determined, and then sensitivity analysis was performed on these metabolites. Heterogeneity between single nucleotide polymorphisms was evaluated by Cochran's Q test. Horizontal pleiotropy was assessed through the application of the MR-Egger intercept method and the MRPRESSO method. The causal effect of osteoporosis on plasma metabolites was also evaluated using the inverse variance weighted method. Additionally, pathway analysis was conducted to identify potential metabolic pathways involved in the regulation of osteoporosis.Results Primary analysis and sensitivity analysis showed that 77 and 61 plasma metabolites had a causal relationship with osteoporosis from the GWAS data in the GCST90038656 and GCST90044600 datasets, respectively. Five common metabolites were identified via intersection. X-13684 levels and the glucose-to-maltose ratio were negatively associated with osteoporosis, whereas glycoursodeoxycholate levels and arachidoylcarnitine(C20) levels were positively associated with osteoporosis(all P < 0.05). The relationship between X-11299 levels and osteoporosis showed contradictory results(all P < 0.05). Pathway analysis indicated that glycine, serine, and threonine metabolism, valine, leucine, and isoleucine biosynthesis, galactose metabolism, arginine biosynthesis, and starch and sucrose metabolism pathways were participated in the development of osteoporosis.Conclusion We found a causal relationship between plasma metabolites and osteoporosis. These results offer novel perspectives with important implications for targeted metabolite-focused interventions in the management of osteoporosis.

Effects of dietary protein/energy ratio on growth performance,carcass trait,meat quality,and plasma metabolites in pigs of different genotypes

Background： The protein/energy ratio is important for the production performance and utilization of available feed resources by animals. Increased protein consumption by mammals leads to elevated feed costs and increased nitrogen release into the environment. This study aimed to evaluate the effects of dietary protein/energy ratio on the growth performance, carcass traits, meat quality, and plasma metabolites of pigs of different genotypes. Methods： Bama mini-pigs and Landrace pigs were randomly assigned to two dietary treatment groups （Chinese conventional diet with low protein/energy ratio or National Research Council diet with high protein/energy ratio; n = 24 per treatment） in a 2 x 2 factorial arrangement. Blood and muscle samples were collected at the end of the nursery, growing, and finishing phases. Results： We observed significant interactions （P 〈 0.05） between breed and diet for total fat percentage, intramuscular fat （IMF） content, protein content in biceps femoris （BF） muscle, and plasma urea nitrogen （UN） concentration in the nursery phase; for average daily gain （ADG）, average daily feed intake （ADFI）, dry matter, IMF content in psoas major （PM） muscle, and plasma total protein and albumin concentrations in the growing phase; and for drip loss and plasma UN concentration in the finishing phase. Breed influenced （P 〈 0.05） growth performance, carcass traits, and meat quality, but not plasma metabolites. Throughout the trial, Landrace pigs showed significantly higher （P 〈 0.0_5） ADG, ADFI, dressing percentage, lean mass rate, and loin-eye area than did Bama mini-pigs, but significantly lower （P 〈 0.0.5） feed/gain ratio, fat percentage, backfat thickness, and IMF content. Dietary protein/energy ratio influenced the pH value, chemical composition of BF and PM muscles, and plasma activities of glutamic-pyruvic transaminase and gamma-glutamyl transpeptidase, and plasma concentration of UN. Conclusions： Compared with Landrace pigs, Bama mini-pigs showed slower growth and lower carcass performance, but had better meat quality. Moreover, unlike Landrace pigs, the dietary protein/energy ratio did not affect the growth performance of Bama mini-pigs. These results suggest that, in swine production, low dietary protein/energy ratio may be useful for reducing feed costs and minimizing the adverse effects of ammonia release into the environment.

Changes in feed intake, nutrient digestion, plasma metabolites, and oxidative stress parameters in dairy cows with subacute ruminal acidosis and its regulation with pelleted beet pulp

The objectives of this study were to 1） determine the variation of nutrient digestion, plasma metabolites and oxidative stress parameters triggered by induced subacute ruminal acidosis （SARA）; and 2） evaluate the ability of pelleted beet pulp （BP） as a replacement for ground corn to alleviate SARA. Eight Holstein-Friesian cows were fed four diets during four successive17 day periods： 1） total mixed ration （TMR） containing 0% finely ground wheat （FGW） （WO）; 2） TMR containing 10% FGW （W10）; 3） TMR containing 20% FGW （W20）; and 4） TMR containing 10% BP as a replacement for 10% ground corn （BP10）. The SARA induction protocol reduced the mean ruminal pH from 6.37 to 5.94, and the minimum ruminal pH decreased from 5.99 to 5.41 from baseline to challenge period. Mean ruminal pH increased from 5.94 to 6.05, and minimum daily ruminal pH increased from 5.41 to 5.63, when BP was substituted for corn. The apparent digestibility of nutrients was not affected by the dietary treatments, except that the digestibility of neutral detergent fibre （NDF） and acid detergent fibre （ADF） was reduced in cows fed the W20 diet compared with cows fed the W0 and W10 diets, and cows fed the BP10 diet had higher NDF and ADF digestibility than the cows fed the W20 diet. Cows fed the W20 diet had a lower plasma concentration of 13-hydroxybutyrate （BHBA）, non-esterified fatty acids （NEFA）, cholesterol, triglyceride, and total antioxidative capacity （TAC）, and a higher plasma concentration of glucose, insulin, malonaldehyde （MDA）, super oxygen dehydrogenises （SOD）, and glutathione peroxidase （GSH-Px） than cows fed the W0 diet. Substitution of BP for corn increased concentrations of plasma BHBA and TAC, but decreased concentrations of plasma MDA. Our results indicate that reduction of fibre digestion; the concomitant increase of plasma glucose and insulin; the decrease of plasma BHBA, NEFA, cholesterol, and triglyceride; and changes of plasma oxidative stress parameters are highly related to SARA induced by W20 diets. These variables may be alternative candidates for SARA diagnosis. We also suggest that the substitution of BP for corn could reduce the risk of SARA, increase fibre digestion, and improve the antioxidant status in dairy cows.

Effects of dietary forage to concentrate ratio and wildrye length on nutrient intake, digestibility, plasma metabolites, ruminal fermentation and fecal microflora of male Chinese Holstein calves

Twenty-eight male, weaned Chinese Holstein calves（（156.8±33.4） kg） were used to investigate the effects of dietary forage to concentrate ratio（F：C） and forage length on nutrient digestibility, plasma metabolites, ruminal fermentation, and fecal microflora. Animals were randomly allocated to four treatments in a 2×2 factorial arrangement： whole-length forage（WL） with low F：C（50：50）; WL with high F：C（65：35）; short-length forage（SL） with high F：C（65：35）; and SL with low F：C（50：50）. Chinese wildrye was used as the only forage source in this trial. The grass in the SL treatments was chopped using a chaff cutter to achieve small particle size（-50% particles 〉19 mm）. Dry matter intake（DMI） and organic matter（OM） intake was increased by increasing both F：C（P〈0.01） and forage length（FL）（P〈0.05）, while acid detergent fiber（ADF） and neutral detergent fiber（NDF） intakes were only increased by increasing the F：C（P〈0.01）. The digestibility of NDF was increased as the FL increased（P〈0.01）, and it was also affected by interaction between F：C and FL（P〈0.05）. Cholesterol（CHO）（P〈0.01）, leptin（LP）（P〈0.05）, and growth hormone（GH）（P〈0.01） concentrations in plasma were increased as dietary F：C increased. A significant increase in plasma triglyceride（TG）（P〈0.01）, insulin（INS）（P〈0.05）, and GH（P〈0.01） levels was observed with decreasing dietary FL. Ruminal p H values of calves fed with low F：C diets were significantly lower than those in high F：C treatment（P〈0.05）. Increasing the F：C enhanced ruminal acetic acid（P〈0.05） and acetic acid/propionic acid（P〈0.01）. Fecal Lactobacillus content was significantly higher, while Escherichia coli and Salmonella contents were significantly lower in WL and high F：C groups（P〈0.05）. Lower fecal scores（higher diarrhea rate） were observed in calves fed with SL hay compared to WL hay（P〈0.05）. Denatured gradient gel electrophoresis（DGGE） bands and richness index（S） were significantly affected by the interaction between F：C and FL（P〈0.05）, under high F：C, band numbers and richness index from WL group were higher than that from SL group（P〈0.05）, whereas there were no differences between WL andSL groups under low F：C（P〉0.05）. Microflora similarity was 50–73% among the different treatments. It is concluded that the WL with high F：C（65：35） diet is suitable for weaned calves.

Production performance and plasma metabolite concentrations of broiler chickens fed low crude protein diets differing in Thr and Gly

The aim of the study was to test the interaction between Thr and Gly in low crude protein(CP)diets in 7 to 28 d broilers on production performance and plasma metabolites.A total of 2,040 broilers were allocated to 17 treatments.A positive control(PC)diet(20.5%CP)was formulated to be adequate in dietary Thr and Gly.A negative control(NC)diet(18.5%CP,deficient in Thr and Gly)was supplemented with crystalline L-Thr and Gly to obtain a 4 Thr×4 Gly design.Dietary Thr was tested at an apparent faecal digestibility(AFD)Thr-to-Lys ratio,which was 55%,58%,61%or 64%,and dietary Gly was tested at an AFD(Gly+Ser)-to-Lys ratio,which was 135%,142%,149%or 156%.Plasma samples were collected at 28 d.The low CP diet,formulated at 64%Thr and 156%Gly,resulted in a higher body weight gain(BWG)(P<0.01)and similar feed conversion ratio(FCR)as the high CP treatment(PC).FCR was improved(P<0.001)by L-Thr supplementation.Quadratic response to dietary Thr was significant for feed intake(FI),BWG and FCR(P<0.01).A near-significant interaction for Thr×Gly was observed for FI and BWG(Plinear=0.091 and P=0.074,respectively).Gly did not affect production performance.An interaction between Thr×Gly on plasma free AA level was observed(P<0.05).Free AA concentration in plasma linearly decreased with increase in AFD Thr-to-Lys ratio,and increased with increase in AFD(Gly+Ser)-to-Lys ratio.Plasma uric acid concentration was higher in PC than in all of the other diets,and plasma triglyceride concentration was decreased by L-Thr supplementation,but not by Gly.In conclusion,Gly was not limiting for growth at low dietary CP level unless Thr was deficient,showing that adequate amounts of Thr in broiler diets can overcome marginal supply of Gly and Ser and allow reduction of dietary CP from 20.5%to 18.5%for broilers from 7 to 28 d of age.

Physiological but not morphological adjustments along latitudinal gradients in a human commensal species,the Eurasian tree sparrow

Human commensal species take advantage of anthropogenic conditions that are less likely to be challenged by the selective pressures of natural environments.Their morphological and physiological phenotypes can therefore dissociate from habitat characteristics.Understanding how these species adjust their morphological and physiological traits across latitudinal gradients is fundamental to uncovering the eco-physiological strategies underlying coping mechanisms.Here,we studied morphological traits in breeding Eurasian tree sparrows(ETSs;Passer montanus)among low-latitude(Yunnan and Hunan)and middle-latitude(Hebei)localities in China.We then compared body mass;lengths of bill,tarsometatarsus,wing,total body,and tail feather;and baseline and capture stress-induced levels of plasma corticosterone(CORT)and the metabolites including glucose(Glu),total triglyceride(TG),free fatty acid(FFA),total protein,and uric acid(UA).None of the measured morphological parameters varied with latitude except in the Hunan population,which demonstrated longer bills than those in other populations.Stress-induced CORT levels significantly exceeded baseline levels and decreased with increasing latitude,but total integrated CORT levels did not vary with latitude.Capture stress-induced significantly increased Glu levels and decreased TG levels,independent of site.However,the Hunan population had significantly higher baseline CORT,baseline and stress-induced FFA levels,but lower UA levels,which differed from other populations.Our results suggest that rather than morphological adjustments,physiological adjustments are mainly involved in coping mechanisms for middle-latitude adaptation in ETSs.It is worth investigating whether other avian species also exhibit such dissociation from external morphological designs while depending on physiological adjustments.