Protein Utilization Research Articles

Comprehensive utilization of Aspergillus niger waste mycelium based on physically induced autolysis: Preparation of chitosan and alternative nitrogen source

In industrial fermentation of citric acid, vast quantities of mycelium from Aspergillus niger (MAN) are produced, with an annual output of 20 million tons in China alone. The disposal of waste MAN poses a significant challenge and signifies a missed opportunity for resource recovery. In this study, physically induced autolysis (PIA) was employed to utilize the MAN proteins and chitosan. The results indicated abundant soluble proteins and sugars were released into the supernatant, while the cell fragments can be used to prepare chitosan. Under optimized autolytic conditions, the rate of total protein utilization of MAN was 78.7 %, and the rate of reserved chitin in the cell fragment was 93.8 %, outperforming other techniques. These findings were consistently reproducible in larger-scale experiments. Chitosan was prepared from cell fragments that eliminated the need for acid treatment and reduced alkali utilization by 50–67 %, thanks to PIA's effective removal of proteins from cell fragments. The rate of total sugar utilization, including both the content of the soluble sugars from autolysate and prepared chitosan, was 75.42 % of the total sugar of MAN. Additionally, the autolysate was proved to be an alternative nitrogen source in the fermentation media for microbial production of 2, 3-butanediol and citric acid, yielding concentrations comparable or superior to traditional sources. Thus, our findings not only achieved high component utilization rates but also facilitated environmentally friendly chitosan preparation, indicating the potential for innovative approaches to convert waste mycelium into valuable resources and advance the industry towards more efficient practices.

TLR2/TLR5 Signaling and Gut Microbiota Mediate Soybean-Meal-Induced Enteritis and Declined Growth and Antioxidant Capabilities in Large Yellow Croaker (Larimichthys crocea)

Soybean meal, renowned for its high yield, cost efficiency, and protein richness, serves as a pivotal plant-based alternative to fish meal. However, high soybean meal inclusion in Larimichthys crocea diets is linked to enteritis and oxidative damage, with unknown mechanisms. Our study aims to elucidate the molecular basis of soybean-meal-induced enteritis and its impact on intestinal microbiota in L. crocea. To this end, four isonitrogenous and isolipidic diets with varying soybean meal levels (0% FM, 15% SBM15, 30% SBM30, and 45% SBM45) were administered to L. crocea for 8 weeks. The results indicated that the SBM30 and SBM45 treatments significantly hindered fish growth, digestive efficiency, and protein utilization. Furthermore, high soybean meal levels (SBM30 and SBM45) activated intestinal Toll-like receptors (TLR2A, TLR2B, TLR5, and TLR22), stimulating C-Rel and mTOR protein expression and elevating ERK phosphorylation. This led to increased pro-inflammatory cytokine production (IL-1β, IL-6, and TNF-α) and decreased anti-inflammatory cytokine expression (IL-4/13A, IL-4/13B, and TGF-β), suggesting a potential signaling pathway for soybean-meal-induced enteritis. Furthermore, enteritis induced by high soybean meal levels led to oxidative damage, evident from increased MDA levels and decreased antioxidant enzyme activities (SOD and CAT). The SBM30 and SBM45 treatments increased Firmicutes and Bacteroidetes abundance in fish gut microbiota, while Proteobacteria abundance decreased. This microbiota shift may enhance soybean meal nutrient utilization, yet high soybean meal concentrations still impair growth. A soybean-meal-rich diet promotes harmful bacteria like Rhodococcus and depletes probiotics like Ralstonia, increasing disease risks. L. crocea has limited tolerance for soybean meal, necessitating advanced processing to mitigate anti-nutritional factors. Ultimately, exploring alternative protein sources beyond soybean meal for fish meal replacement is optimal for L. crocea.

A review on protein utilization and its interactions with carbohydrate and lipid from a molecular perspective in aquaculture: An implication beyond growth.

The world's increasing need for protein faces challenges in aquaculture production. New applications and tools will need to be added at every stage of the manufacturing line to attain this expansion sustainably, safely, and effectively. Utilizing experimental methods to increase aquatic animal production has become more common as aquatic biotechnology has advanced. High-throughput omics technologies have been introduced to address these issues, including transcriptomic, metabolomic, proteomic, and genomes. But it also faces many difficulties, like other food manufacturing industries. One of the best and most durable approaches to address these issues is probably to understand nutritional requirements and modify diet based on need. Molecular approaches are a subset of multiomics technology. Previously, most of the published work was devoted to the biochemical aspects of protein-lipid interactions in biological systems. In this review, we explore this idea and highlight various works that fall under the umbrella of nutrigenomics, with a particular emphasis on protein utilization and its interactions with carbohydrates and lipids.

Deciphering the mechanism underlying poor aqueous solubility of extracted quinoa proteins

This study aimed to decipher the mechanisms underlying poor solubility of quinoa proteins by investigating the form of quinoa proteins dispersed in water and how protein-protein interactions influenced the kinetic stability of proteins in the dispersions. Specifically, the relative solubility and the forms of quinoa proteins in 1–5 w/w% protein dispersions were determined by separating proteins via centrifugation and/or ultrafiltration. The kinetic stability of quinoa proteins in the supernatants over a 3-week storage period was characterized by determining the changes of concentration, composition and physicochemical properties of quinoa proteins and predicting protein-protein interactions. The results showed that quinoa proteins existed mainly as differently-sized protein aggregates in the dispersions, leading to low relative solubility. The coagulation of protein aggregates in the supernatants caused severe precipitation during the first week of storage whereas they were disassociated simultaneously. With further storage, the remaining proteins in the supernatants reached kinetic stability, which was contributed by stronger electrostatic repulsion and lower surface hydrophobicity. Moreover, 11S globulin and 2S albumin were precipitated and solubilized together during storage, which was ascribed to intermolecular interactions driven by multiple sites between 11S globulin and/or 2S albumin. This study lays a foundation for extensive utilization of quinoa proteins.

Diagnostic utility of plasma translocator protein 18 kDa (TSPO) in sepsis: A case-control study.

Translocator protein 18 kDa (TSPO) is a mitochondrial membrane protein that is involved in inflammation, oxidative stress, and steroidogenesis. TSPO may be a marker of inflammatory responses in the brain and other organs, but there have been few studies of the potential clinical significance of measuring the circulating TSPO concentration, especially in patients with sepsis. In this study, we compared the circulating TSPO concentrations of patients with sepsis and healthy controls to investigate the utility of plasma TSPO for the diagnosis of sepsis. Patients with sepsis admitted to the intensive care unit of Hiroshima University Hospital between January 2020 and April 2024 were enrolled. Plasma samples were collected from patients within 24 hours of admission and also from healthy volunteers, and their plasma TSPO concentrations were compared. Receiver operating characteristic analysis was used to evaluate the usefulness of plasma TSPO concentration for the diagnosis of sepsis. We also investigated the relationships of TSPO concentration with the severity of sepsis, complications, and prognosis of the patients. Eighty subjects (52 patients and 28 controls) were included in this study. The plasma TSPO concentrations of the patients with sepsis were significantly lower than those of the healthy controls (0.094 vs 0.25 ng/mL, P < .001), and receiver operating characteristic analysis generated an area under the curve of 0.81 (95% confidence interval: 0.72-0.91). In patients with sepsis, the TSPO concentration was not associated with the severity of sepsis, complications, or prognosis. Plasma TSPO may be a useful biomarker for the diagnosis of sepsis.

Effect of Melia azedarach seed mediated nano-ZnO on growth performance, protein utilisation efficiency, haematology and nutritional status in pigs

The current study was conducted to investigate the effect of Melia azedarach seed-mediated ZnO nanoparticles on growth performance, protein utilisation efficiency, haematology and nutritional status in pigs. A total of 48 pigs were allocated to the following six treatments replicated 8 times: Negative Control (NC, No antibiotic), Treatment 2: Positive control (PC) given a conventional antibiotic (Oxytetracycline, 40 mg/kg feed); Treatment 3: Nano-ZnO 300 mg/L (N300ZnO), Treatment 4: Group given 150 mg/L Melia azedarach seed mediated nano-ZnO (N150MA), Treatment 5: Group given 300 mg/L Melia azedarach seed mediated nano-ZnO (N300MA), Treatment 6: Group given 450 mg/L Melia azedarach seed mediated nano-ZnO (N450MA). The experiment was conducted over 7 weeks. Melia azedarach seed-mediated ZnO nanoparticles had no significant effect on growth performance apart from average daily feed intake (ADFI) with treatment 3 having the highest value. It significantly affected protein consumption and growth efficiency but not protein efficiency ratio and specific growth rate. Melia azedarach seed-mediated ZnO nanoparticles had no significant impact on nutritional parameters, serum minerals apart from phosphorus which can negatively affect renal functioning.

UHPLC-QQQ-MS/MS method for the simultaneous quantification of 18 amino acids in various meats.

Amino acids are an essential source of human protein, and their content and composition are the main factors determining food protein utilization rate. Determining amino acids is essential in the component analysis of food. Therefore, a groundbreaking technique was developed utilizing ultra-high performance liquid chromatography interfaced with a triple quadrupole mass spectrometer (UHPLC-QQQ-MS/MS) for concurrently quantifying 18 amino acids across various types of meat. According to the test results, it can be known that the average content of glutamate (2.03 × 104 ± 3.94 × 103 μg/g in pig feet) was the highest in all meat samples, and the content of aspartate (0.0945 ± 0.0950 μg/g in pork) was the lowest, which was not detected in some samples such as beef and lean meat. Orthogonal partial least-squares discrimination analysis (OPLS-DA) showed: (1) 13 amino acids (arginine, valine, serine, alanine, lysine, glycine, asparagine, methionine, proline, threonine, glutamate, phenylalanine, and leucine, VIP > 1) were used as characteristic amino acids between pork and pig feet; (2) serine, threonine, alanine, histidine, asparagine, and arginine (VIP > 1) were used as signature amino acids in different components of pork (lean meat, fat, and pigskin); (3) asparagine, glutamate, histidine, tyrosine, and valine (VIP > 1) were considered as signature amino acids in different types of meats (pork, mutton, beef, and chicken). This study provides a new UHPLC-QQQ-MS/MS method for the determination of amino acid content in meat and also provides data support for the comprehensive evaluation of the nutritional value of foods containing amino acids.

High-level secretory expression and characterization of an acid protease in Komagataella phaffii and its application in soybean meal protein degradation

Acid proteases play a crucial role in the industrial enzyme market, but low yield limits their widespread application. In this study, we focused on enhancing the secretory expression level of an acid protease (AopepA) from Aspergillus oryzae in Komagataella phaffii through stepwise genetic modification strategies. These included the co-expression of endoplasmic reticulum secretion-associated factors, overexpression of eukaryotic translation initiation factors, knockout of the β-1,3-glucanosyltransferase gene, disruption of the hypoxic heme-dependent repressor gene, and co-expression of the hemoglobin gene. After these modifications, protease activity increased by 4.2-fold, reaching 536.6 U/mL in a shaking flask. The engineered strain produced protease activity of up to 17,392.0 U/mL with a protein concentration of 44.6 g/L in a 5 L fermenter, representing the highest secretory expression level of acid proteases in K. phaffii ever reported. The optimal conditions of AopepA were pH 3.0 and 50 °C. AopepA demonstrated broad hydrolysis activity towards various protein substrates. It efficiently degraded soybean meal proteins into low molecular weight (Mw < 1 kDa, accounting for 82 %) oligopeptides to enhance protein utilization. This study provides valuable insights into improving the secretory expression of acid proteases in K. phaffii and identifies a suitable acid protease for enhancing soybean meal protein utilization.

Prognostic utility of heart-type fatty acid binding protein in patients with cardiac amyloidosis

Abstract Background Heart-type fatty acid binding protein (H-FABP) is a biomarker that has been shown to provide long-term prognostic information in patients with heart failure (HF), yet its role in patients with cardiac amyloidosis (CA) is unknown. Purpose We aim to investigate the association of H-FABP levels with prognosis in patients with CA. Methods Consecutive patients diagnosed with cardiac amyloidosis from October 2015 to May 2022 at Heart Failure Center, of our hospital were enrolled. Patients were categorized into 2 categories based on H-FABP levels measured in the baseline blood sample: &amp;lt; or = 7 ng/ml (low H-FABP), &amp;gt;7 ng/ml (high H-FABP). Univariate and multivariate Cox models were used to identify predictors of survival. Kaplan-Meier analysis was performed to compare survival between patients with low or high H-FABP. Results Overall, ninety-five patients were included. H-FABP was elevated (&amp;gt;7 ng/ml) in 37 patients (39%). Patients with high H-FABP were more likely to present with higher levels of high-sensitivity troponin-T (hs-TNT)(0.17ng/mL vs. 0.11ng/mL, p =0.01) and higher levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP) (18098.76pg/mL vs. 6211.69pg/mL, p&amp;lt;0.001). Patients were followed for an average follow-up period of 440 days, in which 53 (56%) patients died. Univariate and multivariate analysis demonstrated that H-FABP levels (HR 1.48, 95% CI 1.03-2.13, per 1-SD greater) was independently associated with prognosis, even after adjusting for NT-proBNP, hs-TNT and renal function. Conclusions Elevation of H-FABP is associated with an increased risk of death in patients with cardiac amyloidosis. H-FABP can serve as a novel biomarker independent of other established predictors for prognosis in cardiac amyloidosis.Kaplan-Meier curves for overall survival

Effects of insect meals on fish digestibility, blood parameters, and economic performance: a meta-analysis

Abstract This review examines the use of insect meal (IM) as a substitute for fishmeal (FM) in fish feeds within the past decade. While global interest in this alternative is growing, research on its effects on fish digestibility, blood parameters, and economic performance has been limited. Meta-analysis on the apparent digestibility coefficient revealed a negative effect summary on dry matter (−0.92) and crude lipid (−0.51), although the difference between the two groups was relatively minor. However, a positive effect summary was reported in the apparent digestibility of crude protein (0.12), suggesting a potential advantage in the utilization of protein by fish fed with IM-containing diets. Whereas meta-analysis on fish blood parameters demonstrates varying effects summary, necessitating further research. Analysis of economic performance revealed an overall better economic performance in the IM diet with an effect summary of −0.08 for feed cost and 0.00 for the economic profit index. Overall use of IM in aquafeed shows promise in improving feed quality and fish performance, potentially becoming a sustainable alternative to traditional FM. Recommendations include exploring IM’s impact on fish blood parameters through more investigations. Shifting focus from traditional measures to markers directly linked with fish health and immune response can provide more precise insights. Moreover, exploring various components of IM, such as lipids and functional elements like chitin, through simplified breakdown methods, can significantly enhance our comprehension of their nutritional value. Analyzing how each part influences fish health could pave the way for a sustainable and efficient alternative to FM in aquaculture.

Inflammatory Signals Across the Spectrum: A Detailed Exploration of Acute Appendicitis Stages According to EAES 2015 Guidelines.

Background: In this retrospective study, we evaluate the diagnostic utility of C-reactive protein (CRP) and leucocyte count within the EAES 2015 guidelines for acute appendicitis (AA) in differentiating uncomplicated (UAA) from complicated AA (CAA). Methods: Conducted at a tertiary care center in Germany, the study included 285 patients over 18 years who were diagnosed with AA from January 2019 to December 2021. Patient data included demographics, inflammatory markers, and postoperative outcomes. Results: CRP levels (Md: 60.2 mg/dL vs. 10.5 mg/dL; p < 0.001) and leucocyte count (Md: 14.4 Gpt/L vs. 13.1 Gpt/L; p = 0.016) were higher in CAA. CRP had a medium diagnostic value for detecting CAA (AUC = 0.79), with a cutoff at 44.3 mg/L, making it more likely to develop CAA. Leucocyte count showed low predictive value for CAA (AUC = 0.59). CRP ≥ 44.3 mg/L was associated with a higher risk of postoperative complications (OR: 2.9; p = 0.002) and prolonged hospitalization (OR: 3.5; p < 0.001). Conclusions: CRP, within the context of the EAES classification, presents as a valuable diagnostic marker to distinguish CAA from UAA, with a higher risk of postoperative complications and hospitalization. Leucocyte count showed low diagnostic value for the identification of CAA.

New strategies to classify canine pleural effusions and the diagnostic value of acute phase proteins, amylase, and adenosine deaminase in pleural exudates.

In dogs, simplified Light's criteria can discriminate transudates from exudates. Other tests used in human medicine are pleural effusion cholesterol (CHOLPE) and butyrylcholinesterase [BChEPE], the pleural effusion/serum ratio of these analytes (CHOLratio and BChEratio), and the serum albumin minus pleural effusion albumin gradient (SEAG). We aimed to assess the diagnostic accuracies of different biomarkers in dogs with pleural effusion in differentiating exudates from transudates. Secondarily, we evaluated the potential diagnostic utility of pleural effusion acute phase proteins, amylase, and adenosine deaminase in discriminating causes of exudative effusions. Cross-sectional study including 68 client-owned dogs with pleural effusion. There were 48 exudates (10 septic, 16 neoplastic, 9 hemorrhagic, and 13 classified as other exudates) and 20 transudates. All the variables analyzed, except SEAG, were significantly different between exudates and transudates. Using the cut-off values adopted in human literature, accuracies for CHOLPE, CHOLratio, BChEPE, and BChEratio were between 82.35% and 85.29%; all values were significantly lower compared with the previously published simplified Light's criteria accuracy (i.e., 98%, p < .001 for all comparisons). We found the accuracy of the simplified Light's criteria to be similar to what has been previously reported (95.59%, p = .238). Paraoxonase-1 (PON-1PE) activity and the pleural effusion/serum paraoxonase-1 ratio (PON-1ratio) were significantly lower in exudative neoplastic effusions than in exudative hemorrhagic (p = .004 and p = .001) and septic (p = .004 and p < .001) effusions. Simplified Light's criteria were the best method for discriminating transudates from exudates, and a low PON-1PE activity and PON-1ratio in exudative effusions may suggest an underlying neoplasia.

Screening appropriate lactic acid bacteria as adjunct starter to enhance characteristic flavor and sensory profiles of Huangjiu (Chinese rice wine)

Lactic acid bacteria (LAB) are crucial for enhancing Huangjiu flavor, yet no standardized criteria exist for selecting LAB strains for this purpose. This study aimed to investigate the suitability of screening appropriate LAB strains as adjunct starters for Huangjiu fermentation. Twenty-five LAB strains were assessed for their stress resistance and fermentation characteristics. Co-fermentation with Saccharomyces cerevisiae was conducted to explore its role in flavor development, as this yeast greatly contribute to desirable flavor profiles. Notably, Lactiplantibacillus plantarum AR1018, AR1026 and Weissella confusa AR1038 significantly reduced 3-methyl-1-butanol production while increasing levels of ethyl hexanoate and ethyl octanoate. These strains were selected for further analysis of their impact on the physicochemical properties, volatile organic compounds, and sensory attributes of Huangjiu. The LAB starters enhanced total acidity, sugar and protein utilization and ester synthesis, while reducing ethanol and alcohol levels. AR1018 achieved a 16.51% reduction in 3-methyl-1-butanol, whereas AR1038 exhibited a 27.32% increase in ester content compared to the control. Principal component analysis revealed distinct flavor characteristic between Huangjiu produced with AR1018 and AR1038, and sensory evaluation confirmed enhanced desirable attributes, especially for AR1038. This study highlights the potential of LAB starters in improving the flavor quality of rice wine and similar fermented beverages.

Prone to loss: Senescence-regulated protein degradation leads to lower protein extractability in aging tomato leaves

The utilization of proteins extracted from tomato (Solanum lycopersicum) leaves as cost-effective resources for human consumption or animal feed has gained interest. Thus, increasing protein extractability from tomato leaves became a new breeding target. However, the genetic factors influencing this trait remains poorly understood. In this study, we analyzed changes in leaf protein content, protein composition, and extraction yield across developmental stages, which are vegetative growth, flowering, fruit-forming, and mature fruit. Moreover, tomato gene expression across developmental stages was also studied, to identify genes underlying variability in leaf protein extraction. Protein extraction yield decreased from 0.51 g/g to 0.01 g/g leaf protein from the vegetative to mature stage. However, total protein content inferred with Dumas combustion analysis did not change over the developmental stages tested, while the protein-to-peptide ratio decreased significantly. To further analyze potential causes underlying the decline of protein-to-peptide ratio, the enzymatic activity of proteases – i.e. the enzymes responsible for protein degradation – and the expression of genes encoding these enzymes was studied along plant development. The overall specific activity of proteases did not change significantly throughout plant development. On the contrary, the gene expression of distinct members of the aspartic, cysteine, and subtilase protease families increased. Overall, our findings suggest that extraplastidic protein degradation likely underlies the protein degradation observed during senescence. In the future, the reduction of the activity of extraplastidic proteases through biotechnology could represent an effective strategy to develop tomato varieties with improved protein extraction yields.

Rosa roxburghii trattresidue: A novel feed resource for cattle indicated by the non-deleterious performance and blood metabolites.

Rosa roxburghii tratt residue (RRTR) is a regional and uncommon byproduct in Guizhou. Little information is available on whether RRTR can be used as feed for ruminants. In this study, the feasibility of using RRTR as a new feed resource for cattle was investigated by chemical composition analysis, in vitro gas production (Trial 1) and animal feeding experiments (Trial 2). In trial 1, compared to the commonly used fruit residues, RRTR had a proximate nutrient level; the in vitro gas production curve and dynamics fell within the normal range. In trial 2, 16 cattle were allocated to the control and treatment groups, with 8 replicates of 1 cattle each. Cattle in the control group were fed a basal diet without RRTR, while those in the treatment group were fed a diet containing 30% RRTR to replace 30% whole corn silage in basal diet. Feeding RRTR had little effect on the growth performance of the control and treatment cattle (P > 0.05). The feed-to-gain ratio was greater in the treatment group than in the control group throughout the trial period (P < 0.05). The plasma urea levels in the treatment group were lower (P < 0.05) than that in control group, and the levels of other plasma biochemical metabolites were not different between the two groups of cattle (P > 0.05). The in vivo rumen fermentation parameters did not differ between the control and treatment groups (P > 0.05). Our findings indicate that RRTR has a nutritional profile (crude protein, neutral detergent fiber, acid detergent fiber, and crude fiber) similar to that of commonly used fruit residues (such as apple, pineapple, and citrus residue et al.); improves plasma protein utilization efficiency; and has no negative impact on growth performance, albeit with limited effects on feed conversion; blood metabolites, and rumen fermentation parameters in cattle. Accordingly, we conclude, based on the above-mentioned result, that RRTR can serve as a novel feed alternative resource when considering the affordability and as a practical choice for low-cost diets for cattle.

Diagnostic Utility of Canine C-Reactive Protein, Haptoglobin, and 25-Hydroxyvitamin-D in Dogs with Nasal Cavity Disease

In this prospective blinded study, canine C-reactive protein (c-CRP), haptoglobin (HPT), and 25-hydroxyvitamin-D (25(OH)D) were investigated for their diagnostic value in 55 dogs with nasal cavity disease (ND). After comprehensive diagnostics including a culture-dependent microbiological examination (ME) of nasal swabs, 17 dogs were excluded due to additionally detected systemic diseases or steroid pre-treatment. Included were 25 dogs with malignant ND (13 carcinomas and 12 sarcomas) and 30 dogs with benign ND (7 benign tumors, 13 idiopathic rhinitis (IR), and 10 others), as well as 10 controls. In none of the 72 dogs with ND was primary bacterial rhinitis diagnosed. Although within the reference interval, compared to the controls, c-CRP was significantly higher in dogs with ND in general and in every subgroup except for benign tumors. Serum HPT concentrations were not different among groups. Compared to the controls, 25(OH)D concentrations were significantly lower (p = 0.041) in malignant ND and sarcomas (p = 0.025). Despite pre-treatment with antibiotics (40/54; 74.1%), in 23/51 (45%) dogs, the ME was positive. Cultivated bacteria did not differ significantly between nasal diseases. The serum markers were not significantly different regarding the positivity of ME. In conclusion, the investigated serum markers were not clinically useful for the reliable detection of canine ND, as was the ME. Because of the low number of dogs with IR and positive or negative ME, further studies regarding c-CRP are needed in a larger group of IR dogs without concomitant diseases to reliably evaluate its utility in IR dogs with suspected secondary bacterial nasal infection.

Effects of different forms of amino acid supplementation on the performance and intestinal barrier function of laying hens fed a low-protein diet

The aim of this study was to investigate the effects of low-protein diets and the sustained release of synthetic amino acids (AA) on the performance, intestinal barrier function and nitrogen excretion of laying hens. Two hundred eighty-eight 39-week-old Hyline brown laying hens of were randomly divided into 3 groups with 8 replicates per group. The crude protein level in the control group (CON) was 16%, the crude protein levels in the crystal AA supplement group (LCP-CAA) and microencapsulated AA group (LCP-MAA) were both 13%, and the AA levels in the LCP-CAA and LCP-MAA groups were consistent with that in the CON group. The experiment lasted 12 wk, and production performance was assessed weekly. The FCR and ADFI values were significantly greater for the LCP-CAA group than for the CON and LCP-MAA groups (P < 0.05). Two hours after feeding, His levels were significantly greater in the LCP-CAA group than in the LCP-MAA group (P < 0.05); 4 h after feeding, the contents of Met, Thr, Leu and Val were significantly greater in blood from the LCP-MAA group (P < 0.05); 6 h after feeding, Trp, Ile and Arg levels were significantly greater in the LCP-MAA group (P < 0.05). The chylase content significantly decreased in the duodenum of the LCP-CAA group (P < 0.05), and the chylase and trypsin were contents increased in the ileum of the LCP-MAA group (P < 0.05). In the LCP-MAA group, significantly increased mRNA expression levels of Occludin, ZO-1 in duodenum; Occludin, ZO-1, y+LAT1 in jejunum; and ZO-1 in ileum were detected at 8 and 12 weeks (P < 0.05). The fecal nitrogen content significantly decreased in the low protein diet group (P < 0.01). In conclusion, reducing dietary crude protein levels and supplementing with microencapsulated AAs can improve intestinal barrier function, promote digestive enzyme secretion, increase the expression of AA transporters, improve dietary protein utilization efficiency, and reduce nitrogen emission in laying hens.

Potential utility of RSAD2 transcript and protein in early detection of pregnancy in buffaloes.

This study investigates a novel early pregnancy marker in water buffaloes, focusing on RSAD2 mRNA expression, known to be upregulated by interferon-tau (IFNT) during pregnancy. While RSAD2 is primarily recognized for its antiviral effect, we hypothesized its role as a conceptus-induced component in regulating pregnancy in buffaloes. Given its differential expression compared to other IFNT-induced genes in cows, RSAD2 may serve as a biomarker for early pregnancy detection in buffaloes. RNA, cDNA, and plasma samples were obtained from archived samples collected before insemination (d0) and at d20, d25 and d40 after insemination. Twelve RNA samples, having optimal optical density and concentration, from six pregnant and six non-pregnant buffaloes were selected. The cDNA was analyzed to measure the abundance of RSAD2 mRNA using real-time quantitative PCR (RT-qPCR) and plasma for protein expression analysis using Western blot. The RT-qPCR analysis showed a transcript of RSAD2 increased significantly by 7-fold and 6-fold on d20 and d25, compared to both d0 and d40 in the pregnant group only. At d20, the sensitivity of RSAD2 was 100% and the specificity was 83.3%, and at d25-d both the sensitivity and specificity was 100%, indicating low incidences of misdiagnosing early pregnancy in buffaloes. In the non-pregnant group, RSAD2 expression remained low and did not change after insemination. Western blot analysis revealed an immunoreactive RSAD2 protein band. Densitometry analysis of the RSAD2-specific protein band, based on gray mean value, showed significantly increased expression of RSAD2 at d25 compared to d0 in the pregnant group. In conclusion, these results indicated that RSAD2 expressions at both the mRNA and protein levels show promising potential for detecting pregnancy at d25 post-insemination.

Exploring novel antioxidant cyclic peptides in corn protein hydrolysate: Preparation, identification and molecular docking analysis

Antioxidant cyclic peptides were successfully identified from a corn protein hydrolysate. Hydrolysate by Alcalase + Flavourzyme showed the highest cyclic peptide purity (48.36 ± 1.81 %) and higher antioxidant activities compared with other hydrolysate. The success of peptide cyclization in hydrolysate was demonstrated by thermogravimetric analysis and thin-layer chromatography (TLC) analysis. Thermogravimetric analysis showed that the thermal stability of hydrolysate after cyclization was significantly increased, which was related to the formation of cyclic peptides. Peptides with molecular weight less than 1000 Da accounted for more than 80 % in hydrolysate after cyclization. After separation using gel silica chromatography and semi-preparative reverse phase high performance liquid chromatography (RP-HPLC), 22 novel antioxidant cyclic peptides were identified by ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS) and orbitrap-tandem mass spectrometry (Orbitrap-MS/MS). Synthetic cyclic peptides with the same sequence were synthesized and characterized for their antioxidant activity. Molecular docking suggested that the free radical molecules could bind with the cyclic backbone and side chain of cyclic peptides through hydrogen bonding, hydrophobic interaction as well as electrostatic interaction. This study has important implications for the high-value utilization of corn protein and new cyclic peptides drugs or functional food development.

The impact of achieving remission in inflammatory bowel disease on plasma matrix -carboxyglutamate protein levels

Matrix -carboxyglutamic acid (MGLA) protein is a vitamin K dependent peptide which contributes to the immunomodulatory activity of mesenchymal stromal cells. There is a possible association between MGLA protein and inflammatory bowel disease (IBD) which is divided into Crohn’s disease (CD) and ulcerative colitis (UC). However, little is known about the clinical utility of MGLA protein in IBD patients. This study aimed to assess the impact of achieving remission on the serum MGLA protein levels in IBD patients. This prospective observational study included 60 newly diagnosed IBD patients. All patients were subjected to full clinical, laboratory, radiological, and histopathological assessment of IBD at baseline and six months after initiating treatment. Serum MGLA protein level was assessed using an enzyme-linked immunosorbent assay. There were 29 (48.3%) UC cases and 31 (51.67%) CD cases. We observed a significant decrease in serum MGLA protein levels after 6 months of treatment compared to pretreatment values in UC patients (120.490 ± 26.273 vs. 26.320 ± 17.378 nmol/L, p&lt;0.001) and CD patients (125.576 ± 28.208 vs. 28.520 ± 18.443 nmol/L, p&lt;0.001). Serum MGLA protein levels were significantly higher in non-remittent patients compared to remittent UC patients before treatment (142.556 ± 17.096 vs. 110.560 ± 23.659 nmol/L, p&lt;0.001) and after six months of treatment (51.222 ± 4.410 vs. 15.114 ± 3.302 nmol/L, p&lt;0.001). Serum MGLA protein levels were significantly higher in non-remittent patients compared to remittent CD patients before treatment (150.727 ± 7.198 vs. 111.743 ± 25.718 nmol/L, p&lt;0.001) and after six months of treatment (52.182 ± 5.269 vs. 15.506 ± 4.475 nmol/L, p&lt;0.001). This response was irrespective of the therapeutic modality. In conclusion, achievement of remission in IBD patients resulted in a significant decrease in serum MGLA protein levels.