Protein Products Research Articles

Identification and characterization of terpene synthase genes accounting for volatile terpene emissions in the flower of Paeonia lactiflora

Emergence of floral scents marked a significant milestone in biological development of plant, letting out olfactory signals that enable them to induce pollinators. In this research, molecular and metabolomics methodologies were used to explore terpene synthase (TPS) genes, which are responsible for synthesis of floral fragrance in Paeonia lactiflora ‘Wu Hua Long Yu’. Analyses on transcriptome data identified 11 PlTPS genes, among them 7 genes were isolated, and the functions of 5 genes were confirmed. PlTPS1 was identified as a single-product enzyme; PlTPS4, 8 and 9 were identified as bifunctional enzymes, capable of recognizing farnesyl pyrophosphate and geranyl pyrophosphate; PlTPS6 generated multiple products by recognizing farnesyl pyrophosphate. Enzymatic products of these PlTPS proteins closely matched the volatile terpenes emitted by flowers, revealing the relation between PlTPS genes and the release of such volatile compounds. Functions of five PlTPS genes in herbaceous peony were further elucidated through virus-induced gene silencing experiments, as well as overexpression in tobacco. PlTPS1 catalyzes geraniol production, while PlTPS4 catalyzes linalool. PlTPS6 is responsible for producing two sesquiterpenes, caryophyllene and α-humulene. PlTPS8, with the most varied products, mainly catalyzes the production of α-pinene, linalool, and germacrene D. Based on these findings, graphical models summarizing the biosynthesis of floral volatile terpenes in herbaceous peony have been proposed. The characterization of PlTPS genes has elucidated their specific roles in the synthesis of herbaceous peony fragrance, opening up possibilities for introducing or enhancing floral scents in herbaceous peony and other plants.

Effect of fat replacement in high internal phase emulsions constructed by high temperature saccharification of grafted proteins on gel properties and flavor profiles of sausages

In order to mitigate the risk of cardiovascular diseases associated with excessive saturated fatty acid intake, utilizing high internal phase emulsions (HIPEs) as a substitute for animal fat in producing high-quality fat-substituted meat products is an ideal approach. This study involves the preparation of glycosylation products of egg white protein (EWP) through saccharification at high temperatures in the presence of fructooligosaccharides (FO). The resulting glycation products of EWP were employed to create colloidal particles, forming HIPEs, which were further utilized to induce the formation of HIPEs gels (HIPEs-Gs). The study investigated the effects of substituting different ratios (25%, 50%, 75%, and 100%) of animal fat with HIPEs and HIPEs-Gs on the gel properties and flavor characteristics of sausages. Results showed that, compared to the control group, substituting fat with HIPEs significantly improved the gel properties, cooking yield, and G' of sausages, while excessive HIPEs-Gs substitution yielded negative effects. Low-field nuclear magnetic resonance results also demonstrated that adding HIPEs improved water and oil distribution in the sausage batter, enhancing protein's binding capacity with water. Scanning electron microscope revealed that HIPEs substitution led to a denser gel network with smaller pores, effectively "locking in" more water. Analysis of volatile compounds indicated accelerated release of aromatic compounds, alkanes, sulfides, and lipids when fat was substituted with HIPEs and HIPEs-Gs. Electronic tongue analysis suggested that HIPEs-Gs substitution reduced response values for umami and saltiness. In conclusion, compared to HIPEs-Gs, using HIPEs as a fat substitute improves the quality of sausages.

Combined Treatment with Tauroursodeoxycholic Acid and SCD Probiotics Reduces Oxidative Stress in Lung Tissue of Aged Rats

Aging is associated with an increased level of oxidative stress, resulting from an elevated production of reactive oxygen species, which can lead to cellular and tissue damage, particularly in the lungs. This study examined the effects of Tauroursodeoxycholic acid (TUDCA) and SCD Probiotics, both individually and in combination, on oxidative stress markers in the lung tissue of aged Sprague-Dawley rats. The primary objective was to assess the potential of these agents in reducing malondialdehyde (MDA), advanced oxidation protein products (AOPP), and myeloperoxidase (MPO) levels, which are indicative of oxidative damage and inflammation. The results showed that TUDCA significantly decreased MDA and AOPP levels, suggesting its role in maintaining mitochondrial stability and inhibiting apoptotic pathways. SCD Probiotics also demonstrated a reduction in AOPP levels, highlighting their immunomodulatory and antioxidant effects. Furthermore, the combined treatment of TUDCA and SCD Probiotics led to a more pronounced decrease in both MDA and AOPP levels, along with a significant reduction in MPO activity. This suggests a synergistic interaction that enhances the antioxidative and anti-inflammatory properties of the individual treatments. These findings support the therapeutic potential of TUDCA and SCD Probiotics in mitigating oxidative damage in aging lung tissues, proposing that their concurrent use could be an effective strategy against age-related oxidative stress. Further research is warranted to explore these effects across different models and long-term applications.

MiR472 deficiency enhances Arabidopsis thaliana defense without reducing seed production.

After having co-existed in plant genomes for at least 200 million years, the products of microRNA (miRNA) and Nucleotide-Binding Leucine Rich Repeat protein (NLR) genes formed a regulatory relationship in the common ancestor of modern gymnosperms and angiosperms. From then on, DNA polymorphisms occurring at miRNA target sequences within NLR transcripts must have been compensated by mutations in the corresponding mature miRNA sequence. The potential evolutionary advantage of such regulation remains largely unknown and might be related to two non-exclusive scenarios: miRNA-dependent regulation of NLR levels might prevent defense mis-activation with negative effects on plant growth and reproduction; or reduction of active miRNA levels in response to pathogen derived molecules (PAMPS and silencing suppressors) might rapidly release otherwise silent NLR transcripts for rapid translation and thereby enhance defense. Here, we used Arabidopsis thaliana plants deficient for miR472 function to study the impact of releasing its NLR targets on plant growth and reproduction and on defense against the fungal pathogen Plectospharaella cucumerina. We show that miR472 regulation has a dual role, participating both in the tight regulation of plant defense and growth. MIM472 lines, with reduced active miR472, are more resistant to pathogens and, correlatively, have reduced relative growth compared to wild-type plants although the end of their reproductive phase is delayed, exhibiting higher adult biomass and similar seed yield as the wild-type. Our study highlights how negative consequences of defense activation might be compensated by changes in phenology and that miR472 reduction is an integral part of plant defense responses.

Dynamic responses of human lung innate and adaptive immune cells highlight the roles of genes at asthma risk loci.

The lung is a unique immunological niche with diverse immune cell types. The effects of stimulation through innate and adaptive immune receptors on human lung immune cells has largely been extrapolated from studies of blood immune cells. While multiple immune cell types and many genes have been implicated as contributing to asthma, the dynamics of these in human lung immune cells following activation will yield insights into asthma pathogenesis and lung immunity more broadly. Human lung immune cells from 6 donors were isolated. Mixed leukocytes were treated separately with lipopolysaccharide (LPS), F(ab)2-anti-human-IgM/IgG + IL4 and anti-CD3/CD28 for 4 and 18 hours and underwent single cell RNA sequencing (scRNAseq). Lung immune cell types were annotated, and gene expression compared across conditions. Genes at prior asthma-associated genetic loci were characterized across cell types, treatments and timepoints. Expression of non-classical class II genes associated with asthma, HLA-DQA2 and HLA-DQB2, and their protein products was characterized with immunohistochemistry. We characterized gene expression in 116,697 lung immune cells. Cell-, treatment-, and timepoint-specific effects on gene expression were detected in all lung immune cell populations. Correlation of gene expression between lung and blood lymphocyte populations decreased following stimulation. Among the genes that were differentially expressed, 97 receptor:ligand pairs had changes with treatments. 96.0% of genes at asthma risk loci demonstrated differential expression in at least one cell type and at least one treatment. B cells were the cell type with the highest expression of HLA-DQA2 and HLA-DQB2 which increased with anti-IgM/IgG treatment and the HLA-DQB2 protein was identified in lung B cells from a donor with asthma. Human lung immune activation elicits a broad range of cellular responses that deviate from those of blood immune cells and are relevant to asthma. Lung B cells expressing HLA-DQA2 and HLA-DQB2 appear to be involved in a novel antigen presentation pathway that contributes to asthma risk.

High moisture extrusion of pulse proteins: Texture, structure, and in vitro digestion characteristics of extrudates

Pulse proteins are promising components for plant-based food development, yet their diverse subunit compositions, influenced by source origin, result in variable extrusion properties. This study utilized high moisture extrusion (60% moisture content) to prepare extrudates from four pulse proteins: soy, pea, chickpea and mung bean protein isolates (SPI, PPI, CPI, and MPI, respectively). The texture, structure, and in vitro digestion characteristics of extrudates were examined. The results showed that SPI and PPI were mainly composed of 7S and 11S globulins, while CPI and MPI were mainly composed of 7S globulins, and 11S globulins, respectively. SPI extrudates exhibited significantly lower hardness, chewiness, and transverse and longitudinal fracture stresses, while CPI extrudates exhibited a higher anisotropy index. SPI extrudates with a higher β-sheet content (39.08%) exhibited denser structures. The gastrointestinal digestibility (86.65%) of CPI extrudates was higher than other groups. Correlation analysis showed that 7S/11S ratio was positively correlated with the melt viscosity, while negatively correlated with the anisotropy index and in vitro digestibility. This study highlights the distinct extrusion properties of pulse proteins based on their unique subunit composition. The results provide insights for selecting appropriate protein sources and developing plant-based protein products with tailored textural attributes and desirable digestibility.

Cardioprotective action of apocynin in isoproterenol‐induced cardiac damage is mediated through Nrf‐2/HO‐1 signaling pathway

AbstractThis investigation evaluated the therapeutic benefit of apocynin in isoproterenol (ISO)‐induced cardiac damage in rats. ISO‐administered male Wistar rats were treated with apocynin for 2 weeks. Blood plasma and left ventricle of heart tissues were collected and analyzed for oxidative stress‐related parameters such as malondialdehyde (MDA), advanced oxidation protein product (AOPP), and nitric oxide (NO). The activities of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and catalase were also measured. The gene expressions of oxidative stress‐related proteins such as Nrf‐2, HO‐1, and HO‐2 in cardiac tissues were also measured. In silico studies like molecular docking and molecular dynamics were also performed to detect how apocynin interacts with NADPH and nitric oxide synthase at the molecular level. This investigation revealed significant elevation of serum transferase enzymes and creatinine kinase‐Muscle Brain (CK‐MB) activities in ISO‐administered rats compared to the control. Apocynin effectively normalized the serum transferases and CK‐MB activities in the blood of ISO‐stressed rats. Moreover, ISO‐induced elevations of MDA, NO, and AOPP levels were also suppressed by apocynin treatment. Consistently, apocynin restored the reduced SOD and catalase activities in ISO‐administered rats. This restoration of enzyme activity might be due to the increased expression of Nrf‐2 and HO‐1 and reduced expression of iNOS and TNF‐α in ISO‐administered rats. Histological analysis revealed that apocynin treatment ameliorated the mononuclear cell adherence and fibrosis in the cardiac tissue of ISO‐administered rats. Computational studies also support the experimental findings. This study demonstrates that apocynin prevents ISO‐induced cardiac injury not only by preventing inflammation but also by empowering the antioxidant defense system.

Capsinoids Increase Antioxidative Enzyme Activity and Prevent Obesity-Induced Cardiac Injury without Positively Modulating Body Fat Accumulation and Cardiac Oxidative Biomarkers.

Capsinoids are potential antioxidant agents capable of reducing oxidative damage and the resulting complications triggered by obesity. Thus, this study aimed to investigate the effects of capsinoids on adiposity and biomarkers of cardiac oxidative stress in obese rats induced by a high-fat diet. Male Wistar rats were exposed to a high-fat diet for 27 consecutive weeks. After the characterization of obesity (week 19), some of the obese animals began to receive capsinoids (10 mg/kg/day) by orogastric gavage. Adiposity and comorbidities were assessed. In the heart, remodeling, injury, and biomarkers of oxidative stress were determined. The treatment did not reduce obesity-induced adiposity but was efficient in reducing cholesterol levels. Capsinoid treatment did not cause a difference in heart and LV mass, despite having reduced troponin I concentrations. Furthermore, capsinoids did not reduce the increase in the advanced oxidation of protein products and carbonylated proteins caused by obesity in cardiac tissue. In addition, obese rats treated with capsinoids presented high levels of malondialdehyde and greater antioxidant enzyme activity compared to untreated obese rats. In conclusion, treatment with capsinoids increases antioxidative enzyme activity and prevents obesity-induced cardiac injury without positively modulating body fat accumulation and cardiac oxidative biomarkers.

Full-cycle, multidisciplinary and systematic management of citrin deficiency

Professor Takeyori Saheki's team at Kagoshima University, Japan, published a paper in Nature Genetics in June 1999, pinpointing the pathogenic gene for adult-onset type Ⅱ citrullinemia as SLC25A13 and naming the protein product encoded by this gene as citrin. Over the past 25 years, the researches have made positive progress on the pathophysiological mechanism, clinical phenotype, molecular diagnosis, treatment, and prognosis of citrin deficiency (CD) as an autosomal recessive genetic disease. Currently, three age-dependent clinical phenotypes of CD have been found, namely neonatal intrahepatic cholestasis caused by citrin deficiency, failure to thrive and dyslipidemia caused by citrin deficiency, and adult-onset type Ⅱ citrullinemia. Although relevant internal medicine drugs are being researched and developed while liver transplantation has been used for the treatment of CD patients, scientific dietary therapy remains the foundation, core, and key for the management of this disease. Furthermore, CD management involves the full life cycle of patients, requiring the joint efforts of basic and clinical medicine as well as systematic articulation at multi-levels, such as the parents, family, and society. By full-cycle, multidisciplinary, and systematic management, it is an achievable goal for CD patients to learn, work, and live in a healthy manner.

Impact of egg white protein on mycoprotein gel: Insights into rheological properties, protein structure and Molecular interactions

The mechanisms behind Mycoprotein (MYC) formation, especially the interactions with egg white proteins (EWP), are not fully understood despite the widespread use of fungal proteins. This study investigates the formation of MYC-EWP composite gels using physicochemical and structural analyses, supported by molecular simulations. At pH 7, EWP adsorbs onto the MYC mycelium surface via hydrogen bonding and electrostatic interactions, with a binding energy of −21.97 kcal/mol. Increasing EWP concentration enlarges particle size in the suspension. Upon heating, the gel's microstructure shows irregular fibrous structures with EWP filling the spaces within the mycelium. The texture properties improve with higher EWP levels, resulting in a denser gel structure. These findings offer key insights into MYC-EWP interactions, clarifying the gelation process and providing valuable guidance for optimizing fungal protein products.

Positive Intervention of Distinct Peptides in Clostridioides difficile Infection in a Mouse Model

Clostridioides difficile infection (CDI) is a common healthcare-associated infection and the leading cause of gastroenteritis-related deaths worldwide. To investigate the effects of peptide composition of different protein products on CDI, we analyzed and compared the peptide sequences and compositions from Engraulis japonicus and Glycine max using Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). An animal model of CDI was also established to investigate the potential therapeutic effects of these peptides in vivo. The peptide compositions of E. japonicus and G. max differed, with only 11% of the peptide sequences being identical. Oral administration of the tested peptides could reduce intestinal inflammation, repair the intestinal barrier, increase the proportion of beneficial bacteria, and reduce the proportion of harmful bacteria, providing a therapeutic effect against CDI. However, the peptides may differ considerably in some aspects. E. japonicus peptides were superior to G. max peptides in promoting colon epithelial cell proliferation and repairing tight intestinal cell junctions. Interestingly, the two sources of peptides have different effects on the cecal microbiome. E. japonicus peptides can effectively restore the diversity and richness of intestinal microbiota, while G. max peptides have poor regulatory effects on the intestinal microbiota structure. Overall, E. japonicus peptides showed better results than G. max peptides in treating CDI. This study supports the potential treatment of CDI with natural peptides and promotes the development of specialty foods for CDI enteritis. Clostridioides difficile infection (CDI) is a common healthcare-associated infection and the leading cause of gastroenteritis-related deaths worldwide. To investigate the effects of peptide composition of different protein products on CDI, we analyzed and compared the peptide sequences and compositions from Engraulis japonicus and Glycine max using Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). An animal model of CDI was also established to investigate the potential therapeutic effects of these peptides in vivo. The peptide compositions of E. japonicus and G. max differed, with only 11% of the peptide sequences being identical. Oral administration of the tested peptides could reduce intestinal inflammation, repair the intestinal barrier, increase the proportion of beneficial bacteria, and reduce the proportion of harmful bacteria, providing a therapeutic effect against CDI. However, the peptides may differ considerably in some aspects. E. japonicus peptides were superior to G. max peptides in promoting colon epithelial cell proliferation and repairing tight intestinal cell junctions. Interestingly, the two sources of peptides have different effects on the cecal microbiome. E. japonicus peptides can effectively restore the diversity and richness of intestinal microbiota, while G. max peptides have poor regulatory effects on the intestinal microbiota structure.

Production and characterisation of protein isolates from Chilean granado beans (Phaseolus vulgaris L.)

SummaryGranado beans, widely cultivated in Chile, offer plant‐based proteins with functional benefits for healthier food formulations. These legumes have a 28% protein content (dry basis) with peptides called lectins that have the potential of reducing type‐II diabetes. Our lab has developed a membrane‐based method to produce protein isolates from oilseeds. This method was adapted for granado beans. The process involves alkaline extraction at pH 10, followed by ultrafiltration, diafiltration, acid precipitation at pH 4 and freeze‐drying. The resulting precipitated protein isolate contains 81% protein, comparable to other legumes. However, the acid‐soluble protein isolate (SPI) has a protein content of only 53% due to co‐recovery of high molecular weight carbohydrates. The total phenolic compound contents of the protein products were low resulting in both products having an attractive light colour with no astringent taste. The characterisation of the SPI by High‐Performance Liquid Chromatography found that it contained about 40% starch and other carbohydrates. To increase the protein content of SPI to ~90%, further purification steps to remove the carbohydrates are necessary.

Studying the protein complex of flax cake obtained from the seeds of a new variety of kazakh selection

In the Republic of Kazakhstan, pressing of flax seeds (squeezing) is most often used, therefore cake is more common than meal and its resulting physical reserves exceed meal several times. The biochemical characteristics of the protein complex of flaxseed have not been sufficiently studied, which, in our opinion, limits the use of protein products obtained from flax seeds such as flaxseed cake in the production of compound feed for farm animals and poultry. The article presents studies of the protein complex and amino acid composition of flaxseed cake of the Kazakhstan variety “Kostanay 11”. Flaxseed cake contains 36,5% protein, the fat content is 7,5%. The fractional composition of flaxseed cake was compared with the flax varieties “Northern” and “Golden” grown in the Republic of Kazakhstan. The amino acid composition of flaxseed cake was compared with other cakes used in feeding farm animals and poultry. The purpose of the study is to study the fractional composition of the protein complex of flaxseed cake from flax variety "Kostanay 11". A characteristic feature of the protein complex of flax seeds is the high total content of water- and salt-soluble proteins – albumins 29,2% and globulins 16,8%, which are considered well digestible. Flaxseed cake from flax variety "Kostanay 11" in its amino acid value is equal to the known cakes common in feed production and is second only to soybean cake. This allows us to assert that flaxseed cake obtained from the seeds of the “Kostanay 11” variety remained a highly digestible and highly nutritious product in terms of protein absorption, suitable for its inclusion in the diets of all types of farm animals and poultry, starting with feed for young animals.

Anisakis extracellular vesicles elicit immunomodulatory and potentially tumorigenic outcomes on human intestinal organoids

BackgroundAnisakis spp. are zoonotic nematodes causing mild to severe acute and chronic gastrointestinal infections. Chronic anisakiasis can lead to erosive mucosal ulcers, granulomas and inflammation, potential tumorigenic triggers. How Anisakis exerts its pathogenic potential through extracellular vesicles (EVs) and whether third-stage infective larvae may favor a tumorigenic microenvironment remain unclear.MethodsHere, we investigated the parasite's tumorigenic and immunomodulatory capabilities using comparative transcriptomics, qRT-PCR and protein analysis with multiplex ELISA on human intestinal organoids exposed to Anisakis EVs. Moreover, EVs were characterized in terms of shape, size and concentration using classic TEM, SEM and NTA analyses and advanced interferometric NTA.ResultsAnisakis EVs showed classic shape features and a median average diameter of around 100 nm, according to NTA and iNTA. Moreover, a refractive index of 5–20% of non-water content suggested their effective biological cargo. After treatment of human intestinal organoids with Anisakis EVs, an overall parasitic strategy based on mitigation of the immune and inflammatory response was observed. Anisakis EVs impacted gene expression of main cytokines, cell cycle regulation and protein products. Seven key genes related to cell cycle regulation and apoptosis were differentially expressed in organoids exposed to EVs. In particular, the downregulation of EPHB2 and LEFTY1 and upregulation of NUPR1 genes known to be associated with colorectal cancer were observed, suggesting their involvement in tumorigenic microenvironment. A statistically significant reduction in specific mediators of inflammation and cell-cycle regulation from the polarized epithelium as IL-33R, CD40 and CEACAM1 from the apical chambers and IL-1B, GM-CSF, IL-15 and IL-23 from both chambers were observed.ConclusionsThe results here obtained unravel intestinal epithelium response to Anisakis EVs, impacting host’s anthelminthic strategies and revealing for the first time to our knowledge the host-parasite interactions in the niche environment of an emerging accidental zoonosis. Use of an innovative EV characterization approach may also be useful for study of other helminth EVs, since the knowledge in this field is very limited.Graphical

Fluorescent In Situ Hybridization Chain Reaction for RNA in the Drosophila Embryonic and Larval Central Nervous System.

In the Drosophila nerve cord, much is known about the generation of neurons from neuronal stem cells. Over the lifetime of a neuron, the cumulative expression of genes within that neuron determines its fate. Furthermore, gene expression in mature neurons determines their functional characteristics. It is therefore useful to visualize neural gene expression, which is often done via staining with antibodies to a protein of interest. In cases where there is no antibody to a desired gene product, or when it is useful to detect RNA rather than protein products, fluorescent in situ hybridization chain reaction for RNA (HCR RNA-FISH, or HCR for this protocol) can be used to detect and quantify RNA expression. RNA molecules reside predominantly in the cell soma, so HCR can facilitate determining neuron identity because somata position within the nerve cord is stereotyped across animals. HCR provides high-amplitude, high-fidelity signals. In principle, HCR can be broken down into a detection/hybridization stage and an amplification stage. During detection/hybridization, a probe set hybridizes to multiple sequences within a target gene. In the amplification step, concatemerized fluorescent hairpins bind to the hybridized probes. This two-step process increases the specificity of the fluorescent signal and helps reduce the likelihood of background fluorescence compared to traditional in situ hybridization techniques where the hybridizing probe itself contains the fluorescent signal. Here, we describe a protocol for using HCR to study gene expression in the Drosophila embryonic and larval nerve cord. We also describe how to combine HCR with immunofluorescence staining.

The anti-angiogenic, anti-inflammatory and anticoagulant potential of a polysaccharide extracted from the brown alga Cystoseira humilis

The present study aimed to evaluate the properties of a sulfated polysaccharide (PSCH) extracted from the brown alga Cystoseira humilis. We analyzed the chemical composition, anti-angiogenesis activities, anticoagulant, and anti-inflammatory effects of the PSCH, using various techniques encompassing UV–visible spectroscopy, fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (X-RD), scanning electron microscopy (SEM), steric exclusion chromatography (SEC) and high-performance liquid chromatography coupled to refractive index detector (HPLC-RID). The study found that PSCH was a sulfated heteropolysaccharide consisting of glucuronic acid, arabinose, galacturonic acid, xylose and fructose, and has a semi-crystalline nature with a molecular weight of about 379.44 kDa. The PSCH contains 41.77 % carbohydrates, 12.33 % proteins, and 4.59 % sulfates. The PSCH fraction has anti-angiogenic potential highlighted through assays on the chorioallantoic membrane, where vascular branching was inhibited by about 41.65 % and 61.38 % at concentrations of 50 and 100 μg/egg, respectively, compared to the untreated group. PSCH displayed an anti-edematous effect in vivo by reducing malondialdehyde and advanced oxidation protein products in paw tissue after five hours of carrageenan injection. When rats were co-treated with PSCH, there was evidence of a significant increase in antioxidant enzyme activity (superoxide dismutase, glutathione peroxidase, and glutathione) in edematous paw tissues compared to untreated rats. Furthermore, PSCH significantly influenced hematological parameters and the rate of anti-inflammatory serum protein fractions compared to the inflamed group. Interestingly, PSCH may exhibit anticoagulant properties by extending activated partial thrombosis time, thrombin time and prothrombin time. The present study has shown that Cystoseira humilis polysaccharides have antioxidant, anti-angiogenic, anti-inflammatory, and anti-coagulant properties, making them suitable for therapeutic purposes.

PSVI-14 Effects of poor maternal nutrition during gestation on first generation offspring oxidative status in skeletal muscle

Abstract Poor maternal nutrition (restricted- and over-nutrition) during gestation negatively impacts offspring pre- and post-natal growth. Previous work has identified decreased muscle mass and increased adiposity, leading to poor quantity and quality of animal protein products. Alterations in oxidative status, such as the imbalance of antioxidant activity and the production of free radicals, can result in oxidative stress damaging lipid membranes, proteins, and DNA. Changes in oxidative status may contribute to the poor growth outcomes observed in offspring from poorly nourished dams. Therefore, the objective of this research was to determine the effects of poor maternal nutrition during gestation on the oxidative status in offspring skeletal muscle in sheep. We hypothesized that offspring of restricted- and over-fed ewes would have decreased antioxidant activity and increased products of oxidative stress in skeletal muscle. To test this hypothesis, multiparous Dorset ewes (n = 46) pregnant with twins were fed 100% (CON), 60% (RES), or 140% (OVER) of total nutrient requirements (NRC) from d 30 of gestation until parturition. Following parturition, ewes were fed a control diet throughout lactation. At d 282 of age semitendinosus (STN) samples were collected from rams at necropsy, and at d 284 of age muscle biopsies from STN were collected from ewes. Data were analyzed in SAS using PROC MIXED with fixed effects of maternal diet and offspring sex. Data were considered significant at P ≤ 0.05. The activity of the endogenous antioxidant superoxide dismutase (SOD) was decreased by 45% and 33% in RES and OVER offspring, respectively, compared with CON (P ≤ 0.01). Offspring sex did not alter STN SOD activity (P = 0.35). Muscle glutathione peroxidase (GPx) activity was decreased in RES offspring by 20% compared with CON (P ≤ 0.04), but neither differed from OVER offspring (P ≥ 0.24). Ewes had 34% greater GPx activity compared with rams (P < 0.0001). Offspring from RES and OVER ewes had 77% and 87% greater concentrations of malondialdehyde (MDA), a marker of lipid peroxidation, when compared with CON (P < 0.0001). Ewes had 29% greater MDA concentrations compared with rams (P = 0.05). Protein carbonyl (PC), a marker of protein oxidation, concentrations were 36% and 38% greater in RES and OVER offspring, respectively, compared with CON (P ≤ 0.01). The decrease in antioxidant activities and increase in products of protein and lipid oxidation in offspring from poorly nourished dams suggests dysregulation in skeletal muscle oxidative status that may contribute to the negative consequences observed in offspring. Future work should focus on determining the mechanistic role of skeletal muscle oxidative stress in the decreased quantity and quality of animal protein products and identifying potential strategies to mitigate alterations in oxidative status as a result of poor maternal nutrition during gestation.

205 An autosomal recessive mutation in PYGM causes myophosphorylase deficiency in composite cattle

Abstract Between 2020 and 2022, eight calves (6 heifers, 2 bulls) in a Nebraska herd (composite Simmental, Red Angus, Gelbvieh) experienced exercise intolerance during extended physical activity. These calves exhibited an inability to keep pace with the rest of the herd, with some eventually succumbing to physical collapse from which they did not recover. Increased creatine kinase, an indication of muscle degeneration, was confirmed in biopsy and necropsy samples. Available sire pedigrees revealed a common paternal ancestor within 2 to 4 generations in all the affected calves. Dam pedigrees were unavailable; however, the herd retained replacement females that occasionally shared common ancestors with breeding bulls. Thus, a de novo autosomal recessive mutation was hypothesized as the cause of exercise intolerance in these calves. Genomic data (100K SNP) from 6 affected calves and 715 herd mates were initially used for a whole genome-wide association study, pinpointing a significant region on chromosome 29. Whole-genome sequencing of 2 affected calves combined with 145 other sequenced cattle led to the identification of a nonsense mutation in the PYGM gene, also located on chromosome 29. The mutation, confirmed to be present in the skeletal muscle transcriptome, was predicted to produce a premature stop codon. The protein product of PYGM, myophosphorylase, has a crucial role in metabolizing glycogen stored within skeletal muscle to generate energy for muscle cells. As a result, the recessive genotype led to greater (P < 0.05) glycogen concentrations in affected calves (n = 2) compared with heterozygous animals (n = 3) and wild-type controls (n = 2). Immunohistochemistry and label-free quantitative proteomics analysis confirmed the absence of the PYGM protein product in skeletal muscle. Myophosphorylase is needed for efficient glycogen utilization, which is crucial for producing high-quality beef. In the 2 affected calves harvested, increased skeletal muscle glycogen persisted postmortem, resulting in increased pH and dark-cutting beef, which is associated with a negative consumer perception and economic losses to the industry. In addition to identifying the cause of exercise intolerance and welfare concerns in these cattle, this study is among the first to identify a specific genetic mutation associated with dark-cutting beef. Carriers of the mutation did not display any discernible differences in meat quality or measures of animal well-being. While cattle heterozygous for the mutation may not immediately impact the beef industry, identifying carriers is paramount for implementing proactive selection and breeding strategies to prevent the production of affected calves.

Striving for clarity in language about gene expression.

What do we mean when we say 'gene expression'? In the decades following Crick's 1958 central dogma of molecular biology, whereby genetic information flows from DNA (genes) to RNA (transcripts) to protein (products), we have learned a great deal about DNA, RNA, proteins, and the ensuing phenotypic changes. With the advent of high-throughput technologies (1990s), molecular biologists and computer scientists forged critical collaborations to understand the vast amount of data being generated, rapidly escalating gene expression research to the 'omics' level: entire sets of genes (genomes), transcribed RNAs (transcriptomes), and synthesized proteins (proteomes). However, some concessions came to be made for molecular biologists and computer scientists to understand each other-one of the most prevalent being the increasingly widespread use of 'gene' to mean 'RNAs originating from a DNA segment'. This loosening of terminology, we will argue, creates ambiguity and confusion. We propose guidelines to increase precision and clarity when communicating about gene expression, most notably to reserve 'gene' for the DNA template and 'transcript' for the RNA transcribed from that gene. Striving to use perspicuous terminology will promote rigorous gene expression science and accelerate discovery in this highly promising area of biology.

In vitro protein digestive properties and peptidomic characterization of five whole component plant protein beverages using a pepsin-pancreatin model

Plant protein has received great attention for its potential nutrition and health benefits. In this study, the digestive properties and peptidomics of whole component plant protein beverages from soybean, chick pea, pea, oat, and quinoa were characterized using an in vitro pepsin-pancreatin digestion model. The study found that quinoa beverage had a higher in vitro protein digestibility (80.49 %), followed by pea, soybean, chick pea, and oat beverage (79.25–70.89 %). Additionally, quinoa and soybean beverage showed comparably higher in vitro digestible indispensable amino acid score (DIAAS) of 81–97, while chick pea and pea beverage had lower in vitro DIAAS of 58–79 due to low concentration of sulfur-containing AA (SAA). Meanwhile, peptides with molecular weight (MW) below 1 kDa occupied more than 93 % of the digested fraction in quinoa beverage, which might result in its higher DPPH scavenging activity, ABTS scavenging activity and ACE inhibitory activity with the IC50 value of 2.80, 0.20 and 0.90 mg/mL, respectively. Peptidomics characterization combined with rapid screening by PeptideRanker and BIOPEP database further suggested that cereals, especially oat and quinoa, released potentially bioactive peptides with a higher percentage (30.89 % and 22.81 %, respectively) compared to legumes like soybean and chick pea (13.30–18.52 %). Additionally, the identified active peptides were more likely to be derived from globular proteins. The methods and data here provided a reference for a better understanding of the digestive properties and nutritional recommendations of whole component plant protein product.