Body Protein Research Articles

Signaling of GPR17 in Hepatic and Renal Tissues

G protein-coupled receptors (GPCRs) are the largest class of proteins in the human body and are considered to be excellent targets for drug discovery. However, the vast majority of GPCRs are understudied. Using a large-scale TaqMan array screen on both male and female C57BL6 murine livers, we identified GPR17 as one of the most highly expressed orphan receptors within the liver (ΔCt = 19.9, normalized to GAPDH). GPR17 expression in the liver was confirmed using reverse transcriptase polymerase chain reaction (RT-PCR) and was also identified in murine whole kidney from both males and females indicating that GPR17 RNA expression is not sex-dependent. While GPR17 has known roles in oligodendrocyte maturation and myelination in the central nervous system, its functions within the liver and kidney are undetermined. To investigate its physiological functions, we started by uncovering its localization within the liver and kidney and identifying its ligand profile. A series of RNAScope in-situ hybridization assays were performed to localize GPR17 within hepatic tissue. Gpr17 mRNA was found within hepatocytes indicating that its highest expression is in the parenchymal cells of the liver. To identify possible activators for GPR17, we cloned the coding region into both flag-tagged PME18s and the Presto-Tango vectors. Live-cell surface labeling of HEK293T expressing the GPR17 constructs confirmed robust surface expression on the plasma membrane facilitating ligand screening. GPR17 downstream signaling has yet to be studied. Thus, we used the PRESTO-TANGO assay, an exogenous chemical ligand screening assay that relies on a luminescence readout to indicate -arrestin recruitment and receptor activation. A large, non-biased chemical screen of over 500 physiological compounds was tested for activation of GPR17. To date, we identified seven exogenous activators of GPR17: 3-[methyl(1,2,3,4- tetrahydronaphthalen-2-yl)amino]pyrazine-2-carbonitrile, 4-(4-fluorophenyl)-5-methyl-N-(2-methyl-1-oxo-1,2,3,4-tetrahydroisoquinolin-7-yl)thiophene-2-carboxamide,({1-[(2-chlorophenyl)methyl]-3,5-dimethyl-1H-pyrazol-4-yl}methyl)(methyl)[(1-methyl-1H-imidazol-2-yl)methyl]amine,N,N-dimethyl-6-[(5-phenyl-1,3-oxazol-2-yl)methyl]-5H,6H,7H,8H-pyrido[4,3-d]pyrimidin-4-amine,4-phenyl-1-[6-(pyrrolidin-1-yl)-9H-purin-9-yl]butan-2-ol, 2-(3-fluorophenyl)-3-methyl-1-{[1,2,4]triazolo[4,3-a]pyridine-6-carbonyl}pyrrolidine, and 2,6,6-trimethyl-N-({2-methyl-4H,5H,6H,7H-pyrazolo[1,5-a]pyrimidin-6-yl}methyl)-4,5,6,7-tetrahydro-1-benzofuran-4-amine. Ongoing studies are aimed at narrowing down the in vivo activators of GPR17 to ultimately shed light on the physiological functions that this receptor plays in the kidney and liver. NIH/NCATS. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effects of abomasal infusion of soybean or sunflower phospholipids on nutrient digestibility and milk production in lactating dairy cows.

The fatty acid (FA) and phospholipid composition of dietary lecithin may influence FA digestibility and milk production in cattle. Eight multiparous Holstein cows (99.4 ± 9.2 d in milk [DIM]; 48.9 ± 3.8 kg milk/d) were enrolled in a 3 × 3 incomplete Latin square design with 3 treatments provided as continuous abomasal infusates spanning 14-d experimental periods: water (CON), soybean phospholipids (SOY; 74.5 g of deoiled soy lecithin), or sunflower phospholipids (SUN; 133.5 g of hydrolyzed sunflower lecithin). Cows were fed the same diet, which contained (% dry matter) 27.0% neutral detergent fiber (NDF), 15.6% crude protein (CP), 26.2% starch, and 5.87% FA. Treatments did not modify body weight, milk fat, protein, or lactose contents, or the efficiency of producing energy-corrected milk. Cows infused with SUN had greater milk yields than those receiving SOY or CON treatments. Cows infused with SUN had higher total solids, protein, and lactose yields than cows receiving the SOY or CON treatments. Sunflower phospholipids enhanced feed efficiency (milk yield/dry matter intake) relative to SOY or CON. Treatment did not affect intakes or apparent total-tract digestibilities for NDF, CP, starch, or 16-carbon (16C) FA. Cows receiving SUN had greater total FA and 18-carbon (18C) FA intakes than SOY or CON, but treatments did not impact their digestibility. Milk FA composition was modified by treatment. Cows receiving SUN had a greater concentration of polyunsaturated FA and lower concentrations of saturated FA and monounsaturated FA in milk relative to SOY or CON. In conclusion, the abomasal infusion of SUN improved milk production and milk FA composition, indicating potential benefits for dairy cow nutrition and milk quality.

Impact of urban diets on the nutritional physiology of mealworms

Mealworms ( Tenebrio Molitor), the larval stage of yellow mealworm beetles, are a popular feeder insect for birds, amphibians, reptiles, fish, and even human populations throughout the world. As such, the goal of this work was to understand how the diet of mealworms impacts their nutritional quality as variations in quality can impact the animals (and humans) that consume them. We hypothesized that the nutritional quality of mealworms would be altered by dietary intake with urban substrates resulting in reduced nutritional quality. We divided 500 mealworms among each of the following diets designed to model substrates available in urban vs rural, more natural areas: 100% wheat germ (control); 100% Styrofoam; mixture of soil, grasses, and leaves from urban lawns; mixture of soil, grasses and leaves from rural lawns; mixture of wheat germ + carrots; natural organic fertilizer; or natural organic fertilizer with turf builder. The mealworms were maintained at room temperature and the diets were replaced weekly to prevent spoilage and to remove mealworm wastes. Once a week for three weeks, mealworms were sampled and frozen. Data were analyzed by two-way ANOVA with time and substrate as factors. After 3 weeks, mealworms housed in wheat germ + carrots weighed significantly more than all other groups (p<0.05), whereas those housed in styrofoam or urban lawn substrates weighed significantly less at week 3 as compared to week 1 (p<0.01). The urban lawn substrate resulted in greater molting and the highest number of pupae, but also the greatest mortality among the substrates. Moreover, mealworms provided urban lawn weighed less at week 3 as compared to week 1 (p=0.006). Mealworms maintained on wheat germ had significantly greater total protein content (p<0.05) and the greatest increase in protein content from week 0 to week 3 as compared to any other diet (p<0.05). Glucose content, on the other hand, was significantly lower in all groups at week 3 compared to week 0 (p<0.05). Similar trends were seen in total soluble sugars as mealworms housed in styrofoam, urban lawn, rural lawn, natural fertilizer, and fertilizer with turf builder had lower levels of total soluble sugars at week 3 compared to baseline (p<0.03) whereas those fed wheat germ, with or without carrots, had the highest content (vs WG: p≤0.03; vs WGCAR p<0.05). Interestingly, mealworms housed in wheat germ had higher oxidized lipoproteins compared to all other groups (p<0.05) whereas those fed natural fertilizer or wheat germ and carrots had the lowest level of oxidative stress compared to baseline (p<0.05). Together, these variations support our hypothesis that the nutritional quality of mealworms is altered by dietary intake. In general, urban substrates reduced the nutritional content (body mass, protein, and sugar content) of mealworms, which may negatively impact the health and behavior of predators that rely on such prey for food. This study was funded in part by a Jumpstart Grant awarded to RL by the Graduate and Professional Student Association, Arizona State University. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

CHMP2A regulates broad immune cell-mediated antitumor activity in an immunocompetent in vivo head and neck squamous cell carcinoma model

BackgroundNatural killer (NK) cells are key effector cells of antitumor immunity. However, tumors can acquire resistance programs to escape NK cell-mediated immunosurveillance. Identifying mechanisms that mediate this resistance enables us...

Fasting Metabolic Profiles are Cell-Autonomous in Elephant Seal Primary Muscle Cells

Northern elephant seals ( Mirounga angustirostris) undergo prolonged terrestrial fasts during energetically intensive life stages including breeding, molting, and postnatal development. Fasting-tolerant vertebrates generally extend the second phase of fasting, during which body mass declines but homeostasis is preserved through reliance on lipid fuel sources and sparing of body protein stores. In elephant seals, the postnatal fast coincides with development of the extensive body oxygen stores required to support extended breath-hold diving, as well as an increased capacity for apnea-related metabolic suppression. Studies in grizzly bears, another large-bodied fasting-tolerant mammal, have identified both serum-derived and intrinsic cellular regulation of the adipocyte metabolic phenotype. However, it is unknown whether these mechanisms extend across species, tissue, or fasting type (active vs inactive). Thus, we derived proliferative primary skeletal muscle cells (myoblasts) from weaned elephant seal pups; biopsies that yielded cells were collected at both early (~1 week) and late (~8 weeks) points in the postnatal fast. We found that total ATP production rate was greater in myoblasts collected from animals late in the fast (27% increase over early, p=0.08). Throughout the fast, myoblasts relied predominantly on glycolytic rather than oxidative ATP production (p<0.001), and this reliance increased as the fast progressed (p<0.014). Together, these data suggest that metabolic reliance is at least partially cell-autonomous in elephant seal myoblasts. KNA is supported by an NSF Graduate Research Fellowship (DGE 1752814 & 2146752) and a UC Berkeley Fellowship. JPV-M is supported by a NIH/NIGMS grant (R35GM146951). Research was funded by UC Berkeley startup funds and the Winkler Family Foundation. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A mini Review on Analytical and Bioanlytical Techniques in Study of Drugs-Protein Interaction

The choice of a suitable analytical technique for the identification of a drug or a combination of drug mainly depends on the sample matrix complexity and the analyte concentration. The appropriate choice of an analytical method is based on properties of studied analyte such as intrinsic structure and physicochemical properties etc. The extensive development of the pharmaceutical field requires more meticulous analytical methods for the control of various diseases by means of drugs. The aim of this review article was to study the drug protein interaction with respect to various analytical methods which are applied in research and innovative field of Pharmacy. The crucial factor involved and affects the binding of drugs with Body protein is distribution as well as bioactivity of drugs in the body. In biological science and medicine, the quantitative analysis of protein in biological liquids is of great importance. The aim of the review was therefore to summarize all available examples and information about the use of current analytical applications on various categories of drugs in their dosage forms and in biological samples. The different pharmaceutical active compounds, which have been determined using different methods, are summarized for the future reference of researcher.

CHMP5 attenuates osteoarthritis via inhibiting chondrocyte apoptosis and extracellular matrix degradation: involvement of NF-κB pathway

BackgroundOsteoarthritis (OA), the most common joint disease, is linked with chondrocyte apoptosis and extracellular matrix (ECM) degradation. Charged multivesicular body protein 5 (CHMP5), a member of the multivesicular body, has been reported to serve as an anti-apoptotic protein to participate in leukemia development. However, the effects of CHMP5 on apoptosis and ECM degradation in OA remain unclear.MethodsIn this study, quantitative proteomics was performed to analyze differential proteins between normal and OA patient articular cartilages. The OA mouse model was constructed by the destabilization of the medial meniscus (DMM). In vitro, interleukin-1 beta (IL-1β) was used to induce OA in human chondrocytes. CHMP5 overexpression and silencing vectors were created using an adenovirus system. The effects of CHMP5 on IL-1β-induced chondrocyte apoptosis were investigated by CCK-8, flow cytometry, and western blot. The effects on ECM degradation were examined by western blot and immunofluorescence. The potential mechanism was explored by western blot and Co-IP assays.ResultsDownregulated CHMP5 was identified by proteomics in OA patient cartilages, which was verified in human and mouse articular cartilages. CHMP5 overexpression repressed cell apoptosis and ECM degradation in OA chondrocytes. However, silencing CHMP5 exacerbated OA chondrocyte apoptosis and ECM degradation. Furthermore, we found that the protective effect of CHMP5 against OA was involved in nuclear factor kappa B (NF-κB) signaling pathway.ConclusionsThis study demonstrated that CHMP5 repressed IL-1β-induced chondrocyte apoptosis and ECM degradation and blocked NF-κB activation. It was shown that CHMP5 might be a novel potential therapeutic target for OA in the future.

Influence of pH and lipid membrane on the liquid–liquid phase separation of wheat γ-gliadin in aqueous conditions

Protein body (PB) formation in wheat seeds is a critical process influencing seed content and nutritional quality. In this study, we investigate the potential mechanisms governing PB formation through an in vitro approach, focusing on γ-gliadin, a key wheat storage protein. We used a microfluidic technique to encapsulate γ-gliadin within giant unilamellar vesicles (GUVs) and tune the physicochemical conditions in a controlled and rapid way. We examined the influence of pH and protein concentration on LLPS and protein-membrane interactions using various microscopy and spectroscopy techniques. We showed that γ-gliadin encapsulated in GUVs can undergo a pH-triggered liquid–liquid phase separation (LLPS) by two distinct mechanisms depending on the γ-gliadin concentration. At low protein concentrations, γ-gliadins phase separate by a nucleation and growth-like process, while, at higher protein concentration and pH above 6.0, γ-gliadin formed a bi-continuous phase suggesting a spinodal decomposition-like mechanism. Fluorescence and microscopy data suggested that γ-gliadin dense phase exhibited affinity for the GUV membrane, forming a layer at the interface and affecting the reversibility of the phase separation.

Establishment of a prognostic risk model for osteosarcoma and mechanistic investigation.

Objective: To investigate the immune mechanism of osteosarcoma (OS)-specific markers to mitigate bone destruction in the aggressive OS, prone to recurrence and metastasis. Methods: Gene expression patterns from the Gene Expression Omnibus (GEO) database (GSE126209) were analyzed using weighted gene co-expression network analysis (WGCNA), protein-protein interaction (PPI) analysis, least absolute shrinkage and selection operator (LASSO) modeling, and survival analysis to identify charged multivesicular body protein 4C (CHMP4C). Subsequently, its role in regulating the immune system and immune cell infiltration was explored. CHMP4C expression and signaling molecules in OS were assessed in osteosarcoma cell lines (MG63, U2OS, HOS) and hFOB1.19 cells using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The impact of CHMP4C upregulation and interference on OS-related signaling molecules in MG63 cells was studied. Functional validation of CHMP4C in MG63 OS cells was confirmed through cell counting Kit-8 (CCK-8), transwell, and colony formation assays. In vivo experiments were conducted using Specific Pathogen Free (SPF)-grade male BALB/C nude mice for OS xenograft studies. Results: Based on the gene expression profiles analysis of six osteosarcoma samples and six normal tissue samples, we identified 1,511 upregulated DEGs and 5,678 downregulated DEGs in normal tissue samples. A significant positive correlation between the "yellow-green" module and OS was found through WGCNA analysis. Expression levels of CHMP4C, phosphorylated Glycogen Synthase Kinase 3β (p-GSK3β), and β-catenin were notably higher in U2OS, HOS, and MG63 OS cells than in hFOB1.19 human osteoblasts. Overexpressing CHMP4C in MG63 OS cells upregulated CHMP4C, p-GSK3β, and β-catenin while downregulating GSK3β, leading to increased proliferation and migration of MG63 cells. Conversely, interrupting CHMP4C had the opposite effect. High expression of CHMP4C significantly accelerated the growth of OS in nude mice, resulting in substantial upregulation of CHMP4C, p-GSK3β, and β-catenin expression and suppression of Glycogen Synthase Kinase 3β (GSK3β) expression in OS tissues. Conclusion: CHMP4C may serve as a specific immunomodulatory gene for OS. Its activation of the Wnt/β-catenin signaling pathway, mainly by increasing the phosphorylation echelon of GSK3β, promotes the invasion and spread of OS.

CCT6A Regulates Cervical Cancer Cell Glycolysis and Proliferation under Hypoxic Conditions via the Telomerase Cajal Body Protein 1/Telomerase Reverse Tranase.

Cervical cancer is one of the most common female malignancies worldwide, a hypoxic microenvironment usually causes enhanced viability and glycolytic capacity of cervical cancer cells. The aim of this study was to investigate the association between chaperonin containing t-complex polypeptide 1 subunit 6A (CCT6A) and cell proliferation as well as hypoxic glycolysis. Bioinformatics analysis was conducted to explore the association between cervical cancer and its partner protein under hypoxic conditions, namely, CCT6A. Subsequently, the expression of CCT6A was silenced, and the effects of CCT6A silencing on cervical cancer cell proliferation, cell cycle, glycolysis-related proteins, and telomerase activity were examined. Bioinformatics analysis was performed to investigate the expression of CCT6A in cervical cancer under hypoxic conditions. The expression of CCT6A was silenced in cervical cancer cells, HeLa and SiHa, to study its effects on cell proliferation and hypoxic glycolysis. The localization of telomerase activity-related proteins, telomerase CTAB1 and TERT, was detected using immunofluorescence, and their interaction was assessed using immunoprecipitation. A cellular hypoxia model was established, and the products of the glycolysis reaction were detected. A nude mouse tumor model was constructed, and the changes in glycolysis-related proteins in tumor tissues were examined using Western blot, while Ki67 expression in tumor tissues was evaluated by immunohistochemistry. Telomerase activity was found to be enhanced in CCT6A-silenced cervical cancer cells, along with an increase in telomerase TCAB1 and TERT protein binding associated with telomerase activity. Additionally, the proportion of cells in the Gap 2/mitosis (G2/M) stage and the 5-ethynyl-2'-deoxyuridine (EdU) positivity rate were decreased in CCT6A-silenced cells, indicating a reduction in cell proliferation. The expression of cell cycle-related proteins, including cyclin E, CCNA2, and cyclin-dependent kinase 2 (CDK2), was suppressed. Furthermore, under a hypoxic environment, silencing CCT6A led to a significant reduction in cell viability and downregulation of glycolysis-related proteins, such as lactate dehydrogenase A (LDHA) and hexokinase 2 (HK2). Mechanistically, silencing CCT6A may reduce telomerase activity by inhibiting the TCAB1/TERT interaction. Additionally, TERT was found to activate the promoter region of the HK2 gene, and inhibition of TERT activity reduced the transcriptional level of HK2. The study primarily explored the involvement of CCT6A in cervical cancer, yet it did not account for the myriad of other elements potentially influencing cell proliferation and glycolysis. It is essential to recognize that cervical cancer's etiology is multifaceted, shaped by an array of genetic variations, environmental influences, and protein interactions. Silencing CCT6A could effectively attenuate the upregulation of cell proliferation and glycolytic function mediated by TCAB1/TERT in cervical cancer cells.

MSRB2 Ameliorates H2O2-induced Chondrocyte Oxidative Stress and Suppresses Apoptosis in Osteoarthritis

ABSTRACT Background Chondrocyte oxidative stress and apoptosis are critical factors contributing to the pathogenesis of osteoarthritis (OA). Methionine sulfoxide reductase B2 (MSRB2) is a mitochondrial protein that protects cells from oxidative stress and is involved in apoptosis. This study aimed to investigated the expression of MSRB2 in articular cartilage tissues and elucidated its effect on H2O2-stimulated chondrocytes. Methods Human chondrocytes were cultured in Dulbecco’s modified Eagle’s medium (DMEM)/F12. MSRB2 overexpression in chondrocytes was achieved by transfecting with an MSRB2 overexpression plasmid. Western blot, quantitative RT-PCR, Immunofluorescence staining, and TUNEL assay were employed in this study. Results MSRB2 expression was found to be reduced in OA patients. Furthermore, overexpression of MSRB2 in H2O2-induced chondrocytes mitigated apoptosis and enhanced cell viability. Elevated MSRB2 expression diminished chondrocyte ROS contents, decreased cytochrome C (Cyc) in the cytoplasm, and regulated mitochondrial membrane potential to maintain mitochondrial homeostasis. Interestingly, knockdown of charged multivesicular body protein 5 (CHMP5) led to a decreased inMSRB2 expression in chondrocytes. Additionally, protein levels of CHMP5 and MSRB2 were reduced in H2O2-stimulated chondrocytes, and silencing CHMP5 reduced MSRB2 expression. Knockdown of CHMP5 increased cleaved caspase-3 expression in H2O2-induced chondrocytes and elevated TUNEL-positive chondrocytes. Conclusion MSRB2 decreased in OA, and overexpression of MSRB2 alleviated oxidative stress and apoptosis of chondrocyte.

The ARPKD Protein DZIP1L Regulates Ciliary Protein Entry by Modulating the Architecture and Function of Ciliary Transition Fibers.

Serving as the cell's sensory antennae, primary cilia are linked to numerous human genetic diseases when they malfunction. DZIP1L, identified as one of the genetic causes of human autosomal recessive polycystic kidney disease (ARPKD), is an evolutionarily conserved ciliary basal body protein. Although it has been reported that DZIP1L is involved in the ciliary entry of PKD proteins, the underlying mechanism remains elusive. Here, an uncharacterized role of DZIP1L is reported in modulating the architecture and function of transition fibers (TFs), striking ciliary base structures essential for selective cilia gating. Using C. elegans as a model, C01G5.7 (hereafter termed DZIP-1) is identified as the sole homolog of DZIP1L, which specifically localizes to TFs. While DZIP-1 or ANKR-26 (the ortholog of ANKRD26) deficiency shows subtle impact on TFs, co-depletion of DZIP-1 and ANKR-26 disrupts TF assembly and cilia gating for soluble and membrane proteins, including the ortholog of ADPKD protein polycystin-2. Notably, the synergistic role for DZIP1L and ANKRD26 in the formation and function of TFs is highly conserved in mammalian cilia. Hence, the findings illuminate an evolutionarily conserved role of DZIP1L in TFs architecture and function, highlighting TFs as a vital part of the ciliary gate implicated in ciliopathies ARPKD.

Biochemical and Histoarchitectural evaluation of 4-Vinylcyclohexane induced ovarian cancer against Alpinia purpurata (Vieill). K. Schum

ObjectiveAlpinia purpurata is being studied for its potential to treat various conditions, including diabetes, arthritis, and tuberculosis. This research explores the impact of Alpinia purpurata (Vieill). K. Schum on ovarian cancer induced by 4-vinyl cyclohexane in Wistar albino rats. Materials and methodsFive sets of 100–120 g Wistar albino rats were assembled. Group 1 was the control group. Group 2 received intraperitoneal injection of 4-vinyl cyclohexane (80 mg/kg) for one month. Group 3 Treated similarly to Group 2 subsequently for the next two months treated with oral administration of A. purpurata ethyl acetate leaf extract (200 mg/kg body weight) Group 4 Treated similarly to Group subsequently for the next two months treated with Standard Drug Cisplatin (5 mg/kg body weight) weekly twice intraperitoneally. Group 5 acquired daily oral A. purpurata leaf extract (200 mg/kg) for two months. The rats were euthanized after the experiment under light chloroform anesthesia. Ovary and liver samples were obtained for lipid peroxidation, anti-oxidants, membrane-bound enzymes, tumor indicators, and histological investigation. ResultsOver a 60-day period, rats were given an ethyl acetate extract of A. purpurata at a dose of 200 mg/kg, which lead to in a substantial (p < 0.05) increase in body protein content, as well as enzyme levels. Furthermore, the use of the ethyl acetate extract significantly (p < 0.05) recovered the altered lipid peroxidation activities in the ovarian tissues of both control and experimental rats to near-normal levels. These data imply that the extract has the capacity to quench free radicals, indicating possible anticancer effects. ConclusionThe results suggested that, the ethyl acetate extract of A. purpurata exhibited significant antitumor activity against 4-vinyl cyclohexane induced ovarian cancer bearing rats.

Dietary supplementation of ornithine improved the growth, immune response and acute NH3 stress tolerance of Pacific white shrimp, Penaeus vannamei

Ornithine is an arginine family amino acid and is not incorporated into protein. This study evaluated its role in growth, immune response and acute NH3 stress tolerance of Penaeus vannamei. Six diets with graded levels (0.6, 0.9, 1.2, 1.5, 1.8, 2.1 g kg−1) of L-ornithine were formulated, and fed the shrimp for 8 weeks. The growth of shrimp was significantly improved with the dietary ornithine levels increased from 0.6 to 1.5 g kg−1. Feed efficiency, protein efficiency ratio and protein productive value showed a similar trend to the growth performance of shrimp. Whole body protein and muscle lipid of shrimp were significantly increased in shrimp fed 1.8 g kg−1 ornithine compared to those fed 0.6 g kg−1 ornithine. The contents of several amino acids were increased with the dietary ornithine increased to 1.5 g kg−1. Elevated levels of ornithine increased the Nitric Oxide (NO) content and suppressed arginase activity in the hemolymph. The relative mRNA expression of superoxide dismutase, heat shock protein 70, dorsal and caspase 3 were increased with the dietary addition of ornithine, while the mRNA level of arginase was decreased with the increasing dietary ornithine levels. After 9 h acute NH3 stress, mortality of shrimp reduced with the increasing dietary ornithine levels. The relative mRNA levels of superoxide dismutase, heat shock protein 70, dorsal and catalase were increased with the dietary addition of ornithine after the acute NH3 stress. The second-order regression analysis of weight gain and survival after acute stress revealed that the suitable ornithine content in the shrimp diet was 1.4–1.5 g kg−1.

Post-prandial tracer studies of protein and amino acid utilisation: what can they tell us about human amino acid and protein requirements?

Nitrogen balance (NB), the principal methodology used to derive recommendations for human protein and amino acid requirements, has been widely criticised, and calls for increased protein and amino acid requirement recommendations have been made, often on the basis of post-prandial amino acid tracer kinetic studies of muscle protein synthesis, or of amino acid oxidation. This narrative review considers our knowledge of the homeostatic regulation of the FFM throughout the diurnal cycle of feeding and fasting and what can and has been learnt from post-prandial amino acid tracer studies, about amino acid and protein requirements. Within the FFM, muscle mass in well fed weight-stable adults with healthy lifestyles appears fixed at a phenotypic level within a wide range of habitual protein intakes. However homoeostatic regulation occurs in response to variation in habitual protein intake, with adaptive changes in amino acid oxidation which influence the magnitude of diurnal losses and gains of body protein. Post-prandial indicator amino acid oxidation (IAAO) studies have been introduced as an alternative to NB and to the logistically complex 24 h [13C-1] amino acid balance studies, for assessment of protein and amino acid requirements. However, a detailed examination of IAAO studies shows both a lack of concern for homeostatic regulation of amino acid oxidation and major flaws in their design and analytical interpretation, which seriously constrain their ability to provide reliable values. New ideas and a much more critical approach to existing work is needed if real progress is to be made in the area.

Protein requirements may be lower on a training compared to rest day but are not influenced by moderate training volumes in endurance trained males.

The impact of training volume on protein requirements in endurance trained males was investigated with indicator amino acid oxidation (IAAO) methodology on a recovery day (REST) or after a 10 or 20 km run while consuming a single suboptimal protein intake (0.93g/kg/day). Phenylalanine excretion (F13CO2; inverse proxy for whole body protein synthesis) was greatest and phenylalanine net balance was lowest on REST compared to post-exercise recovery with no difference between training volumes. Single point F13CO2 was indistinguishable from past IAAO studies using multiple protein intakes. Our results suggest that protein requirements may be greatest on recovery days but are not influenced by moderate training volumes in endurance athletes.

Nutrition status and incidence of malnutrition in home enteral nutrition patients: A cross sectional study

Home enteral nutrition (HEN) is a long-term, life-sustaining nutrition therapy for patients unable to consume sufficient food orally. Patients rely on a prescribed, manufactured product to provide their full nutrient requirements, although some patients may have supplementary oral intake. Prescribed enteral nutrition is used as a treatment for malnutrition, but may, in the long-term, cause poor nutrition status. This study aimed to investigate the nutrition status (energy, protein, vitamin D, and selenium) and malnutrition incidence in long-term HEN patients in the Counties Manukau region. In this cross-sectional study, 42 adults on HEN for 4+ weeks under the care of Te Whatu Ora Health New Zealand were analysed. Participants’ enteral and oral feeding regimes were tracked using patient records and five non-consecutive 24-hour recalls. Biochemical markers, body mass index (BMI), body composition (BIA), and nutrition focussed physical findings were evaluated using reference standards and the Global Leadership Initiative on Malnutrition (GLIM) malnutrition criteria(1). Independent t-tests and Mann-Whitney tests compared participants based on their enteral and supplementary oral intakes and adherence to their enteral prescription. Dependent t-tests and Wilcoxon tests evaluated nutrients contributions from various feeding methods and sources. Over half (54.7%, n = 23) relied exclusively on enteral nutrition, but 60% did not achieve their full energy prescription. Compared to requirements based on the Oxford equation and 1g/kg of body weight, energy and protein intake was low in 20% of all participants, mean intake of these participants was 1,242 ± 183 kcal and 57.5 ± 13.5 g respectively. Participants with full enteral intake had a significantly higher vitamin D intake (14.9 µg, P&lt;0.05) than those with supplementary oral intake (11.2 µg, P&lt;0.05). However, those with oral intake had significantly higher intake of selenium, energy, and all the macronutrients than those with sole enteral intake. Vitamin D and selenium intakes were significantly greater in participants obtaining their full prescription than those that did not. No participants had low vitamin D or selenium blood concentrations, however 40% and 38.1% respectively were high. There was a significant relationship between meeting their energy prescription and high plasma selenium. Low BMI, mid arm muscle circumference, and fat free mass index were observed in 47.5%, 40.5%, and 44.8% of participants respectively. This was not statistically significant between groups. Fat mass and waist circumference were significantly higher in participants on full enteral nutrition. According to the GLIM malnutrition criteria, 62.5% (n = 25) of all participants were malnourished. In conclusion, while HEN patients maintain good vitamin D and selenium status, energy and protein malnutrition are evident. The types of food consumed by those with oral intake may be responsible for the differences in nutritional status. Further attention to prescription adherence and nutritional balance from HEN and oral intake is necessary for this vulnerable group.

Enhancement of Growth Performance and Body Coloration Through The Inclusion of Torch Ginger Powder in The Feed of Red Tilapia (Oreochromis sp.)

Red tilapia (Oreochromis sp.) is a highly produced aquaculture fish among freshwater species. Coloration is an important factor in determining consumer preferences. Therefore, an 8-week experiment was conducted to enhance the growth performance and body coloration through the inclusion of torch ginger in the feed of red tilapia. Five experimental feeds were prepared from commercial feed (crude protein: 34%, crude lipid: 5%) containing torch ginger powder at 2.5, 5.0, 7.5, and 10% (T1, T2, T3, &amp; T4) inclusion levels while feeding without torch ginger act as a control (T0). Red tilapia (Initial body weight: 0.4±0.1g; Initial total length: 4.0±0.1cm) were distributed at stocking number 20 fish/tank into fiberglass tanks with a capacity of 80 L and triplicates for each treatment. The fish were fed with experimental feeds until satiation. The growth of fish was significantly higher (P&lt;0.05) when the fish fed with T1, T2, and T3 compared to control treatment T0. Moreover, the feed intake was significantly increased (P&lt;0.05) when the torch ginger powder was added to the experimental feeds. In addition, the FCR was significantly better (P&lt;0.05) when the fish fed on T1, T2, and T3 compared to T0. The inclusion of torch ginger powder in the feed did not significantly affect (P&lt;0.05) the body protein and lipids of the experimental fish. The increment of redness was significantly higher (P&lt;0.05) when the fish fed with T1, T2, T3, and T4 compared to T0. However, it showed a declining trend of redness when the inclusion of torch ginger increased to 5.0% and above. Thus, the present study suggested that the inclusion of torch ginger powder improved the performance and enhanced the body coloration of juvenile red tilapia. However, a further increase of torch ginger powder by more than 2.5% did not help in the enhancement of the body coloration.

One-year trajectories of nutritional status in perimenopausal women: a community-based multi-centered prospective study

BackgroundNutritional status is a modifiable factor associated with perimenopausal women’s health and quality of life. Assessing body composition indicators helps to comprehensively understand nutritional status compared with using body mass index (BMI) only. However, few published studies measured the trends in body composition among perimenopausal women.ObjectivesTo assess the one-year trajectory of the nutritional status of perimenopausal women and to explore its influential factors.MethodsA community-based observational study with 3-wave repeated measurements at 6-month intervals was carried out. The nutritional status indicators include weight, body mass index (BMI), and body composition variables. Bioelectrical impedance analysis was used to assess body composition. Repeated measures ANOVA and Chi-square test were used to calculate the changes in nutritional status and generalized estimating equations were performed to explore their influential factors.Results2760 participants completed the study. Increasing trajectories in weight (from 56.05 ± 7.55 to 57.02 ± 7.60), fat mass (from 17.99 ± 4.80 to 20.49 ± 4.90), and waist-hip ratio (from 0.86 ± 0.04 to 0.91 ± 0.15) were found (P < 0.001). Decreasing trajectories in skeletal muscle (from 20.30 ± 2.38 to 19.19 ± 2.46), protein level (from 7.39 ± 0.79 to 7.06 ± 0.81), and total body water (from 27.87 ± 2.92 to 27.00 ± 3.01) were found (P < 0.001). Being married/unmarried with a partner and without negative life events were associated with higher total body water, skeletal muscle, and protein level, while negatively associated with fat mass and waist-hip ratio. Age was positively associated with fat mass (P < 0.001). Participants with junior high school education were prone to increased fat mass (P = 0.018) compared with those holding primary school education and below. A per capita monthly income of 1500 to 3000 Yuan was associated with higher total body water, skeletal muscle, and protein level (P < 0.001) compared with a per capita monthly income of less than 1500 Yuan.ConclusionWorsening nutritional status exists in perimenopausal women, which is characterized by increased weight, fat mass, and waist-hip ratio, and decreased skeletal muscle, total body water, and protein level. For greater efficiency, precision nutritional interventions are needed, and recipients should be classified into different risk levels based on their sociodemographic background.

Identification and Analysis of Expression Patterns of the Caleosin Genes in Hickory (Carya cathayensis Sarg.)

The deciduous tree hickory (Carya cathayensis) holds economic significance in China due to its high oil content, particularly in unsaturated fatty acids. Oil bodies are crucial for storing triacylglycerol (TAG), with caleosin serving as a predominant oil body protein that aids in oil body formation and stability maintenance. Our study utilized bioinformatics techniques to identify caleosin genes within Carya cathayensis, Carya illinoinensis, and Juglans regia. Three caleosin genes were discovered in the genomes of Carya cathayensis, Carya illi-noinensis, and Juglans regia. These genes encode hydrophilic proteins. Additionally, all caleosin proteins feature a single Ca2+-binding EF-hand, a conserved “proline knot” motif, and a C-terminal hydrophilic region with four potential phosphorylation sites. The caleosin proteins in Carya cathayensis consist of α-helix, β-corner, extended chain, and random curl structures. Cis-acting elements related to stress response and hormone signaling were identified in Carya cathayensis, Carya illinoinensis, and Juglans regia, with distinct cis-acting elements implicated in seed-specific regulation in Carya cathayensis. Additionally, subcellular localization analysis confirmed that CcaCLO1 and CcaCLO2 were localized within oil bodies. Transcriptome analysis and quantitative real-time polymerase chain reaction (qRT-PCR) data demonstrated a significant up-regulation of CcaCLO1 expression during the developmental stages of the Carya cathayensis embryo. Furthermore, qPCR findings indicated that caleosins from Carya cathayensis were responsive to salt stress, with a significant up-regulation of CcaCLO1 following exposure to salt stress treatment. Consequently, caleosin genes in Carya cathayensis, Carya illinoinensis, and Juglans regia share similar physicochemical characteristics and conserved motifs. Specifically, CcaCLO1 in Carya cathayensis primarily responds to embryo development and salt stress. These findings offer foundational insights for future investigations into the regulatory mechanisms of oil accumulation and response to salt stress in hickory.