Protein Mass Research Articles

Diet-induced impairment of skeletal muscle and adipose tissue metabolic homeostasis and its prevention by probiotic administration.

Western dietary pattern is one of the main contributors to the increased risk of obesity and chronic diseases, through oxidative stress and inflammation, that are the two key mechanisms targeting metabolic organs, such as skeletal muscle and adipose tissue. The chronic exposure to high levels of dietary fatty acids can increase the amount of intramyocellular lipids in skeletal muscle, altering glucose homeostasis and contributing to a reduction in mitochondrial oxidative capacity. Probiotic administration is a promising approach as preventive strategy to attenuate metabolic damage induced by Western diet. Here, we investigated the beneficial effect of Limosillactobacillus reuteri DSM 17938 on the inflammatory state and oxidative balance in the skeletal muscle and adipose tissue of adult rats fed a western diet for 8weeks, focusing on the role of skeletal muscle mitochondria. Limosillactobacillus reuteri DSM 17938 administration protected the skeletal muscle from mitochondrial dysfunction and oxidative stress, preventing the establishment of inflammation and insulin resistance. Interestingly, a further beneficial effect of the probiotic was exerted on body composition, favoring the deposition of protein mass and preventing adipose tissue hypertrophy and inflammation. These results open the possibility for the use of this probiotic in therapeutic approaches for nutrition-related diseases.

Unveiling Aquatic Organism Health Through eProteins: A Contemporary Perspective

Environmental DNA (eDNA) analysis has transformed our understanding of aquatic ecosystems, but traditional methods often lack insights into organismal health. The emerging field of environmental protein (eProtein) analysis offers a novel approach to monitoring the physiological status of aquatic organisms. Recent advancements in proteomic technologies have enabled the detection and characterization of stress-responsive proteins in water samples, providing valuable insights into organismal health and environmental stressors. This perspective paper explores the potential of eProtein analysis for monitoring aquatic organism health, disease dynamics and reproductive cycles. Methodological advancements in protein extraction and mass spectrometry have enhanced the sensitivity and specificity of eProtein analysis, facilitating comprehensive molecular profiling and biomarker identification. By integrating eProtein analysis into aquaculture management and environmental monitoring, researchers can proactively manage fish health, mitigate disease outbreaks and safeguard aquatic ecosystems. Future research directions include refining extraction methods, establishing standardized protocols and leveraging interdisciplinary collaborations to maximize the potential of eProtein analysis for aquatic research and conservation.

End-To-End Automated Intact Protein Mass Spectrometry for High-Throughput Screening and Characterization of Bispecific and Multispecific Antibodies.

Bispecific antibodies (bsAbs) and multispecific antibodies (msAbs) represent a promising frontier in therapeutic antibody development, offering unique capabilities not achievable with traditional monoclonal antibodies. Despite their potential, significant challenges remain due to their increased molecular complexity. One prominent challenge is the correct assembly of light and heavy chains, as improper pairing leads to mispaired or incompletely assembled species that lack therapeutic efficacy and possess undesired properties, impairing the developability, manufacturability, and safety. There is a critical need for rapid, sensitive analytical tools to monitor and control these undesired species and ensure the quality assessment of bsAbs and msAbs. To address this need, we present a novel high-throughput, format-agnostic intact mass workflow that significantly enhances the efficiency of detecting and quantifying biotherapeutic products and related impurities. This workflow integrates automated sample preparation, novel high-resolution rapid mass detection powered by SampleStream-MS, and an advanced data analysis pipeline. It offers increased throughput and data quality while substantially reducing analysis turnover time and labor. This was demonstrated in a pilot program where ∼800 multispecific antibodies were processed in 10 working days. The article details the evaluation and validation of our method, demonstrating its repeatability and intermediate precision in terms of measurement accuracy and relative quantification of various product-related species. We underscore the transformative potential of this end-to-end high-throughput workflow in expediting bispecific and multispecific antibody discovery, optimizing production processes, and ensuring high-quality development and manufacturing for therapeutic antibodies.

Strategies to verify equimolar peptide release in mass spectrometry-based protein quantification exemplified for apolipoprotein(a).

Quantitative protein mass spectrometry (MS) is ideally suited for precision diagnostics and for reference standardization of protein analytes. At the Leiden Apolipoprotein Reference Laboratory we apply MS strategies to obtain detailed insight into the protein-to-peptide conversion in order to verify that quantifier peptides are not partly concealed in miscleaved protein backbone. Apolipoprotein(a) (apo(a)) was digested in a non-optimal manner to enhance the number of miscleaved peptides that were identified by high resolution liquid chromatography tandem-MS measurements. The protein-to-peptide conversion was carefully mapped with specific attention for miscleaved peptides that contain an apo(a) quantifier peptide. Four different isotopologues of each apo(a)-quantifier peptide were applied to evaluate linearity of internal peptide standards during measurement of specific real-life samples. Two apo(a) quantifier peptides that were concealed in two different miscleaved peptides were included into a multiple reaction monitoring list in our targeted MS-based apo(a) quantifications to alert for potential protein digestion discrepancies. The presence of miscleaved peptides could be ruled out when applying our candidate reference measurement procedure (RMP) for apo(a) quantification. These data further corroborate the validity of our apo(a) candidate RMP as higher order method for certification of commercial Lp(a) tests that is endorsed by theInternational Federation of Clinical Chemistry and Laboratory Medicine. MS-based molecular detection and quantification of heterogeneous apo(a) proteoforms will allow manufacturers' transitioning from confounded lipoprotein(a) [Lp(a)] mass levels into accurate molar apo(a) levels.

Combining Quantitative Proteomics and Interactomics for a Deeper Insight into Molecular Differences between Human Cell Lines.

In modern biomedical research, cultivable cell lines are an indispensable tool, and the selection of cell lines that exhibit specific functional profiles is often critical to success. Cellular functional pathways have evolved through the selection of protein intra- and intermolecular interactions collectively referred to as the interactome. In the present work, quantitative in vivo protein cross-linking and mass spectrometry were used to probe large-scale protein interactome differences among three commonly employed human cell lines, namely, HEK293, MCF-7, and HeLa cells. These data illustrated highly reproducible quantitative interactome levels with R2 values larger than 0.8 for all biological replicates. Proteome abundance levels were also measured using data-independent acquisition quantitative proteomics methods. Combining quantitative interactome and proteome information allowed the visualization of cell type-specific interactome changes mediated by proteome level adaptations and independently regulated interactome changes to gain deeper insight into possible drivers of these changes. Among the largest detected alterations in protein interactions and conformations are changes in cytoskeletal proteins, RNA-binding proteins, chromatin remodeling complexes, mitochondrial proteins, and others. Overall, these data demonstrate the utility and reproducibility of quantitative cross-linking to study system-level interactome variations. Moreover, these results illustrate how combined quantitative interactomics and proteomics can provide unique insight into cellular functional landscapes.

Condition states in anchovy (Engraulis encrasicolus) and sardine (Sardina pilchardus) revealed by energy and proximate composition relationships.

Energy content has long been proposed as a fundamental, integrated, and reliable indicator of the condition of individuals as it reflects past bioenergetics and influences future life-history traits. There is a direct biochemical link between energy density and body composition described by four main compounds in fish (protein, lipid, ash, and water), with proteins and lipids being the sources of energy. If relationships between water content, or lipid content, and energy density have been well described in relative terms, the absolute mass variations in the proximate composition have been overlooked and thus their interpretation is often equivocal. In our study, based on a large and unique dataset on the proximate composition and energy density of anchovy (Engraulis encrasicolus) and sardine (Sardina pilchardus) from sampling in the Bay of Biscay and the English Channel, we aimed to better explain the patterns between water content and other proximate components or energy density, based on the dynamics of proteins, lipids, and water absolute masses. For the first time, we defined good, intermediate, and poor condition states in wild fish, based on water content, corresponding to the different dynamics of lipids and proteins in the metabolism of individuals. Anchovy and sardine exhibited remarkably similar patterns of variation in the compounds and in the limits between the condition states with respect to water content. Those patterns revealed that water mass remained constant for a given fish size whatever its condition state, and that variability in water content only resulted from the variation in lipid and protein masses. Furthermore, the differential dynamics of proteins and lipids, with only lipids mobilized in the good condition state, only proteins in the poor condition state, and both proteins and lipids in the intermediate condition state, elucidates the nonlinear pattern observed in the relationship between energy density and water content. Overall, our results highlight the importance of monitoring the intraspecific variations in water content to predict the proximate composition and energy content in small pelagic fish and better assess individual and population conditions in changing ecosystems as well as to better parameterize bioenergetic models.

Immunocapture mass spectrometry: macroscopic history, recent trends and future prospects.

Literature on immunological pretreatment and mass spectrometry was searched in the Web of Science database. The collected data were utilized for bibliometric analysis and literature review to identify trends in this field. Immunological pretreatment initially began in chemical toxicology using gas chromatography mass spectrometry and research interests later shifted to biologics pharmacokinetics, biomarkers, anti-drug antibodies and diagnosis. The key advantage of using mass spectrometry was the ability to distinguish molecular subtypes. Future research interests are expected to focus on a broader range of analyte types (such as intact protein mass spectrometry) and methodological improvements, including better protein digestion and microfluidics, aiming for faster, less labor-intensive and more reliable analysis through multiplexing and automation.

M6A RNA METHYLATION AS A THERAPEUTIC TARGET IN HIGH GRADE GLIOMA

Abstract AIMS High grade glioma (HGG) is a devastating brain cancer that has poor prognostic outcomes. Very recently, the field of epitranscriptome has emerged as a novel druggable target for multiple diseases, including cancers. In the present study, we sought to investigate the functional importance of N6-methyladenosine (m6A) RNA methylation in the pathogenesis and drug resistance of paediatric and adult high grade glioma. METHOD To identify the expression of key m6A effectors, RT-qPCR and immunofluorescence were undertaken in patient derived HGG cell lines, primary low passage HGG cells and human tissues. The expression of these effectors was modified using siRNA-mediated knockdown and small molecules to identify their effects on key transcription factors. The consequences of modification on the ability of the cells to form neurospheres and invade was also assessed. Developing chemotherapy-resistant cell lines is undertaking to assess the effect of m6A RNA levels on the ability of HGG cells to adapt to environmental changes. Measuring the level of m6A-modified transcripts were achieved by developing LC-MS/MS and MeRIP (methylated RNA immunoprecipitation) qPCR techniques. RESULTS We found that key m6A effectors such as methyltransferase like 3 (METTL3), Wilms tumor 1-associating protein (WTAP) and fat mass and obesity-associated (FTO) are expressed at significantly higher level in paediatric and adult HGG models than in control at both gene and protein levels. Furthermore, these effectors regulate the expression of key transcription factors in self-renewal, cell cycle regulation and tumorigenesis. siRNA-mediated depletion of m6A effectors and inhibition of METTL3 using small molecules such as STM2457 and STM3006 reduce the capacity of cells to form neurospheres and invade as well as improving the sensitivity of HGG cells to chemotherapeutic agents. CONCLUSION Our findings suggest that m6A effectors are potential pro-oncogenic factors that, if druggable, will represent an advanced avenue in HGG therapy.

Veterinary and sanitary assessment of milk quality in black-and-white cows fed with extruded compound feeds in North Kazakhstan

Background: Today, one of the most effective ways to increase the digestibility of nutrients is extrusion processing. Raw materials are exposed to high temperatures and pressure, which leads to qualitative and nutritional transformations. The study aims to identify the effect of extruded compound feeds on cow productivity and the qualitative composition of milk.Methods: Based on the chemical analysis of milk, the following indicators were studied: fat mass fraction, protein mass fraction, casein, lactose, dry matter, skimmed milk residue, urea, density, titrated acidity, active acidity, and somatic cells. An organoleptic evaluation of milk was carried out according to the following indicators: color, smell, taste, appearance, and consistency. Once a month, a control milking was carried out to account for productivity.Results: The EG's raw milk had higher casein and lactose content compared to the CG, with EG II showing the highest levels. Additionally, milk from the EG contained more solids, indicating better nutritional and technological properties, with solid content being 3.8-5.7% higher than in the CG.Conclusion: Based on the obtained results, it was found that milk productivity had increased by 10-20%, the fat mass fraction by 7-10%, the protein mass fraction by 5.1-6.5%, the amount of dry matter by 3.8-5.7%, and lactose by 1.1-4.3%.Keywords: Extruded Compound Feeds; Milk Productivity; Black-And-White Breed; Fat; Protein

Toward Absolute Quantification of Soluble Proteins via Proton Nuclear Magnetic Resonance Spectroscopy: Total Protein Concentration in Blood Plasma.

For absolute protein quantification using nuclear magnetic resonance (NMR) spectroscopy, we considered proteins as homopolymers and effective amino acid (AA) residues (AAREff) as monomer units. For diverse classes of proteins, we determined the AAREff molecular weight as 111.5 ± 3.2 Da and the number of hydrogens per AA as 7.8 ± 0.2. Their ratio of 14.3 ± 0.3 (g/LP)/(mol/LH) remains constant across various protein classes and is equivalent to Kjeldahl's nitrogen-to-protein conversion constant of 5.78 ± 0.29 gN/gP. By analogy to the Kjeldahl method, we suggest that the total integral of a 1H NMR solution protein spectrum could be used for total protein quantification. We synthesized low-resolution protein spectra from the weighted sums of individual AA spectra and compared them with experimental spectra. In the methyl region, the ratio of the protein mass to the total number of protons in the synthetic spectra (corrected for the chemical shift mismatch) was ∼1 (mg/mL)/mM, which agrees with an earlier reported experimental ratio for urine (1.05 ± 0.06 (mg/mL)/mM). For human blood plasma, in the methyl region, we found empirical ratios of 1.115 ± 0.006 (mg/mL)/mM (using 96 patient samples) and 1.121 ± 0.011 (mg/mL)/mM for the NIST plasma standard. This numerical agreement points to universal conversion constants, i.e., protein mixtures with unknown compositions could be quantified without the need for calibration standards by measuring the millimolar proton concentration within the methyl region of the NMR spectrum using the same conversion constant.

Covalent modification of protein by chemical probe in living cells for structural and interaction studies.

Cellular activities are predominantly carried out by proteins that can dynamically adopt different structural conformations and differentially interact with other biomolecules according to cellular needs. Chemical probes are small molecules used to selectively interact and modulate the activities of specific proteins to study their functions such as the validation of potential drug targets. The remarkable performance of AlphaFold algorithms in the prediction of protein structures has pivoted interest toward elucidating the intracellular dynamics of protein structural conformation where covalent modification of proteins by chemical probes could be used to shed light upon. However, due to the barrier to entry by cell membrane and the general unfavorable reactive conditions of the intracellular environment, most studies using reactive chemical probes are still conducted on purified proteins and cell lysates. Nevertheless, recent progresses have been made in designing chemical probes with improved membrane permeability, stability and reactivity. This paper surveys the literature on recent advancements in membrane-permeable chemical probes and their applications with protein mass spectrometry for the intracellular studies of protein structural conformations and biomolecular interactions.

Effects of different intensity resistance training on fat level, energy intake, and appetite among overweight and obese female college students

This study examines how different intensity resistance exercises impact fat, energy intake, and appetite regulation in overweight and obese female college students in Jiangsu Province, China. This study used a randomized controlled trial (RCT) design that investigates resistance training intensity-target result causation. The study determines the impact of High-Intensity Interval Training (HIIT) and Moderate-Intensity Resistance Training (MIRT) on body composition, fat level/body composition, and muscle mass them. In Jiangsu Province, China, 515 overweight and obese female college students aged 18–25 were sampled. Intervention groups were assigned randomly to campus recruits based on demographic and anthropometric. This study demonstrated significant correlations between resistance training intensity, body composition, diet, and appetite regulation in overweight and obese colleagues. MIRT and HIIT outperformed Control in weight loss and diet and used valid measures like DEXA (Dual-Energy X-ray Absorptiometry), blood tests and food diaries to regulate appetite hormones. Increased protein and muscle mass minimize resistance exercise intensity-metabolic consequences. The study analyzes these impacts to show how exercise intensity, food, and metabolic health are interconnected. These findings help build evidence-based obesity prevention and treatment strategies for this demographic. Healthcare providers, legislators, and university stakeholders use this study to tackle college student obesity. The results suggest that campus wellness initiatives should focus on resistance exercise and diet to combat obesity. This research promotes exercise science and health behavior change theory for cross-cultural lifestyle interventions.

Bacterial ghosts engineered with lipidated antigens as an adjuvant-free vaccine for Chlamydia abortus

Bacterial ghosts (BGs) provide novel vaccine delivery platforms because of their inherent adjuvant properties and efficient antigen delivery capabilities. However, effective engineering strategies are required to modify them for different antigens. In this study, the Escherichia coli (E. coli) ghost was modified by using a lpp’-ompA chimera, a widely used bacterial surface display vector, with a protective antigen macrophage infectivity potentiator (MIP) of Chlamydia abortus (C. abortus), and its protective effect was evaluated in a mouse model. The MIP fusion protein accumulated at 1.2% of the ghost total protein mass and a significant portion of the protein was modified into lipoproteins upon translocation to the BG surface. Lipidated MIP-modified recombinant E. coli ghosts (rECG-lpp’-MIP) effectively promoted antigen-presenting cells (APCs) uptake of antigens and stimulated APCs activation in vivo and in vitro. Immunization with rECG-lpp’-MIP and no adjuvant induced intense specific humoral responses as well as Th1-biased cellular immune responses, which significantly improved the efficiency of C. abortus infection clearance in mice and reduced pathological damage to the uterus. In summary, this study demonstrates that recombinant E. coli ghosts modified with lipidated antigens could help to develop an effective C. abortus vaccine and aid in the development of a universal adjuvant-free vaccine platform.

B-090 Efficient Hydrophilic Surface Coating with Low Concentrations of Bovine Gamma Globulins

Abstract Background Bovine Gamma Globulin (BGG) is a high purity immunoglobulin fraction of bovine serum. BGG is used as a blocker to prevent non-specific binding in immunoassays and, due to the mixture of immunoglobulins, its blocking capabilities differ from traditional blockers like Bovine Serum Albumin (BSA). Our objective was to evaluate the adsorption characteristics of BGG to In vitro diagnostic relevant hydrophilic surfaces compared to BSA. A bead-based immunoassay was utilized to quantify the mass of protein coating a bead surface and atomic force microscopy (AFM) to analyze physical properties of the protein-surface layer. Methods The mass of BGG bound to hydrophilic beads (Dynabeads M-270 Carboxylic Acid) was determined across varying concentrations ranging from 0.1% to 1.0% in pH 7.0 phosphate buffer. Coated beads were heated in 1x SDS loading dye to dissociate bound protein, then ran on a nonreducing SDS-PAGE gel. The total protein was measured using densitometry against a standard curve, using ImageJ, and analyzed with GraphPad Prism. This methodology was also used for BSA. AFM was used to image a topographical view of BGG binding to a silica surface. Samples were prepared at concentrations from 0.1% to 1.0% in phosphate buffer at pH 7.0. Followed by a series of washes to remove non-bound molecules, the surface was allowed to dry, then scratched with an AFM cantilever. The surface was then analyzed for roughness and thickness of BGG adsorption for each concentration. Results BGG and BSA have notable differences in their coating properties at a concentration of 1.0%. In the bead assay, the protein-surface binding is enhanced ∼4.8-fold for BGG compared to BSA. Interestingly, even at 0.1% BGG bound ∼0.8-fold more than BSA at 1.0%. Despite a 6-fold increase in mass bound to the bead surface with 1.0% BGG compared to 0.1%, AFM revealed a 1.3-fold and 1.8-fold decrease in height and roughness at the lower concentration, while maintaining effective coating. These results suggest that higher BGG concentrations result in thicker, rougher surface coating. This could lead to steric hindrance and obstruction of binding sites, affecting assay performance. Therefore, a lower concentration of BGG may lead to better surface coating and prevent reduced sensitivity and accuracy within immunoassays. Conclusions The bead binding assay demonstrates that BGG displays surface adsorption with comparable or improved results with lower concentrations compared to BSA. At 0.1%, BGG exhibits effective binding to a hydrophilic surface equivalent to that achieved by BSA at 1.0%. Additionally, AFM data suggest that even at 0.1%, BGG effectively coats the silica surface and potentially reduces steric hinderance compared to BGG at 1%. While BSA remains a reliable choice for many applications, our findings suggest that BGG can be an effective blocker for immunoassay applications. The choice of a suitable blocker varies depending on factors such as assay high background, sensitivity, and specificity. By tailoring the choice of blocker to the specific requirements of the assay, researchers can optimize assay performance by improving the accuracy and the reliability of the assay.

Greater energy surplus promotes body protein accretion in healthy young men: A randomized clinical trial

Background & aimsCaloric overfeeding combined with adequate protein intake increases not only body fat mass but also fat-free mass. However, it remains unclear whether the increase in fat-free mass due to overfeeding is associated with an increase in total body protein mass. We evaluated the hypothesis that overfeeding would promote an increase in total body protein mass. MethodsIn our randomized controlled trial, 23 healthy young men were fed a diet equivalent to their energy requirements with a +10 % energy surplus from protein alone or a +40 % energy surplus (+10 % from protein, +30 % from carbohydrate) for 6 weeks. We estimated total body protein mass by a four-compartment model using dual-energy X-ray absorptiometry, deuterium dilution, and hydrostatic underwater weighing. ResultsThe 40 % energy surplus over 6 weeks significantly increased body protein mass compared to baseline by 3.7 % (0.44 kg; 95 % confidence interval [CI], 0.21–0.67 kg; P = 0.003); however, the 10 % energy surplus did not result in a significant change (0.00 kg; 95 % CI, −0.38–0.39 kg; P = 0.980). A significant interaction between intervention duration (time) and energy surplus (group) was observed for total body protein mass (P = 0.035, linear mixed-effects model), with a trend toward a significant difference in total body protein mass gain between groups (P = 0.059, Wilcoxon rank sum test). The increase in body protein mass due to the energy surplus was correlated with an increase in fat mass (r = 0.820, p = 0.002). ConclusionsA higher energy intake was found to promote an increase in body protein mass in healthy men consuming excess protein, suggesting the importance of energy surplus in body protein accumulation. This effect of energy surplus may be related to factors such as increased body fat mass and the associated secretion of adipokines. Trial registrationThe trial was registered with the University Hospital Medical Information Network Clinical Trial Registry as UMIN000034158.

Fabrication and characterisation of whey–pea protein‐based emulsion gels induced by transglutaminase cross‐linking

Food industry has partially replaced animal proteins with plant‐derived alternatives, and effects of combining these two types on properties of food systems need to be elucidated. This study fabricated and characterised emulsion gels of mixed whey and pea protein, varying protein mass ratios from 100:0 to 0:100 and oil volumes from 30% to 45%, induced by transglutaminase cross‐linking. An increase in pea protein led to larger particle size and reduced surface charge in the solutions. Emulsion droplet size ranged from 0.60 to 15.17 μm, increasing with higher levels of pea protein and oil. Mixed protein gels exhibited significantly greater hardness compared to gels with individual proteins. Water‐holding capacity of pure whey protein emulsion gels exceeded 90%, while the value remained above 80% when pea protein was less than 50%. Partially replacing whey protein with pea protein may have potential applications in food products such as salad dressings and cheese.

Association Between Serum Levels of Vitamin D and Biochemical Markers Among Hematopoietic Stem Cell Transplantation Candidates: A Cross-Sectional Study

Background: Although vitamin D has been known as an effective substance in bone homeostasis, recent studies indicated a number of other biological properties attributed to vitamin D. Patients, who are candidates for hematopoietic stem cell transplantation (HSCT), were shown to be at high risk of vitamin D deficiency. Objectives: This study aimed at exploring the association between serum levels of vitamin D and biochemical markers among HSCT candidates. Methods: Totally, 214 patients, aged 18 to 65 years, were recruited in the current cross-sectional study. Within 24 hours of admission to the Bone Marrow Transplant ward, baseline clinical and demographic characteristics of study participants, serum levels of vitamin D, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), hemoglobin, albumin, total protein, CRP-albumin ratio (CAR), and body mass index (BMI) were assessed. Participants were divided into 4 groups based on their serum vitamin D levels: subjects with deficient, insufficient, sufficient, and optimal levels of vitamin D. Results: Across the 4 defined categories of serum vitamin D levels, there was no significant difference in terms of BMI, laboratory parameters, inflammatory factors, and biochemical markers. This lack of significant variation remained in both unadjusted and adjusted models. Conclusions: These observations indicate a lack of significant association between serum vitamin D levels and BMI, inflammatory factors, and biochemical markers in individuals undergoing evaluation for HSTC.

Effects of fasting on golden pompano Trachinotus ovatus: Physiological and biochemical responses

In practice, golden pompano (Trachinotus ovatus) reared in offshore cages may experience short-term fasting during extreme weather conditions, yet their physiological and biochemical responses remain poorly understood. To investigate the responses of this species to fasting, golden pompano weighing 138.0 ± 2.1 g were deprived of food for 0, 7, 14, 21 and 28 days, followed by assessments of body mass, somatic parameters, proximate composition, muscle glycogen, intestinal digestive enzymes, muscle amino acid and fatty acid profiles, nutritional value, and muscle texture and water holding capacity. Lower condition factor, hepatosomatic index, viscerosomatic index, hepatic glycogen, and intestinal activities of amylase, lipase and trypsin were found in the fasted fish than in the non-fasted fish, while a sharp decline in these parameters occurred on the first week of fasting. Fresh mass, dry mass, whole body and muscle contents of crude protein and lipid tended to decrease with prolonged fasting, while no statistical differences were found in these parameters between fish fasted for 7 days and the non-fasted fish. Furthermore, the decline rate of whole body crude lipid was consistently higher than that of whole body crude protein throughout the different stages of the 28-day fasting period, with the exception of the second week of fasting. No statistical differences were found in the total essential amino acids (∑EAA), non-essential amino acids (∑NEAA), ratio of ∑EAA to∑NEAA, chemical score, amino acid score, and essential amino acid index in the fasted fish compared to the non-fasted fish of group 0d. Fasting altered the muscle profile of SFA, MUFA and PUFA, sometimes reducing muscle springiness (groups 7d and 14d) and hardness (group 21d). Compared to non-fasted fish, fish fasted for 7 and 14 days exhibited statistically lower or equal drip loss, water loss rate, storage losses, and frozen exudate rate, whereas higher cooked meat rate. The above results indicate that during the 28-day fasting period, the visceral fat and hepatic glycogen are initially metabolized, followed by body proteins and lipids, to maintain the normal metabolism in fasted fish. Fasting has beneficial effects on the nutritional value of amino acids and fatty acids in muscle, although it modifies the muscle SFA, MUFA and PUFA composition and sometimes has negative effects on muscle texture. Considering the balance between body weight loss and flesh quality improvement, the duration of fasting for cultured golden pompano prior to harvesting should not exceed 14 days.

Liver-specific Coxsackievirus and adenovirus receptor deletion develop metabolic dysfunction–associated fatty liver disease

Metabolic dysfunction–associated fatty liver disease (MAFLD) is a common liver disease associated with obesity and is caused by the accumulation of ectopic fat without alcohol consumption. Coxsackievirus and adenovirus receptor (CAR) are vital for cardiac myocyte-intercalated discs and endothelial cell-to-cell tight junctions. CAR has also been reported to be associated with obesity and high blood pressure. However, its function in the liver is still not well understood. The liver of obese mice exhibit elevated CAR mRNA and protein levels. Furthermore, in the liver of patients with non-alcoholic steatohepatitis, CAR is reduced in hepatocyte cell–cell junctions compared to normal levels. We generated liver-specific CAR knockout (KO) mice to investigate the role of CAR in the liver. Body and liver weights were not different between wild-type (WT) and KO mice fed a paired or high-fat diet (HFD). However, HFD induced significant liver damage and lipid accumulation in CAR KO mice compared with WT mice. Additionally, inflammatory cytokines transcription, hepatic permeability, and macrophage recruitment considerably increased in CAR KO mice. We identified a new interaction partner of CAR using a protein pull-down assay and mass spectrometry. Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3C (APOBEC3C) demonstrated a complex relationship with CAR, and hepatic CAR expression tightly regulated its level. Moreover, Apolipoprotein B (ApoB) and Low-density lipoprotein receptor (LDLR) levels correlated with APOBEC3C expression in the liver of CAR KO mice, suggesting that CAR may regulate lipid accumulation by controlling APOBEC3C activity. In this study, we showed that hepatic CAR deficiency increased cell-to-cell permeability. In addition, CAR deletion significantly increased hepatic lipid accumulation by inducing ApoB and LDLR expression. Although the underlying mechanism is unclear, CARs may be a target for the development of novel therapies for MAFLD.

Prognostic Value of Performance Status, Albumin, and CRP in Last-Line Chemotherapy for Pancreatic vs. Other Gastrointestinal Cancers-Simple Tools Matter.

Patients with advanced gastrointestinal cancers often receive chemotherapy near the end of life (EoL), raising concerns about overtreatment. The PALLiON trial, a cluster-randomized trial, assessed the impact of a complex intervention on frequency of EoL treatment; the intervention involved palliative care referrals and the use of PROMs. The present secondary analysis evaluated the prognostic value of baseline performance status (PS), albumin (alb), C-reactive protein (CRP), and body mass index (BMI) for overall survival, comparing pancreatic (PAN, n = 189) vs. other gastrointestinal cancer patients (GI, n = 286). Baseline PS, alb, CRP, mGPS (modified Glasgow prognostic score), and BMI were analyzed using Cox regression. Adjusted for age, sex, and hospital size, PS ≥ 2 and alb < 35 g/L predicted shorter survival in both PAN and GI cancers, while CRP > 10 predicted shorter survival only in GI cancers. In PAN, PS ≥ 2 predicted a 78.4% higher probability of shorter survival, and mGPS 2 predicted a 68.7% higher probability. In GI, mGPS 2 predicted a 70.8% higher probability, whereas PS was not significant. BMI did not improve predictive models. PS ≥ 2 and low albumin are strong predictors of short survival in PAN, whereas increased CRP and low albumin (mGPS 2) are predictors in GI.