Protein Concentration Research Articles

Effect of high-pressure homogenization optimized by response surface methodology on the techno-functional properties of protein concentrate isolated from date seed

The study investigates the impact of high-pressure homogenization (HPH) optimized using response surface methodology (RSM) on the techno-functional properties of protein concentrate isolated from date seeds. The protein concentrate was subjected to HPH at various pressures (50, 100, and 150 MPa) and protein concentrations (1 %, 2 %, and 3 %) to optimize solubility, emulsifying activity, and antioxidant properties. RSM indicated optimal conditions at 150 MPa and 1 % protein concentration, resulting in significant enhancements in solubility (14.10 % to 43.69 %) and antioxidant activity (60.5 TE/g to 71.67 TE/g). The emulsifying activity index (EAI) increased from 11.92 m²/g to 22.29 m²/g, though the emulsion stability index (ESI) slightly decreased from 17.63 min to 16.15 min. Besides, HPH improved oil-binding capacity (1.73 g/g to 3.02 g/g) while reducing water-binding capacity (2.76 g/g to 1.38 g/g). Structural analysis revealed that HPH caused partial protein unfolding and aggregation, indicated by changes in FTIR spectra, reduced fluorescence intensity, and increased surface sulfhydryl content (1.58 µmol/g to 2.65 µmol/g). These findings highlight HPH as an effective method to enhance the functional properties of date seed protein concentrate for potential applications in the food industry. In addition, the current study can lay a foundation for future HPH applications in other protein-rich by-products such as soybean cake, rapeseed cake, wheat bran, and others.

Long-term monocyte activation after coronary artery bypass grafting: an exploratory prospective observational study

Major surgery such as coronary artery bypass grafting (CABG) is associated with an increased post-operative risk of atherosclerotic cardiovascular events. Cells of the innate immune system can adopt a long-lasting pro-inflammatory and atherogenic phenotype after brief exposure to exogenous or endogenous inflammatory stimuli, a process called “trained immunity”. We hypothesized that the surgery-induced inflammation leads to sustained alterations in monocyte function, which promote the subsequent occurrence of cardiovascular events. Blood from 13 patients undergoing elective CABG was obtained before, 3-7 days (median 4) after, and 6-8 weeks (median 6) weeks after surgery. At 3-7 days postoperatively, circulating C-reactive protein (CRP) concentration, leukocyte counts and ex vivo Peripheral Blood Mononuclear Cell (PBMC) IL-6, TNFα and IL-1Ra production after stimulation (with various inflammatory stimuli) were significantly increased. Simultaneously, there was a reduction in monocyte HLA-DR expression. 6-8 weeks after CABG there was an ongoing systemic pro-inflammatory state with higher CRP concentrations, increased stimulated ex vivo PBMC IL-6 production, changes in monocytes subsets, and a higher expression of CCR2 on monocytes compared to baseline. In conclusion, CABG induces a persistent systemic inflammatory reaction with a sustained activated monocyte phenotype. This might contribute to the increased atherosclerotic cardiovascular event risk observed in cardiac surgery patients.

Nutritional therapy among adult patients with severe burns: A retrospective observational study.

The objective of this study was to examine the effects of nutritional therapy in adult patients with severe burns. Sixty adult patients with severe burns were enrolled. Data on nutritional intake through enteral nutrition (EN) or parenteral nutrition (PN) on days 7, 14, 21, and 28 post-injury were collected. Patients were divided into target and non-target groups according to whether their energy or protein intake reached the target. Age, length of ventilation, and total bilirubin (TBIL), albumin (ALB), prealbumin (pALB), and C-reactive protein (CRP) concentrations of patients were recorded. The percentage of protein targets with protein delivery was lower than that of energy target with energy delivery. The ratio of PN protein to total protein was lower than that of PN energy to total energy on days 7, 14, 21, and 28 (p<0.001, p<0.001, p=0.001, and p=0.003, respectively). Compared to the non-target group on day 21, the target group was younger, had lower TBIL on day 7, higher ALB and pALB on day 21, and lower CRP on day 14 (p=0.025, p=0.021, p=0.028, p=0.029, and p=0.049, respectively). Multivariate logistic regression analysis showed that older age and longer ventilation were independent risk factors in patients who did not meet the nutritional target on day 21 (p=0.026 and p=0.043, respectively). The protein intake of adult patients with severe burns was low. Compared to the non-target group, the target group had better laboratory test results. Older age and longer ventilation were independent risk factors for patients not meeting the nutritional target.

Bacterial nanocellulose production: Improvement in productivity and properties via a sustainable medium

High production cost is a significant barrier to commercial bacterial nanocellulose (BNC) production. This study addresses this issue using a low-cost molasses and cheese whey medium via Gluconacetobacter hansenii. The one-factor-at-a-time method investigated the effect of critical factors on BNC production, including total sugar and total protein concentrations (g/L), initial pH, and additives such as ethanol and acetic acid (%(v/v)). The productivity in the HS medium was 0.125 g/L/day, while the low-cost medium without additives achieved a productivity of 0.5275 g/L/day. Although the addition of ethanol decreased the productivity, the inclusion of 0.4 %(v/v) acetic acid increased the productivity to 0.64 g/L/day. Using the low-cost medium with acetic acid led to a 40-fold reduction in production costs compared to the HS medium. Furthermore, transitioning from HS media to the low-cost medium resulted in BNC with thicker fibres, a higher crystallinity index (%) and improved mechanical properties. The ratio of Iα/ Iβ in BNC produced in HS media decreased from 1.68 to 1.09 in the low-cost medium. The thermal properties of BNC produced in the low-cost medium also showed slight improvements compared to those in the HS medium. The degree of polymerization significantly increased from 1096 to 1457 in the low-cost medium compared to HS media. These findings highlight the potential of using a low-cost medium to produce BNC with enhanced properties, offering a sustainable and cost-effective alternative to conventional plastics.

Long-term prognosis and related factors of neonatal and small infants meningitis caused by streptococcus agalactiae

Objective: To explore the long-term prognosis and related factors of neonates and infants meningitis caused by Streptococcus agalactiae. Methods: The Clinical data of neonates and small infants with Streptococcus agalactiae meningitis admitted to the Children's Hospital of Zhengzhou University from January 2015 to May 2022 were retrospectively collected, and long-term follow-up of neurological development was conducted for the patients, with a follow-up period of ≥18 months. According to long-term neurological prognosis indicators, the patients were divided into good prognosis group and poor prognosis group, and the multivariate logistic regression model was used to analyze the factors affecting the long-term prognosis of the patients. Results: A total of 86 children were included, with 50 males and 36 females, and their age ranged from 3 to 85 (29.7±18.5) days. The follow-up duration was 20 to 78(47.2±17.0) months, and the age at the end of follow-up was 1.8 to 6.6(4.2±1.3) years. Among them, there were 54 cases in good long-term prognosis group and 32 cases in poor prognosis guoup. Infectious shock [56.3%(18/32) vs 13.0%(7/54), P<0.001], hypoproteinemia [46.9%(15/32) vs 22.2%(12/54), P=0.017], surgical intervention for neurological complications [31.0%(13/32) vs 7.4%(4/54), P=0.003], abnormal neurological examination at discharge [37.5%(12/32) vs 7.4%(4/54), P<0.001], and abnormal cranial imaging at discharge [53.1%(13/32) vs 7.4%(4/54), P<0.001] were higher in the poor prognosis group than those in the good prognosis group. The peripheral blood WBC count [M(Q1, Q3)] [4.7(3.4, 7.6)]×109/L at the onset of the disease was lower in the poor prognosis group than that in the good prognosis group [10.1(3.8, 14.1)]×109/L, and the cerebrospinal fluid protein concentration [3.0(2.1, 3.8)] g/L was higher in the poor prognosis group than that in the good prognosis group [2.5(1.9, 2.8)] g/L (both P<0.05). The multivariate logistic regression model found that peripheral blood WBC<4.0×109/L (OR=5.501,95%CI:1.430-21.170,P=0.013), infectious shock (OR=4.600,95%CI:1.008-21.004,P=0.047), neurological complications undergoing surgical intervention (OR=8.961,95%CI:1.792-44.812,P=0.008), and abnormal head imaging at discharge (OR=12.636,95%CI:2.502-63.829,P=0.002) were related factors for poor long-term prognosis in neonatal and infants meningitis caused by Streptococcus agalactiae. Conclusions: Neonates and infants meningitis caused by Streptococcus agalactiae have a high incidence of long-term adverse prognosis. Peripheral blood WBC<4.0×109/L, infectious shock, neurological complications undergoing surgical intervention, and abnormal cranial imaging at discharge are related factors for poor long-term prognosis.

Comparative analysis of nutritional and mineral compositions of Farfantepenaeus notialis and Macrobrachium vollenhovenii for dietary planning and food industry applications

AbstractThis study compares the nutritional composition of two commercially important crustaceans, Farfantepenaeus notialis and Macrobrachium vollenhovenii, to assess their dietary suitability. Proximate and mineral analyses were performed on dried samples of each species, evaluating fat, protein, ash, moisture, and mineral concentrations. Results reveal significant differences. F. notialis showed higher fat (1.79% ± 0.001%) and protein (6.69% ± 0.2%) compared to M.vollenhovenii (fat: 1.54% ± 0.001%; protein: 5.34% ± 0.4%), making it a more energy-dense protein source. Conversely, M. vollenhovenii had higher ash (5.38% ± 0.002%) and moisture (22.12% ± 0.004%), with higher calcium (256.24 mg/g ± 0.74) and potassium (71.02 mg/g ± 0.76) content, key for bone health and cardiovascular function. F. notialis was richer in iron (5.18 mg/g ± 0.03), useful for preventing iron deficiency. Both species had substantial zinc levels (~ 0.8 mg/g), essential for immune health. T-tests confirmed significant differences in nutritional parameters and minerals. The findings highlight the benefits of including both species in diets for improved nutritional diversity, especially in regions with limited food options. Graphical Abstract

FGF8 protects against polymicrobial sepsis by enhancing the host's anti-infective immunity

Abstract Background Sepsis is characterized by a life-threatening syndrome caused by an unbalanced host response to infection. Fibroblast Growth Factor 8 (FGF8) has been newly identified to play important roles in inflammation and innate immunity, but its role in host response to sepsis is undefined. Methods A cecal ligation and puncture (CLP) -induced mouse sepsis model was established to evaluate the immunomodulatory function of FGF8 during sepsis. The underlying molecular mechanisms were elucidated by cell models using relevant molecular biology experiments. The clinical value of FGF8 in the adjuvant diagnosis of sepsis was evaluated using clinical samples. Results FGF8 protein concentrations were elevated in CLP-induced septic mice compared to controls. In vivo, FGF8 blockade using anti-FGF8 antibody significantly increased mortality and bacterial burden and was paralleled by significantly aggravated tissue injury after CLP. Therapeutic administration of recombinant FGF8 (rFGF8) improved the bacterial clearance and mortality of septic mice in a FGFR1-dependent manner. In vitro, FGF8 directly enhanced bacterial phagocytosis and killing of macrophages by enhancing the phosphorylation of the ERK1/2 signaling pathway, which could be abrogated with the ERK1/2 pathway inhibitor U0126. Clinically, serum FGF8 levels in both adult and pediatric patients with sepsis from ICU were significantly higher than those in healthy controls. Conclusions These results present a previously unrecognized role of FGF8 in improving survival of sepsis by enhancing host immune defense. Therefore, targeting FGF8 may provide new strategies for the diagnosis and immunotherapy of sepsis.

Effects of Hyperbaric Oxygen Therapy in Treatment on Diverse Diseases

INTRODUCTION: Hyperbaric oxygen therapy (HBOT) indeed holds significant therapeutic potential across various medical conditions, primarily by increasing the concentration of oxygen delivered to tissues under pressure. Saturating tissues with oxygen allows for the effective alleviation of underlying hypoxia, facilitating healing, tissue repair and mechanisms of action leads to reducing concentration of pro-inflammatory acute phase proteins, interleukins, and cytokines. Conversely, HBOT while also boosting levels of growth factors and other cytokines that promote angiogenesis. REVIEW METHODS: The article was complied by analyzing data from PubMed and Google Scholar regarding effects of hyperbaric therapy. THE STATE OF KNOWLEDGE: Studies indicate that HBOT provides numerous opportunities in the treatment of extensive diseases acress various systems. Neurodegenerative disorders involve progressive nerve cell damage leading to motor or cognitive decline, exacerbated by oxidative stress and inflammation. HBOT has shown promise in delaying disease onset by improving mitochondrial dysfunction in motor neuron disease. In case of intestinal obstruction HBOT reduces intestinal gas volume, diameter of obstructed loops, and improves intestinal contractility. Significant benefits have been observed in conditions such as Crohn's disease and ulcerative colitis. Hyperbaric therapy presents potential benefits for the infertility, improving the environment for blastocyst implantation and pregnancy. CONCLUSION: There is no doubt that HBOT has a beneficial impact on treatment of many systems such as the nervous, cardiovascular, digestive, and skeletal. This innovative approach of hyperbaric therapy extends to selected disorders such as neurodegenerative diseases, Crohn's disease, ulcerative colitis, and infertility.

From Beer to Cheese: Characterization of Caseinolytic and Milk-Clotting Activities of Proteases Derived from Brewer’s Spent Grain (BSG)

This study explores the extraction and characterization of proteolytic enzymes from brewer’s spent grain (BSG) and their potential as sustainable coagulants in the dairy industry. BSG samples from various beer types (Blonde Ale, IPA, Kölsch, Honey, and Porter) were obtained from two artisanal breweries in Mar del Plata, Argentina. Optimization of caseinolytic activity (CA) and protein extraction was conducted using a Plackett–Burman design, followed by a Box–Behnken design. Optimal protein concentration was achieved at intermediate pH and high temperature, while CA peaked at pH 8.0. The specific caseinolytic activity (SCA) varied among the extracts, with BSG3 showing the highest activity (99.6 U mg−1) and BSG1 the lowest (60.4 U mg−1). Protease inhibitor assays suggested the presence of aspartic, serine, metallo, and cysteine proteases. BSG3 and BSG4 showed the highest hydrolysis rates for α-casein (70% and 78%). For κ-casein, BSG1, BSG2, and BSG3 demonstrated moderate activity (56.5%, 49%, and 55.8), while BSG4 and BSG5 exhibited the lowest activity. Additionally, the milk-clotting activity (MCA) of BSG extracts was comparable to plant-based coagulants like Cynara cardunculus and Ficus carica. These findings highlight the potential of BSG-derived proteases as alternative coagulants for cheese production, offering a sustainable link between the brewing and dairy industries.

Toxic effects of chlorpyrifos on biochemical composition, enzyme activity and gill surface ultrastructure of three species of small fishes from India.

The effects of chlorpyrifos, a frequently detected organophosphate in aquatic ecosystems, on biochemical (protein and glycogen) contents and oxidative enzyme activities (catalase and lipid peroxidation) in liver, muscle and gill tissues of three freshwater fish Trichogaster fasciata, Mystus vittatus and Heteropneustes fossilis were evaluated after 21-day exposure to 1 and 10% of 96h LC50 of this pesticide, which were 1.63 and 16.3µg L-1; 5.87 and 58.7µg L-1 and 2.12 and 21.2µg L-1, respectively. On comparing with control, significant reductions in protein concentration were found in liver, muscle and gill of the three fishes treated with both higher as well as lower concentrations of the pesticide except in gill of M. vittatus and liver of H. fossilis treated with the lower concentrations. Glycogen content reductions were significant in the liver and muscle of the fishes, as well as gill tissue of T. fasciata treated with the two pesticide concentrations. Significant elevations of catalase activity were found in liver of the three fishes treated with the higher concentrations, in muscle tissues of both T. fasciata and M. vittatus treated with both the concentrations and in gills of the three fishes except H. fossilis treated with the lower concentration of the pesticide. Significant elevations of lipid peroxidation level were also found in liver of all the three fish species treated with the higher concentrations, in the muscle tissue of M. vittatus as well as in the gill of T. fasciata and H. fossilis treated with both the concentrations of the pesticide. Chlorpyrifos exposed gill ultrastructure of T. fasciata, M. vittatus and H. fossilis revealed concentration-dependent effects of the pesticide on gill surface ultrastructure which include distortion of primary and secondary lamellae, deterioration of pavement cell and microridge structures, extrusion of red blood cells (RBCs), secretion of mucous layer on filament, sloughing of primary lamellae and clumping of secondary lamellae. The present study parameters could serve as useful biomarkers for evaluating the risk of pesticide toxicity to fish. These findings also point out the possible health risks to the consumers of these fish captured from contaminated water bodies.

Impact of primary sequence changes on the self-association properties of mammalian cystathionine beta-synthase enzymes.

Cystathionine beta-synthase (CBS) is an evolutionarily conserved enzyme that plays a key role in mammalian sulfur amino acid biochemistry, mutations in which are the cause of classical homocystinuria (HCU), an inborn error of metabolism. Although there is agreement in the literature that CBS is a homomultimer, its precise structure is a source of confusion. Here, we performed a series of experiments examining the quaternary structure of various wild-type and mutant CBS enzymes using a combination of native gel electrophoresis, in situ activity assays, analytical ultracentrifugation, and gel filtration. Our data show that recombinantly expressed and purified full-length wild-type human CBS enzyme (hCBS) and HCU-causing variants (p.P422L, p.I435T, and p.R125Q CBS) form high molecular weight assemblies that are consistent with the properties expected of a filament. The filament is enzymatically active, and its size is sensitive to protein concentration. This behavior contrasts sharply with hCBS enzymes containing small deletions within the Bateman domain, which form stable tetramers and octamers regardless of concentration. Examination of liver lysates from humans and mice confirms the existence of enzymatically active high molecular weight aggregates in vivo, but also shows that these aggregates are specific to human CBS and do not occur in mice. Molecular modeling using AlphaFold2 suggests that these experimentally observed differences may be explained by subtle differences in the interaction mediated by the Bateman domains. Our results show that small differences in amino acid sequence can cause large differences in the size and shape of CBS multimers.

Probing the Interplay of Protein Self-Assembly and Covalent Bond Formation in Photo-Crosslinked Silk Fibroin Hydrogels.

Covalent crosslinking of silk fibroin via native tyrosine residues has been extensively explored; however, while these materials are very promising for biomedical, optical, soft robotics, and sensor applications, their structure and mechanical properties are unstable over time. This instability results in spontaneous silk self-assembly and stiffening over time, a process that is poorly understood. This study investigates the interplay between self-assembly and di-tyrosine bond formation in silk hydrogels photo-crosslinked using ruthenium (Ru) and sodium persulfate (SPS) with visible light. The effects of silk concentration, molecular weight, Ru/SPS concentration, and solvent conditions are examined. The Ru/SPS system enables rapid crosslinking, achieving gelation within seconds and incorporating over 90% of silk into the network, even at very low protein concentrations (≥0.75%wt/v). A model emerges where silk self-assembly both before and after crosslinking affects protein phase separation, mesoscale structure, and dynamic changes in the hydrogel network over time. Silk concentration has the greatest impact on hydrogel properties, with higher silk concentration hydrogels experiencing two orders of magnitude increase in stiffness within 1 week. This new understanding and ability to tune hydrogel properties and dynamic stiffening aids in developing advanced materials for 4D biofabrication, sensing, 3D cancer models, drug delivery, and soft robotics.

Determining the rate-limiting processes for cell division in Escherichia coli.

A critical cell cycle checkpoint for most bacteria is the onset of constriction when the septal peptidoglycan synthesis starts. According to the current understanding, the arrival of FtsN to midcell triggers this checkpoint in Escherichia coli. Recent structural and in vitro data suggests that recruitment of FtsN to the Z-ring leads to a conformational switch in actin-like FtsA, which links FtsZ protofilaments to the cell membrane and acts as a hub for the late divisome proteins. Here, we investigate this putative pathway using in vivo measurements and stochastic cell cycle modeling at moderately fast growth rates. Quantitatively upregulating protein concentrations and determining the resulting division timings shows that FtsN and FtsA numbers are not rate-limiting for the division in E. coli. However, at higher overexpression levels, they affect divisions: FtsN by accelerating and FtsA by inhibiting them. At the same time, we find that the FtsZ numbers in the cell are one of the rate-limiting factors for cell divisions in E. coli. Altogether, these findings suggest that instead of FtsN, accumulation of FtsZ in the Z-ring is one of the main drivers of the onset of constriction in E. coli at faster growth rates.

Abstract B022: Characterizing invasiveness in CCCR cells: Role of miR-100, miR-125b and EVs

Abstract Introduction: Extracellular vesicles have been found to be important regulators of intercellular communication between cancer cells. EV cargo varies greatly between different cell types, the composition of which provides important insights into the role of donor and recipient cells. Cetuximab Resistant Colorectal Cancer (CCCR) cells upregulate miR-100 and miR-125b and select them as cargo for their respective secreted EVs. These miRNAs are responsible for altered gene expression in the CCCR cells and can also be functionally transferred as part of EVs between donor and recipient cells. Bioinformatic analysis identified Cingulin (CGN) mRNA as one of the possible targets of miR-100 and miR-125b. The action of miRNAs leads to degradation of CGN mRNA ultimately decreasing cellular concentration of the functional protein. Absence of CGN has been associated with increased invasiveness and metastatic potential in the cancer cells. We carried out immunofluorescent imaging of CCCR cells which revealed downregulation of CGN in CCCR cells compared to the parental Cetuximab sensitive Colorectal Cancer (CC) cells. Similar effects were observed in recipient cells that obtained the miRNAs as part of EVs originating from CCCR donor cells. This project helps elucidate the roles of miR-100 and miR-125b in CCCR cells and how EVs facilitate their functional transfer between nearby cancer cells. Methods: CCCR cell lines were derived from parental CC cells in the lab of Dr. Robert Coffey by iterative selection of cetuximab resistance after growth in 3D. CRISPR/Cas9 technology was used to knockout miR-100 and miR-125b in CCCR cells. Transwell co-culture experiments were carried out to analyze EV transfer of miRNAs. Transfer was monitored by immunofluorescence using antibodies against CGN. Cells were stained and imaged as Z stacks under the 63x Immersion setting. ImageJ was used to analyze the stacks and CGN concentrations between the cells. For quantification of immunofluorescence, additional Z stack images were obtained from an inverted microscope and fluorescence digital camera. Images were analyzed using Fiji software to obtain specific CGN concentrations between the cells. Results and Discussion: Immunofluorescent imaging revealed statistically significant downregulation of CGN in CCCR cells. The protein was also downregulated in the recipient knockout cells as part of the Transwell co-culture experiment with CCCR donor cells. CGN is a tight junction protein and its absence confers increased invasiveness in 3D growth with enhanced metastasis. Decreased CGN concentration in the miR-100 and miR-125b enriched EV recipient knockout cells is indicative of functional transfer of the two miRNAs between cancer cells. These distinctive cellular dynamics could be harnessed to develop potential cancer therapies that provide a better prognosis and limit the metastatic potential of targeted cancer cells. Citation Format: Muhammad Shameer, Hannah M Nelson, Shimian Qu, Liyu Huang, Kevin C Corn, Sydney N Chapman, Nicole L Luthcke, Sara A Schuster, Tellie D Stamaris, Lauren A Turnbull, Lucas L Guy, Xiao Liu, Danielle L Michell, Elizabeth M Semler, Kasey C Vickers, Qi Liu, Jeffrey L Franklin, Alissa M Weaver, Marjan Rafat, Robert J Coffey, James G Patton. Characterizing invasiveness in CCCR cells: Role of miR-100, miR-125b and EVs [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr B022.

A Proteomics-Based Approach for Prediction of Different Cardiovascular Diseases and Dementia.

Many studies have explored whether individual plasma protein biomarkers improve cardiovascular disease risk prediction. We sought to investigate the use of a plasma proteomics-based approach in predicting different cardiovascular outcomes. Among 51 859 UK Biobank participants (mean age, 56.7 years; 45.5% male) without cardiovascular disease and with proteomics measurements, we examined the primary composite outcome of fatal and nonfatal coronary heart disease, stroke, or heart failure (major adverse cardiovascular events), as well as additional secondary cardiovascular outcomes. An exposome-wide association study was conducted using relative protein concentrations, adjusted for a range of classic, demographic, and lifestyle risk factors. A prediction model using only age, sex, and protein markers (protein model) was developed using a least absolute shrinkage and selection operator-regularized approach (derivation: 80% of cohort) and validated using split-sample testing (20% of cohort). Their performance was assessed by comparing calibration, net reclassification index, and c statistic with the PREVENT (Predicting Risk of CVD Events) risk score. Over a median 13.6 years of follow-up, 4857 participants experienced first major adverse cardiovascular events. After adjustment, the proteins most strongly associated with major adverse cardiovascular events included NT-proBNP (N-terminal pro B-type natriuretic peptide; hazard ratio [HR], 1.68 per SD increase), proADM (pro-adrenomedullin; HR, 1.60), GDF-15 (growth differentiation factor-15; HR, 1.47), WFDC2 (WAP four-disulfide core domain protein 2; HR, 1.46), and IGFBP4 (insulin-like growth factor-binding protein 4; HR, 1.41). In total, 222 separate proteins were predictors of all outcomes of interest in the protein model, and 86 were selected for the primary outcome specifically. In the validation cohort, compared with the PREVENT risk factor model, the protein model improved calibration, net reclassification (net reclassification index +0.09), and c statistic for major adverse cardiovascular events (+0.051). The protein model also improved the prediction of other outcomes, including ASCVD (c statistic +0.035), myocardial infarction (+0.023), stroke (+0.024), aortic stenosis (+0.015), heart failure (+0.060), abdominal aortic aneurysm (+0.024), and dementia (+0.068). Measurement of targeted protein biomarkers produced superior prediction of aggregated and disaggregated cardiovascular events. This study represents an important proof of concept for the application of targeted proteomics in predicting a range of cardiovascular outcomes.

Optical characterization of molecular interaction strength in protein condensates.

Biomolecular condensates have been identified as a ubiquitous means of intracellular organization, exhibiting very diverse material properties. However, techniques to characterize these material properties and their underlying molecular interactions are scarce. Here, we introduce two optical techniques - Brillouin microscopy and quantitative phase imaging (QPI) - to address this scarcity. We establish Brillouin shift and linewidth as measures for average molecular interaction and dissipation strength, respectively, and we used QPI to obtain the protein concentration within the condensates. We monitored the response of condensates formed by FUS and by the low-complexity domain of hnRNPA1 (A1-LCD) to altering temperature and ion concentration. Conditions favoring phase separation increased Brillouin shift, linewidth, and protein concentration. In comparison to solidification by chemical crosslinking, the ion-dependent aging of FUS condensates had a small effect on the molecular interaction strength inside. Finally, we investigated how sequence variations of A1-LCD, that change the driving force for phase separation, alter the physical properties of the respective condensates. Our results provide a new experimental perspective on the material properties of protein condensates. Robust and quantitative experimental approaches such as the presented ones will be crucial for understanding how the physical properties of biological condensates determine their function and dysfunction.

The association between adipokines dysregulation and the occurrence of atrial fibrillation and obese patients, is it relevant?

Atrial fibrillation (AF) is the most common arrhythmia, affecting approximately 33.5 millions of people worldwide. Unfortunately, the prevalence of this arrhythmia will increase within the following two decades, resulting in a higher mortality rate and a higher economic burden for public health services. Obesity, specifically central obesity, plays an essential role in developing AF by increasing pericardial fat and epicardial adipose tissue thickness, generating a chronic inflammation state where dysregulation of the serum concentration of several proinflammatory proteins occurs and indirectly promotes AF. Therefore, recent research has focused on analyzing the circulating concentration of different molecules, including pro and anti-inflammatory adipokines, and their association with AF. Herein, we review several studies addressing the association of adipokines with the onset or recurrence of AF to establish such association as a potential biomarker for the prevention or adequate treatment of this arrhythmia. We concluded that the insight into this topic is very controversial and needs further research.

Features оf Changes in the Level of Neurotransmitters and Markers of Neurodegeneration in the Brain of Rats with Experimental Parkinson’s Disease Against the Background of Course Therapy with Steroid Glycoalkaloids

Parkinson’s disease is a chronic neurodegenerative disease, one of the pathobiochemical manifestations of which is an imbalance of neurotransmitters in the brain. The aim of this work is to study the effect of glycoalkaloids on changes in the level of neurotransmitters and markers of brain neurodegeneration in rats with experimental Parkinson’s disease. Parkinson’s disease was modeled in animals by intracerebral administration of rotenone. The studied glycoalkaloids were administered orally at a dose of 0.06 mg / kg, a course of 28 days after pathology modeling. The study found that the use of the analyzed glycoalkaloids did not affect the concentration of dopamine and serotonin, and also significantly (p 0.05) reduced the level of ACh by 35.9% and increased the level of GABA by 55.1%, reduced the concentration of S100β protein by 68.6% and increased the level of BDNF by 52.4%.

Associations Between Plasma Levels of NLRP3 Protein, Interleukin-1 Beta and Features of Acute ST-Elevation Myocardial Infarction

Background. Phenotyping inflammation in ST-elevation myocardial infarction (STEMI) is a challenge for modern cardiology. NLRP3 inflammasome is a proven predictor of adverse outcomes in cardiovascular disease, but its specificity in stratifying inflammatory activity in patients with myocardial infarction (MI) has not been demonstrated. The aim of this paper is to describe the levels of NLRP3 protein and IL-1β concentrations and their changes in dynamics and associations with clinical, laboratory and instrumental characteristics of patients with STEMI. Methods. A total of 45 patients with STEMI were enrolled. Concentrations of NLRP3 and IL-1β were evaluated in arterial and venous EDTA blood from the infarct-related coronary and peripheral arteries and veins on days 1, 3 and 7 after MI. Results and Conclusions. The concentrations of markers were higher on the first day after MI with a maximum decrease on the third day. The levels of both markers in venous plasma correlated with those in arterial blood, allowing their routine determination in venous plasma on the first day after MI. IL-1β levels correlated directly with the wall motion index and inversely with left ventricular ejection fraction and stroke volume, which characterize the potential contribution to adverse myocardial remodeling. There were two multidirectional trends in changes in NLRP3 and IL-1β levels during hospitalization. Initially higher levels with a gradual decrease by day 7 were associated with a longer duration of myocardial ischemia and higher plasma troponin I levels. Further evaluation of the long-term outcomes of MI will allow identifying inflammatory factors that input to the development of secondary major adverse cardiac events and will provide a new step in the understanding of inflammatory phenotyping.

Disinfection effect of ozonated water on SARS-CoV-2 in the presence of salivary proteins.

Ozonated water is expected to be an effective disinfectant for SARS-CoV-2 present on environmental fomites; however, ozone is consumed by organic substances, resulting in attenuation of its effect. SARS-CoV-2 present in saliva can contaminate environmental surfaces; therefore, it is essential to understand the effect of organic substances in saliva on the disinfectant properties of ozonated water. To assess organic factors in saliva and the extent to which they diminish the effect of ozonated water on SARS-CoV-2. Ozonated water was exposed to salivary organic factors and residual ozone concentrations were measured. SARS-CoV-2 was exposed to a salivary factor and virus inactivation by ozonated water was measured. Amylase and mucin consumed ozone in a concentration-dependent manner. Urea did not. Ozonated water appeared to inactivate SARS-CoV-2 within 30 sec. The amount of inactivated SARS-CoV-2 decreased as the protein concentration increased. Virus inactivation was stronger by 1.5 mg/L ozonated water than by 0.5 mg/L ozonated water. This study suggests that the salivary amylase and mucin decay ozone in a concentration-dependent manner, thereby attenuating the disinfection properties of ozonated water for SARS-CoV-2. An increase of the initial amount of ozone can ameliorate the disinfection effect of ozonated water on SARS-CoV-2. Ozone consumption should be taken into consideration for virus infection control. These results provide fundamental information about the effect of ozonated water when used to decontaminate surfaces harbouring SARS-CoV-2 in saliva.