Plasma Protein Concentration Research Articles

Metabolic and Intestinal Morphometric Responses of Nile Tilapia Fed Diets Containing Soybean and Protease

Proteases facilitate the breakdown of components associated with antinutritional factors. This study evaluated the effects of protease supplementation in Nile tilapia diets on metabolic and intestinal responses. Three diets were tested: soybean meal SM1 (fish meal FM:SM = 1:1), SM2 (1:2), and SM3 (1:3), based on the protein content. All the diets were without or with protease (0.44 g/kg). The fish were randomly allocated to 18 tanks/49 days. In SM3 without protease, an increase in the villus height and width, as well as the number of goblet cells, was observed. Protease supplementation increased the villus height compared to diets without the enzyme. The SM1 with protease resulted in a reduction in the digestive somatic index, but significantly increased the hepatosomatic and intestinal quotient indices. In SM2 with protease, reductions in the amino acid (AA) content and aspartate aminotransferase activity in the liver were observed. In the muscles, supplementation increased the AA concentrations. Higher plasma AA and albumin concentrations were detected in the SM2 and SM3 groups with protease. A significant interaction was found, with SM3 without protease showing lower plasma protein and AA concentrations. In SM2 with protease, higher levels of albumin and globulin were recorded. Protease in soybean meal diets improved the intestinal health and metabolism in tilapia.

C-Reactive Protein, International Normalized Ratio, and Fibrinogen in Diagnostic Scale of Complicated Acute Appendicitis

Background/Objectives: Differentiating complicated acute appendicitis (CA) and uncomplicated acute appendicitis (UC) is essential to guide clinical management. While CA requires urgent surgical management, UC can be treated with antibiotic therapy in selected cases. However, accurate identification of CA remains a clinical challenge. This study aimed to identify factors associated with CA and to develop a diagnostic severity scale. Methods: In this retrospective study, we included 132 adult patients (>16 years) with a confirmed postsurgical diagnosis of appendicitis, of whom 52 had CA and 80 had UA. Signs, symptoms, comorbidities, laboratory values, and ultrasonographic findings were evaluated to determine predictive factors and construct a diagnostic scale. Results: The factors most significantly associated with CA were elevated plasma concentrations of C-reactive protein (>7.150 mg/dL), fibrinogen (481.5 mg/dL), International Normalized Ratio (INR) (>1.150), and the presence of free fluid periappendicular. The combination of these factors within one scale showed an area under the curve (AUC) of 0.84, with a sensitivity of 78.75% and a specificity of 82.69%. Conclusions: Serum C-reactive protein concentration, fibrinogen, and INR can be employed individually or as part of a scale as important indicators in diagnosing CA.

Application of Principal Component Analysis as a Prediction Model for Feline Sporotrichosis.

Sporotrichosis is a worldwide zoonotic disease that is spreading and causing epidemics in large urban centers. Cats are the most susceptible species to develop the disease, which could cause significant systemic lesions. The aim was to investigate and to identify predictive indicators of disease progression by correlations between the blood profile (hematological and biochemical analytes) and cutaneous lesion patterns of 70 cats diagnosed with Sporothrix brasiliensis. The higher occurrence in male cats in this study could be related to being non-neutered and having access to open spaces. Principal component analysis (PCA) with two principal components, followed by binary logistic regression, and binary logistic regression analysis, with independent variables and backward elimination modeling, were performed to evaluate hematological (n = 56) and biochemical (n = 34) analytes, including red blood cells, hemoglobin, hematocrit, leukocytes, segmented neutrophils, band neutrophils, eosinophils, lymphocytes, monocytes, total plasma protein, albumin, urea, creatinine, and alanine aminotransferase. Two logistic regression models (PCA and independent variables) were employed to search for a predicted model to correlate fixed (isolated) and disseminated cutaneous lesion patterns. Total plasma protein concentration may be assessed during screening diagnosis as it has been recognized as an independent predictor for the dissemination of cutaneous lesion patterns, with the capability of serving as a predictive biomarker to identify the progression of cutaneous lesions induced by S. brasiliensis infections in cats.

The impact of measurement techniques and common effectors on red blood cell aggregation

Background Red blood cell aggregation is largely influenced by hematocrit and plasma protein content with aggregation increasing as hematocrit and fibrinogen increase. Aggregation measurement techniques include light-transmission or laser-backscatter, and the impact of measurement technique on aggregation measurements with respect to changes in hematocrit and plasma protein is not well understood. Objective The objective of this study was to evaluate aggregation measurement techniques with respect to common effectors of red blood cell aggregation. Methods We obtained whole blood samples from 10 healthy participants and varied hematocrit, plasma albumin, and plasma fibrinogen concentration in a sample of healthy male and female participants. We then measured red blood cell aggregation using a Myrenne Aggregometer (light-transmission aggregometer) and a Laser Optical Rotational Red Cell Analyzer (laser-backscatter aggregometer). Results For Myrenne, aggregation increased from 30% to 40% hematocrit, then declined at 50% hematocrit. Measurement in the LORRCA demonstrated an increase as hematocrit increased from 30% to 40%, and again increased at 50%, which differed from the light transmission method. Red blood cell aggregation increased with increased fibrinogen concentration, but did not change significantly as albumin concentration increased. Conclusions Red blood cell aggregation is expected to increase as hematocrit increases, but when light transmission aggregometry is used, aggregation decreases as hematocrit increases past 40%. Each measurement technique has its own set of methodological strengths and weaknesses.

Neonates exposed to HIV but uninfected exhibit an altered gut microbiota and inflammation associated with impaired breast milk antibody function

BackgroundInfants exposed to HIV but uninfected have altered immune profiles which include heightened systemic inflammation. The mechanism(s) underlying this phenomenon is unknown. Here, we investigated differences in neonatal gut bacterial and viral microbiome and associations with inflammatory biomarkers in plasma. Further, we tested whether HIV exposure impacts antibody-microbiota binding in neonatal gut and whether antibodies in breast milk impact the growth of commensal bacteria.ResultsNeonates exposed to HIV but uninfected (nHEU) exhibited altered gut bacteriome and virome compared to unexposed neonates (nHU). In addition, HIV exposure differentially impacted IgA-microbiota binding in neonates. The relative abundance of Blautia spp. in the whole stool or IgA-bound microbiota was positively associated with plasma concentrations of C-reactive protein. Finally, IgA from the breast milk of mothers living with HIV displayed a significantly lower ability to inhibit the growth of Blautia coccoides which was associated with inflammation in nHEU.ConclusionnHEU exhibits profound alterations in gut bacterial microbiota with a mild impact on the enteric DNA virome. Elevated inflammation in nHEU could be due to a lower capacity of breast milk IgA from mothers living with HIV to limit growth the of gut bacteria associated with inflammation.BDUn311MBkuDfDvveFNavEVideo

Brain endothelial permeability, transport, and flow assessed over 10 orders of magnitude using the in situ brain perfusion technique

BackgroundCerebral blood flow normally places a limit on the magnitude of brain vascular permeability (P) that can be measured in vivo. At normal cerebral blood flow, this limit falls at the lower end of lipophilicity for most FDA-approved CNS drugs. In this study, we report on two methods that can be used to overcome this limitation and measure brain vascular permeability values that are up to ~1000 times higher using the in situ brain perfusion technique.MethodsRat brain was perfused with physiological saline at increased flow rate and in the presence of various concentrations of plasma protein, serum albumin or alpha-acid glycoprotein. Plasma protein was added to the saline perfusion fluid to lower extraction into the measurable range using the Crone Renkin “diffusion-flow” equation to calculate brain PoS.ResultsCerebrovascular Po was determined for 125 solutes, of which 78 showed little or no evidence of active efflux transport. Fifty of the solutes were in the lipophilicity zone (Log Poct 1–5) of most FDA-approved CNS drugs. Care was taken to ensure the integrity of the brain vasculature during perfusion and to measure flow accurately using markers that had been verified for the flow rates. The results showed a linear relationship between Log Po and Log Poct over ~10 orders of magnitude with values for diazepam, estradiol, testosterone, and other agents that exceed prior published values by fivefold to 200-fold.ConclusionsThe results show that brain vascular permeability can be measured directly in vivo for highly lipophilic solutes and the PS values obtained match reasonably with that predicted by the Crone-Renkin flow diffusion equation with care taken to validate the accuracy for the component measurements and with no need to invoke “enhanced” or “induced” dissociation.

Prediction of Major Adverse Cardiovascular Events in Patients with Peripheral Artery Disease Using Circulating Immunomodulatory Proteins.

Background/Objectives: The leading cause of death for people with peripheral artery disease (PAD) is major adverse cardiovascular events (MACE), including heart attacks and strokes. However, research into biomarkers that could help predict MACE in patients with PAD has been limited. Immunomodulatory proteins are known to significantly influence systemic atherosclerosis, suggesting they could be useful prognostic indicators for MACE in patients with PAD. In this study, we evaluated a broad panel of immunomodulatory proteins to identify those linked to MACE in individuals with PAD. Methods: We conducted a prognostic study involving a prospectively recruited cohort of 406 patients consisting of 254 with PAD and 152 without PAD. At the baseline, we measured the plasma concentrations of 17 circulating immunomodulatory proteins and followed the cohort for two years. The primary outcome was 2-year MACE, a composite of myocardial infarction, stroke, or death. Plasma protein concentrations were compared between patients with PAD with and without 2-year MACE using Mann-Whitney U tests. We further examined the prognostic potential of differentially expressed proteins through a Cox proportional hazards analysis, determining their independent associations with 2-year MACE while controlling for all the baseline demographic and clinical characteristics, including the existing coronary artery and cerebrovascular diseases. Additionally, A Kaplan-Meier analysis was performed to evaluate the 2-year freedom from MACE in patients with low versus high levels of the differentially expressed proteins based on the median plasma concentrations. Results: The mean age of the cohort was 68.8 years (SD 11.1), with 134 patients (33%) being female. During the two-year follow-up, 63 individuals (16%) developed MACE. The following proteins were significantly elevated in patients with PAD who experienced 2-year MACE compared to those who did not: galectin-1 (0.17 [SD 0.06] vs. 0.10 [SD 0.07] pg/mL, p = 0.012), alpha-1-microglobulin (16.68 [SD 7.48] vs. 14.74 [SD 6.71] pg/mL, p = 0.019), and galectin-9 (0.14 [SD 0.09] vs. 0.09 [SD 0.05] pg/mL, p = 0.033). The Cox proportional hazards analysis indicated that these three proteins were independently associated with 2-year MACE after adjusting for all the baseline demographic and clinical factors: galectin-1 (HR 1.45 [95% CI 1.09-1.92], p = 0.019), alpha-1-microglobulin (HR 1.31 [95% CI 1.06-1.63], p = 0.013), and galectin-9 (HR 1.35 [95% CI 1.02-1.78], p = 0.028). Over the two-year follow-up, patients with higher levels of galectin-1, galectin-9, and alpha-1-microglobulin had a lower freedom from MACE. Additional analysis showed that these three proteins were not significantly associated with 2-year MACE in patients without PAD. Conclusions: Among the 17 immunomodulatory proteins evaluated, galectin-1, galectin-9, and alpha-1-microglobulin were found to be independently and specifically associated with 2-year MACE in patients with PAD. Assessing the plasma concentrations of these proteins can aid in risk stratification for MACE in patients with PAD, helping to inform clinical decisions regarding multidisciplinary referrals to cardiologists, neurologists, and vascular medicine specialists. This information can also guide the aggressiveness of medical management, ultimately improving cardiovascular outcomes for patients with PAD.

Identification and Evaluation of Angiogenesis-Related Proteins That Predict Major Adverse Cardiovascular Events in Patients with Peripheral Artery Disease.

The most common cause of death in patients with peripheral artery disease (PAD) are major adverse cardiovascular events (MACEs), including myocardial infarction (MI) and stroke. However, data on biomarkers that could be used to help predict MACEs in patients with PAD to guide clinical decision making is limited. Angiogenesis-related proteins have been demonstrated to play an important role in systemic atherosclerosis and may act as prognostic biomarkers for MACEs in patients with PAD. In this study, we evaluated a large panel of angiogenesis-related proteins and identified specific biomarkers associated with MACEs in patients with PAD. We conducted a prognostic study using a prospectively recruited cohort of 406 patients (254 with PAD and 152 without PAD). Plasma concentrations of 22 circulating angiogenesis-related proteins were measured at baseline, and the cohort was followed for 2 years. The primary outcome was 2-year MACEs (composite of MI, stroke, or death). Plasma protein concentrations were compared between PAD patients with and without 2-year MACEs using Mann-Whitney U tests. Differentially expressed proteins were further investigated in terms of their prognostic potential. Specifically, Cox proportional hazards analysis was performed to determine the independent association between differentially expressed proteins and 2-year MACEs, controlling for all baseline demographic and clinical characteristics, including existing coronary artery disease and cerebrovascular disease. Kaplan-Meier analysis was conducted to assess 2-year freedom from MACEs in patients with low vs. high levels of the differentially expressed proteins based on median plasma concentrations. The mean age of the cohort was 68.8 (SD 11.1), and 134 (33%) patients were female. Two-year MACEs occurred in 63 (16%) individuals. The following proteins were significantly elevated in PAD patients with 2-year MACEs compared to those without 2-year MACEs: endostatin (69.15 [SD 58.15] vs. 51.34 [SD 29.07] pg/mL, p < 0.001), angiopoietin-like protein 4 (ANGPTL4) (0.20 [SD 0.09] vs. 0.12 [SD 0.04] pg/mL, p < 0.001), and ANGPTL3 (51.57 [SD 21.92] vs. 45.16 [SD 21.90] pg/mL, p = 0.001). Cox proportional hazards analysis demonstrated that these three proteins were independently associated with 2-year MACEs after adjusting for all baseline demographic and clinical characteristics: endostatin (HR 1.39 [95% CI 1.12-1.71] p < 0.001), ANGPTL4 (HR 1.35 [95% CI 1.08-1.68], p < 0.001), and ANGPTL3 (HR 1.35 [95% CI 1.12-1.63], p < 0.001). Over a 2-year follow-up period, patients with higher levels of endostatin, ANGPTL4, and ANGPTL3 had a lower freedom from MACEs. Supplementary analysis demonstrated that these three proteins were not significantly associated with 2-year MACEs in patients without PAD. Among a panel of 22 angiogenesis-related proteins, endostatin, ANGPTL4, and ANGPTL3 were identified to be independently and specifically associated with 2-year MACEs in patients with PAD. Measurement of plasma concentrations of these proteins can support MACE risk stratification in patients with PAD, thereby informing clinical decisions on multidisciplinary referrals to cardiologists, neurologists, and vascular medicine specialists and guiding aggressiveness of medical treatment, thereby improving cardiovascular outcomes in patients with PAD.

Physiological Stress Response and Oxidative Status in Tambaqui (Colossoma macropomum) Fed Diets Supplemented with Selenium.

The present study aimed to evaluate the physiological responses to transport stress in juvenile tambaqui (Colossoma macropomum) fed a diet supplemented with hydroxy-selenomethionine (OH-SeMet; Selisseo®, Adisseo) and determine through stress biomarkers whether selenium supplementation could reduce the impact of transport stress on tambaqui resilience. Juvenile fish (15.71 ± 1.90 g) were randomly distributed into five experimental groups; one fed a basal diet without inclusion of OH-SeMet (0.0 mg Se/kg) and four other groups fed supplemented diets with selenium in the form of OH-SeMet at concentrations of 0.3, 0.6, 0.9, and 1.2 mg Se/kg over 75 days. After this period, the fish were transported for 4 h in plastic bags (60 L), and sampling took place before transport (baseline), immediately after transport, and 24 h later. The effects of dietary OH-SeMet in tambaqui challenged by transport were studied in terms of antioxidative status (superoxide dismutase, catalase, glutathione peroxidase, glutathione s-transferase, and reduced glutathione); stress markers (serum cortisol, blood glucose, ionic and hematological parameters), and immunological status (respiratory burst activity and total protein, A/G ratio). Following transport, we observed an elevation in cortisol and blood glucose concentrations, accompanied by hemoconcentration, as evidenced by elevated hematocrit, erythrocyte count, hemoglobin concentration, and mean corpuscular hemoglobin concentration (MCHC). Additionally, there was a decrease in immune response, indicated by reduced respiratory activity of leukocytes and an osmotic imbalance, which culminated in hypochloremia, increased plasma protein concentration, and a rise in red blood cell volume, all of which were measured 24 h after the stressor was applied. Twenty-four hours after transport, the antioxidant defense system was activated, and no recovery of homeostasis was observed for most of the parameters analyzed. The dietary inclusion of selenium did not alleviate the hormonal, metabolic, hematological, and ionic changes caused by transport, but at a concentration of 0.9 mg Se/kg, it was able to increase the activity of enzymes related to oxidative stress (catalase and superoxide dismutase, mainly) in tambaqui juveniles submitted to this challenge.

Misfolding of Transthyretin In Vivo is Controlled by the Redox Environment and Macromolecular Crowding

Transthyretin (TTR) amyloidosis is a progressive disorder characterized by peripheral neuropathy, autonomic dysfunction, and cardiomyopathy. The precise mechanism by which TTR misfolds and forms fibrils in vivo remains incompletely understood, posing challenges to the development of effective therapeutics. In this study, we reveal that the recently identified non-native pathological species (NNTTR), which is enriched in the plasma of ttr-val30met gene carriers, exhibits strong amyloidogenic properties, making it a promising therapeutic target. Notably, we demonstrate that NNTTR formation is dependent on an intermolecular disulfide bond and can be promoted by oxidative conditions while being effectively suppressed by reducing agents. The formation of this disulfide bond is incompatible with the native TTR fold, thereby necessitating structural flexibility. We further show that this required flexibility can be constrained using tetramer-stabilizing drugs, thereby suppressing NNTTR formation. Interestingly, the flexibility is also hindered by macromolecular crowding, and NNTTR formation is strongly suppressed by the high protein concentration in plasma. This suppression is released upon dilution, which thus promotes NNTTR formation in areas with lower protein content, highlighting a potential link to the interstitial space, brain, and vitreous body of the eye, where TTR-amyloid is frequently observed. In summary, we demonstrate that NNTTR displays strong amyloidogenic features, underscoring its potential as a therapeutic target. We identify the redox environment and macromolecular crowding as key modulatory factors. Our findings propose a mechanistic explanation for TTR misfolding and suggest a novel therapeutic approach.

In vitro Effect of Low-level Lasers on Proteomic Concentration in Human Blood Plasma Using 375 nm and 650 nm Lasers

Abstract Background: As a continuation of earlier laboratory research and its findings, we are studying the effects of biostimulation and alteration on human blood plasma to improve blood circulation in blood vessels, treat some infections, and treat various diseases, including blood protein-related ones. Methods: Blood samples were collected through venipuncture into tubes containing, ethylenediaminetetraacidic as an anticoagulant from healthy adult donors, and plasma was separated from blood components. Blood plasma samples were irradiated for varying periods (5, 10, 15, and20) min. Before and after irradiation, total protein and albumin concentrations were calculated using 375 nm and 650 nm lasers. Using a spectrophotometer, the concentration of total protein and albumin was determined for each sample. Results: At the (375 and 650) nm laser wavelength and exposure durations of (5, 10, 15, and 20) min, it was observed that the total protein concentration had significant differences between pre- and postirradiation probate value (P = 0.05, P = 0.05, P = 0.05, and P = 0.05, respectively). It was observed that the total protein and albumin concentrations had significant differences between pre- and postirradiation. In addition, the results demonstrate that the concentration of total protein and albumin decreases more significantly at a laser wavelength of 650 nm compared to a laser wavelength of 375 nm at times of (5 and 10) min. Conclusions: Our results clearly indicate that low-level lasers with different wavelengths of ( 375 , 630) nm both affect the concentration of total protein and albumin in human blood plasma, which can contribute to the treatment of many pathological conditions in the future.

Proteomic and genomic evaluation of plasma protein biomarkers for atrial fibrillation

Abstract Background Atrial fibrillation (AF) is a common arrhythmia and an important cause of ischemic stroke. Several plasma proteins have been suggested as biomarkers for the risk of AF and subsequent stroke in previous studies, but validation in independent cohorts is needed. Large-scale integration of proteomics with genetic and clinical data holds potential for refinement of disease prediction and risk stratification. Purpose This study aimed to evaluate previously suggested plasma protein biomarkers of AF and ischemic stroke using plasma protein measurements in a large cohort. Methods Ten tentative protein biomarkers previously associated with AF or ischemic stroke in AF patients, were evaluated in a nested case-control study comprising 49,757 individuals from the UK biobank (UKB). Plasma proteins were measured by normalized protein expression on the Olink platform. Outcomes were incident AF and ischemic stroke. The proteins were evaluated in Cox regression models adjusted for sex, age, and clinical risk factors. To assess whether the proteins were independent predictors, they were evaluated in sensitivity analyses with additional adjustment for NT-proBNP levels. Polygenic risk scores were calculated for plasma protein concentrations using the PRS-CS-auto algorithm and tested for association with AF or ischemic stroke in an independent subset of the UKB cohort without plasma protein measurements (n=379,392). Results All ten plasma proteins were associated with incident AF in the primary model, and eight were associated with incident ischemic stroke (P-values &amp;lt; 0.0025 for all). Adjusted for NT-proBNP levels, the proteins interleukin-6, GDF15, angiopoietin 2, and Troponin-I remained independently associated with AF, and the proteins GDF15, Cystatin-C, interleukin-6, ADAMTS13, and FGF23 remained independently associated with ischemic stroke. Polygenic risk scores for NT-proBNP and angiopoietin 2 were associated with increased rates of AF. No polygenic scores were significantly associated with ischemic stroke. Conclusion Large-scale proteomic evaluation validated ten plasma proteins associated AF and found eight to be associated with ischemic stroke. Seven proteins were associated with AF or ischemic stroke independently of NT-proBNP levels. Additional genomic evaluation provided evidence of associations between incident AF and genetically predicted plasma levels of angiopoietin 2 and NT-proBNP. As proteomic analyses of the plasma proteome become cheaper and more readily available, these findings could contribute to future risk stratification and selection of individuals for AF screening programs.Proteins associated with AFProteins associated with ischemic stroke

Deep phenotyping of pulmonary edema and pulmonary vascular permeability in COVID-19 ARDS.

Clinical monitoring of pulmonary edema due to vascular hyperpermeability in acute respiratory distress syndrome (ARDS) poses significant clinical challenges. Presently, no biological or radiological markers are available for quantifying pulmonary edema. Our aim was to phenotype pulmonary edema and pulmonary vascular permeability in patients with coronavirus disease 2019 (COVID-19) ARDS. Transpulmonary thermodilution measurements were conducted in 65 patients with COVID-19 ARDS on the day of intubation to determine the extravascular lung water index (EVLWi) and pulmonary vascular permeability index (PVPi). In parallel, ventilatory parameters, clinical outcomes, the volume of lung opacity measured by chest computed tomography (CT), radiographic assessment of lung edema (RALE) score by chest radiography, and plasma proteomics (358 unique proteins) were compared between tertiles based on the EVLWi and PVPi. Regression models were used to associate EVLWi and PVPi with plasma, radiological, and clinical parameters. Computational pathway analysis was performed on significant plasma proteins in the regression models. Patients with the highest EVLWi values at intubation exhibited poorer oxygenation parameters and more days on the ventilator. Extravascular lung water strongly correlated with the total volume of opacity observed on CT (r = 0.72, P < 0.001), whereas the PVPi had weaker associations with clinical and radiological parameters. Extravascular lung water did not correlate with the RALE score (r = 0.15, P = 0.33). Plasma protein concentrations demonstrated a stronger correlation with PVPi than with EVLWi. The highest tertile of PVPi was associated with proteins linked to the acute phase response (cytokine and chemokine signaling) and extracellular matrix turnover. In the clinical setting of COVID-19 ARDS, pulmonary edema (EVLWi) can be accurately quantified through chest CT and parallels deterioration in ventilatory parameters and clinical outcomes. Vascular permeability (PVPi) is strongly reflected by inflammatory plasma proteins.NEW & NOTEWORTHY This study is unique in that it phenotypes pulmonary edema in COVID-19 ARDS using various clinical parameters and biomarkers. First, there is a noteworthy tipping point in the amount of pulmonary edema at which ventilatory and clinical parameters deteriorate. Second, chest CT gives a good approximation of the amount of pulmonary edema. Finally, pulmonary vascular permeability is strongly reflected by inflammatory plasma proteins.

Plasma proteomic signatures of liver steatosis and fibrosis in people living with HIV: a cross-sectional study

Insights into the mechanisms driving metabolic dysfunction-associated steatotic liver disease (MASLD) in people living with HIV (PLHIV) remain limited. Plasma proteomics holds promise for biomarker discovery and the elucidation of biological mechanisms. We performed cross-sectional analyses on data from 1036 virally suppressed PLHIV using antiretroviral treatment (ART) from the Dutch multi-centre 2000HIV cohort. Participants underwent transient elastography to assess liver steatosis (controlled attenuation parameter (CAP)≥263dB/m) and -fibrosis (liver stiffness measurement (LSM)≥7.0kPa). Plasma protein concentrations (n=2367) (Olink® Explore Panel) were compared between PLHIV with vs. without liver steatosis and PLHIV with vs. without fibrosis. Enriched pathways (using GO, KEGG and Reactome libraries) and correlations with clinical characteristics wereassessed, and analyses were stratified by BMI category. In addition, concentrations of 242 proteins werecompared between individuals ("controls") with and without liver steatosis (ratio of methylene:methylene and water >5.6% on magnetic resonance spectroscopy) from a separate cohort (300-OB), all having a BMI >26kg/m2. Steatosis and fibrosis were associated with 67/2367 (2.2%) and 17/2367 (0.7%) differentially expressed proteins (DEP), respectively, enriched in mostly metabolic pathways. Immunoglobulin superfamily member 9 (IGSF9) was amongst the top DEP associated with both steatosis and fibrosis. Stratifying by BMI revealed 8/2367 DEP associated with steatosis in lean- and 12/2367 DEP in overweight/obese individuals, with two shared DEP (IGSF9 and GHR). Conversely, protein signatures of overweight/obese PLHIV (32/242 DEP) and overweight/obese HIV-uninfected individuals (32/242 DEP) exhibited substantial overlap with 16 shared DEP. Notably, DEP correlated with HIV characteristics in lean individuals but not in overweight/obese PLHIV. Lean and overweight/obese PLHIV exhibit distinct proteomic signatures associated with liver steatosis, with the former being more strongly correlated with HIV-specific factors and ART. In addition, we identified a protein, IGSF9, strongly related to liver fibrosis and steatosis across BMI categories. The 2000HIV study is funded by ViiV Healthcare.

Proteomic analysis of plasma-derived extracellular vesicles: pre- and postprandial comparisons

Extracellular vesicles (EVs) are key in intercellular communication, carrying biomolecules like nucleic acids, lipids, and proteins. This study investigated postprandial characteristics and proteomic profiles of blood-derived EVs in healthy individuals. Twelve participants fasted overnight before baseline assessments. After consuming a controlled isocaloric meal, EVs were isolated for proteomic and flow cytometric analysis. Plasma triacylglyceride levels confirmed fasting completion, while protein concentrations in plasma and EVs were monitored for postprandial stability. Proteomic analysis identified upregulated proteins related to transport mechanisms and epithelial/endothelial functions postprandially, indicating potential roles in physiological responses to nutritional intake. Enrichment analyses revealed vesicle-related pathways and immune system processes. Flow cytometry showed increased expression of CD324 on CD9+CD63+CD81+ large extracellular vesicles postprandially, suggesting an epithelial origin. These findings offer valuable insights into postprandial EV dynamics and their potential physiological significance, highlighting the need for stringent fasting guidelines in EV studies to account for postprandial effects on EV composition and function.

Sub-chronic Toxicological Evaluation of Drinking Water from Internally Displaced Persons (IDP) Camp in Wassa, Federal Capital Territory, Abuja, Nigeria

The number of Internally Displaced Persons (IDPs) camps has increased recently in Nigeria due to insurgency and natural disasters such as floods. This study aimed to evaluate the sub-chronic toxicological effects of the source of the drinking water in Wassa camp, Abuja, Nigeria. Forty-eight (48) mice were randomized into four (4) groups of twelve (12) mice each and orally administered different concentrations (5, 10, and 20 mL/kg body weight respectively) of the water sample from Wassa IDP for fourteen (14) days. The alteration was noticed in the liver and kidney indices of mice by a significant increase (p&lt;0.05) in kidney-body weight ratio, kidney, liver and plasma protein concentrations, and a considerable decrease (p&lt;0.05) in albumin, creatinine, uric acid and total bilirubin concentrations, 14-days post-treatment, particularly at a higher dose. Gamma-glutamyl transaminase in the liver and aspartate transaminase in both liver and kidney significantly (p&lt;0.05) increased in mice with a resultant significant decrease (p&lt;0.05) in the serum, 14 days post-treatment at 20 mL/kg bwt. There was no significant difference (p&gt;0.05) observed in serum triglycerides, very-low-density-lipoprotein cholesterol concentrations at all doses 14-days post-treatment compared with the control but the concentrations of serum low-density-lipoprotein cholesterol and total cholesterol significantly reduced (p&lt;0.05) at 20 mL/kg bwt 14-days post-treatment. The drinking water altered some liver-function indices while the cardiac-function indices were less affected. Therefore, prolonged consumption of the water may not be relatively safe. The provision of clean drinking water for the Wassa IDP camp will prevent adverse biochemical reaction reactions from prolonged water consumption.

The measurement of plasma fructosamine as a diagnostic tool to improve the interpretation of plasma glucose and proteins in alpacas (Vicugna pacos).

This study assesses the use of fructosamine as a diagnostic tool for hyperglycemia in alpacas in view of their sensitivity to stress and susceptibility to conditions like lipid mobilization syndrome. Plasma fructosamine, like in diagnosing diabetes in cats and dogs, can reveal long-term blood glucose trends, differentiating stress-induced spikes from persistent diabetic hyperglycemia. In 125 alpacas presented as patients of a veterinary clinic, plasma glucose and fructosamine concentrations were compared for correlations with findings of the general clinical examination, laboratory parameters, demographic data, and a behavioral stress assessment processed by using principal component analysis. Hyperglycemia was observed on admission of 71% (89/125) of the animals. This was significantly associated with a higher concentration of serum cortisol and a higher behavioral stress scoring. Fructosamine above the reference limit was detected in only 15% (13/89) of the hyperglycemic individuals. In addition to a positive correlation of fructosamine to glucose concentration, positive relationships with different plasma proteins were detected. A relationship to stress parameters was not observed. These findings underscore stress as a significant trigger for hyperglycemia in alpacas and suggest fructosamine as a valuable parameter for distinguishing between stress-induced and diabetic hyperglycemia. However, the dependence of fructosamine formation on total plasma protein concentration should be considered to avoid misinterpretation.

Methylglyoxal mediates the association between 2-hour plasma glucose and HbA1c with inflammation: The Maastricht Study.

Glucose excursions in persons with diabetes may drive chronic inflammation. Methylglyoxal (MGO) is formed from glucose, is elevated in persons with diabetes, and is a potent glycating agent linked with inflammation. We investigated whether glucose excursions are associated with low-grade inflammation and whether MGO mediates this association. We used data from The Maastricht Study, an extensive phenotyping study into the etiology of type 2 diabetes and its complications. Data of 3017 participants, who underwent an oral glucose tolerance test and where data on MGO levels and inflammation were available, were used. Linear regression analyses, adjusted for potential confounders, evaluated associations between fasting plasma glucose (FPG), 2-hours plasma glucose (2h-PG) and HbA1c and low-grade inflammation (stdβ, [95% confidence interval]), calculated from plasma concentrations of C-reactive protein, serum amyloid A, interleukin-6, interleukin-8, tumor necrosis factor and soluble intercellular adhesion molecule-1. Mediation analyses investigated whether MGO mediated these associations. 2h-PG (0.172 [0.110; 0.234]) and HbA1c (0.148 [0.101; 0.196]), but not FPG (0.049 [-0.002; 0.100]), were associated with low-grade inflammation. 2h-PG and HbA1c were also associated with 2h-MGO (0.471 [0.407; 0.534], and 0.244 [0.195; 0.294], respectively). Furthermore, 2h-MGO was independently and positively associated with low-grade inflammation (0.078 [0.037, 0.120]). 2h-MGO mediated 23% of the association between 2h-PG and inflammation, and 16% of the association between HbA1c and inflammation. MGO mediates the association between post-load glucose excursions and HbA1c with inflammation, providing evidence for a role of postprandial MGO formation to hyperglycemia-induced low-grade inflammation.

An overview of interpretability of two models of unbound fraction that are used in combination with the well-stirred model for predicting hepatic clearance of drugs

Hypothetical and experimental models of unbound fraction have been proposed to facilitate predicting the hepatic clearance (CLH) of drugs from values of intrinsic clearance for the unbound drug (CLint-in vitro-unbound) and the well-stirred model (WSM). The hypothetical model (fu-adjusted) is adjusting the unbound fractions determined in plasma in vitro to estimate the maximum unbound fractions at the hepatocytes if each drug-protein complex in plasma becomes fully dissociated at the membrane by any albumin (ALB)-facilitated hepatic uptake mechanism. The model of fu-adjusted is also adjusting the unbound fraction for a pH gradient effect across the membrane. Alternatively, the new experimental model (fu-dynamic) measures the unbound fractions resulting to the dynamic dissociation kinetics from proteins in the presence of plasma and a liver enzyme in an in vitro assay. The objective of this study was to conduct an in-depth analysis of previous CLH predictions made with these unbound fractions in a companion manuscript. Furthermore, a new dataset on transporter substrates was also included in this study. Finally, the physiological basis of fu-adjusted has been redefined to extend its applicability with more drugs. In this case, there are lower concentrations of binding proteins in liver versus plasma that could also explain the higher unbound fractions for that organ. The outcomes associated to additional analyses pointed out that fu-adjusted, again, generally provided the most accurate predictions of CLH because fu-dynamic has generated superior biases of underpredictions or overpredictions. For slowly metabolized drugs bound to ALB, fu-dynamic was definitively less accurate than fu-adjusted. For other drug properties, fu-dynamic fared better but it was still not generally more accurate than fu-adjusted. Furthermore, experimental values of fu-dynamic were sometimes incoherent. For example, drugs bound to alpha-acid glycoprotein (AGP) did not follow the principle of fu-dynamic (i.e., values of fu-dynamic did not correlate with values of CLint-in vitro-unbound) by contrast to those drugs bound to ALB. Therefore, the current experimental setting for fu-dynamic might be unsuitable in some circumstances. Overall, this study confirmed that calculated values of fu-adjusted were as accurate as experimental values of fu-dynamic and can even be more accurate. A guidance on which unbound fraction to use in the WSM is also provided.

The role of protein corona in advancing plasma proteomics.

The protein corona, a layer of biomolecules forming around nanoparticles in biological environments, critically influences nanoparticle interactions with biosystems, affecting pharmacokinetics and biological outcomes. Initially, the protein corona presented challenges for nanomedicine and nanotoxicology, such as nutrient depletion in cell cultures and masking of nanoparticle-targeting species. However, recent advancements have highlighted its potential in environmental toxicity, proteomics, and immunology. This viewpoint focuses on leveraging the protein corona to enhance the depth of plasma proteome analysis, addressing challenges posed by the high dynamic range of protein concentrations in plasma. The protein corona simplifies sample preparation, enriches low-abundance proteins, and improves proteome coverage. Innovations include using diverse nanoparticles and spiking small molecules to increase the number of quantified proteins. Reproducibility issues across core facilities necessitate standardized protocols. Moreover, top-down proteomics enables proteoform-specific measurements, providing deeper insights into protein corona composition. Future research should aim at improving top-down proteomics techniques and integrating protein corona studies and proteomics for personalized medicine and advanced diagnostics.