Protein Concentration Research Articles

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

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. 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. 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.

X-ray Reflectivity Probing the Structural Evolution of Sunflower Proteins Adsorbed at the Air-Water Interface.

The study delves into the adsorption of sunflower proteins at the air/water interface using specular X-ray reflection. The research involved fitting models of the protein films to the reflectivity data, resulting in detailed images of the X-ray scattering length density profiles perpendicular to the air/water interface. The sunflower protein isolate that is examined consists of multiple components, and the study proposes a transition from a 1-slab model to a 4-slab model to represent the changing layer structure over time. This transition is significant as it reflects the increasing complexity of the protein film as more proteins adsorb at the interface. Initially, sunflower proteins form a monolayer at the air/water boundary, consisting of a protein-rich, hydrophobic portion closest to the interface and a more diffuse, hydrophilic portion extending into the bulk aqueous phase. The structural changes at the interface over time depend on the bulk protein concentration in the solution. For solutions at relatively low concentrations (C ≤ 0.5 g/L), a lower amount of adsorption results in a larger, more extensive interface area for each species and a thinner protein adsorption layer. The overall thickness of a saturated monolayer is approximately 100 Å, which is close to the maximum dimension of sunflower globulins, with the thickness of the corresponding hydrophobic portion being about 20 Å. For solutions at relatively high concentrations (C ≥ 1.0 g/L), even after forming a saturated monolayer, structural evolution continues within the experimental time frame, occurring on both hydrophilic and hydrophobic sides. Additional proteins from the bulk diffuse toward the interface, forming an extra layer in the water phase and causing an increase in the overall thickness. Furthermore, a distinct sublayer develops next to the air phase, indicating a further structuration of the hydrophobic portion.

Cytokine and growth factor correlation networks associated with morbidities in extremely preterm infants

BackgroundCytokines and growth factors (GF) have been implicated in the development of retinopathy of prematurity (ROP) and bronchopulmonary dysplasia (BPD). We hypothesize that even small coordinated changes in inflammatory proteins or GFs may reveal changes in underlying regulating mechanisms that do not induce obvious changes in concentration of individual proteins. We therefore applied correlation network analysis of serum factors to determine early characteristics of these conditions.MethodsConcentrations of 17 cytokines and five GFs were measured and analysed in blood samples from cord blood, on day one and during the following month in 72 extremely preterm infants. Spearman’s correlation networks distinguishing BPD and severe ROP patients from non-affected were created.ResultsMost cytokine concentrations correlated positively with each other and negatively with GFs. Very few individual cytokines differed between patients with and without ROP or BPD. However, networks of differently correlated serum factors were characteristic of the diseases and changed with time. In ROP networks, EPO, G-CSF and IL-8 (cord blood), BDNF and VEGF-A (first month) were prominent. In BPD networks, IL-1β, IGF-1 and IL-17 (day one) were noted.ConclusionsNetwork analysis identifies protein signatures related to ROP or BPD in extremely preterm infants. The identified interactions between serum factors are not evident from the analysis of their individual levels, but may reveal underlying pathophysiological mechanisms in the development of these diseases.

Macronutrient concentrations in human milk beyond the first half year of lactation: a cohort study

ObjectiveHuman milk composition is dynamic. While extensive research has focused on its macronutrient concentrations during the first 6 months of lactation, limited research exists for extended lactation periods. This study...

Abstract 4136448: A Novel Cardiomyocyte Targeting Peptide Enhances Calcium Handling

Introduction: Intracellular calcium handling plays a central role in cardiac contractility and relaxation. Genes such as the Ca 2+ -ATPase pump (SERCA2a) and the Dihydropteridine reductase (DHPR) enzyme are especially important in the cycling of intracellular calcium. As prior research has established a link between treatment with our cardiac targeting peptide (CTP: a 12-amino acid, cardiomyocyte-specific cell penetrating peptide) and upregulation of calcium-handling genes such as SERCA2a and DHPR, we hypothesized that treatment with our novel hybrid cardiac targeting peptide (hCTP: positions #3 and 11 replaced with the D-enantiomer form of the amino acids) will improve intracellular calcium handling in human cardiomyocytes (CMCs) by increasing expression of key calcium handling genes instrumental in sequestering calcium in diastole and lowering cytosolic levels. Methods: To test hCTP’s calcium handling abilities, CMCs were plated on day 1 and allowed to settle for 24 hours, after which they received daily treatment with various concentrations of hCTP (1-25µM) for three days. On day 5, cells underwent either intracellular calcium measurements using the FLIPR 6 Calcium assay or were prepared for western blotting. Western blotting was performed to determine the protein concentrations of SERCA2a and DHPR, as compared to a GAPDH control. To test hCTP’s intrinsic calcium handling abilities, various concentrations of hCTP were mixed with a constant amount of elemental calcium dissolved in water. After 20 min at 37 o C, the concentration of free Ca 2+ was measured with an ABL90 Flex Plus radiometer. Results: In CMCs, treatment with hCTP led to a decrease in cytosolic calcium in a dose dependent fashion; concentrations as low as 1µM led to a significant decrease in cytosolic calcium (p< 0.0001 for all groups). Western blots performed on the treated CMCs revealed an increase in expression of both DHPR and SERCA2a, normalized to GAPDH control. Outside of the cellular environment, hCTP did not appear to display inherent calcium sequestring properties, suggesting that improvements in calcium handling are due to upregulation of gene expression. Conclusion: hCTP appears to lower cytosolic calcium and improve calcium handling through upregulation of SERCA2a and DHPR. Our data indicates that hCTP is worth pursuing as a stand-alone, cardio-selective therapy for cardiomyopathies involving calcium mishandling.

Bactericidal effect of far ultraviolet-C irradiation at 222 nm against bacterial peritonitis.

Far ultraviolet-C irradiation at 222 nm has potent bactericidal effects against severe infections such as peritonitis, with minimal cytotoxicity. Bacterial peritonitis due to bowel perforation is a serious condition with high mortality despite current treatments. This study investigated the safety and efficacy of intraperitoneal far ultraviolet-C irradiation at 222 nm. In vitro experiments optimized the fluid conditions for bacterial or protein concentrations prior to in vivo evaluation. In vivo efficacy was assessed in a rat peritonitis model induced by Escherichia coli, measuring intra-abdominal bacterial concentration, blood cytokine levels, and mortality rates. Safety was evaluated by analyzing cyclobutane pyrimidine dimers as markers of DNA damage in five abdominal organs: stomach, small intestine, colon, liver, and spleen. Statistical analyses employed parametric methods for normally distributed data and non-parametric methods for data without normality. Optimal in vitro conditions included 106 CFU/mL bacteria, 0.5 mW/cm2 irradiation, and 10-3 mg/mL protein. In the rat model, far ultraviolet-C irradiation at 222 nm significantly decreased intra-abdominal bacteria, reduced blood cytokines (interleukin-1β and interleukin-6), and elevated survival rates from 20% to 60%, compared to lavage alone. The formation of cyclobutane pyrimidine dimers was significantly lower with 222 nm irradiation than with 254 nm, suggesting reduced DNA damage. These findings indicate that far ultraviolet-C irradiation at 222 nm, when combined with lavage, represents a promising therapeutic strategy for bacterial peritonitis, providing effective bacterial reduction and a favorable safety profile. Further research is needed to verify these findings and investigate long-term safety and potential clinical applications.

Comprehensive Proteomics Profiling Identifies Circulating Biomarkers to Distinguish Hypertrophic Cardiomyopathy from Other Cardiomyopathies with Left Ventricular Hypertrophy.

Background: Distinguishing hypertrophic cardiomyopathy (HCM) from other cardiomyopathies with left ventricular hypertrophy (LVH), such as hypertensive LVH, transthyretin amyloid cardiomyopathy (ATTR-CM), and aortic stenosis (AS), is sometimes challenging. Using plasma proteomics profiling, we aimed to identify circulating biomarkers and dysregulated signaling pathways specific to HCM. Methods: In this multicenter case-control study, plasma proteomics profiling was performed in cases with HCM and controls with hypertensive LVH, ATTR-CM, and AS. Two-thirds of patients enrolled earlier in each disease group were defined as the training set, and the remaining one-third as the test set. Protein concentrations in HCM were compared with those in hypertensive LVH (comparison 1), ATTR-CM (comparison 2), and AS (comparison 3). Candidate proteins that meet the following 2 criteria were selected: (1) Higher abundance in HCM throughout all 3 comparisons or lower abundance in HCM throughout all 3 comparisons with univariable P<0.05 and |log2(fold change)| >0.5 in both the training and test sets and (2) Independently associated with HCM with multivariable P<0.05 after adjusting for clinical parameters significantly different between HCM and controls. Using the selected candidate proteins, a logistic regression model to distinguish HCM from controls was developed in the training set and applied to the test set. Finally, pathway analysis was performed in each comparison using proteins with different abundance. Results: Overall, 4,979 proteins in 1,415 patients (HCM, n=879; hypertensive LVH, n=331; ATTR-CM, n=169; AS, n=36) were analyzed. Of those, 5 proteins were selected as candidate proteins. The logistic regression model with these 5 proteins had an area under the receiver-operating-characteristic curve of 0.86 (95% CI 0.82-0.89) in the test set. The MAPK and HIF-1 pathways were dysregulated in HCM throughout the 3 comparisons. Conclusions: This study identified circulating biomarkers that distinguish HCM from other cardiomyopathies with LVH independently from confounders and revealed signaling pathways associated with HCM.

Cicer arietinum phytosome ameliorates hepatosteatosis via downregulation of fatty acid synthase and stearoyl-CoA desaturase 1 in rats

BackgroundHepatosteatosis is considered a universal problematic health due to bad lifestyle. Thereby, the current study evaluates the influence of the Cicer arietinum polyunsaturated fatty acids (CAP) and newly synthesized C. arietinum polyunsaturated fatty acids phytosome (CAPP) against non-alcoholic fatty liver disease (NAFLD) persuaded through a high-fat diet (HFD) in addition to tamoxifen (TAM) in male albino rats. Forty-eight rats were separated into eight groups (6 rats/group). Rats of the control group were administered distilled water for 45 consecutive days, while phosphatidyl choline (PC), CAP, and CAPP groups administered distilled water (15 days), afterward administered PC, CAP, and CAPP, respectively (500 mg/kg b.wt), orally for 30 days. All the previous groups fed normal diet for the 45 days, while NAFLD rats feed HFD for 45 days and receive TAM (200 mg/kg b.wt, i.p) daily for 15 days, followed by administration of vehicle, PC, CAP, and CAPP orally for another 30 days.ResultsHepatosteatosis was appraised biochemically by significant increase in the concentrations of serum AST, ALT, γGT, LDH, ALP, total bilirubin, total lipid, triglycerides, fatty acid synthase (FAS), stearoyl-CoA desaturase 1 (SCD-1), and LDL-cholesterol, as well as hepatic total lipids and triglycerides. In addition, a significant decline in serum total protein, albumin, and HDL-cholesterol concentrations was observed in comparison with the control group. NAFLD induces oxidative stress by noteworthy increase in hepatic MDA, H2O2, and meaningful reduction in hepatic GSH, SOD, GST, GRD, and CAT levels as compared with the corresponding control group. Liver histological changes were noted in the NAFLD group as compared to the control. Interestingly, CAP and CAPP treatments modulate the abnormal effects of NAFLD in all the previous parameters. For the histological changes caused by NAFLD, the liver tissue appeared nearly normal after the treatment with CAP and CAPP.ConclusionCAP and CAPP administration may have a potential role in alleviating hepatosteatosis. This may relate to its downregulation against FAS, SCD-1, and oxidative stress.

Stimulation of Saliva Affects the Release of Aroma in Wine: A Study of Microbiota, Biochemistry, and Participant Origin.

Saliva influences the release of aroma in the oral cavity. The composition of human saliva varies depending on stimulation and host's origin; however, the compositional differences of saliva and their influences on aroma release have not been fully evaluated. In this study, we recruited 30 healthy adults (15 Australians and 15 Chinese) and collected saliva samples at three stages: before, during, and after stimulation. Salivary samples were characterized by the flow rate, total protein concentration, esterase activity, microbiome composition by full-length 16S rRNA gene sequencing, and the ability to release aroma from wine by headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). Differences in salivary composition and specific wine volatiles were found between Australian and Chinese participants and among the three stimulation stages. Significant correlations between the relative abundance of 3 bacterial species and 10 wine volatiles were observed. Our results confirm the influence of participant's geographic origin and stimulation on the salivary composition, highlighting the role of salivary components, especially salivary bacteria, on the release of aroma from wine.

Fe/Co-modified Enteromorpha bio-hydrochar enhanced anaerobic digestion of chicken manure with sulfadimethazine: focusing on synergistic mechanism and microbial community succession

Residual antibiotics in chicken manure may interfere with the stability of anaerobic digestion (AD) and inhibit resource utilization efficiency. In this study, we aimed to enhance chicken manure bio-methanation in AD with high-concentration of sulfamethazine (SMZ) by adding metal (Fe, Co)-modified Enteromorpha-based hydrochar (Co-HC, Fe-HC). The results showed that Fe-HC and Co-HC increased the degree of acidogenesis by 1.25 times and 1.58 times, respectively. The maximum protein concentration in EPS was increased by 47.64% and 72.5% after adding Fe-HC and Co-HC. However, only Co-HC demonstrated notable improvements in both methane production and SMZ removal efficiency. Electrochemical analysis showed that Co-HC possessed a richer variety of oxygen and nitrogen functional groups, along with superior electron exchange capabilities compared to Fe-HC. Furthermore, microbiological assessments revealed that Co-HC enriched syntrophic bacteria (such as Syntrophomonas and Mesotoga), facilitating direct interspecies electron transfer (DIET) and subsequently enhancing biomethane production. The abundance of genes involved in electron transfer increased significantly with Co-HC, with a maximum increase of 75.86% in Co1.5-HC treatment. Additionally, the elimination of antibiotic resistance genes (sul1, sul2) increased by 65.66% in the Co1.5-HC treatment. This study offers a theoretical foundation and empirical support for the synergistic improvement of livestock and poultry manure containing high antibiotic concentrations, thereby helping to overcome challenges posed by recalcitrant substances.Graphical

Spectroscopic and Molecular Docking Studies on the Influence of Inulin on the Interaction of Sophoricoside with Whey Protein Concentrate.

The influence of inulin on the interaction of sophoricoside (Sop) with whey protein concentrate (WPC) was investigated using various spectroscopic methods, including fluorescence spectroscopy (intrinsic fluorescence, synchronous fluorescence, and three-dimensional fluorescence), ultraviolet-visible (UV-Vis) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and molecular docking. Sop was found to quench the intrinsic fluorescence of WPC by a static mechanism, both with and without the addition of inulin, and to enhance the antioxidant capacity of the protein. The addition of inulin slightly increased the binding distance between WPC and Sop, while reducing the number of binding sites from two to one. Non-covalent interactions, predominantly van der Waals forces and hydrogen bonding, were maintained between Sop and the protein. Synchronous fluorescence spectroscopy revealed that Sop prevents the exposure of hydrophobic groups on tryptophan residues, leading to increased surface hydrophilicity of the WPC complex. This aligns with the decreased protein surface hydrophobicity measured by 8-Anilino-1-naphthalenesulfonic acid (ANS) binding assays. With inulin, the overall hydrophobicity of the protein was lower than in the system without inulin, suggesting that both inulin and Sop improve the solubility of WPC. Three-dimensional fluorescence spectral analysis showed a reduction in fluorescence intensity and a red shift in the presence of both Sop and inulin. FTIR spectroscopy indicated a slight increase in the secondary structure ordering of WPC following the addition of both Sop and inulin, suggesting structural stabilization under heating conditions. Molecular docking highlighted the potential for hydrogen bond formation between Sop and WPC.

Canadian Organic Wheat Breeding with On-farm Selection: A Case Study using Landrace and Modern Parents

Participatory plant breeding (PPB) has the potential to be an alternative to a centralized breeding model for niche markets that are historically underserved. Our objectives were to evaluate the performance of two parental cultivars from a Canadian PPB program: a modern spring wheat (Triticum aestivum L.) cultivar (5602HR) and a landrace (Red Fife) - with the progeny of the cross when selected by two farmers located 1800 km apart (denoted Farm1 and Farm2) and evaluate progeny differences from each other under organic conditions. Red Fife was 18 cm taller, matured 5 days later, had greater lodging susceptibility, greater seed mass, and 3.5% lower protein concentration than 5602HR. Farmer genotypes were similar to 5602HR in protein concentration and lodging severity, and similar to Red Fife in plant height (14 cm taller than 5602HR) and seed mass. Farmer genotypes did not yield differently from either parent. Farmer genotypes did not differ from each other in most parameters measured, however, Farm1 had greater lodging resistance under high fertility conditions than Farm2 despite similar plant height. This research provides a proof of concept for the role that farmer selectors can play in selecting for positive traits for organic production and provide insight into organic farmers’ preferences. The research also demonstrated that using an older landrace (ie., Red Fife) allowed positive features such as tall stature to be incorporated into the resulting progeny.

Pharmacodynamic effects of semorinemab on plasma and CSF biomarkers of Alzheimer's disease pathophysiology.

Semorinemab, an anti-tau monoclonal antibody, was assessed in two Phase II trials for Alzheimer's disease (AD). Plasma and cerebrospinal fluid (CSF) biomarkers provided insights into the drug's potential mechanism of action. Qualified assays were used to measure biomarkers of tau, amyloidosis, glial activity, neuroinflammation, synaptic function, and neurodegeneration from participant samples in Tauriel (NCT03289143) and Lauriet (NCT03828747) Phase II trials. Plasma phosphorylated Tau 181 (pTau181) and CSF chitinase-3-like protein 1 (YKL-40) increased following semorinemab treatment in both studies. In Lauriet, increasing plasma glial fibrillary protein (GFAP) concentrations stabilized with semorinemab, while this was not observed in Tauriel. Other AD pathophysiology biomarkers showed no consistent response to semorinemab. Increases in CSF YKL-40 suggest that semorinemab may stimulate microglia activation in the presence of AD-associated Tau pathology, but not in healthy controls. Stabilization of plasma GFAP in Lauriet indicates a possible impact on reactive gliosis in mild-to-moderate AD. Tauriel ClinicalTrials.gov Identifier: NCT03289143. Lauriet ClinicalTrials.gov Identifier: NCT03828747. Phase 1 ClinicalTrials.gov Identifier: NCT02820896. AD pathophysiology biomarkers were measured to assess the mechanism of action. Semorinemab increased CSF YKL-40 in participants with AD but not in healthy controls. Semorinemab possibly stabilized plasma GFAP in the Lauriet trial. Semorinemab treatment may activate microglia and moderate reactive gliosis.

Insights into larval development and protein biochemical alterations of Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae) following Beauveria bassiana and Solanum lycopersicum treatments

BackgroundThe polyphagous notorious pest, black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), cause significant production losses due to its distinctive feeding and hiding behavior, making it particularly challenging to control it with conventional methods. Therefore, sustainable agriculture demands more effective and environmentally safe pest control solutions. This study aimed to investigate the toxicity of two insecticide alternatives, the entomopathogenic fungus (EPF) Beauveria bassiana and Solanum lycopersicum extract (Tomato plant crude extract, TPCE), using two bioassay methods: the poisoned bait method and the leaf dipping method. In addition, the impact of these biological tools on larval development and protein profiles was evaluated.ResultsThe bait application of both tested materials exhibited higher toxicity than the leaf dipping method, as indicated by the toxicity index. The LC50 values for B. bassiana were 1.6 × 10⁸ and 1.8 × 10⁶ conidia ml−1 using the leaf dipping method and poisoned baits method, respectively. For TPCE, the LC50 values were 4.35 and 1.51 mg ml−1 for the same methods, respectively. In addition, sublethal concentrations of both materials altered the larval and pupal durations. B. bassiana significantly reduced the concentration of larval hemolymph protein. A maximum of 12 protein bands in the control sample, with molecular weights (Mw) ranging between 35 and 120 kDa, were detected by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). In B. bassiana-treated larvae, ten bands were detected with Mw ranging from 35 to 120 kDa. At least seven bands were detected in TPCE-treated larvae, with Mw ranging from 35 to 97 kDa.ConclusionsThe findings of this study can be integrated into management programs for A. ipsilon. In addition, the availability of B. bassiana and TPCE in Egypt and their cost-effectiveness as insecticide alternatives support their use in the management programs of this critical pest. These methods are particularly effective when applied in bait form.Graphical

Dengue Fever in Pregnant Women with a Favorable Outcome

Dengue fever is a viral infectious disease, it manifests itself from mild to severe severity, in some cases to fatal outcomes. The number of cases of dengue fever has been increasing in recent years. The results of many studies have also shown adverse effects on pregnant infected women. This article describes the clinical course of Dengue fever in pregnant women, which occurred in moderate severity and without complications. Dengue fever in a pregnant woman manifested itself in the form of complaints of pain in the epigastric region, bleeding gums and petechial hemorrhages within 1 day. She had a high fever for 5 days before going to the hospital. Upon admission, her body temperature was 37 °C, blood pressure was 110/80 mmHg, pulse rate was 80 beats per minute, respiratory rate was 20 beats per minute. She had mild dehydration, normal breathing and heartbeat. The height of the bottom corresponds to the gestation period of 31 weeks, and the fetal heart rate was 144 beats per minute. Petechiae measuring at least 1-2 m in diameter were found around her face, forearms and both basal areas. In a laboratory study, the results showed changes in the number of platelets in the form of thrombocytopenia, leukocytopenia and in the biochemical analysis of blood by increasing the concentration of C-reactive protein and the detection of antibodies in serum during serological analysis. Although many studies have shown that Dengue during pregnancy increases the risk of preeclampsia, hemorrhagic dengue fever (HLD), fetal distress, premature birth, cesarean section and maternal mortality and intrauterine development delay and stillbirth consequences. This clinical case concludes with favorable outcomes. But the physiological effects of dengue fever during pregnancy (for both mother and child), depending on the trimester, as well as research and treatment methods require further study, especially in the region of endemic zones and India.

Maternal BMI During Lactation Is Associated with Major Protein Compositions in Early Mature Milk.

The present study identified multiple proteins in early mature milk and explored the correlation between protein compositions in HM and maternal BMI during lactation. A total of 70 mothers giving birth to single-term infants from four representative cites were enrolled in this research. Milk samples were collected between 9 and 11 a.m. to avoid the influence of circadian rhythms. The concentration of total protein in the milk samples was determined using the Bradford method, and the concentrations of α-lactalbumin, lactoferrin, osteopontin, αs-1 casein, β-casein, and κ-casein, butyrophilin, periodic acid Schiff 6/7, fatty acid-binding protein, and xanthine oxidoreductase in the milk samples were measured through a previously published method using ultra-performance liquid chromatography coupled with mass spectrometry. A semi-structured questionnaire investigation and body measurements were carried out by trained investigators to collect the information of subjects. In the univariate models, the concentrations of TP (r = 0.306), α-La (r = 0.260), LF (r = 0.371), OPN (r = 0.286), and αS1-CN (r = 0.324) were all positively and significantly correlated with maternal BMI. In the models' adjusted covariates, the concentrations of TP (Lg β = 7.4 × 10-3), LF (Lg β = 19.2 × 10-3), αS1-CN (Lg β = 8.2 × 10-3) and the proportion of LF (β = 0.20%) were positively correlated with continuous maternal BMI changes. TP concentrations in the HM of obese mothers were higher than in the other three groups (Lg β: 66.7 × 10-3~140.5 × 10-3), α-La concentrations were higher than in the underweight and normal groups (Lg β: 94.4 × 10-3~145.7 × 10-3), and OPN concentrations were higher than in the overweight groups (Lg β = 103.6 × 10-3). The concentrations of LF (Lg β: -298.2 × 10-3~-191.0 × 10-3), OPN (Lg β: -248.9 × 10-3~-145.3 × 10-3), and αS1-CN (Lg β: -160.7 × 10-3~-108.3 × 10-3) in the HM of underweight mothers were lower than those in the other three groups. β-CN concentrations were lower than normal (Lg β = -125.1 × 10-3) and obese groups (Lg β = -165.7 × 10-3), κ-CN concentrations were lower than the overweight (Lg β = -132.5 × 10-3) and obese groups (Lg β = -147.9 × 10-3), and the proportion of LF was lower than that of the overweight (β = -2.80%) and obese groups (β = -2.52%). The proportion of LF in normal mothers was lower than that in the overweight group (β = -1.15%). No statistically significant associations between four MFGM proteins and maternal BMI were determined as the equation models could not be fitted (p for F-test < 0.05). Obese mothers had higher concentrations of multiple protein components than other groups, while underweight mothers had lower concentrations. The association between BMI and protein compositions may be more pronounced for certain protein types.

Implemention of Innovative Process Analytical Technologies to Characterize Critical Quality Attributes of Co-Formulated Monoclonal Antibody Products.

Characterizing co-formulated monoclonal antibodies (mAbs) poses significant challenges in the pharmaceutical industry. Due to the high structural similarity of the mAbs, traditional analytical methods, compounded by the lengthy method development process, hinder product development and manufacturing efficiency. There is increasing critical need in the pharmaceutical industry to streamline analytical approaches, minimizing time and resources, ensuring a rapid clinical entry and cost-effective manufacturing. This study investigates the application of process analytical technologies (PAT) to address such challenges. Our investigation introduces two complementary technologies, on-line ultra-performance liquid chromatography (online UPLC) and multimode fluorescence spectroscopy (MMFS), as potential PAT tools tailored for characterizing critical quality attributes (CQA) in co-formulated mAb products. Specifically, the CQAs under evaluation include the total protein concentration of the mAbs within the co-formulation and the ratio of mAb A to mAb B. Online UPLC enables direct and automated measurement of the CQAs through physical separation, while MMFS determines them in a non-destructive and more swift manner based on chemometric modeling. We demonstrate these technologies' comparable performance to conventional methods, alongside substantial benefits such as reduced analytical turnaround time and decreased laboratory efforts. Ultimately, integrating them as innovative PAT tools expedites the delivery of therapeutic solutions to patients and enhances manufacturing efficiency, aligning with the imperative for swift translation of scientific discoveries into clinical benefits.

Acute phase responses in clinically healthy multiparous Holsteins with and without calcium dysregulation during the early postpartum period

Patterns of calcium dysregulation resulting in low total serum calcium concentrations (tCa) at 4 DIM, known as dyscalcemia, commonly occur in multiparous Holsteins. Dyscalcemia is associated with risk of disease, decreased production, and poor reproductive performance. Inflammation is well-documented early in lactation and is associated with similarly suboptimal outcomes. The acute phase response produces markers and mediators of inflammation; therefore, the objective of our case-control study was to evaluate postpartum patterns of 3 positive acute phase proteins in cows with and without dyscalcemia. We hypothesized that dyscalcemic cows would experience more activated inflammation than eucalcemic cows and that inflammation would precede dyscalcemia diagnosis. Multiparous Holstein cows at 2 commercial dairy farms in central New York were enrolled from a parent study based on tCa at 4 DIM, at a 2:1 ratio of eucalcemic (tCa >2.3 mmol/L; n = 32) to dyscalcemic cows (tCa <2.2 mmol/L; n = 16). Blood was collected 1 to 3 d before parturition and once every 24 h postpartum through 4 DIM. Samples were analyzed for 3 acute phase proteins, serum amyloid A (SAA), haptoglobin and lipopolysaccharide binding protein (LBP). Patterns of protein concentrations in blood over time were compared using linear mixed effects models including fixed effects of calcium status group, time, parity group, farm, relevant 2-way interactions, and the random effect of cow. Overall, dynamics of acute phase proteins showed that dyscalcemic cows experienced increased acute phase responses compared with eucalcemic cows, and that these responses preceded dyscalcemia diagnosis at 4 DIM. Dyscalcemic cows had elevated concentrations of SAA beginning at 2 DIM (eucalcemic: mean = 13.88 µg/mL, 95% CI = 11.34 to 16.99 µg/mL; dyscalcemic: mean = 32.95 µg/mL, 95% CI = 24.55 to 44.21 µg/mL). and continuing through 4 DIM (eucalcemic: mean = 8.14 µg/mL, 95% CI = 6.66 to 9.95 µg/mL; dyscalcemic: mean = 30.01 µg/mL, 95% CI = 22.60 to 39.83 µg/mL). Haptoglobin concentrations were also elevated in the blood of dyscalcemic cows from 2 DIM (eucalcemic: mean = 0.39 g/L, 95% CI = 0.31 to 0.49 g/L; dyscalcemic: mean = 1.11 g/L, 95% CI = 0.79 to 1.56 g/L) through 4 DIM (eucalcemic: mean = 0.27 g/L, 95% CI = 0.21 to 0.34 g/L; dyscalcemic: mean = 1.65 g/L, 95% CI = 1.19 to 2.28 g/L). Concentrations of LBP exhibited a different pattern with a small difference between groups at 3 DIM (eucalcemic: mean = 4.67 µg/mL, 95% CI = 4.02 to 5.42 µg/mL; dyscalcemic: mean = 7.91 µg/mL, 95% CI = 6.49 to 9.63 µg/mL) that became larger at 4 DIM (eucalcemic: mean = 4.88 µg/mL, 95% CI = 4.22 to 5.64 µg/mL; dyscalcemic: mean = 10.79 µg/mL, 95% CI = 8.84 to 13.17 µg/mL). Our work supports the hypothesis that dyscalcemia and inflammatory activity are associated in dairy cows under naturally occurring postpartum conditions. While the causal structure of this relationship remains unknown, improved understanding of inflammation and dyscalcemia may provide insight into mechanisms by which some cows experience maladaptation during the early postpartum period.

SARS-CoV-2 and HSV-1 Induce Amyloid Aggregation in Human CSF Resulting in Drastic Soluble Protein Depletion.

The corona virus (SARS-CoV-2) pandemic and the resulting long-term neurological complications in patients, known as long COVID, have renewed interest in the correlation between viral infections and neurodegenerative brain disorders. While many viruses can reach the central nervous system (CNS) causing acute or chronic infections (such as herpes simplex virus 1, HSV-1), the lack of a clear mechanistic link between viruses and protein aggregation into amyloids, a characteristic of several neurodegenerative diseases, has rendered such a connection elusive. Recently, we showed that viruses can induce aggregation of purified amyloidogenic proteins via the direct physicochemical mechanism of heterogeneous nucleation (HEN). In the current study, we show that the incubation of HSV-1 and SARS-CoV-2 with human cerebrospinal fluid (CSF) leads to the amyloid aggregation of several proteins known to be involved in neurodegenerative diseases, such as APLP1 (amyloid β precursor like protein 1), ApoE, clusterin, α2-macroglobulin, PGK-1 (phosphoglycerate kinase 1), ceruloplasmin, nucleolin, 14-3-3, transthyretin, and vitronectin. Importantly, UV-inactivation of SARS-CoV-2 does not affect its ability to induce amyloid aggregation, as amyloid formation is dependent on viral surface catalysis via HEN and not its ability to replicate. Additionally, viral amyloid induction led to a dramatic drop in the soluble protein concentration in the CSF. Our results show that viruses can physically induce amyloid aggregation of proteins in human CSF and result in soluble protein depletion, thus providing a potential mechanism that may account for the association between persistent and latent/reactivating brain infections and neurodegenerative diseases.

Covalent Attachment of Functional Proteins to Microfiber Surfaces via a General Strategy for Site-Selective Tetrazine Ligation.

Surface modification of materials with proteins has various biological applications, and hence the methodology for surface modification needs to accommodate a wide range of proteins that differ in structure, size, and function. Presented here is a methodology that uses the Affinity Bioorthogonal Chemistry (ABC) tag, 3-(2-pyridyl)-6-methyltetrazine (PyTz), for the site-selective modification and purification of proteins and subsequent attachment of the protein to trans-cyclooctene (TCO)-functionalized hydrogel microfibers. This method of surface modification is shown to maintain the functionality of the protein after conjugation with proteins of varying size and functionalities, namely, HaloTag, NanoLuc luciferase (NanoLuc), and fibronectin type III domains 9-10 (FNIII 9-10). The method also supports surface modification with multiple proteins, which is shown by the simultaneous conjugation of HaloTag and NanoLuc on the microfiber surface. The ability to control the relative concentrations of multiple proteins presented on the surface is shown with the use of HaloTag and superfolder GFP (sfGFP). This application of the ABC-tagging methodology expands on existing surface modification methods and provides flexibility in the site-selective protein conjugation methods used along with the rapid kinetics of tetrazine ligation.