Search

Rationale: Accurate measurement of trace compounds in blood samples is important in clinical diagnosis and life science. Ambient ionization mass spectrometry, however, suffers from the matrix effect when dealing with complex samples such as blood. Therefore, it is important to reduce the matrix effects in blood samples. Methods: A low-cost and disposable Teflon tube was used as a platform to precipitate the protein in blood. The analytes are extracted into organic solvent, and the precipitated protein can be adsorbed by the chromatography paper inserted. Therefore, the Teflon tube after precipitation can be directly subjected to paper spray ionization mass spectrometry, achieving one-step analysis of blood. Results: High sensitivity and satisfactory stability were achieved for pharmaceuticals, acids, and endogenic metabolites in blood. The absolute signal intensities of characteristic product ions of the tested analytes were 8–20 times higher after protein precipitation than those obtained using paper spray. Detection limits and quantitative performance were evaluated for three drugs: carbamazepine, metformin, and tioconazole. In addition, the limits of detection and quantitation were improved 9–14- and 8–12-fold, respectively. Conclusions: Protein precipitation coupled to paper spray with a tube and then to mass spectrometry was successfully achieved and applied in the one-step analysis of trace compounds in blood samples. The experimental results showed that this method was sensitive, stable, convenient, and economic for the direct analysis of blood. [ABSTRACT FROM AUTHOR]

Workplace‐collected blood spots deposited on filter paper were analysed with multiplexed affinity‐based protein assays and found to be suitable for proteomics analysis. The protein extension assay (PEA) was used to characterize 92 proteins using 1.2 mm punches in repeated samples collected from 20 workers. Overall, 97.8% of the samples and 91.3% of the analysed proteins passed quality control. Both within and between spot correlations using six replicates from the same individual were above 0.99, suggesting that comparable levels are obtained from multiple punches from the same spot and from consecutive spots. Protein levels from dried blood and wet serum from the same individuals were compared and the majority of the analysed proteins were found to be significantly correlated. These results open up for simplified sample collection of blood in field conditions for proteomic analysis, but also highlight that not all proteins can be robustly measured from dried whole blood. [ABSTRACT FROM AUTHOR]

Rationale: To open up new ways for matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS)‐based patient screening, blood serum is the most preferred specimen because of its richness in patho‐physiological information and due to ease of collection. To overcome deleterious freeze/thaw cycles and to reduce high costs for shipping and storage, we sought to develop a procedure which enables MALDI‐MS protein profiling of blood serum proteins without the need for serum freezing. Methods: Blood sera from patients/donors were divided into portions which after pre‐incubation were fast frozen. Thawed aliquots were deposited on filter paper discs and air‐dried at room temperature. Intact serum proteins were eluted with acid‐labile detergent‐containing solutions and were desalted by employing a reversed‐phase bead system. Purified protein solutions were screened by MALDI‐MS using standardized instrument settings. Results: MALDI mass spectra from protein solutions which were eluted from filter paper discs and desalted showed on average 25 strong ion signals (mass range m/z 6000 to 10,000) from intact serum proteins (apolipoproteins, complement proteins, transthyretin and hemoglobin) and from proteolytic processing products. Semi‐quantitative analysis of three ion pairs: m/z 6433 and 6631, m/z 8205 and 8916, as well as m/z 9275 and 9422, indicated that the mass spectra from either pre‐incubated fast‐frozen serum or pre‐incubated dried serum spot eluted serum contained the same information on protein composition. Conclusions: A workflow that avoids the conventional cold‐chain and yet enables the investigation of intact serum proteins and/or serum proteolysis products by MALDI‐MS profiling was developed. The presented protocol tremendously broadens the clinical application of MALDI‐MS and simultaneously allows a reduction in the costs for storage and shipping of serum samples. This will pave the way for clinical screening of patients also in areas with limited access to health care systems, and/or specialized laboratories. [ABSTRACT FROM AUTHOR]

Background: Hospital transfusion services order blood products to satisfy orders and maintain inventory levels during unexpected periods of increased blood demand. Surplus inventory may outdate before being allocated to a recipient. Blood product outdating is the largest contributor to blood wastage. Study Design: A province‐wide redistribution program was designed and implemented to redistribute near‐outdate plasma protein and related blood products from low‐usage to high‐usage hospitals. Program operations and details are described in this paper. Two transport container configurations were designed and validated for transport of all blood products. A cost‐analysis was performed to determine the effectiveness of this redistribution program. Results: A total of 130 hospital transfusion services contributed at least one near‐outdate blood product for redistribution between January 2012 and March 2020. These services redistributed 15,499 products through 3412 shipments, preventing the outdating of $17,570,700 CAD worth of product. Program costs were $14,900 for shipping and $30,000 for staffing. Failed time limits or non‐compliance with packing configurations resulted in $388,200 worth of blood products (97 shipments containing 816 products) being discarded. Courier transport delays was the most common reason (42/97; 43%) for transport failure. Conclusion: Redistributing near‐outdate blood products between hospitals is a feasible solution to minimize outdating. Despite heterogeneity of Canadian blood product inventory, all products (each with unique storage and transport requirements) were successfully redistributed in one of two validated and simple containers. Total operation costs of this program were small in comparison to the $17.6 million in savings associated with preventing the discard of outdated products. [ABSTRACT FROM AUTHOR]

Deep learning systems are typically designed to perform for a wide range of test inputs. For example, deep learning systems in autonomous cars are supposed to deal with traffic situations for which they were not specifically trained. In general, the ability to cope with a broad spectrum of unseen test inputs is called generalization. Generalization is definitely important in applications where the possible test inputs are known but plentiful or simply unknown, but there are also cases where the possible inputs are few and unlabeled but known beforehand. For example, medicine is currently interested in targeting treatments to individual patients; the number of patients at any given time is usually small (typically one), their diagnoses/responses are still unknown, but their general characteristics (such as genome information, protein levels in the blood, and so forth) are known before the treatment. We propose to call deep learning in such applications targeted deep learning. In this paper, we introduce a framework for targeted deep learning, and we devise and test an approach for adapting standard pipelines to the requirements of targeted deep learning. The approach is very general yet easy to use: It can be implemented as a simple data‐preprocessing step. We demonstrate on a variety of real‐world data that our approach can indeed render standard deep learning faster and more accurate when the test inputs are known beforehand. [ABSTRACT FROM AUTHOR]

Introduction: Hereditary factor X deficiency is a rare bleeding disorder, with limited treatment options. This paper describes the approach to pre‐clinical development and characterization of a high‐purity plasma‐derived factor X concentrate, to achieve orphan drug marketing authorization for the treatment of hereditary factor X deficiency. Methods: A chromatographic process was developed, to purify factor X from human plasma for fractionation. The product was characterized using in vitro, in vivo and ex vivo tests for potency, purity, thrombogenicity, immunogenicity, toxicity and stability. Results: The production process complied with good pharmaceutical manufacturing practice. It achieved 6000‐fold purification and 100‐fold concentration of the factor X protein compared to human plasma. The factor X protein was 94%–96% pure. Other residual plasma proteins were well below levels in plasma, minimizing potential interference in hemostasis after therapeutic administration. Effective virus‐reduction during manufacture, and the absence of thrombogenicity, toxicity and immunogenic potential were confirmed, addressing the main safety concerns historically associated with plasma‐derived therapeutics. The freeze‐dried product remained stable between +2°C and +30°C for at least three years. After reconstitution with water for injections, the factor X activity was maintained for at least 48 h at +18°C to +22°C. Conclusion: Targeted pre‐clinical development of the first highly‐purified concentrate to treat hereditary factor X deficiency is described. Following international guidelines for nonclinical safety testing, particular strategies were adopted for unmodified products derived from human blood plasma. This approach may also be relevant to the development of other ultra‐orphan medicinal products. [ABSTRACT FROM AUTHOR]

The purpose of this paper is to fabricate novel nanoparticles (NPs) from a single disulfide bond‐bridged block copolymer poly(hydroxyethyl methacrylate)‐S‐S‐polycaprolactone (PHEMA‐S‐S‐PCL). The novel biomaterial was synthesized by ring‐opening polymerization and reversible addition–fragmentation chain transfer polymerization. The cargo‐free NPs were fabricated with the solvent evaporation method, and studies on NPs' characterizations were carried out. The hydrogen nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy spectra confirmed the synthesis of PHEMA‐S‐S‐PCL copolymer. Thermo‐gravimetric analysis curves indicated that the obtained PHEMA‐S‐S‐PCL copolymer had good thermostability. Transmission electron microscopy and dynamic light scatter results suggested that the cargo‐free NPs were in round shapes with an average diameter of 103.6 ± 0.12 nm. The low critical micelle concentration of cargo‐free NPs (7.9 × 10−4 mg/ml) indicated that these NPs would keep their spherical shapes after being attenuated by abundant liquid (e.g., blood or body fluid). Furthermore, these NPs showed high stability at the presence of bovine serum albumin. Therefore, it could be speculated that these NPs would not be absorbed by proteins in blood, and they could be used as a candidate carrier for drug delivery. [ABSTRACT FROM AUTHOR]

The urgent global public health need presented by severe acute respiratory syndrome‐coronavirus 2 (SARS‐CoV‐2) has brought scientists from diverse backgrounds together in an unprecedented international effort to rapidly identify interventions. There is a pressing need to apply clinical pharmacology principles and this has already been recognized by several other groups. However, one area that warrants additional specific consideration relates to plasma and tissue protein binding that broadly influences pharmacokinetics and pharmacodynamics. The principles of free drug theory have been forged and applied across drug development but are not currently being routinely applied for SARS‐CoV‐2 antiviral drugs. Consideration of protein binding is of critical importance to candidate selection but requires correct interpretation, in a drug‐specific manner, to avoid either underinterpretation or overinterpretation of its consequences. This paper represents a consensus from international researchers seeking to apply historical knowledge, which has underpinned highly successful antiviral drug development for other viruses, such as HIV and hepatitis C virus for decades. [ABSTRACT FROM AUTHOR]

Background and Objective: Azurocidin is a neutrophil-derived protein in gingival crevicular fluid (GCF) which, according to relevant studies, might correlate with periodontal disease. The aim of the present study was to evaluate azurocidin as a potential biomarker for chronic periodontitis., Material and Methods: One hundred and one patients participated in the study, divided into two groups. Forty-eight were included in the periodontally healthy group (HP) and fifty-three in the chronic periodontitis group (CP). Clinical indices included probing depth (PD), recession (REC), clinical attachment level (CAL), bleeding on probing (BOP) and plaque (PL). Pooled GCF samples were collected with paper strips, freezed in liquid nitrogen (-196°C), stored at -80°C, and the levels of azurocidin were analyzed with ELISA. Values were transformed and expressed for comparisons in pg/30 s sample. Statistical comparisons were performed using non-parametric tests (Mann-Whitney) at the 0.05 level. Furthermore, the diagnostic accuracy of the procedure was assessed with receiver operator characteristic curves (ROC), areas under the curve (AUC), and the Youden's J Index calculated., Results: Demographic data were comparable between the two groups. Clinical parameters and the levels of azurocidin were statistically significantly higher in the CP group when compared to the HP group (Mann-Whitney test, P < .05). Quantitative data from ELISA demonstrated a high diagnostic accuracy of azurocidin, with AUC calculated higher than 0.9 at the 0.000 level., Conclusion: Azurocidin in GCF is a promising biomarker for periodontal disease. The results of the present study agree with previous studies in the literature showing an up-regulated trend in the levels of azurocidin in periodontitis patients., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

There is a laboratory and clinical need to know the impact of direct oral anticoagulants (DOACs) on diagnostic tests to avoid misinterpretation of results. Although the regulatory labelling documents provide some information about the influences of each DOAC on diagnostic tests, these are usually limited to some of the most common tests and no head to head comparison is available. In this paper, we report the impact of DOACs on several thrombophilia tests, including assessment of antithrombin, protein S and protein C activity assays, detection of activated protein C resistance and assays used for lupus anticoagulant. Results are compared and discussed with data obtained from literature. The final goal of this comprehensive review is to provide practical recommendations for laboratories to avoid misdiagnosis due to oral direct factor Xa (FXa) or IIa (FIIa) inhibitors. Overall, oral direct FXa (apixaban, betrixaban, edoxaban and rivaroxaban) and FIIa (dabigatran) antagonists may affect clot‐based thrombophilia diagnostic tests resulting in false‐positive or false‐negative results. An effect on FIIa‐based thrombophilia diagnostic tests is observed with dabigatran but not with anti‐FXa DOACs and conversely for FXa‐based thrombophilia diagnostic tests. No impact was observed with antigenic/chromogenic methods for the assessment of protein S and C activity. In conclusion, interpretation of thrombophilia diagnostic tests results should be done with caution in patients on DOACs. The use of a device/chemical compound able to remove or antagonize the effect of DOACs or the development of new diagnostic tests insensitive to DOACs should be considered to minimize the risk of false results. [ABSTRACT FROM AUTHOR]

Mutations in the X‐linked phosphatidylinositol glycan, class A gene (Pig‐a) lead to loss of glycosylphosphatidylinositol (GPI) anchors and GPI‐anchored proteins from the surface of erythrocytes and other mammalian cells. The Pig‐a gene mutation assay quantifies in vivo gene mutation by immunofluorescent labeling and flow cytometry to detect the loss of GPI‐anchored proteins on peripheral blood erythrocytes. As part of the regulatory acceptance of the assay, a public database has been created that provides detailed information on Pig‐a gene mutation assays conducted in rats and mice. A searchable version of the database is available through a website designed and hosted by the University of Maryland School of Pharmacy. Currently, the database contains only mouse and rat data, but it is anticipated that it will expand to include data from other species, including humans. A major purpose in developing the database was to aid in the preparation of a Retrospective Performance Analysis and Detailed Review Paper required for Organisation for Economic Co‐operation and Development Test Guideline acceptance. We anticipate, however, that it also will be useful for accessing and comparing Pig‐a data to data from other assays and for conducting quantitative assessments of Pig‐a gene mutation responses. Environ. Mol. Mutagen., 60:759–762, 2019. © 2019 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) have emerged as a promising tool for clinical liquid biopsy. However, the identification of EVs derived from blood samples is hindered by the presence of abundant plasma proteins, which impairs the downstream biochemical analysis of EV‐associated proteins and nucleic acids. Here, we employed optimized asymmetric flow field‐flow fractionation (AF4) combined with density cushion ultracentrifugation (UC) to obtain high‐purity and intact EVs with very low lipoprotein contamination from human plasma and serum. Further proteomic analysis revealed more than 1000 EV‐associated proteins, a large proportion of which has not been previously reported. Specifically, we found that cell‐line‐derived EV markers are incompatible with the identification of plasma‐EVs and proposed that the proteins MYCT1, TSPAN14, MPIG6B and MYADM, as well as the traditional EV markers CD63 and CD147, are plasma‐EV markers. Benefiting from the high‐purity of EVs, we conducted comprehensive miRNA profiling of plasma EVs and nanosized particles (NPs), as well as compared plasma‐ and serum‐derived EVs, which provides a valuable resource for the EV research community. Overall, our findings provide a comprehensive assessment of human blood EVs as a basis for clinical biopsy applications. [ABSTRACT FROM AUTHOR]

The contribution of electrospun nanofibrous membranes (e.NFMs) in the biosensing platforms opens up a new prospect for the invention of faster and more sensitive analytical devices. In this paper, we utilized e.NFM of polyethersulfone (PES) as a solid substrate for the protein immobilization through two different approaches: physical and covalent. Scanning electron microscopy (SEM) and Fourier‐transform‐infrared (FTIR) tests were performed to study the effect of plasma treatment on protein immobilization efficacy. Moreover, taking advantage of ELISA technique, the influence of different parameters, namely, nanofibers diameter, membrane thickness, plasma treatment time, an incubation time of ethyl‐3‐(3‐dimethylaminopropyl)‐carbodiimide/N‐hydroxysuccinimide (EDC/NHS), and their ratio on antibody immobilization efficacy through two mentioned approaches, was also assessed. [ABSTRACT FROM AUTHOR]

In transfusional iron overload, extra-hepatic iron distribution differs, depending on the underlying condition. Relative mechanisms of plasma non-transferrin bound iron ( NTBI) generation may account for these differences. Markers of iron metabolism (plasma NTBI, labile iron, hepcidin, transferrin, monocyte SLC40A1 [ferroportin]), erythropoiesis (growth differentiation factor 15, soluble transferrin receptor) and tissue hypoxia (erythropoietin) were compared in patients with Thalassaemia Major ( TM), Sickle Cell Disease and Diamond-Blackfan Anaemia ( DBA), with matched transfusion histories. The most striking differences between these conditions were relationships of NTBI to erythropoietic markers, leading us to propose three mechanisms of NTBI generation: iron overload (all), ineffective erythropoiesis (predominantly TM) and low transferrin-iron utilization ( DBA). [ABSTRACT FROM AUTHOR]

Nanostructured surfaces are effective at physically killing bacterial cells, highlighting their prospective application as biomaterials. The benefits of application of mechano‐bactericidal nanostructures as an alternative to chemical functionalisation are well documented, however, the effects of protein adsorption are not well understood. In this work, theoretical and experimental analyses are conducted by studying the adsorption of human serum proteins (HSP) to nanosheet titanium (Ti) and its subsequent effect on the mechano‐bactericidal efficacy toward Staphylococcus aureus and Pseudomonas aeruginosa cells. The nanosheet pattern exhibits enhanced antibiofouling behaviour mantaining high bactericidal efficiency toward both Gram‐negative and Gram‐positive cells in the presence of adsorbed HSP. To ascertain the immunomodulatory response, S. aureus cells are introduced to protein‐conditioned Ti nanosheet surfaces prior to introducing RAW 264.7 macrophages. On the pre‐infected nanostructured surfaces, macrophages exhibit wound healing behaviour with superior activation of M2‐like macrophage polarization and secretion of anti‐inflammatory cytokines. By contrast, macrophages attached to infected smooth surfaces activated the M1‐like polarized phenotype via the high expression of pro‐inflammatory cytokines, indicating persistent inflammation. The outcomes of this work demonstrate the suitability of Ti nanosheets as a potential biomaterial surface whereby the mechano‐bactericidal activity is not compromised by HSP adsorption and, furthermore, positively influenced an anti‐inflammatory immune response. [ABSTRACT FROM AUTHOR]

Background: To date, most existing models for predicting neuromyelitis optica spectrum disorder (NMOSD) are based primarily on clinical characteristics. Blood‐based NMOSD severity and prognostic predictive immune‐ and inflammation‐related biomarkers are needed. We aimed to investigate the associations between plasma inflammatory biomarkers and relapse and attack severity in NMOSD. Methods: This two‐step, single‐center prospective cohort study included discovery and validation cohorts. We quantified 92 plasma inflammatory proteins by using Olink's proximity extension assay and identified differentially expressed proteins in the relapse group (relapse within 1 year of follow‐up) and severe attack group. To define a new molecular prognostic model, we calculated the risk score of each patient based on the key protein signatures and validated the results in the validation cohort. Results: The relapse prediction model, including FGF‐23, DNER, GDNF, and SLAMF1, predicted the 1‐year relapse risk. The severe attack prediction model, including PD‐L1 and MCP‐2, predicted the severe clinical attack risk. Both the relapse and severe attack prediction models demonstrated good discriminative ability and high accuracy in the validation cohort. Conclusions: Our discovered biomarker signature and prediction models may complement current clinical risk stratification approaches. These inflammatory biomarkers could contribute to the discovery of therapeutic interventions and prevent NMOSD progression. [ABSTRACT FROM AUTHOR]

Superficial, systemic microcirculations, distinct from the pulmonary circulation, supply the mucosae of human nasal and conducting airways. Non‐injurious, inflammatory challenges of the airway mucosa cause extravasation without overt mucosal oedema. Instead, likely reflecting minimal increases in basolateral hydrostatic pressure, circulating proteins/peptides of all sizes are transmitted paracellularly across the juxtaposed epithelial barrier. Thus, small volumes of extravasated, unfiltered bulk plasma appear on the mucosal surface at nasal and bronchial sites of challenge. Importantly, the plasma‐exuding mucosa maintains barrier integrity against penetrability of inhaled molecules. Thus, one‐way epithelial penetrability, strict localization, and well‐controlled magnitude and duration are basic characteristics of the plasma exudation response in human intact airways. In vivo experiments in human‐like airways demonstrate that local plasma exudation is also induced by non‐sanguineous removal of epithelium over an intact basement membrane. This humoral response results in a protective, repair‐promoting barrier kept together by a fibrin‐fibronectin net. Plasma exudation stops once the provisional barrier is substituted by a new cellular cover consisting of speedily migrating repair cells, which may emanate from all types of epithelial cells bordering the denuded patch. Exuded plasma on the surface of human airways reflects physiological microvascular‐epithelial cooperation in first line mucosal defense at sites of intact and regenerating epithelium. [ABSTRACT FROM AUTHOR]

One of the key pharmacokinetic properties of most small molecule drugs is their ability to bind to serum proteins. Unbound or free drug is responsible for pharmacological activity while the balance between free and bound drug can impact drug distribution, elimination, and other safety parameters. In the hepatic impairment (HI) and renal impairment (RI) clinical studies, unbound drug concentration is often assessed; however, the relevance and impact of the protein binding (PB) results is largely limited. We analyzed published clinical safety and pharmacokinetic studies in subjects with HI or RI with PB assessment up to October 2022 and summarized the contribution of PB results on their label dose recommendations. Among drugs with HI publication, 32% (17/53) associated product labels include PB results in HI section. Of these, the majority (9/17, 53%) recommend dose adjustments consistent with observed PB change. Among drugs with RI publication, 27% (12/44) of associated product labels include PB results in RI section with the majority (7/12, 58%) recommending no dose adjustment, consistent with the reported absence of PB change. PB results were found to be consistent with a tailored dose recommendation in 53% and 58% of the approved labels for HI and RI section, respectively. We further discussed the interpretation challenges of PB results, explored treatment decision factors including total drug concentration, exposure–response relationships, and safety considerations in these case examples. Collectively, comprehending the alterations in free drug levels in HI and RI informs treatment decision through a risk‐based approach. [ABSTRACT FROM AUTHOR]

Introduction: Recent evidence recommends the use of biologics in patients with severe uncontrolled type 2 chronic rhinosinusitis with nasal polyp (CRSwNP) owing to its propensity for recurrence after functional endoscopic sinus surgery (FESS). Among the type 2 biologics used for the treatment of nasal polyps, dupilumab (Dupi, anti‐IL‐4) exhibited superior efficacy and safety in indirect comparison studies. Objective: This study aimed to evaluate the objective and subjective outcomes of patients with CRSwNP treated with and without adjuvant Dupi therapy after FESS. Methods: Adult patients with type 2 CRSwNP who underwent FESS with adjuvant Dupi after surgery were enrolled. A matched control group without adjuvant Dupi therapy were recruited during the same period. All patients underwent nasal endoscopy and completed the sinonasal outcome test‐22 questionnaire evaluations at baseline and 3 months after surgery. Results: A total of 10 patients who received postoperative adjuvant therapy with Dupi and 20 patients who underwent surgery only were included. Patients with add‐on Dupi therapy had significantly higher eosinophil cationic protein levels in the serum, eosinophil counts in peripheral blood, prevalence of asthma, and nasal polyp score at baseline. Both treatments were effective in reducing the patient's symptoms by SNOT‐22 at 3 months postoperatively. However, patients with adjuvant Dupi therapy exhibited significantly better endoscopic scores than those with surgery only (p =.022). Conclusion: Surgery plays an important role in treating patients with CRSwNP, and adjuvant Dupi use may facilitate objective mucosal recovery postoperatively. Level of Evidence: 4. [ABSTRACT FROM AUTHOR]

Summary: Objectives: The aim of this paper is to test whether α‐defensins and bacterial/permeability‐increasing protein were related to obesity and cardiovascular risk factors in prepubertal children. Methods: Plasma α‐defensins and bacterial/permeability‐increasing protein, body mass index (BMI), waist circumference, systolic blood pressure (SBP), carotid intima media thickness (cIMT), HOMA‐IR and HMW‐adiponectin were assessed. Results: In a cross‐sectional study (N = 250), higher α‐defensins concentrations were positively associated with BMI, waist, SBP, cIMT, HOMA‐IR and negative correlated with HMW‐adiponectin (all between r = 0.191 and r = 0.377, p ≤ 0.01 and p ≤ 0.0001). Conversely, plasma bacterial/permeability‐increasing protein concentrations presented inversed associated with the same parameters (all between r = −0.124 and r = −0.329; p ≤ 0.05 and p ≤ 0.0001). In a longitudinal study (N = 91), α‐defensins at age 7 were associated with BMI (β = 0.189, p = 0.002; model R2 = 0.847) and waist (β = 0.241, pthinsp;= 0.001; model R2 = 0.754) at age 10. Conclusions: α‐Defensins and bacterial/permeability‐increasing protein may be the markers of childhood obesity. Increased concentrations of α‐defensins may predict BMI and abdominal fat deposition in children. [ABSTRACT FROM AUTHOR]

Blood is generated in very large volumes as a by-product in slaughterhouses all around the world. On the one hand, blood generation presents a serious environmental issue because of its high pollutant capacity; however, on the other hand, blood has the potential to be collected and processed to generate high-added-value food ingredients based on its exceptional nutritive value and its excellent functional properties. In this paper, we review the current state of the art for blood processing, from collection to final recovery of protein isolates, the functional properties of blood, impact of processing on functional properties, and potential applications as food ingredients. Furthermore, future challenges are outlined for this underutilized and abundant product from the meat industry. [ABSTRACT FROM AUTHOR]

Summary: The plant hormone auxin and its directional intercellular transport play a major role in diverse aspects of plant growth and development. The establishment of auxin gradients requires the asymmetric distribution of members of the auxin efflux carrier PIN‐FORMED (PIN) protein family to the plasma membrane. An endocytic pathway regulates the recycling of PIN proteins between the plasma membrane and endosomes, providing a mechanism for dynamic localisation.N‐Ethylmaleimide‐sensitive factor adaptor protein receptors (SNAP receptors, SNAREs) mediate fusion between vesicles and target membranes and are classed as Q‐ or R‐SNAREs based on their sequence. We analysed gain‐ and loss‐of‐function mutants, dominant‐negative transgenics and localisation of the Arabidopsis R‐SNARE VAMP714 protein to understand its function.We demonstrate that VAMP714 is essential for the insertion of PINs into the plasma membrane, for polar auxin transport, root gravitropism and morphogenesis. VAMP714 gene expression is upregulated by auxin, and the VAMP714 protein co‐localises with endoplasmic reticulum and Golgi vesicles and with PIN proteins at the plasma membrane.It is proposed that VAMP714 mediates the delivery of PIN‐carrying vesicles to the plasma membrane, and that this forms part of a positive regulatory loop in which auxin activates a VAMP714‐dependent PIN/auxin transport system to control development. [ABSTRACT FROM AUTHOR]

Introduction: Intervertebral disc degeneration and Modic change are the main spinal structural changes associated with chronic low back pain (LBP). Both conditions are thought to manifest local inflammation and if inflammatory proteins translocate to the blood circulation could be detected systemically. The work here assesses whether the presence of disc degeneration is associated with detectable blood level changes of five inflammatory markers and whether chronic LBP is associated with these changes. Materials and Methods: Two hundred and forty TwinsUK cohort participants with both MRI disc degeneration grade and Modic change extent, and IL‐6, IL‐8, IL‐8 TNF, and CX3CL1 protein blood concentration measurements were included in this work. Linear mixed effects models were used to test the association of blood cytokine concentration with disc degeneration score and Modic change volumetric score. Association of chronic LBP status from questionnaires with disc degeneration, Modic change, and cytokine blood concentration was also tested. Results: No statistically significant association between disc degeneration or Modic change with cytokine blood concentration was found. Instead, regression analysis pointed strong association between cytokine blood concentration with body mass index for IL‐6 and with age for IL‐6 and TNF. Mild association was found between IL‐8 blood concentration and body mass index. Additionally, LBP status was associated with Modic change volumetric score but not associated with any cytokine concentration. Conclusions: We found no evidence that Modic change and disc degeneration are able to produce changes in tested blood cytokine concentration. However, age and body mass index have strong influence on cytokine concentration and both are associated with the conditions studied which may confound associations found in the literature. It is then unlikely that cytokines produced in the disc or vertebral bone marrow induce chronic LBP. [ABSTRACT FROM AUTHOR]

Protein glycosylation responds sensitively to disease states. It is implicated in every hallmark of cancer and has recently started to be considered as a hallmark itself. Changes in N‐glycosylation microheterogeneity are more dramatic than those of protein expression due to the non‐template nature of protein glycosylation. This enables their potential use in serum‐based diagnostics. Here, we perform glycopeptidomics on serum from patients with oropharyngeal squamous cell carcinoma (OPSCC), compared to controls and comparing between cancers based on etiology (human papilloma virus‐ positive or negative). Using MS2, we then targeted glycoforms, significantly different between the groups, to identify their glycopeptide compositions. Simultaneously we investigate the same serum proteins, comparing whether N‐glycosylation changes reflect protein‐level changes. Significant glycoforms were identified from proteins such as alpha‐1‐antitrypsin (SERPINA1), haptoglobin, and different immunoglobulins. SERPINA1 had glycovariance at 2 N‐glycosylation sites, that were up to 35 times more abundant in even early‐stage OPSCCs, despite minimal differences between SERPINA1 protein levels between groups. Some identified glycoforms' fold changes (FCs) were in line with serum protein level FCs, others were less abundant in early‐stage cancers but with great variance in higher‐stage cancers, such as on immunoglobulin heavy constant gamma 2, despite no change in protein levels. Such findings indicate that glycovariant analysis might be more beneficial than proteomic analysis, which is yet to be fruitful in the search for biomarkers. Highly sensitive glycopeptide changes could potentially be used in the future for cancer screening. Additionally, characterizing the glycopeptide changes in OPSCC is valuable in the search for potential therapeutic targets. [ABSTRACT FROM AUTHOR]

Glycidol fatty acid esters that are present in foods are degraded in vivo to the animal carcinogen glycidol, which binds to the N‐terminal valine of hemoglobin (Hb) to form N‐(2,3‐dihydroxypropyl)valine (diHOPrVal) adducts. The existence of other chemicals that are converted to glycidol is unknown. To determine the effect of different exposure conditions on the formation of diHOPrVal adducts, several glycidol‐related chemicals (3‐monochloropropane‐1,2‐diol; 3‐MCPD, epichlorohydrin, glyceraldehyde, acrylic acid, and 1,2‐propanediol) were evaluated using in vitro and in vivo (single/repeated dose) methods. In vitro, the reaction of 3‐MCPD or epichlorohydrin with human Hb produced 17% and 0.7% of diHOPrVal, as compared to equimolar glycidol, respectively. Following a single administration of glycidol‐related compounds to ICR mice, diHOPrVal formation was observed only in the epichlorohydrin‐treated group after day 5 of exposure. After 14 days of repeated dosing, the amounts of diHOPrVal produced by epichlorohydrin and 3‐MCPD in vivo were <1% of diHOPrVal produced by an equal molar concentration of glycidol. Furthermore, glyceraldehyde group produced 0.2% of diHOPrVal at the same molar concentration of glycidol equivalents, in which diHOPrVal formation could not be confirmed by the in vitro assay. The results indicate the usefulness of diHOPrVal as an exposure marker for glycidol; however, the contribution of its formation in vivo by exposure to various chemicals will be necessary to validate and interpret the results. [ABSTRACT FROM AUTHOR]

The hydrogen evolution reaction (HER), catalysed by proteins at mercury electrodes and reflected in chronopotentiometric stripping peak H, provides a label-free and reagentless analytical technique that is sensitive to protein structure. Here we show how the kinetic isotope effect affected the HER catalysed by the protein bovine serum albumin (BSA). We found that the deuteron bond, which is stronger than that of a proton, contributed to less effective transport of deuterons mediated by BSA at the HgjD2O interface, and enhanced structural stability of the surface-attached native BSA in D2O solution. A structural transition was also observed in the surface-attached urea-denatured BSA, and is probably due to the destabilisation of some secondary structural remnants retained by the 17 SSbonds. Because the catalytically active groups involved in proton or deuteron transfer in native proteins are often exposed towards solutions and their protons exchange almost instantly, no signs of H/D exchange were observed in native BSA using peak H under the given conditions. [ABSTRACT FROM AUTHOR]

Background: During sepsis, higher plasma cell-free hemoglobin (CFH) levels portend worse outcomes. In sepsis models, plasma proteins that bind CFH improve survival. In our canine antibiotic-treated Staphylococcus aureus pneumonia model, with and without red blood cell (RBC) exchange transfusion, commercial human haptoglobin (Hp) concentrates bound and compartmentalized CFH intravascularly, increased CFH clearance, and lowered iron levels, improving shock, lung injury, and survival. We now investigate in our model how very high CFH levels and treatment time affect Hp's beneficial effects.Materials and Methods: Two separate canine pneumonia sepsis Hp studies were undertaken: one with exchange transfusion of RBCs after prolonged storage to raise CFH to very high levels and another with rapidly lethal sepsis alone to shorten time to treat. All animals received continuous standard intensive care unit supportive care for 96 hours.Results: Older RBCs markedly elevated plasma CFH levels and, when combined with Hp therapy, created supraphysiologic CFH-Hp complexes that did not increase CFH or iron clearance or improve lung injury and survival. In a rapidly lethal bacterial challenge model without RBC transfusion, Hp binding did not increase clearance of complexes or iron or show benefits seen previously in the less lethal model.Discussion: High-level CFH-Hp complexes may impair clearance mechanisms and eliminate Hp's beneficial effect during sepsis. Rapidly lethal sepsis narrows the therapeutic window for CFH and iron clearance, also decreasing Hp's beneficial effects. In designing clinical trials, dosing and kinetics may be critical factors if Hp infusion is used to treat sepsis. [ABSTRACT FROM AUTHOR]

Introduction: Severe COVID‐19 management is still challenging. Having a laboratory factor to predict the severity of a patient's condition can be very useful in how to approach each patient. There have been studies concentrating on the correlation between serum C‐reactive protein (CRP) level and COVID‐19 severity but we aim to reach a threshold for CRP in disease severity determination. Methods: We conducted a thorough search on PubMed, Web of Science and Google Scholar databases from early 2019 to October 2021, and 323 studies were assessed for eligibility in three phases. We used the Newcastle‐Ottawa Scale to examine the validity of the studies. The t‐test was applied for the CRP level cutoffs. Results: Eventually, 11 articles and 1615 patients were included in this systematic review. Our analysis evaluated combined mean, median, and standard deviation of severe patients' CRP to be respectively 73.37, 53.80, and 47.936. Based on the combined mean, 75 mg/dL was suggested as an initial threshold for baseline CRP in hospitalized patients for developing severe conditions. Conclusion: This study recommends that COVID‐19 patients with on‐admission serum CRP levels of 75 mg/dL and more are likely associated with severe conditions. Thus, anti‐inflammatory agents and further following may be helpful in these patients. [ABSTRACT FROM AUTHOR]

Aims: Patients with heart failure (HF) exhibit poor prognosis, which is further deteriorated by pulmonary hypertension (PH), with negative impact on morbidity and mortality. As PH due to left HF (LHF‐PH) is among the most common causes of PH, there is an urge according to the 2021 European Society of Cardiology HF guidelines to find new biomarkers that aid in prognostication of this patient cohort. Given the role of tumour necrosis factor‐alpha (TNF‐α) in HF progression, we aimed to investigate the prognostic value of plasma proteins related to TNF‐α in patients with LHF‐PH, in relation to haemodynamic changes following heart transplantation (HT). Methods and results: Twenty TNF‐α‐related plasma proteins were analysed using proximity extension assay in healthy controls (n = 20) and patients with LHF‐PH (n = 67), before and 1 year after HT (n = 19). Plasma levels were compared between the groups, and the prognostic values were determined using Kaplan–Meier and Cox regression analyses. Plasma levels of lymphotoxin‐beta receptor (LTBR), TNF receptor superfamily member 6B (TNFRSF6B), and TNF‐related apoptosis‐inducing ligand receptors 1 and 2 (TRAIL‐R1 and TRAIL‐R2, respectively) were higher in LHF‐PH pre‐HT vs. controls (P < 0.0001), as well as higher in pre‐HT vs. post‐HT (P < 0.001). The elevated pre‐HT levels of LTBR, TNFRSF6B, TRAIL‐R1, and TRAIL‐R2 decreased towards the levels of healthy controls after HT. Higher preoperative levels of LTBR, TNFRSF6B, TRAIL‐R1, and TRAIL‐R2 in LHF‐PH were associated with worse survival rates (P < 0.002). In multivariate Cox regression models, each adjusted for age and sex, LTBR, TNFRSF6B, TRAIL‐R1, and TRAIL‐R2 predicted mortality (P < 0.002) [hazard ratio (95% confidence interval): 1.12 (1.04–1.19), 1.01 (1.004–1.02), 1.28 (1.14–1.42), and 1.03 (1.02–1.04), respectively]. Conclusions: Elevated pre‐HT plasma levels of the TNF‐α‐related proteins LTBR, TNFRSF6B, TRAIL‐R1, and TRAIL‐R2 in LHF‐PH decreased 1 year after HT, displaying a normalization pattern towards the levels of the healthy controls. These proteins were also prognostic, where higher levels were associated with worse survival rates in LHF‐PH, providing new insight in their potential role as prognostic biomarkers. Larger studies are warranted to validate our findings and to investigate their possible pathobiological mechanisms in LHF‐PH. [ABSTRACT FROM AUTHOR]

Proteomic approaches have unique advantages in the identification of biological pathways that influence physical frailty, a multifactorial geriatric syndrome predictive of adverse health outcomes in older adults. To date, proteomic studies of frailty are scarce, and few evaluated prefrailty as a separate state or examined predictors of incident frailty. Using plasma proteins measured by 4955 SOMAmers in the Atherosclerosis Risk in Community study, we identified 134 and 179 proteins cross‐sectionally associated with prefrailty and frailty, respectively, after Bonferroni correction (p < 1 × 10−5) among 3838 older adults aged ≥65 years, adjusting for demographic and physiologic factors and chronic diseases. Among them, 23 (17%) and 82 (46%) were replicated in the Cardiovascular Health Study using the same models (FDR p < 0.05). Notably, higher odds of prefrailty and frailty were observed with higher levels of growth differentiation factor 15 (GDF15; pprefrailty = 1 × 10−15, pfrailty = 2 × 10−19), transgelin (TAGLN; pprefrailty = 2 × 10−12, pfrailty = 6 × 10−22), and insulin‐like growth factor‐binding protein 2 (IGFBP2; pprefrailty = 5 × 10−15, pfrailty = 1 × 10−15) and with a lower level of growth hormone receptor (GHR, pprefrailty = 3 × 10−16, pfrailty = 2 × 10−18). Longitudinally, we identified 4 proteins associated with incident frailty (p < 1 × 10−5). Higher levels of triggering receptor expressed on myeloid cells 1 (TREM1), TAGLN, and heart and adipocyte fatty‐acid binding proteins predicted incident frailty. Differentially regulated proteins were enriched in pathways and upstream regulators related to lipid metabolism, angiogenesis, inflammation, and cell senescence. Our findings provide a set of plasma proteins and biological mechanisms that were dysregulated in both the prodromal and the clinical stage of frailty, offering new insights into frailty etiology and targets for intervention. [ABSTRACT FROM AUTHOR]

β2‐microglobulin (β2‐m) is a plasma protein derived from physiological shedding of the class I major histocompatibility complex (MHCI), causing human systemic amyloidosis either due to persistently high concentrations of the wild‐type (WT) protein in hemodialyzed patients, or in presence of mutations, such as D76N β2‐m, which favor protein deposition in the adulthood, despite normal plasma levels. Here we describe a new transgenic Caenorhabditis elegans (C. elegans) strain expressing human WT β2‐m at high concentrations, mimicking the condition that underlies dialysis‐related amyloidosis (DRA) and we compare it to a previously established strain expressing the highly amyloidogenic D76N β2‐m at lower concentrations. Both strains exhibit behavioral defects, the severity of which correlates with β2‐m levels rather than with the presence of mutations, being more pronounced in WT β2‐m worms. β2‐m expression also has a deep impact on the nematodes' proteomic and metabolic profiles. Most significantly affected processes include protein degradation and stress response, amino acids metabolism, and bioenergetics. Molecular alterations are more pronounced in worms expressing WT β2‐m at high concentration compared to D76N β2‐m worms. Altogether, these data show that β2‐m is a proteotoxic protein in vivo also in its wild‐type form, and that concentration plays a key role in modulating pathogenicity. Our transgenic nematodes recapitulate the distinctive features subtending DRA compared to hereditary β2‐m amyloidosis (high levels of non‐mutated β2‐m vs. normal levels of variant β2‐m) and provide important clues on the molecular bases of these human diseases. [ABSTRACT FROM AUTHOR]

Diet can affect the inflammatory state of the body. Accordingly, the dietary inflammatory index (DII) has been developed to quantify the inflammatory properties of food items. This study sought to investigate the association between dietary inflammation index (DII) and the odds ratio of elevated CRP (E‐CRP) through a systematic review and meta‐analysis study. The International electronic databases of PubMed, Web of Science (ISI), and Scopus were searched until May 2023 to find related articles. From 719 studies found in the initial search, 14 studies, with a total sample size of 59,941 individuals, were included in the meta‐analysis. The calculated pooled odds ratio (OR) of E‐CRP in the highest DII category was 1.39 (95% CI: 1.06, 1.14, test for heterogeneity: p =.63, and I2 =.0%) in comparison with the lowest DII category. Also, the results of this study showed that each unit increase in DII as a continuous variable generally elicited a 10% increase in the odds of E‐CRP (OR 1.10, 95% CI 1.06, 1.14, test for heterogeneity: p =.63, and I2 =.0%). Subgroup meta‐analyses showed that there is a higher E‐CRP odds ratio for the articles that reported energy‐adjusted DII (E‐DII) instead of DII, the studies that measured CRP instead of hs‐CRP, and the studies that used 24‐h recall instead of FFQ as the instrument of dietary intake data collection. Individuals with a higher DII were estimated to have higher chances of developing elevated serum CRP. This value was influenced by factors such as the participants' nationality, instruments of data collection, methods used to measure inflammatory biomarkers, study design, and data adjustments. However, future well‐designed studies can help provide a more comprehensive understanding of the inflammatory properties of diet and inflammatory serum biomarkers. [ABSTRACT FROM AUTHOR]

Within the last decade new technologies have been developed and implemented which employ light, often in the presence of a photosensitizer, to inactivate pathogens that reside in human blood products for the purpose of transfusion. These pathogen reduction technologies attempt to find the proper balance between pathogen kill and cell quality. Each system utilizes various chemistries that not only impact which pathogens they can inactivate and how, but also how the treatments affect the plasma and cellular proteins and to what degree. This paper aims to present the various chemical mechanisms for pathogen reduction in transfusion medicine that are currently practiced or in development. [ABSTRACT FROM AUTHOR]

Surface nanotopograpy has been recognized as an important regulator of cellular responses including those of immune cells, the latter being of particular importance for implantable materials since these can determine biomaterial fate. In this paper, evidence is provided that the scale of surface nanotopography modulates the conformation of attached serum proteins, which in turn controls immune cell adhesion and activation. Model surfaces of tailored nanotopography of heights of 16, 38, and 68 nm are created by covalent immobilization of gold nanoparticles to an oxazoline‐rich plasma polymer film. This strategy not only produces surfaces of tailored nanofeature density but allows control of the outermost surface chemistry. Circular dichroism spectroscopy and Mac‐1 positive THP‐1 monocytes studies demonstrate distinct protein unfolding patterns, which upregulate or downregulate the expression of proinflammatory cytokines and cells attachment. The findings presented in this paper shed light on the missing relationship between surface nanotopography, protein unfolding, and the immune response. On the other hand, this work demonstrates the possibility to use specifically tailored surface nanotoporaphy scales to modulate and achieve desired immune responses. This study provides the missing link between surface nanotopography, protein unfolding, and immune responses. Precisely engineered surfaces demonstrate nanotopography size‐dependent fibrinogen unfolding and exposure of hidden peptide sequences, which activate the Mac‐1 receptor of inflammatory cells. Nanofeatures of ≈38 nm lead to minimal protein unfolding and reduced expression of proinflammatory cytokines. [ABSTRACT FROM AUTHOR]

Keywords: allergens; food allergy; IgE; IgG4; peanut EN allergens food allergy IgE IgG4 peanut 1835 1836 2 07/14/20 20200701 NES 200701 To the Editor, Mose et al1 reopened the discussion on bioavailability of digestion-stable food allergens in human. Their paper elegantly shows that peanut allergen can be detected in serum, minutes to hours following peanut consumption by healthy subjects. [Extracted from the article]

Background: Attainment of adequate transfer of passive immunity (TPI) is critical to health of calves; however, studies comparing available tools for measurement of TPI in individual beef animals are limited. Objectives: To report agreement between 4 tests evaluating individual TPI status in beef calves. Animals: One hundred ninety‐six beef calves born to cows and heifers presenting for calving management or dystocia. Methods: Retrospective study to assess serum immunoglobulin (IgG) concentrations via turbidimetric immunoassay (TI), gamma‐glutamyl transferase (GGT), serum total protein (TP), and single radial immunodiffusion (RID; reference standard). Test agreement was evaluated using Passing‐Bablok regression, Bland‐Altman analysis, Cohen's kappa, and receiver operating characteristic (ROC) curves with and without covariate adjustment to determine optimal thresholds. Results: Correlation between RID and test results varied: TI, ρ = 0.757; TP, ρ = 0.715; GGT: ρ = 0.413. For the TI compared to RID, regression analysis identified a constant (intercept = −0.51 [CI: −2.63, 3.05]) and proportional (slope = 1.87 [CI: 1.69, 2.08]) bias. Based on ROC, TI concentrations of ≤9.89 and ≤13.76 g/L, and TP concentrations of ≤5.5 and ≤6.0 g/dL, indicated IgG concentrations <18.0 and <25.0 g/L, respectively. Conclusions and Clinical Importance: Within this cohort of calves, TI demonstrated the best correlation with RID; however, significant bias was identified which led to frequent underestimation of IgG concentration. Serum total protein demonstrated less correlation with RID but had less misclassification than TI. Both TI and TP demonstrated less correlation for calves that received colostrum replacement prompting clinical awareness of colostrum type when evaluating individual TPI in beef calves. [ABSTRACT FROM AUTHOR]

During skeletal development, bone growth and mineralization require transport of substantial amounts of calcium, while maintaining very low concentration. How an organism overcomes this major logistical challenge remains mostly unexplained. To shed some light on the dynamics of this process, cryogenic focused ion beam‐scanning electron microscopy (cryo‐FIB/SEM) is used to image forming bone tissue at day 13 of a chick embryo femur. Both cells and matrix in 3D are visualized and observed as calcium‐rich intracellular vesicular structures. Counting the number of these vesicles per unit volume and measuring their calcium content based on the electron back‐scattering signal, the intracellular velocity at which these vesicles need to travel to transport all the calcium required for the mineral deposited in one day within the collagenous tissue can be estimated. This velocity at 0.27 µm s−1 is estimated, which is too large for a diffusion process and rather suggests active transport through the cellular network. It is concluded that calcium logistics is hierarchical and based on several transport mechanisms: first through the vasculature using calcium‐binding proteins and the blood flow, then active transport over tens of micrometers through the network of osteoblasts and osteocytes, and finally diffusive transport over the last one or two microns. [ABSTRACT FROM AUTHOR]

Extracellular vesicles and particles (EVPs) are secreted by organs across the body into different circulatory systems, including the bloodstream, and reflect pathophysiologic conditions of the organ. However, the heterogeneity of EVPs in the blood makes it challenging to determine their organ of origin. We hypothesized that small (s)EVPs (<100 nmin diameter) in the bloodstreamcarry distinctive protein signatures associated with each originating organ, and we investigated this possibility by studying the proteomes of sEVPs produced by six major organs (brain, liver, lung, heart, kidney, and fat). We found that each organ contained distinctive sEVP proteins: 68 proteins were preferentially found in brain sEVPs, 194 in liver, 39 in lung, 15 in heart, 29 in kidney, and 33 in fat. Furthermore, we isolated sEVPs from blood and validated the presence of sEVP proteins associated with the brain (DPP6, SYT1,DNM1L), liver (FABPL, ARG1, ASGR1/2), lung (SFPTA), heart (CPT1B), kidney (SLC31), and fat (GDN). We further discovered altered levels of these proteins in serum sEVPs obtained from old mice compared to young mice. In sum, we have cataloged sEVP proteins that can serve as potential biomarkers for organ identification in serum and show differential expression with age. [ABSTRACT FROM AUTHOR]

The complexity of cardiovascular diseases (CVDs), which remains the leading cause of death worldwide, makes the current clinical pathway for cardiovascular risk assessment unsatisfactory, as there remains a substantial unexplained residual risk. Simultaneous assessment of a large number of plasma proteins may be a promising tool to further refine risk assessment, and lipoprotein‐associated proteins have the potential to fill this gap. Technical advances now allow for high‐throughput proteomic analysis in a reproducible and cost‐effective manner. Proteomics has great potential to identify and quantify hundreds of candidate marker proteins in a sample and allows the translation from isolated lipoproteins to whole plasma, thus providing an individual multiplexed proteomic fingerprint. This narrative review describes the pathophysiological roles of atherogenic apoB‐containing lipoproteins and the recent advances in their mass spectrometry‐based proteomic characterization and quantitation for better refinement of CVD risk assessment. [ABSTRACT FROM AUTHOR]

The Human Plasma Proteome has always been the most investigated compartment in proteomics‐based biomarker discovery, and is considered the largest and deepest version of the human proteome, reflecting the state of the body in health and disease. Even if efforts have been always dedicated to the refinement of proteomic approaches to investigate more deeply the plasma proteome, it should not be forgotten that also highly abundant plasma proteins, like human serum albumin (HSA), often neglected in these studies, might provide fundamental physiological functions in plasma, and should be better considered. This review summarizes the important roles of HSA in the context of cardiovascular diseases (CVD), and in particular in heart failure. Notwithstanding much attention has been historically directed toward the association of HSA levels and CVD risk, the advances in the field of mass spectrometry research allow also a better characterization of the effects of oxidative modifications that could alter not only the structure but also the function of HSA. [ABSTRACT FROM AUTHOR]

Objective: To retrospectively analyze the efficacy and safety of dupilumab in the treatment of bullous pemphigoid. Methods: From October 2020 to October 2022, the medical records of patients with bullous pemphigoid who were treated with dupilumab in our department were collected retrospectively to analyze the therapeutic effect and changes in laboratory indexes. Results: The records of a total of 11 patients with bullous pemphigoid who were treated with dupilumab was reviewed. Within 2 weeks of the treatment, 10 (90.9%) of the 11 patients had complete or substantial control of the disease. The BPDAI scores of the patients decreased from baseline 113 (62, 181) to 37 (6, 130) at 2 weeks (p =.001) and 4 (0, 37) at 12 weeks after treatment (p <.001). In the 11 patients treated with dupilumab, the relief time of pruritus was 0–3 days (0.5, 7) days, and the pruritus was significantly alleviated after 2 weeks (t = 15.925, p <.001). The DLQI score decreased from (25.5 ± 2.5) before treatment, to (11.8 ± 4.4) at 2 weeks (t = 10.764, p <.001) and (2.1 ± 1.9) at 12 weeks (t = 30.038, p <.001). The patients had high eosinophil counts, high serum IgE levels, low serum total protein levels, and abnormal blood coagulation function. The aforementioned indicators gradually returned to normal after treatment. No adverse reactions occurred during the treatment. Conclusion: Dupilumab can effectively control the condition of bullous pemphigoid, efficiently relieve pruritus symptoms, and is relatively safe. [ABSTRACT FROM AUTHOR]

Lactococcosis, caused by the Gram‐positive bacterium Lactococcus garvieae, is a major concern in rainbow trout (Oncorhynchus mykiss) farms, which are regularly affected by outbreaks especially during the summer/fall months. In these farms, unvaccinated healthy and symptomatic fish can coexist with vaccinated fish. In the present study, innate (leukogram, serum lysozyme activity, peroxidase activity, antiprotease activity, bactericidal activity, total IgM and total proteins), and specific immune parameters (serum antibodies to L. garvieae) were assessed in unvaccinated adult rainbow trout naturally exposed to the pathogen, with or without evidence of clinical signs, or subjected to vaccination. Blood was drawn from all three groups, and blood smears were prepared. Bacteria were found in the blood smears of 70% of the symptomatic fish but not in any of the asymptomatic fish. Symptomatic fish showed lower blood lymphocytes and higher thrombocytes than asymptomatic fish (p ≤.05). Serum lysozyme and bactericidal activity did not vary substantially among groups; however, serum antiprotease and peroxidase activity were significantly lower in the unvaccinated symptomatic group than in the unvaccinated and vaccinated asymptomatic groups (p ≤.05). Serum total proteins and total immunoglobulin (IgM) levels in vaccinated asymptomatic rainbow trout were significantly higher than in unvaccinated asymptomatic and symptomatic groups (p ≤.05). Similarly, vaccinated asymptomatic fish produced more specific IgM against L. garvieae than unvaccinated asymptomatic and symptomatic fish (p ≤.05). This preliminary study provides basic knowledge on the immunological relationship occurring between the rainbow trout and L. garvieae, potentially predicting health outcomes. The approach we proposed could facilitate infield diagnostics, and several non‐specific immunological markers could serve as reliable indicators of the trout's innate ability to fight infection. [ABSTRACT FROM AUTHOR]

Background: The Impella® microaxial blood pumps utilize purge fluid containing heparin to prevent biofouling of internal surfaces. Purge fluid interfaces with blood or blood components at two notable internal locations: (1) 5–8 μm radial gap ("Radial Gap" or "Gap 1") between the motor shaft and bearing, a site accessible by blood proteins or small molecules; and (2) 100 μm axial gap ("Axial Gap" or "Gap 2") between the impeller rotor and bearing, the site of mixing with larger circulating blood components. Despite its efficacy, heparin in the purge fluid complicates overall patient anticoagulation management. Here, we investigate sodium bicarbonate as an alternative to heparin in the purge fluid in a simulated purge gap micro‐environment. Methods: To assess protein stability simulated at Gap 1, human serum albumin (HSA; 40 mg/ml) species were quantified utilizing size exclusion liquid chromatography (SEC‐HPLC) after stirring with purge fluid (5% dextrose in water (D5W) with heparin (25 U/ml) or sodium bicarbonate (25 or 50 mEq/L)) over a 24‐h period. pH measurements were taken immediately prior to stirring. Mixing between blood and purge fluid at Gap 2 was mimicked in vitro utilizing a 60:40 blood: purge fluid ratio. Purge fluid consisted of D5W with or without sodium bicarbonate (25 or 50 mEq/L). Human citrated blood samples were freshly collected with or without the addition of heparin (5 U/ml). Coagulability was determined via thromboelastography (TEG). pH measurements of blood mixtures were taken immediately before and after TEG analysis. Results: Sodium bicarbonate alone or synergistically with heparin was effective in increasing protein stability, increasing pH, and reducing coagulability. In the Gap 1 model, sodium bicarbonate led to preservation of HSA monomer after 24 h mixing, with monomer composing 88.3 ± 2.3% and 88.6 ± 0.9% of total HSA species for 25 or 50 mEq/L sodium bicarbonate, respectively. Only 60.4 ± 4.3% monomer was observed with D5W alone (p < 0.005). HSA aggregates and fragments were evident in heparin and D5W purge mixtures, but absent in sodium bicarbonate (25 and 50 mEq/L). pH of HSA mixtures significantly increased in the presence of sodium bicarbonate. In the Gap 2 model, combined heparin (5 U/ml) and sodium bicarbonate prolonged clotting time (TEG‐ACT), leading to an average increase of 795 ± 275 s (p = 0.04) and 846 243 s (p = 0.03). This trend of reduced coagulability was similarly observed in clot initiation time (R time), clot formation time (K time), and clotting rate (α angle). Blood mixture pH measurements increased with addition of sodium bicarbonate in both heparinized and non‐heparinized blood samples. Conclusion: Sodium bicarbonate in the purge fluid has the potential to significantly increase protein stability and reduce protein denaturation at the Impella® radial gap (Gap 1), while reducing blood coagulation at the Impella® axial gap (Gap 2). The influence of sodium bicarbonate on the biochemical environment of the purge fluid may ensure stable purge flow resistance and play a synergistic or supportive role in the purge gap micro‐environment when used with systemic anticoagulation. [ABSTRACT FROM AUTHOR]

Genome‐wide association studies (GWAS) have identified around 200 loci associated with breast cancer risk. However, protein targets for these loci remain largely unknown. Identifying protein targets and biomarkers can improve the understanding of cancer biology and etiology and identify high‐risk individuals for cancer prevention. In this study, we investigated genetically predicted levels of 1142 circulating proteins with breast cancer risk in 133 384 cases and 113 789 controls of European ancestry included in the Breast Cancer Association Consortium (BCAC). We identified 22 blood protein biomarkers associated with the risk of overall breast cancer at a false discovery rate (FDR) <0.05, including nine proteins encoded by genes located at least 500 kb away from previously reported risk variants for breast cancer. Analyses focusing on 124 encoding genes located at GWAS‐identified breast cancer risk loci found 20 proteins associated with overall breast cancer risk and one protein associated with triple‐negative breast cancer risk at FDR <0.05. Adjustment for the GWAS‐identified risk variants significantly attenuated the association for 13 of these proteins, suggesting that these proteins may be the targets of these GWAS‐identified risk loci. The identified proteins are involved in various biological processes, including glutathione conjugation, STAT5 signaling and NF‐κB signaling pathways. Our study identified novel protein targets and risk biomarkers for breast cancer risk. [ABSTRACT FROM AUTHOR]

Background: Allergen source‐derived proteases are a critical factor in the formation and development of asthma. The cysteine protease activity of house dust mite (HDM) disrupts the epithelial barrier function. The expression of cystatin SN (CST1) is elevated in asthma epithelium. CST1 inhibits the cysteine protease activity. We aimed to elucidate the role of epithelium‐derived CST1 in the development of asthma caused by HDM. Methods: CST1 protein levels in sputum supernatants and serum of patients with asthma and healthy volunteers were measured by ELISA. The ability of CST1 protein to suppress HDM‐induced bronchial epithelial barrier function was examined in vitro. The effects of exogenous CST1 protein on abrogating HDM‐induced epithelial barrier function and inflammation were examined in mice in vivo. Results: CST1 protein levels were higher in sputum supernatants (142.4 ± 8.95 vs 38.87 ± 6.85 ng/mL, P < 0.0001) and serum (1129 ± 73.82 vs 703.1 ± 57.02 pg/mL, P = 0.0035) in patients with asthma than in healthy subjects. The levels were significantly higher in patients with not well‐ and very poorly controlled asthma than those with well‐controlled asthma. Sputum and serum CST1 protein levels were negatively correlated with lung function in asthma. CST1 protein levels were significantly lower in the serum of HDM‐specific IgE (sIgE)‐positive asthmatics than in sIgE‐negative asthmatics. The HDM‐induced epithelial barrier function disruption was suppressed by recombinant human CST1 protein (rhCST1) in vitro and in vivo. Conclusion: Our data indicated that human CST1 protein suppresses asthma symptoms by protecting the asthmatic bronchial epithelial barrier through inhibiting allergenic protease activity. CST1 protein may serve as a potential biomarker for asthma control. [ABSTRACT FROM AUTHOR]

Summary: Pattern recognition receptors (PRRs) are plasma membrane‐localised proteins that sense molecular patterns to initiate pattern‐triggered immunity (PTI). Receptor‐like cytoplasmic kinases (RLCKs) function downstream of PRRs to propagate signal transduction via the phosphorylation of substrate proteins. The identification and characterisation of RLCK‐regulated substrate proteins are critical for our understanding of plant immunity.We showed that SHOU4 and SHOU4L are rapidly phosphorylated upon various patterns elicitation and are indispensable for plant resistance to bacterial and fungal pathogens. Protein–protein interaction and phosphoproteomic analysis revealed that BOTRYTIS‐INDUCED KINASE 1, a prominent protein kinase of RLCK subfamily VII (RLCK‐VII), interacted with SHOU4/4L and phosphorylated multiple serine residues on SHOU4L N‐terminus upon pattern flg22 treatment. Neither phospho‐dead nor phospho‐mimic SHOU4L variants complemented pathogen resistance and plant development defect of the loss‐of‐function mutant, suggesting that reversible phosphorylation of SHOU4L is critical to plant immunity and plant development.Co‐immunoprecipitation data revealed that flg22 induced SHOU4L dissociation from cellulose synthase 1 (CESA1) and that a phospho‐mimic SHOU4L variant inhibited the interaction between SHOU4L and CESA1, indicating the link between SHOU4L‐mediated cellulose synthesis and plant immunity.This study thus identified SHOU4/4L as new components of PTI and preliminarily revealed the mechanism governing SHOU4L regulation by RLCKs. [ABSTRACT FROM AUTHOR]

Aim: To investigate whether co‐ingestion of dietary protein with, or before, carbohydrate may be a useful strategy to reduce postprandial hyperglycaemia in older men with type 2 diabetes (T2D). Materials and Methods: Blood glucose, plasma insulin and glucagon concentrations were measured for 180 minutes following ingestion of a drink containing 30 g of glucose (G; 120 kcal), 30 g of whey protein (120 kcal), 30 g of glucose plus 30 g of whey protein (GP; 240 kcal), or control (~2 kcal) in older men with T2D (n = 10, 77 ± 1 years; 31 ± 1.7 kg/m2) and without T2D (n = 10, 78 ± 2 years; 27 ± 1.4 kg/m2). Mixed model analysis was used. Results: GP versus G markedly reduced the increase in blood glucose concentrations (P <.001) and had a synergistic effect on the increase in insulin concentrations (P <.001), in men both with and without T2D. Glucose concentrations were higher in men with T2D compared with those without T2D, whereas insulin and glucagon concentrations were largely unaffected by the presence of T2D. Gastric emptying was faster in men with T2D than in those without T2D. Conclusions: The ability of whey protein to reduce carbohydrate‐induced, postprandial hyperglycaemia is retained in older men with T2D compared with those without T2D, and whey protein supplementation may be a useful strategy in the prevention and management of T2D in older people. [ABSTRACT FROM AUTHOR]

Understanding the molecular basis of adaptations in coevolving species requires identifying the genes that underlie reciprocally selected phenotypes, such as those involved in venom in snakes and resistance to the venom in their prey. In this regard, California ground squirrels (CGS; Otospermophilus beecheyi) are eaten by northern Pacific rattlesnakes (Crotalus oreganus oreganus), but individual squirrels may still show substantial resistance to venom and survive bites. A recent study using proteomics identified venom interactive proteins (VIPs) in the blood serum of CGS. These VIPs represent possible resistance proteins, but the sequences of genes encoding them are unknown despite the value of such data to molecular studies of coevolution. To address this issue, we analyzed a de novo assembled transcriptome from CGS liver tissue—where many plasma proteins are synthesized—and other tissues from this species. We then examined VIP sequences in terms of three characteristics that identify them as possible resistance proteins: evidence for positive selection, high liver expression, and nonsynonymous variation across CGS populations. Based on these characteristics, we identified five VIPs (i.e., α‐2‐macroglobulin, α‐1‐antitrypsin‐like protein GS55‐LT, apolipoprotein A‐II, hibernation‐associated plasma protein HP‐20, and hibernation‐associated plasma protein HP‐27) as the most likely candidates for resistance proteins among VIPs identified to date. Four of these proteins have been previously implicated in conferring resistance to the venom in mammals, validating our approach. When combined with the detailed information available for rattlesnake venom proteins, these results set the stage for future work focused on understanding coevolutionary interactions at the molecular level between these species. Research Highlights: Using RNA‐sequencing data from liver, we assembled a California ground squirrel transcriptome and used it to identify putative resistance proteins that interact with northern Pacific rattlesnake venom proteins setting the stage for molecular studies of coevolution. [ABSTRACT FROM AUTHOR]