Coagulation Proteins Research Articles

Coagulation Parameters among Individuals with Hepatitis B Infections in Okada, Edo State, Nigeria

Hepatitis B Viral (HBV) infection is a condition of inflammation of the liver. The Liver plays a vital role in the body which includes filtration of the blood, production of coagulation proteins and also as a storage organ. In this study, alteration of some coagulation parameters which include, Prothrombin time (PT), activated partial thromboplastin time (APTT), Platelets counts (PLATS) were determined in sixty (60) known seropositive individuals using standard haemostatic techniques. There was a significant reduction in the mean value (mean values) on platelet counts in the seropositive individuals (p- value) compared to the control group (mean value), also revealed in this study is an increase in the mean-time which was significant of PT among test individuals compared to seronegative controls individuals, there was an increase prothrombin time as well as Activated partial prothrombin time with both having p- values < 0.001. The average PT and APTT were 25.53 and 48.89 respectively. Individuals with Hepatitis B infection are thus liable to thrombocytopenia if not managed.

Association of Complement and Coagulation Pathway Proteins With Treatment Response in First-Episode Psychosis: A Longitudinal Analysis of the OPTiMiSE Clinical Trial.

Treatment response to specific antipsychotic medications is difficult to predict on clinical grounds alone. The current study hypothesizes that the baseline complement pathway activity predicts the treatment response and investigates the relationship between baseline plasma biomarkers with treatment response to antipsychotic medications. Baseline plasma samples were collected from first episode of psychosis patients (n = 243) from a multi-center clinical trial. The participants were treated with amisulpride for 4 weeks. Levels of complement and coagulation proteins at baseline were measured using both data-dependent and data-independent mass spectrometry approaches. The primary outcome was remission status at 4 weeks and the secondary outcomes included change in psychotic and functional symptoms over the period of treatment. In addition, immunoassays were performed at baseline for complement C1R, as well as for activation markers C4a and sC5b-9. The plasma level of complement variant C4A was significantly associated with remission at 4 weeks. Moreover, higher levels of several complement and coagulation pathway proteins were associated with a reduction in psychotic symptoms and an improvement in functioning. Immunoassays showed an association of baseline levels of C1R and C4a as well as complement activation marker sC5b-9 levels with treatment response. The results demonstrated that the response to antipsychotic treatment might be related to pre-treatment levels of plasma complement and coagulation pathway proteins. This is consistent with independent evidence associating immune dysfunction with the pathophysiology of psychosis. Moreover, these results inform the development of novel therapeutic approaches that target the complement system for psychosis.

Coagulation of model infant formulae: Impact on their in vitro dynamic gastric digestion

The rheological properties and the in vitro dynamic gastric digestion behaviours of model infant formulae containing various ratios of casein to whey protein (0:100, 40:60, 60:40 and 80:20) were investigated using a rheometer and an infant human gastric simulator. The pH profiles and the viscoelastic moduli were determined during acidification by glucono-δ-lactone and at different pepsin concentrations (0, 0.3, 1.0, 1.7 and 2.5 U/mL). The infant formula containing whey protein only formed a weak gel at about pH 5. In the presence of pepsin, all casein-containing infant formulae formed gels at pH ∼6.0 and only low pepsin activity (>0.3 U/mL) was required for the gelation. The strength of the gel/coagulum was dependent mainly on the casein content. Coagulation of protein and flocculation of oil droplets, induced by both pepsin proteolysis and the low pH under gastric conditions, were observed in the infant formulae during dynamic digestion in the human gastric simulator. Confocal images showed that the infant formula containing whey protein only formed smaller flocs of aggregated protein and oil droplets, which may have led to their observed faster protein digestion, than the infant formulae containing a combination of casein and whey protein. The casein-dominant infant formulae had greater aggregation than the whey-protein-dominant infant formulae during gastric digestion, which led to their lower rate of casein digestion. These results suggest that the extent of coagulation and the gastric digestion of the proteins in infant formulae can be controlled by the rational selection of the protein components and the ratio of casein to whey protein, such that different nutritional requirements of infants can be met.

Coagulation factor dysregulation in polycystic ovary syndrome is an epiphenomenon of obesity.

Obese women with polycystic ovary syndrome (PCOS) exhibit a hypercoagulable state, with the suggestion that this may be obesity-driven rather than an intrinsic facet of PCOS; however, this has not yet been definitively determined since body mass index (BMI)is so highly correlated with PCOS. Therefore, only a study design where obesity, insulin resistance and inflammation are matched can answer this question. This was a cohort study. Patients Weight and aged-matched nonobese women with PCOS (n = 29) and control women (n = 29) were included. Measurements Plasma coagulation pathway protein levels were measured. Circulating levels of a panel of nine clotting proteins known to differ in obese women with PCOS were determined by Slow Off-rate Modified Aptamer (SOMA)-scan plasma protein measurement. Women with PCOS showed a higher free androgen index (FAI) and anti-Müllerian hormone, but measures of insulin resistance, and C reactive protein (as a marker of inflammation), did not differ between the nonobese women with PCOS and the control women. Seven pro-coagulation proteins (plasminogen activator inhibitor-1, fibrinogen, fibrinogen gamma chain, fibronectin, d-dimer, P-selectinand plasma kallikrein) and two anticoagulant proteins (vitamin K-dependent protein-S and heparin cofactor-II) known to be elevated in obese women with PCOS did not differ from controls in this cohort. This novel data showthat clotting system abnormalities do not contribute to the intrinsic mechanisms underlying PCOS in this nonobese noninsulin resistant population of women with PCOS matched for age and BMI, and without evidence of underlying inflammation, but rather the clotting factor changes are an epiphenomenon coincident with obesity; therefore, increased coagulability is unlikely in these nonobese PCOS women.

CT hyperdense cerebral artery sign reflects distinct proteomic composition in acute ischemic stroke thrombus

BackgroundHyperdense cerebral artery sign (HCAS) is an imaging biomarker in acute ischemic stroke (AIS) that has been shown to be associated with various clinical outcomes and stroke etiology. While prior...

Biomarkers for Lipid and Albumin Metabolism in Hospitalized Patients with Underlying Diseases and Community-Acquired Pneumonia Caused by Bacterial or SARS-CoV-2 Infection.

To look at the differences and similarities in albumin and lipid metabolism in non-severe COVID-19 infection, non-severe community-acquired pneumonia, and severe community pneumonia with underlying diseases, as well as the relationship between albumin and lipid metabolism and inflammatory mediators. This retrospective analysis comprised 253 individuals with bacterial pneumonia and COVID-19 infection (1 May 2021-1 May 2022). Routine blood examination, blood lipid levels, albumin level, C-reactive protein (CRP) levels, coagulation function, cardiac enzymes, liver function, renal function, immunological function, and bacterial culture were also collected. Correlation analysis was performed using Spearman's test for lipid parameter and Inflammatory factors in the blood. Furthermore, the multiple linear regression (MLR) analysis was employed to analyze the multicollinearity in lipidomics data. The statistical analysis was performed using SPSS statistic version 19.0. There were 63 (24.90%) non-severe community-acquired pneumonia patients (NSCAP), 48 (18.97%) severe community-acquired pneumonia patients (SCAP), 112 (44.27%) non-severe COVID-19 infection patients (NSCOV), and 30 (11.86%) healthy volunteers (HV). In all, 45.59% (116/253) of the patients had underlying diseases. Patients with community-acquired pneumonia had lower albumin and cholesterol levels than those with non-severe COVID-19 infection and healthy controls (t = -3.81, -2.09, P = 0.00, 0.04). Albumin, triglyceride, cholesterol, and LDL-C levels in peripheral blood were considerably lower in the SCAP group than in the NSCAP group. Albumin, cholesterol, HDL-C, LDL-C, and aop-A were all inversely connected with CRP in the SCAP with underlying illness group, but cholesterol level was favorably correlated with lymphocyte count (R = 0.36, P = 0.01). Hypoproteinemia, hypotriglyceridemia, and an elevated neutrophil-to-lymphocyte count ratio are all risk factors for severe community-acquired pneumonia. Hypoalbuminemia and abnormal lipid metabolism are important indicators of bacterial infection, especially severe bacterial pneumonia.

ВПЛИВ РІЗНИХ СИЧУЖНИХ ЕНЗИМІВ НА ОСНОВНІ ПОКАЗНИКИ МОЦАРЕЛИ ЗА ТЕХНОЛОГІЇ М’ЯКИХ СИРІВ

У статті викладено результати дослідження впливу різних сичужних ензимів на основні показники моцарели за технології м’яких сирів. Для постановки досліду було сформовано ІІІ групи проб молока (n=5). У контрольній групі проб для зсідання молока використовували сичужний ензим мікробіального походження. У І дослідній групі проб застосовувався ензимний препарат із сичугів молочних телят, екстрагований за методикою Ю.Я. Свириденка. У ІІ дослідній групі застосовували ензимний препарат, який екстрагували із сичугів молочних телят за методикою С.В. Мерзлова. Тривалість зсідання білків при виробництві сиру «Моцарела» із використанням мікробіального ензимного препарату 23 хв., при використанні ензимів І дослідної групи – 27 хв. і при використанні сичужних ензимів ІІ дослідної групи – 33 хв. З огляду на вихід готового сиру «Моцарела» з комерційною метою для отримання більшого прибутку, доцільно використовувати ензим мікробіального походження. Згідно із сенсорним аналізом зразки із використанням телячого сичужного ензиму екстрагованих за методикою С.В. Мерзлова І дослідна група проб характеризувалися вираженим сирним, кисломолочним, без сторонніх присмаків та запахів, властивим м’якому свіжому сиру смаком і запахом; поверхня чиста без механічних ушкоджень, пружна; консистенція мазка, ніжна, в міру щільна; колір білий, рівномірний за всією масою; тісто з наявністю вічок. Найгіршими органолептичними показниками характеризувалися зразки ІІ дослідної групи.

Increased thrombin activatable fibrinolysis inhibitor activity is associated with hypofibrinolysis in dogs with sepsis.

Disorders of coagulation are well-recognized in dogs with sepsis, but data regarding fibrinolysis disorders are limited. We aimed to characterize fibrinolysis in dogs with sepsis compared to healthy controls. We hypothesized that dogs with sepsis would be hypofibrinolytic, and that hypofibrinolysis would be associated with non-survival. This was a prospective observational cohort study. We enrolled 20 client-owned dogs with sepsis admitted to the Cornell University Hospital for Animals and 20 healthy pet dogs. Coagulation and fibrinolytic pathway proteins including antiplasmin activity (AP), antithrombin activity (AT), thrombin activatable fibrinolysis inhibitor activity (TAFI), D-dimer concentration, fibrinogen concentration, and plasminogen activity were measured and compared between groups. Overall coagulation potential, overall fibrinolysis potential, and overall hemostatic potential were calculated from the curve of fibrin clot formation and lysis over time. Compared to healthy controls, dogs with sepsis had lower AT (P = 0.009), higher AP (P = 0.002), higher TAFI (P = 0.0385), and higher concentrations of fibrinogen (P < 0.0001) and D-dimer (P = 0.0001). Dogs with sepsis also had greater overall coagulation potential (P = 0.003), overall hemostatic potential (P = 0.0015), and lower overall fibrinolysis potential (P = 0.0004). The extent of fibrinolysis was significantly negatively correlated with TAFI. No significant differences were observed between survivors and non-survivors. Dogs with sepsis were hypercoagulable and hypofibrinolytic compared to healthy dogs, suggesting potential utility of thromboprophylaxis in this patient population. The association between high TAFI and low overall fibrinolysis potential might provide a potential mechanism for this hypofibrinolysis.

DNA methylation changes and increased mRNA expression of coagulation proteins, factor V and thrombomodulin in Fuchs endothelial corneal dystrophy

Late-onset Fuchs endothelial corneal dystrophy (FECD) is a disease affecting the corneal endothelium (CE), associated with a cytosine-thymine-guanine repeat expansion at the CTG18.1 locus in the transcription factor 4 (TCF4) gene. It is unknown whether CTG18.1 expansions affect global methylation including TCF4 gene in CE or whether global CE methylation changes at advanced age. Using genome-wide DNA methylation array, we investigated methylation in CE from FECD patients with CTG18.1 expansions and studied the methylation in healthy CE at different ages. The most revealing DNA methylation findings were analyzed by gene expression and protein analysis. 3488 CpGs had significantly altered methylation pattern in FECD though no substantial changes were found in TCF4. The most hypermethylated site was in a predicted promoter of aquaporin 1 (AQP1) gene, and the most hypomethylated site was in a predicted promoter of coagulation factor V (F5 for gene, FV for protein). In FECD, AQP1 mRNA expression was variable, while F5 gene expression showed a ~ 23-fold increase. FV protein was present in both healthy and affected CE. Further gene expression analysis of coagulation factors interacting with FV revealed a ~ 34-fold increase of thrombomodulin (THBD). THBD protein was detected only in CE from FECD patients. Additionally, we observed an age-dependent hypomethylation in elderly healthy CE.Thus, tissue-specific genome-wide and gene-specific methylation changes associated with altered gene expression were discovered in FECD. TCF4 pathological methylation in FECD because of CTG18.1 expansion was ruled out.

The Gilded Clot: Review of Metal-Modulated Platelet Activation, Coagulation, and Fibrinolysis.

The processes of blood coagulation and fibrinolysis that in part maintain the physical integrity of the circulatory system and fluidity of its contents are complex as they are critical for life. While the roles played by cellular components and circulating proteins in coagulation and fibrinolysis are widely acknowledged, the impact of metals on these processes is at best underappreciated. In this narrative review we identify twenty-five metals that can modulate the activity of platelets, plasmatic coagulation, and fibrinolysis as determined by in vitro and in vivo investigations involving several species besides human beings. When possible, the molecular interactions of the various metals with key cells and proteins of the hemostatic system were identified and displayed in detail. It is our intention that this work serve not as an ending point, but rather as a fair evaluation of what mechanisms concerning metal interactions with the hemostatic system have been elucidated, and as a beacon to guide future investigation.

Effect of ingestion temperature on the pepsin-induced coagulation and the in vitro gastric digestion behavior of milk

Pepsin-induced protein coagulation occurs in the gastric environment when the milk pH is above the isoelectric point of casein proteins. In this study, the effect of milk temperature (4–48 °C) on the hydrolysis of κ-casein by pepsin and the consequent protein coagulation was studied at pH 6.0 for 120 min. Quantitative determination of the released para-κ-casein showed that both the κ-casein hydrolysis reaction rate constant and the pepsin denaturation rate constant increased with an increase in temperature. The temperature coefficient (Q10) of the specific hydrolysis of κ-casein was calculated to be ∼1.95. The coagulation process was investigated by the evolution of the storage modulus (Gʹ). At higher temperature, the milk coagulated faster but had a lower firming rate and Gʹmax with larger aggregates and voids were observed. The digestion behavior of the milk ingested at 4 °C, 37 °C, or 50 °C was investigated for 240 min in a human gastric simulator, in which the milk temperature increased or decreased to 37 °C (body temperature) over ∼ 60 min. The coagulation of the 4 °C milk was slower than for the 37 °C and 50 °C milk. The curd obtained from the 4 °C milk had a looser and softer structure with a significantly higher moisture content at the initial stage of digestion (20 min) which, in turn, facilitated the breakdown and hydrolysis of the caseins by pepsin. During the digestion, the curd structure became more cohesive, along with a decrease in moisture content. The knowledge gained from this study provides insight into the effect of temperature on the kinetics of pepsin-induced milk coagulation and the consequent digestion behavior.

Extracellular vesicle proteomics and phosphoproteomics identify pathways for increased risk in patients hospitalized with COVID-19 and type 2 diabetes mellitus

Recent studies suggest that extracellular vesicles (EVs) play a role in the pathogenesis of SARS-CoV-2 infection and the severity of COVID-19. However, their role in the interaction between COVID-19 and type 2 diabetes (T2D) has not been addressed. Here, we characterized the circulating EV proteomic and phosphoproteomic landscape in patients with and without T2D hospitalized with COVID-19 or non-COVID-19 acute respiratory illness (RSP). We detected differentially expressed protein and phosphoprotein signatures that effectively characterized the study groups. The trio of immunomodulatory and coagulation proteins C1QA, C1QB, and C1QC appeared to be a central cluster in both the COVID-19 and T2D functional networks. PKCβ appeared to be retained in cells by being diverted from EV pathways and contribute to the COVID-19 and T2D interaction via a PKC/BTK/TEC axis. EV-shuttled CASP3 and ROCK1 appeared to be coregulated and likely contribute to disease interactions in patients with COVID-19 and T2D. Predicted activation of AMPK, MAPK, and SYK appeared to also play important roles driving disease interaction. These results suggest that activated cellular kinases (i.e., PKC, AMPK, MAPK, and SYK) and multiple EV-shuttled kinases (i.e., PKCβ, BTK, TEC, MAP2K2, and ROCK1) may play key roles in severe COVID-19, particularly in patients with comorbid diabetes.

Lactaderhin's C1 domain restricts protein's ensemble as it binds to charged membranes.

Lactadherin's (Lact) binding to phosphotidylserine (PS) enriched cell surfaces is critical for controlling our blood coagulation. Although structural and mutagenesis studies indicate important binding regions of this protein, they are not sufficient to construct a detailed, membrane-bound model especially since they mainly consider only one of its two globular domains, namely C2. We thus use molecular dynamics (MD) simulations and the AlphaFold program to generate an ensemble of Lact encounter complexes with a negatively charged bilayer consistent with biochemical studies. This was accomplished by first using the highly mobile membrane mimetic (HMMM) to efficiently simulate its C2 domain binding to PS lipids. We then converted the HMMM model to standard full-tail lipid membranes to further sample and obtain converged bound structures that we call the “vertical” and “side-lying” ensembles. From these bilayer-bound poses, we infer which one allows for attachment of its other C1 domain and use AlphaFold to reconstruct it and further simulate to capture equilibrium complexes. Modeling revealed that C1 domain restricts the dynamics structural ensemble of Lact bound to membranes. We also report Lact's full dynamic structure in the presence of negatively charged lipids and compare them with other homologous blood coagulation proteins to provide a rationale for its competitive binding. These results shed light on the structural mechanism of blood coagulation by considering its binding in a membrane environment which can serve as a general framework for studying these critical protein-membrane interactions. They also provide a structural basis for further understanding the significance that PS lipids play in regulating blood coagulation processes.

OP02 Distinct biological profiles associated with the risk of short-term relapse and mid/long-term relapse in Crohn’s disease patients stopping infliximab

Abstract Background In Crohn’s disease (CD) patients stopping infliximab (IFX), the risk of short-term relapse (&amp;lt;6 months) and mid/long-term (&amp;gt;6 months) relapse were associated with distinct biological profiles (STORI trial). Herein, we aim to test the external validity of this finding in an independent trial (SPARE). Methods The SPARE trial has included 211 CD patients (from 64 sites in Europe and Australia) in steroid-free remission &amp;gt;6 months, receiving a combined therapy (IFX and immunosuppressant (IS)) &amp;gt;8 months and who were then randomised in three arms: continuing combo, stopping IFX or stopping IS. The arm stopping IFX was used to externally validate our findings generated in the STORI trial. To this end, the measurement of 161 proteins obtained in the baseline serum of STORI was repeated in SPARE (arm stopping IFX) with the same technologies: selected reaction monitoring (SRM, 69 proteins measured in 67 patients) or proximity extension assay (PEA, 92 proteins measured in 63 patients). Associations between serum protein levels and time to relapse (HR: hazard ratio and its associated statistics) were determined by using univariable Cox model in the stratified (relapse &amp;lt;6 or &amp;gt;6 months) and non-stratified cohort. Results The blood proteins depicting the risk of short-term or mid/long-term relapse in STORI and SPARE were selected as shown Fig. 1 (SRM) and Fig. 2 (PEA). The representation of those markers in volcano plots clearly confirmed that short-term and mid/long-term relapsers present distinct biological profiles (Fig. 3-4). This is also corroborated by the fact that only ORM1 was selected as a marker of short-term and mid/long-term relapse (Fig. 1-2). In STORI and SPARE, the risk of short-term relapse was associated (p-value&amp;lt;0.05) with a high serum level of inflammatory markers (IL6, CRP, HPR, ORM1, LRG1, HP, CP, APCS, ITIH3), complement components (C8B, C4B), blood coagulation proteins (F9, SERPIND1), markers of dendritic cells (LAMP3, CLEC4C) and a low serum level of a complement component (MASP1) (Fig. 1-2-3). In STORI and SPARE, the risk of mid/long-term relapse was associated (p-value&amp;lt;0.05) with a high serum level of a complement component (CFB) and a low serum level of a protease inhibitor (SERPINA4), a growth factor (FGF2), an anti-inflammatory cytokine (IL10) (Fig. 1-2-4). For other proteins, STORI and SPARE showed convergent tendencies (p-value&amp;lt;0.01; Fig. 1-2-3-4) or differences (Fig. 1-2). Conclusion We confirm that, after stopping IFX in CD patients, the risk of short-term and mid/long-term relapse are associated with distinct biological profiles. The risk of short-term relapse is deeply associated with residual inflammation while the biological picture depicting the risk of mid/long-term relapse seems less clear.

"iPSC-derived liver organoids and inherited bleeding disorders: Potential and future perspectives".

The bleeding phenotype of hereditary coagulation disorders is caused by the low or undetectable activity of the proteins involved in hemostasis, due to a broad spectrum of genetic alterations. Most of the affected coagulation factors are produced in the liver. Therefore, two-dimensional (2D) cultures of primary human hepatocytes and recombinant overexpression of the factors in non-human cell lines have been primarily used to mimic disease pathogenesis and as a model for innovative therapeutic strategies. However, neither human nor animal cells fully represent the hepatocellular biology and do not harbor the exact genetic background of the patient. As a result, the inability of the current in vitro models in recapitulating the in vivo situation has limited the studies of these inherited coagulation disorders. Induced Pluripotent Stem Cell (iPSC) technology offers a possible solution to overcome these limitations by reprogramming patient somatic cells into an embryonic-like pluripotent state, thus giving the possibility of generating an unlimited number of liver cells needed for modeling or therapeutic purposes. By combining this potential and the recent advances in the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 technology, it allows for the generation of autologous and gene corrected liver cells in the form of three-dimensional (3D) liver organoids. The organoids recapitulate cellular composition and organization of the liver, providing a more physiological model to study the biology of coagulation proteins and modeling hereditary coagulation disorders. This advanced methodology can pave the way for the development of cell-based therapeutic approaches to treat inherited coagulation disorders. In this review we will explore the use of liver organoids as a state-of-the-art methodology for modeling coagulation factors disorders and the possibilities of using organoid technology to treat the disease.

T Cell Receptor Sequences Amplified during Severe COVID-19 and Multisystem Inflammatory Syndrome in Children Mimic SARS-CoV-2, Its Bacterial Co-Infections and Host Autoantigens.

Published hypervariable region V-beta T cell receptor (TCR) sequences were collected from people with severe COVID-19 characterized by having various autoimmune complications, including blood coagulopathies and cardiac autoimmunity, as well as from patients diagnosed with the Kawasaki disease (KD)-like multisystem inflammatory syndrome in children (MIS-C). These were compared with comparable published v-beta TCR sequences from people diagnosed with KD and from healthy individuals. Since TCR V-beta sequences are supposed to be complementary to antigens that induce clonal expansion, it was surprising that only a quarter of the TCR sequences derived from severe COVID-19 and MIS-C patients mimicked SARS-CoV-2 proteins. Thirty percent of the KD-derived TCR mimicked coronaviruses other than SARS-CoV-2. In contrast, only three percent of the TCR sequences from healthy individuals and those diagnosed with autoimmune myocarditis displayed similarities to any coronavirus. In each disease, significant increases were found in the amount of TCRs from healthy individuals mimicking specific bacterial co-infections (especially Enterococcus faecium, Staphylococcal and Streptococcal antigens) and host autoantigens targeted by autoimmune diseases (especially myosin, collagen, phospholipid-associated proteins, and blood coagulation proteins). Theoretical explanations for these surprising observations and implications to unravel the causes of autoimmune diseases are explored.

Insight into the Processing, Gelation and Functional Components of Tofu: A Review

Soybeans are a good source of protein and functional nutrition for humans and are used in the production of many foods, such as soymilk and tofu, which are popular in Asia. Soymilk is a beverage with a milky appearance and is produced by grinding soybeans in water; this raw soymilk is then filtered and heated. Soymilk can be used to produce tofu, a gelatinous food, in a process that generally involves coagulating, forming, and pressing soymilk. Tofu is also a source of functional ingredients, such as isoflavones, that have received much attention for their antioxidant properties. These isoflavones bind to soy protein to form soy protein–isoflavone complexes in tofu. In this review, we examined the processing, gelation and functional components of tofu. First, we introduced novel processing technologies (such as thermal and nonthermal processing) and hydrocolloids that affect the rheological property and texture of tofu. Then, because the coagulation and gelation of soymilk are important steps in the tofu manufacturing process, we also described detailed coagulation and gelation mechanisms of soybean proteins in tofu. Finally, we described the functional compounds and ingredients in tofu. This review provides basic knowledge for the further application of novel processing technology in tofu processing.

Phytase influence on soymilk protein colloid stability studied with thermographic method

Coagulation of soymilk is well studied from a technological point of view. However, physicochemical features of the process of soymilk protein colloidal stability destruction may be of interest both from a purely scientific point of view and for improving production processes. In this study, an attempt was made to analyze the role of phytinates in formation of the colloidal stability of soymilk proteins. A possible role of the dissociation of phosphate groups of phytic acid in the rise of an electric charge stabilizing soy protein micelles in water solution has been suggested. The ability of phytase to cut off the phosphate groups of phytic acid was used to substantiate our assumption. To study the kinetics of coagulation of soymilk proteins, a thermographic method was used. The results of the experiments show that the addition of phytase to soymilk can significantly accelerate its acid coagulation, which is an indirect confirmation of the above assumption.

Preparation of Cryoprecipitate and Cryo-depleted Plasma for Proteomic Research Analysis.

When frozen plasma is slowly thawed in cold conditions (1-6°C), high-molecular-weight plasma proteins precipitate forming a concentrate known as cryoprecipitate. The concentrate is enriched with several important coagulation proteins, including fibrinogen, antihemophilic factor (factor VIII), von Willebrand factor, fibrin stabilizing factor (factor XIII), fibronectin, and small amounts of other plasma proteins. In current medical practice, clinical-grade preparations of cryoprecipitate are used mostly to correct fibrinogen deficiency caused by acute blood loss or due to functional abnormalities of the fibrinogen protein. In the past, cryoprecipitate was used to treat von Willebrand disease and hemophilia A (factor VIII deficiency), but the availability of more highly purified coagulation factor concentrates or recombinant protein preparations has superseded the use of cryoprecipitate for these coagulopathies. Cryo-depleted plasma (also called cryosupernatant) is the plasma supernatant remaining following removal of the cryoprecipitate from frozen-thawed plasma and contains all the remaining soluble plasma proteins. This protocol describes the research-scale preparation of cryoprecipitate and cryo-depleted plasma suitable for proteomic studies and is based on the procedures used to prepare clinical-grade cryoprecipitate.

Expression of the gene encoding blood coagulation factor VIII without domain B in E. coli bacterial expression system.

In this article, we have demonstrated the feasibility of generating an active form of recombinant blood coagulation factor VIII using an E. coli bacterial expression system as a potential treatment for hemophilia type A. Factor VIII (FVIII), an essential blood coagulation protein, is a key component of the fluid phase blood coagulation system. So far, all available recombinant FVIII formulations have been produced using eukaryotic expression systems. Mammalian cells can produce catalytically active proteins with all the necessary posttranslational modifications. However, cultivating such cells is time-consuming and highly expensive, and the amount of the obtained product is usually low. In contrast to eukaryotic cells, bacterial culture is inexpensive and allows the acquisition of large quantities of recombinant proteins in a short time. With this study, we aimed to obtain recombinant blood coagulation factor VIII using the E. coli bacterial expression system, a method not previously explored for this purpose. Our research encompasses the synthesis of blood coagulation factor VIII and its expression in a prokaryotic system. To achieve this, we constructed a prokaryotic expression vector containing a synthetic factor VIII gene, which was then used for the transformation of an E. coli bacterial strain. The protein expression was confirmed by mass spectrometry, and we assessed the stability of the gene construct while determining the optimal growth conditions. The production of blood coagulation factor VIII by the E. coli bacterial strain was carried out on a quarter-technical scale. We established the conditions for isolation, denaturation, and renaturation of the protein, and subsequently confirmed the activity of FVIII.