Background Cirrhosis is associated with an increased frequency of thromboembolic events, particularly in the portal sector. Several hypotheses have been put forward to explain this hypercoagulability state. However, studies assessing the expression of procoagulant microparticles (MP) in cirrhosis are rare.Aims To evaluate the expression of procoagulant MP in cirrhosis using thrombin generation (TG) test and to assess the activity of MP according to the clinical and biological status of patients.Methods 39 cirrhotic patients and 39 control subjects were enrolled in a cross sectional comparative study. The biological explorations were performed at inclusion (The conventional tests of hemostasis, factor VIII (FVIII) and protein C (PC) assay and TG test carried out under sensitive conditions to MP activity and standard conditions).Results The mean age of patients was 65 years. Etiologies were dominated by viral infection (64%). The majority of patients were classified as Child-Pugh stage A (49%) and had decompensated cirrhosis (54%). Portal thrombosis was observed in 9 patients and hepatocellular carcinoma (HCC) was documented in 14 patients. Cirrhotic patients had higher FVIII/PC ratio and lower TG under different experimental conditions than controls. However, by expressing the results of MP dependent TG to the results of TG under optimal condition, patients generated as much thrombin as controls. MP dependent TG increased with the severity, decompensated status and complications of the disease.Conclusion The MP procoagulant activity in cirrhotic patients is comparable to that in controls and increases with disease severity, decompensation, and complications.
Background Human plasma-derived albumin (HA) is a critical therapeutic protein widely used in clinical applications. However, albumin aggregation remains one of the biggest challenges for plasma protein manufacturers worldwide, as it can compromise product stability, efficacy, safety, and yield. Factors that contribute to aggregation include protein concentration, ionic strength, temperature, agitation, pH fluctuations, and stabilizer concentrations during processing. This study examines how varying albumin concentrations affect aggregation under controlled pasteurisation conditions using identical parameters for complete process including agitation, temperature, and stabilizer concentration. Methods Albumin was purified from human plasma using a modified Cohn's fractionation method, yielding Fraction V intermediate paste. This paste underwent a series of ultrafiltration steps followed by pasteurisation at 60 +/- 1 degrees C for 10 hours, a vital virus inactivation process. To maintain consistency, all influencing parameters such as agitation, temperature, and stabilizer concentrations were kept constant. Three experimental designs were employed: (i) analysing polymer and aggregate formation across multiple batches with albumin concentrations between 20% and 28% having similar starting material i.e. Fraction V paste at 1 kg scale, obtained from manufacturing facility from same manufacturing pool of plasma, (ii) assessing the impact of NaCl at different albumin concentrations during pasteurisation, purified from same lot of Fraction V paste at 1 kg scale, and (iii) performing a trend analysis of polymer aggregate percentage on controlled albumin concentrations (about 25%, 23%, and 21%) in a single batch started at 3 kg scale of Fraction V paste where it was divided into three parts and diluted at three different above mentioned concentrations followed by pasteurisation. Aggregation levels were quantified post-pasteurisation. Results The findings indicate that albumin concentrations below 23% resulted in significantly higher polymer and aggregate formation (>6.5%), whereas concentrations above 25% reduced aggregation to <. The presence of NaCl had minimal stabilizing effects, particularly at lower albumin concentrations. The trend analysis further confirmed that higher albumin concentrations mitigate heat-induced aggregation, likely due to molecular crowding effects. Conclusion Albumin concentration plays a crucial role in maintaining structural integrity during pasteurisation, with higher concentrations (>25%) providing enhanced stability. While NaCl contributes to stabilization, it is insufficient at lower albumin concentrations.
Background Vitamin B12 (cobalamin) deficiency can manifest through various nonspecific oral symptoms. Early diagnosis is crucial but challenging due to subtle and progressive clinical features. Laboratory investigations like serum cobalamin levels and mean corpuscular volume (MCV) are used but have limitations. Purpose To assess the reliability of red patches and burning mouth symptoms as oral biomarkers for clinical and subclinical vitamin B12 deficiency, and correlate them with serum cobalamin and MCV values. Methods A sequential diagnostic analysis was conducted on 120 adult subjects (aged 18-45), divided into a control group consisting of asymptomatic subjects with subclinical cobalamin deficiency and a study group with symptomatic subjects with clinical cobalamin deficiency . Oral examinations and laboratory tests for MCV and serum cobalamin levels were performed. Statistical analyses included descriptive statistics, t-tests, and logistic regression. Results Significant differences were found in MCV and cobalamin levels between study and control groups ( P < .001). The presence of red patches and burning mouth symptoms strongly correlated with variations in MCV and cobalamin levels. Logistic regression analysis showed high accuracy (98.3%) in distinguishing between groups based on these markers. Conclusion Oral red patches and burning mouth symptoms, in conjunction with MCV and cobalamin levels, can serve as reliable indicators for identifying clinical cobalamin deficiency. This underscores the importance of monitoring these oral indicators during initial clinical examinations for early detection and management of vitamin B12 deficiency.
Background Transfusion-transmitted infections (TTI) are among the most critical challenges of blood transfusion. Patients with lower health literacy can have adverse consequences for blood transfusion and due to the lack of standard tools for measuring health literacy related to TTI, this study was conducted to develop a standardized tool for assessing Health literacy associated with TTI. Aim This study aimed to develop a standardized tool for assessing Health literacy related to TTI. Material and Methods This study was a cross-sectional analytical study of the tool development and validation type, according to the definitions provided for health literacy, questions were designed in the five areas of search, understanding, evaluation, decision-making, and application. The questionnaire was provided to relevant experts to evaluate the content validity with a qualitative and quantitative approach. After making corrections according to the experts’ opinions, the Content Validity Index (CVI) and the Content Validity Ratio (CVR) were checked. It was given to people who were at least literate in reading and writing levels to check the quantitative and qualitative face validity. Finally, the structural validity of the tool was checked by performing a confirmatory factor analysis and its reliability by calculating Cronbach's Alpha. Results The CVR and CVI had an average of 0.86 and 0.93 respectively. The reliability of the questionnaire was calculated using Cronbach's alpha coefficient of 0.744. In examining the structural validity of the tool, the root mean squared error (RMSE) index was less than 0.0001, the Goodness of Fit Index (GFI) was 0.975, the Adjusted Goodness of Fit Index (AGFI) was 0.957, Bentler and Bonnet's Normed Fit Index (NFI) was 1 and Comparative Fit Index (CFI) was 0.856. Conclusion The tool developed is reliable and valid, and can be used to measure the level of health literacy related to transfusion-transmitted infections.
Background Plasma-derived human C1-esterase inhibitor (C1-INH) concentrates are indicated for the treatment of acute episodes of hereditary angioedema. Intravenous C1-INH concentrate provides rapid relief to patients with acute hereditary angioedema symptoms. Objective To develop a safe and efficient purification process for C1-INH from human plasma. Design A combination of anion exchange and hydrophobic interaction chromatography was used to purify C1-INH from human plasma. Method The cold insoluble fraction (cryoprecipitate) was separated via centrifugation; then, the cryoprecipitate-poor plasma was subjected to different separation techniques to purify C1-INH. First, separation was performed using diethylaminoethyl (DEAE) anion exchange chromatography to capture coagulation factors. Then, C1-INH was purified from the remaining plasma using quaternary amine (a strong anion exchanger) chromatography, followed by DEAE (a weak anion exchanger) chromatography. The final product was purified via hydrophobic interaction chromatography, followed by ultrafiltration to concentrate the purified C1-INH. This process is cost-effective because coagulation factors were captured before C1-INH purification and the waste fraction after the capture stage was diverted to Cohn fractionation to isolate other potential plasma products. Results The final C1-INH preparation had a specific activity of >12 IU/mg protein, which is sufficient for its intended purpose. The purified preparation was characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and coagulation factors were measured in the purified preparation. Furthermore, viral activity decreased via solvent detergent treatment and virus filtration. Conclusion We successfully developed a purification process for C1-INH from human plasma. This process is easily adaptable and has been successfully scaled up to approximately 15-fold. It can be further scaled up for the large-scale production of C1-INH.
