A chromatographically purified human TGF-β1 fraction from virally inactivated platelet lysates

Background and Objectives TGF‐β1 exerts important physiological functions in osteogenesis and chondrogenesis and may be of therapeutic interest. The aim of this work was to develop a scalable purification process of TGF‐β1 from virally inactivated human platelets. Study Design and Methods Apheresis platelet concentrates (N = 12) were solvent/detergent (S/D) treated (1% TnBP/1% Triton X‐45; 31 °C) and the resulting platelet lysates were clarified by oil extraction and centrifugation, then chromatographed on an anion‐exchange DEAE‐Sepharose Fast‐Flow column equilibrated in a PBS buffer, pH 7·5. The column was washed to eliminate unbound proteins and the S/D agents. Bound proteins were eluted using a 1 M NaCl–PBS buffer pH 7·5 (DEAE‐eluate). The content in TGF‐β1, PDGF‐AB, VEGF, IGF‐1, EGF, and b‐FGF was measured by ELISA. Proteins, lipids, and S/D agents were assessed. Protein profile was determined by SDS‐PAGE under reduced or non‐reduced conditions. Results Most proteins, including albumin and immunoglobulins G, A, and M did not bind to the DEAE column as evidenced also by SDS‐PAGE. Essentially all PDGF, VEGF, and IGF were in the breakthrough. The DEAE‐eluate contained close to 60% of the TGF‐β1 at a mean concentration of about 102 ng/ml, whereas EGF, b‐FGF were at about 0·72 and 0·18 ng/ml, respectively. The content in TnBP and Triton X‐45 was <2 ppm. Conclusion A fraction enriched in TGF‐β1 can be prepared from virally inactivated human platelet lysates using an easily scale process. Its interest in regenerative medicine and cell therapy will be evaluated in further studies.     

A comparison of methods of pathogen inactivation of FFP

Introduction Current methods for pathogen inactivation of plasma involve four major processes using solvent–detergent (SD), methylene blue (MB), amotosalen and riboflavin as additives. Three of these methods involve the use of visible or ultraviolet light. Methods A comparison of the four methods was made using publications in Medline, Pubmed, Embase and Biosis to obtain data on the logistics of use, the quality of the plasma proteins and the effectiveness of pathogen inactivation. Results Three of the methods, MB, amotosalen and riboflavin, are designed for use in a blood bank; the SD method is generally applied at a centralized manufacturing centre and involves large plasma pools. All methods result in a reduction in protein values with the per cent retention of FVIII activity in the range of 67–78% and fibrinogen of 65–84%. Protein S and alpha₂‐antiplasmin are lower following solvent–detergent treatment. Alterations in fibrinogen structure have been reported with methylene blue. Discussion Three of the methods are designed for small volume use in a blood bank. All four methods have some effect on the coagulant proteins; however, the final concentrations are within regulated limits. While there is variability in the effectiveness against pathogens, direct comparison is difficult because of the methodologies used. Nonetheless, all are effective in inactivating HIV and other lipid‐enveloped pathogens. Clinical studies on the effectiveness of these products are surprisingly sparse, and no randomized clinical trials have yet been performed with amotosalen or riboflavin plasmas.     

多酶清洗液超声清洗联合过氧化氢等离子低温灭菌处理硬式腹腔镜的效果分析

目的探讨多酶清洗液超声清洗联合过氧化氢等离子低温灭菌处理硬式腹腔镜与辅助器械的效果。方法将2015年8月—2017年7月广州医科大学附属肿瘤医院各科室使用后的140套硬式腹腔镜随机分为试验组(70套)与对照组(70套),其中对照组器械于传统手工清洗后采用多酶清洗液手工刷洗联合过氧化氢等离子低温灭菌予以处理,试验组器械于传统手工清洗后采用多酶清洗液超声清洗联合过氧化氢等离子低温灭菌予以处理;处理后,使用目测法、ATP检测法及咽拭子培养法对比两组器械的清洗及灭菌效果。结果试验组目测清洗合格率为98.6%,对照组目测清洗合格率为84.3%,两组对比采用卡方检验,χ~2=9.115,P0.05,差异具有统计学意义;试验组ATP检测清洗合格率为97.1%,对照组ATP检测清洗合格率为81.4%,两组对比采用卡方检验,χ~2=9.035,P0.05,差异具有统计学意义;咽拭子培养法检测结果显示,两组器械细菌培养结果均为阴性,合格率均达100%,无统计学意义。结论采用多酶清洗液超声清洗联合过氧化氢等离子低温灭菌处理硬式腹腔镜,清洗效果好,灭菌合格率可达100%,适于临床应用。     

烷基苯烷烃脱氢和烷基化装置流程模拟

应用ＨＹＳＩＭ软件对烷基苯联合装置中的烷烃脱氢和烷基化装置进行了全流程模拟计算,并与ＵＯＰ原设计数据作比较,为装置设计积累了数据,对装置的技术改造和优化操作条件提供了全流程模拟平台。     

多酶预处理溶液联合超声对口腔内科器械的清洗效果评价

目的:评价多酶预处理溶液联合超声对口腔内科器械的清洗效果。方法:选取2019年2月~5月我院口腔内科常规使用并进行消毒的器械620支,随机分为两组,分别用多酶预处理溶液(对照组)和多酶预处理溶液联合超声清洗(实验组)处理。比较两组目测清洗合格率以及STF清洗效果检测卡合格率。结果:实验组器械目测合格率以及STF清洗效果检测卡检测合格率均高于对照组(P<0.05)。结论:多酶预处理溶液联合超声清洗对于口腔内科器械具有更加有效和安全的清洗效果。     

异功散加味联合多酶片及双歧杆菌三联活菌散治疗儿童厌食症脾胃气虚证的临床疗效

目的：观察异功散加味联合多酶片及双歧杆菌三联活菌散治疗儿童厌食症脾胃气虚证临床疗效。方法：研究纳入2022年1月至2023年6月河北省中医院收治的儿童厌食症脾胃气虚证患儿112例,根据随机数字表法将其分为对照组与观察组(各56例),对照组患儿采取多酶片及双歧杆菌三联活菌散治疗,观察组患儿在对照组治疗基础上联合异功散加味治疗,研究过程中两组各脱落2例。对比两组患儿临床疗效,治疗前后各组患儿中医证候(主证、次证与舌脉)积分变化,食量恢复正常时间及体质量增加量,治疗前后患儿钙、铁、锌含量变化与血红蛋白及白蛋白水平变化、胃泌素及胃动素、血管活性肠肽、生长抑素等胃肠激素水平变化、神经肽Y、促食欲素及瘦素等血清食欲调节因子水平变化,观察研究过程两组患儿的不良反应发生情况。结果：治疗后对照组患儿的总有效率为85.19%(46/54),观察组总有效率为98.15%(53/54),观察组患儿总有效率明显高于对照组(χ²=5.939,P<0.05)。与对照组儿童比较,观察组儿童食量恢复正常时间更短,体质量增加更多(P<0.05)。与本组治疗前比较,两组患儿中医证候(主证、...     

Pharmacokinetic study of minipooled solvent/detergent‐filtered cryoprecipitate factor VIII

Summary. Eighteen cryoprecipitate minipools, each made of 30 units of low volume, concentrated cryoprecipitate, have been treated by solvent‐detergent and filtration (S/D‐F) in a single‐use CE‐marked bag system. The S/D‐F cryoprecipitate contained a mean of 10.5 IU mL⁻¹ factor VIII (FVIII), 17 mg mL⁻¹ clottable fibrinogen, and >10 IU mL⁻¹ von Willebrand factor ristocetin co‐factor, and anti‐A and anti‐B isoagglutinins were undetectable. The products have been infused in 11 severe (FVIII <1%) haemophilia A patients (mean age: 17.4 years; mean weight: 57.6 kg) at a dose close to 40 IU kg⁻¹. Patients were hospitalized for at least 36 h to determine FVIII recovery, half‐life and clearance. They were also closely monitored for possible adverse events. None of the infused patients demonstrated reactions or adverse events even though they did not receive anti‐allergic drugs or corticosteroids prior to infusion. The mean recovery of FVIII 10 min postinfusion was 69.7%. Mean FVIII half‐life was 14.2 h and clearance was 2.6 mL h⁻¹ kg⁻¹. All patients had a bleeding‐free interval of 8–10 days postS/D‐F cryoprecipitate infusion. The data show that S/D‐F cryoprecipitate FVIII presents a normal pharmacokinetics profile, and support that it could be safely used for the control of acute and chronic bleeding episodes in haemophilia A patients.     

Clusterin expression in follicular dendritic cells associated with prion protein accumulation

Peripheral accumulation of abnormal prion protein (PrP) in variant Creutzfeldt-Jakob disease and some animal models of transmissible spongiform encephalopathies (TSEs) may occur in the lymphoreticular system. Within the lymphoid tissues, abnormal PrP accumulation occurs on follicular dendritic cells (FDCs). Clusterin (apolipoprotein J) has been recognized as one of the molecules associated with PrP in TSEs, and clusterin expression is increased in the central nervous system where abnormal PrP deposition has occurred. We therefore examined peripheral clusterin expression in the context of PrP accumulation on FDCs in a range of human and experimental TSEs. PrP was detected immunohistochemically on tissue sections using a novel highly sensitive method involving detergent autoclaving pretreatment. A dendritic network pattern of clusterin immunoreactivity in lymphoid follicles was observed in association with the abnormal PrP on FDCs. The increased clusterin immunoreactivity appeared to correlate with the extent of PrP deposition, irrespective of the pathogen strains, host mouse strains or various immune modifications. The observed co-localization and correlative expression of these proteins suggested that clusterin might be directly associated with abnormal PrP. Indeed, clusterin immunoreactivity in association with PrP was retained after FDC depletion. Together these data suggest that clusterin may act as a chaperone-like molecule for PrP and play an important role in TSE pathogenesis.     

Toothpaste detergents: a potential source of oral soft tissue damage?

Abstract: Objectives: Toothpastes are thought to be of benefit to cleaning teeth but may also have the potential for soft tissue damage at least on the cellular level by inclusion of detergents in their formulation. The aim of this study was to observe the in vitro response of oral mucosa like cells to toothpaste detergents. Methods: TERT‐1 keratinocytes were exposed to varying concentrations of the detergents Adinol, Sodium Lauryl Sulphate, Tego Betain and Pluronic as well as PBS and culture medium. After 2‐min exposure, cells were washed and incubated in fresh medium for 24 h. Cell death was then spectrophotmetrically measured using an MTT assay. Results: Except for Pluronic, cell viability was markedly reduced for all detergents at all increasing concentrations when compared to the positive medium control. Cells treated with Pluronic were stimulated compared to medium alone. Conclusions: These in vitro data suggest that some detergents may have the potential to cause soft tissue damage in the mouth. Although in vivo, saliva may neutralize such effects. The results for Pluronic suggest a possible oxidative stress response that bears further study.