Modified DALI LDL-apheresis using trisodium citrate anticoagulation plus bicarbonate or lactate-buffered hemofiltration substitution fluids as primers

BACKGROUND: DALI (direct adsorption of lipids) is the first LDL-apheresis technique able to adsorb low-density lipoprotein (LDL) and lipoproteina) directly from whole blood. In the standard procedure, acid citrate dextrose (ACD-A) is used as anticoagulation and the adsorber is rinsed with a specially manufactured priming solution (PS). Using neutral trisodium citrate (TSC) instead of ACD-A might improve the acid-base homeostasis during DALI apheresis; moreover, applying wholesale hemofiltration solutions instead of the special PS might avoid the use of two separate solutions for both priming before and reinfusion after the treatment, thus simplifiying the procedure. AIM: The present study was performed to test the effect of neutral (TSC) anticoagulation and of two different commercially available hemofiltration (HF) priming solutions on the efficacy and biocompatibility of DALI apheresis. MATERIALS AND METHODS: Five hypercholesterolemic chronic DALI patients were treated prospectively, on a weekly or biweekly basis, 3 times each by standard DALI-apheresis (A). by DALI using 4% TSC and bicarbonate-buffered HF BIC35-210 priming (B). as well as by DALI using 4% TSC and lactate-buffered HF 23 priming (C). After the sessions, the extracorporeal circuit (ECC) was rinsed with saline in study arm A and with the corresponding HF solutions in study arms B and C, respectively. RESULTS: Acute LDL-cholesterol reductions in the study arms A/B/C averaged 64/64/63%, for Lp(a) 62/64/62%, respectively (n=15). Clinically, all sessions were essentially uneventful and no clots were observed in the ECC. No major differences were found between the 3 study arms with respect to biocompatibility (elastase, C3a, thrombin-antithrombin, beta-thromboglobulin, bradykinin). CONCLUSION: DALI apheresis using TSC anticoagulation and HF solutions for both priming and reinfusion proved to be as safe and effective as the standard DALI apheresis. These modifications, however, further simplify the procedure.     

Photodynamic therapy-mediated hypericin-loaded nanostructured lipid carriers against vulvovaginal candidiasis

Introduction and Aim: The indiscriminate use and adverse effects of the main conventional antifungal agents compromise the effectiveness of treating vulvovaginal candidiasis (VVC), mainly caused by the species Candida albicans. This study evaluated the effectiveness of photodynamic therapy (PDT) and the in vitro and in vivo anti-candida potential of the hypericin (HYP)-loaded nanostructured lipid carriers (NLC). Materials and Methods: Empty NLC and NLC-HYP were characterized by the dynamic light scattering technique and transmission electron microscopy to evaluate the average particle size distribution and its morphologies. The in vitro inhibition photodynamic effect of the systems was tested to reduce the planktonic viability of C. albicans. The therapeutic assay photodynamic of the systems was performed to treat VVC in mice. Results: Empty NLC and NLC-HYP presented values of average hydrodynamic diameter, polydispersity index, and ζ-potential from 136 to 133 nm, 0.16 to 0.22, and -18 to -30 mV, respectively, on day 30. Microscopically, the systems showed spherical morphologies and nanoscale particles. Furthermore, in the in vitro inhibition assay, the treatment of PDT with NLC-HYP (NLC-HYP+) showed a significant reduction of the C. albicans planktonic viability compared to YNB negative control after 5 min of LED light irradiation. In the in vivo therapeutic assay, the antifungal group (vaginal antifungal cream) and NLC-HYP+ evaluated in the dark and by PDT, respectively, had a significant log₁₀ reduction in fungal burden compared to the infected group on day 8 of the VVC treatment. Conclusion: Due to the in vitro and in vivo anti-candida potential, PDT-mediated systems can be an effective strategy in VVC therapy.     

Effect of 50 Milliliters of 50% Dextrose in Water Administration on the Blood Sugar of Euglycemic Volunteers

Abstract. Objective : To evaluate the effect of administration of 1 ampule of 50% dextrose in water solution (D₅₀W) on serum glucose levels in healthy adult volunteers, the authors set out to determine whether a pre-D₆₀W serum glucose level can be predicted from the ED sample. Methods : This was a prospective, interventional study conducted from the ED of an urban, university-affiliated hospital. All subjects were healthy employee volunteers between 25 and 40 years of age. Baseline serum glucose levels were determined and all subjects were given an IV bolus of 25 grams of 50% dextrose solution. The main outcome measures were post-D₅₀W serum glucose levels (observed) at 5 predetermined time intervals (5 min, 15 min, 30 min, 1 hr, and 2 hr). An expected change in serum glucose was calculated using the volume of distribution formula for glucose. Results : Twenty-five volunteers (17 males and 8 females) participated in the study. The mean baseline serum glucose was 82.3 ± 13.5 mg/dL. The mean post-infusion levels were: 244.4 ± 44.6 mg/dL (5 min), 145.8 ± 52.3 mg/dL (15 min), 88.1 ± 28.8 mg/dL (30 min), 77.6 ± 13.6 mg/dL (60 min), and 83.2 ± 11.4 mg/dL (120 min). Using a mixed-effect regression model, statistically significant increases in serum glucose levels were found at 5 minutes (p < 0.001) and 15 minutes (p < 00001) following administration of D₅₀W. There was a return to baseline serum glucose by 30 minutes. The expected change based on the volume of distribution formula (53.7 ± 34.9) did not correlate with the observed changes at any measured time interval. Conclusion : Without pre-intervention blood drawing by emergency medical services, it is not possible to accurately predict pre-D₅₀W serum glucose levels based on post-D₅₀W glucose levels. The diagnosis of hypoglycemia as the etiology of altered mental status must therefore remain a diagnosis of exclusion. In addition, the return of serum glucose to baseline after 30 minutes suggests the duration of the effect of 1 ampule of D₅₀W. Frequent re-evaluation of the serum glucose levels of suspected or proven hypoglycemic patients after administration of D₅₀W should be considered.     

Heterogeneous glycosylation patterns of human PAI-1 may reveal its cellular origin

The main inhibitor of intravascular fibrinolysis is plasminogen activator inhibitor 1 (PAI-1) which binds to and irreversibly inhibits tissue plasminogen activator (tPA). PAI-1 is present in blood, both in platelets and in plasma, and PAI-1 levels are associated with risk of atherothrombosis. Several tissues express PAI-1 but the source of plasma PAI-1 is not known. We recently found that platelets can de novo synthesize PAI-1 and the amount synthesized in vitro in 24 hours is 35-fold higher than required to maintain normal plasma levels. Recombinant human PAI-1 expressed in different cell types or secreted naturally by human cell lines, exhibit heterogeneous glycosylation patterns. The aim of this study was to investigate the hypothesis that platelets might be the source of plasma PAI-1 and that the cellular source of PAI-1 can be determined by its tissue-specific glycosylation pattern. PAI-1 was isolated from platelets, macrophages, endothelial cells, adipose tissue, as well as plasma from lean and obese subjects. The glycosylation was analyzed by nanoLC-MS/MS. PAI-1 isolated from cell lysates and conditioned media from macrophages, endothelial cells, and adipose tissue expressed heterogeneous glycosylation patterns. By contrast, no glycans were detected on PAI-1 isolated from plasma or platelets from healthy lean individuals. Hence, our data suggest that platelets may be the main source of plasma PAI-1 in lean individuals. Interestingly, plasma PAI-1 from obese subjects had a glycan composition similar to that of adipose tissue suggesting that obese subjects with elevated PAI-1 levels may have a major contribution from other tissues.     

ACD全血及红细胞MAP的质量分析与保存研究

目的　充分了解 ACD全血和红细胞 MAP的质量及其保存效果 ,推广成份输血。方法　 1取 ACD全血及红细胞 MAP各 2人份各分成 6小袋进行 2～ 3 0天的血糖、钾、钠、酸碱度、白细胞、红细胞、血小板与红细胞平均容积的动态观察 ;2结合质控活动 ,对 ACD全血和红细胞 MAP中的血糖、蛋白质、钾、钠、比重、白细胞、红细胞、血小板、红细胞平均容积血、血红蛋白和红细胞压积等进行随机检测与分析。结果　 1红细胞 MAP中的血比重、白细胞、红细胞、血小板、红细胞压积、血红蛋白及血糖明显高于 ACD全血 ,而血浆蛋白、钠离子与酸碱度明显低 ;2血液在 2～ 3 0天的保存期间 ,白细胞、红细胞、血小板、红细胞平均容积均无改变 ;3酸碱度、钾、钠离子随保存期而明显变化 ,钾与钠离子的日变化量在红细胞 MAP比在 ACD全血中明显。结论　全血保存在 ACD或红细胞混悬于 MAP保存液中是安全与可行的 ,红细胞 MAP的内容物与 ACD全血不同 ,红细胞 MAP在失血和贫血病人中的应用值得关注并应予推广。     

Extracellular vesicle characteristics in stored red blood cell concentrates are influenced by the method of detection

Extracellular vesicles (EVs), including microvesicles and exosomes, are small phospholipid vesicles (≤1 μm in diameter) that are present in blood products, accumulate during storage, and have a potential transfusion-related immunomodulatory role. Knowledge of EVs in stored blood products is limited due to the challenges and difficulties in detecting these heterogeneous submicron-sized vesicles. The aim of this study was to assess the impact of different approaches to characterize EVs in stored RBC products. Quantification and size-profiling of EVs in leukoreduced red cell concentrates (RCCs) were examined on day 3, 7, 21, and 42 of storage using tunable resistive plus sensing (TRPS), flow cytometer (FC), and dynamic light scatting (DLS) methods. Using the TRPS method, the concentration of EVs < 200 nm significantly increased throughout storage (p < 0.05). This change in exosome concentration was not detectable with FC or DLS due to limitations in their ability to resolve particles <200 nm and/or accurately determine EV concentration. Both the TRPS and FC demonstrate that the concentration of EVs ≥ 200 nm significantly increases in RCCs by day 42/43 compared to EVs present on day 3 (p < 0.001). As the DLS measures the average size of particles in suspension, only an increase in the zeta-average size was observed during storage. EV size and concentration in RBC products is significantly influenced by the length of storage. Overall, this study shows that combining technologies may be important to improve the characterization and study of EVs in stored RCCs.     

高浓度葡萄糖注射液细菌内毒素的测定

50%的葡萄糖溶液可抑制鲎试剂凝集反应,干扰细菌内毒素的测定,本文使用中空纤维透析器,以双向超滤技术,将细菌内毒素与葡萄糖分离,降低了葡萄糖浓度,消除了葡萄糖对鲎试剂凝集反应的抑制作用,方法实用,操作简便,快速灵敏。     

Laboratory preparation and evaluation of a multiparameter hemocytometry control

A protocol for the laboratory preparation of a multiparameter hemocytometry control is given. Human platelets, stabilized by a basically simplified and inexpensive fixation procedure, are added to our previously described white and red blood cell control. Evaluation of this multiparameter control shows good precision characteristics and acceptable mechanical stability for at least 7 weeks, as measured in the Coulter counter Model S Plus-II. The control can basically contribute to the realization of the essence of internal quality control: continuous self-auditing and continuous attempts at improvement of performance.     

Generation of procoagulant activity by mononuclear phagocytes: optimisation of an in vitro assay

Leucocyte procoagulant activity (PCA) has previously been advocated as a useful measurement of cell-mediated immune reactivity. The assay is, however, susceptible to inter- and intra-experimental variation. This investigation identifies several factors which influence the stability and reproducibility of the test. Major factors which affect the recalcification time of plasma, include plasma lability, medium/buffer pH and both the nature and concentration of the indicator cells used in the assay. C. parvum-induced mouse peritoneal exudate cells have been used as a novel source of mononuclear phagocytes in the generation of PCA. Their sensitivity as indicator cells has been demonstrated by their responsiveness to stimulation by phytomitogen, endotoxin, and lymphokine (macrophage procoagulant-inducing factor). A simple test based on antigen-induced PCA of these cells, has provided an in vitro index of in vivo sensitisation to sheep red blood cells. Optimisation and standardisation of conditions detailed in this report for estimating PCA, renders the assay of value in monitoring lymphocyte and macrophage activation.     

ST-16: A new Bw16 subtype present in Mexican-Americans

Using a large battery of Bw16, w38, and w39 antisera, a new variant of Bw16 has been identified in four unrelated Mexican-American families. The serologi pattern obtained is distinct from that for the Bw38, Bw39, and 8W57 antigens. Absorption studies confirm the existence of this new Bw16 subtype which we term ST-16. ST-16 is Bw6-associated, with antigen frequency estimated to be 2.5% in Mexican-Americans.