An in vitro method for predicting the efficacy of WBC separation using different starch preparations and anticoagulant ratios

BACKGROUND: Separation of blood components depends on a number of factors, including the viscosity of the plasma and the number and size of the various cellular elements. To enhance granulocyte collection, it is common practice to alter the plasma environment by the addition of sedimenting agents such as hydroxyethyl starch. Recently, because of its prolonged persistence in the circulation, the higher-molecular-weight form of hydroxyethyl starch, Hespan (HP), has been replaced by the lower-molecular-weight form, pentastarch (PS). However, the yield appears to be lower. A rapid in vitro approach was used to permit comparison of the efficiency of separation of WBCs by the use of PS and HP and different ratios of anticoagulants that also alter the sedimenting characteristics of blood. STUDY DESIGN AND METHOD: Blood from individual persons was collected into sodium citrate at ratios of 1:8, 1:12, and 1:16. Samples were evaluated either before or after the addition of PS or HP and after centrifugation. RESULTS: The addition of HP increased the sedimentation rate to at least four times that of plasma (10.9 vs. 47.9 mm); PS approximately doubled the rate. Viscosity was altered by the introduction of either starch. These changes (ranging from a rate of 4.2 in HP with a 1:16 anticoagulant to 3.6 in PS with a 1:8 ratio of anticoagulant) reflected the anticipated effects of anticoagulant dilution and carbohydrate addition. Granulocyte recovery was highest, with a 1:12 anticoagulant ratio in all tests with HP producing the greatest yield (HP, 101%; PS, 89%; control, 78%). CONCLUSION: HP is far more effective than its lower-molecular-weight substitute PS in the generation of granulocytes in the buffy coat of whole blood. This method provides a simple, rapid, in vitro approach to evaluating the separating efficiency of solutions.     

Gene therapy using Aβ variants for amyloid reduction

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Cutis laxa,exocrine pancreatic insufficiency and altered cellular metabolomics as additional symptoms in a new patient with ATP6AP1-CDG

Congenital disorders of glycosylation (CDG) are genetic defects in the glycoconjugate biosynthesis. > 100 types of CDG are known, most of them cause multi-organ diseases. Here we describe a boy whose leading symptoms comprise cutis laxa, pancreatic insufficiency and hepatosplenomegaly. Whole exome sequencing identified the novel hemizygous mutation c.542 T > G (p.L181R) in the X-linked ATP6AP1, an accessory protein of the mammalian vacuolar H⁺-ATPase, which led to a general N-glycosylation deficiency. Studies of serum N-glycans revealed reduction of complex sialylated and appearance of truncated diantennary structures. Proliferation of the patient's fibroblasts was significantly reduced and doubling time prolonged. Additionally, there were alterations in the fibroblasts' amino acid levels and the acylcarnitine composition. Especially, short-chain species were reduced, whereas several medium- to long-chain acylcarnitines (C14-OH to C18) were elevated. Investigation of the main lipid classes revealed that total cholesterol was significantly enriched in the patient's fibroblasts at the expense of phophatidylcholine and phosphatidylethanolamine. Within the minor lipid species, hexosylceramide was reduced, while its immediate precursor ceramide was increased. Since catalase activity and ACOX3 expression in peroxisomes were reduced, we assume an ATP6AP1-dependent impact on the β-oxidation of fatty acids. These results help to understand the complex clinical characteristics of this new patient.     

Some examples of the clinical importance of the basic and heparin-induced phospholipase A in human plasma

Phospholipase A activity was determined in human plasma with biological ³²P-labelled phosphatidylethanolamine from rat liver following cardiac operations with the aid of the heart-lung-machine. The basic activity before heparin and the initiation of bypass was 72 nmol β ml^?1 β h^?1 plasma; thereby large differences in the basic activity were detected between cyanotic and acyanotic heart diseases. A sharp increase of phospholipase activity was observed after heparin (a smaller one in intensive care patients with low-dose heparin), and a further increase up to the end of perfusion. Normal values are reached after giving protamine at the end of the operation. An important role for plasma phospholipase in the regulation of physiological and pathophysiological processes may be supposed from these results.     

加兰他敏对APP/PS1转基因小鼠海马区星形胶质细活化及C/EBPβ表达的影响

目的研究加兰他敏对APP/PS1转基因小鼠海马区星形胶质细胞活化、C/EBPβ表达及行为学的影响。方法选取10月龄雄性APP/PS1转基因小鼠20只,随机分为模型对照组(10只)和治疗组(10只),同月龄、同背景的C57BL/6野生型雄性小鼠10只作为正常对照组。治疗组皮下注射加兰他敏溶液5mg/kg,2次/d,连续治疗8周,正常对照组和模型对照组给予皮下注射等量生理盐水。应用Morris水迷宫实验于干预治疗8周后开始测定各组小鼠空间学习记忆能力,连续7d,采用免疫组织化学、免疫荧光及Western-blot方法观察各组小鼠海马区星形胶质细胞活化及C/EBPβ表达水平。结果与正常对照组相比,模型对照组和治疗组小鼠Morris检测第5、6天平均逃避潜伏期延长,穿越平台次数减少(P0.05,P0.05),而治疗组其逃避潜伏期较模型对照组缩短(P0.05),穿越平台次数增多(P0.05);同时治疗组小鼠星形胶质细胞活化被明显抑制,胶质纤维酸性蛋白(GFAP)的阳性表达面积〔(5.003±0.823)%〕及C/EBPβ的表达量(87.711±14.622)较模型对照组〔(7.116±1.040)%,119.920±16.901〕明显减少(P0.05,P0.05)。结论加兰他敏改善APP/PS1转基因AD小鼠的学习记忆能力可能与其抑制星形胶质细胞的活化及C/EBPβ的表达有关。     

Propensity score to evaluate prognosis in pregnancy-associated breast cancer: Analysis from a French cancer network

PurposeTo compare the prognosis of pregnancy associated breast cancer occurring during pregnancy (BCP) to non-pregnancy associated breast cancers (non-BCP) in young women managed at a national expert center.MethodsRetrospective cohort study of a prospective database using propensity score matching (PSM) analysis with known prognostic factors.ResultsWe analyzed data of 49 patients with BCP and 104 with non-BCP diagnosed between 2002 and 2017 at Tenon University Hospital (Paris, France). The BCP tumors were often locally advanced (lymph node metastases in 59%), of high grade (55%) and highly proliferative (67% with Ki67 ≥ 20%). After PSM, breast cancer-free survival (p = 0.45) and breast cancer specific survival (p = 0.81) were similar in the two groups. The recurrence rate was 12% vs 18% (p = 0.45) and the death rate was 6% vs 8% (p = 0.74) for the BCP and non-BCP groups, respectively. No difference in recurrence type was observed between the groups (p = 0.60).ConclusionsAfter PSM for known prognostic factors, the prognosis of BCP patients did not differ from that of young patients with non-BCP.     

一氧化氮和TGF-β1的血浆水平与进展性脑卒中的关系

目的探讨一氧化氮（Nitric oxide,NO）和转化生长因子-β1（Transforming growth factor-β1,TGF-β1）血浆水平与进展性脑卒中（progression stroke,PS）的关系。方法对发病24h内住院经头颅CT和/或MRI检查证实为大脑中动脉（MCA）供血区脑梗死的患者,以斯堪的那维亚卒中量表（SSS）在住院72h内评分较入院时下降2分以上者判定为PS的标准。诊断为PS患者51例。以入院时SSS评分相近的MCA供血区非PS患者为对照组（54例）;在住院48h内空腹抽取静脉血进行NO、TGF-β1测定,并对双侧颈动脉作彩色多普勒检查。结果2组年龄、入院时评分SSS及合并症（高血压病、糖尿病、冠心病、高胆固醇血症史）无显著性差异（P〉0.05）。PS组NO和TGF-β1水平显著低于对照组（P〈0.01）,PS组颈动脉内膜-中膜厚度（inti ma-media thickness,I MT）显著高于对照组（P〈0.05）。结论NO和TGF-β1水平显著下降是PS发生发展的因素之一,脑梗死早期检测其血浆NO和TGF-β1水平有助于预测PS的发生,为临床的治疗决策和预后判断具有重要意义。