BACKGROUND & AIMS: :The hydroxylation of CMP-N-acetylneuraminic acid (CMP-NeuAc) in the formation of CMP-N-glycolylneuraminic acid requires several components which comprise an electron transport system. A protein, which replaces one of the components, was purified to homogeneity from a horse erythrocyte lysate. Based on its partial amino acid sequence and immunological cross-reactivity, this protein was identified as soluble cytochrome b5 lacking the membrane domain of microsomal cytochrome b5. The electron transport system involved in CMP-NeuAc hydroxylation was reconstituted, and then characterized using the purified horse soluble cytochrome b5 and a fraction from mouse liver cytosol. The hydroxylation reaction requires a reducing reagent, DTT being the most effective. Either NADH or NADPH was used as an electron donor, but the activity with NADPH amounted to about 74% of that with NADH. The hydroxylation was inhibited by salts and azide due to interruption of the electron transport from NAD(P)H to cytochrome b5 and in the terminal enzyme reaction, respectively.

背景与目标： ：CMP-N-乙酰基神经氨酸的形成中的CMP-N-乙酰神经氨酸（CMP-NeuAc）的羟基化需要包含电子传输系统的几种组分。从马红细胞裂解物中纯化出一种蛋白质，以取代一种成分。根据其部分氨基酸序列和免疫交叉反应性，该蛋白被鉴定为缺乏微粒体细胞色素b5膜结构域的可溶性细胞色素b5。重建参与CMP-NeuAc羟基化的电子传输系统，然后使用纯化的马可溶细胞色素b5和来自小鼠肝细胞溶质的部分进行表征。羟基化反应需要还原剂，DTT是最有效的。 NADH或NADPH均用作电子供体，但NADPH的活性约为NADH的74％。分别由于电子从NAD（P）H到细胞色素b5的转移以及在末端酶反应中的中断，盐和叠氮化物抑制了羟基化作用。

BACKGROUND & AIMS: The interaction of arginine analogues, which are known to inhibit nitric oxide synthase, with two cationic amino acid transporters of human erythrocytes (systems y+ and y+L) was studied. Arginine and relevant analogues [NG-monomethyl-L-arginine (L-NMMA); NG-monomethyl-D-arginine (D-NMMA) and NG-nitro-L-arginine (L-NOARG)] were found to inhibit labeled lysine influx into intact erythrocytes. As expected, the pattern of inhibition reflected the contribution of the two distinct transport systems. All analogues showed a higher affinity for system y+L than for system y+. The half-saturation (inhibition) constants estimated for systems y+ and y+L (+/- SEM) were (microM)L-arginine, 55.7 +/- 5.4 and 2.4 +/- 0.1; L-NMMA, 151 +/- 13 and 7.5 +/- 0.5; D-NMMA, 2660 +/- 404 and 269 +/- 25; L-NOARG, 9414 +/- 169 and 594 +/- 35. The transport properties of the analogues were investigated using an assay based on the trans-stimulation of lysine efflux. The addition of saturating concentrations of unlabeled analogues to the external medium stimulated efflux of labeled lysine through systems y+L and y+, showing that the analogues can enter the cell through these pathways.

背景与目标： 研究了已知能抑制一氧化氮合酶的精氨酸类似物与人类红细胞的两种阳离子氨基酸转运蛋白（系统y和y L）的相互作用。精氨酸和相关类似物[NG-单甲基-L-精氨酸（L-NMMA）；发现NG-单甲基-D-精氨酸（D-NMMA）和NG-硝基-L-精氨酸（L-NOARG）]可抑制标记的赖氨酸流入完整的红细胞。不出所料，抑制模式反映了两种不同运输系统的贡献。所有类似物对系统y L的亲和力均高于对系统y的亲和力。对于系统y和y L（/ -SEM）估计的半饱和（抑制）常数为（microM）L-精氨酸，55.7 / -5.4和2.4 / -0.1； L-NMMA，151 /-13和7.5 /-0.5； D-NMMA，2660 /-404和269 /-25； L-NOARG，9414 /-169和594 /-35。使用基于赖氨酸外排的反式刺激的分析方法研究了类似物的转运特性。将饱和浓度的未标记类似物添加到外部介质中会刺激标记的赖氨酸通过系统y L和y流出，这表明类似物可以通过这些途径进入细胞。

BACKGROUND & AIMS: BACKGROUND:Neonatal hyperbilirubinaemia remains one of the most common clinical conditions requiring therapeutic intervention. Nevertheless, reliable indicators of bilirubin toxicity are still missing. This prompted us to investigate (a) the progression of cytotoxic events produced by increasing concentrations of bilirubin; (b) the relevance of the membrane lipid package on bilirubin binding to erythrocytes; and (c) the reliability of chloroform extraction compared with albumin extraction to evaluate erythrocyte-bound bilirubin and cytotoxicity. MATERIALS AND METHODS:Morphological alterations, free bilirubin, erythrocyte-bound bilirubin (albumin- and chloroform-extractable), haemolysis and membrane-released lipids, were determined in human erythrocytes at 4 degrees C or 37 degrees C, after 4 h incubation at pH 7.4, with increasing molar ratios of bilirubin to albumin (0.5-5). The reversibility of cytotoxicity by albumin washing was assessed by morphological analysis. RESULTS:Decreased free bilirubin, lower erythrocyte-bound bilirubin concentration by albumin extraction (superficial/non-aggregated bilirubin) and higher values by chloroform extraction (deep/aggregated bilirubin) were observed for 37 degrees C vs. 4 degrees C, at molar ratios > 1. Echinocytosis increased with bilirubin concentration and temperature and was not fully reversed by albumin washing. Haemolysis was already significant at a molar ratio of 1, and was enhanced by temperature at molar ratios 3 and 5 (P < 0.01). The loss of membrane lipids was remarkable at molar ratios > or = 0.5, both at 4 degrees C and 37 degrees C (P < 0.01), although correlation with bilirubin concentration was only significant at 37 degrees C (r = 0.971; P < 0.01). CONCLUSIONS:These results suggest that increased lipid fluidity and high bilirubin concentrations promote membrane bilirubin translocation and toxicity. They also show that albumin is not able to displace the bilirubin located deeply or aggregated within the membrane, which in turn is removed by chloroform. Accordingly, chloroform-extractable rather than albumin-extractable bilirubin is a more accurate parameter to assess erythrocyte-bound bilirubin during severe hyperbilirubinaemia.

背景与目标： 背景：新生儿高胆红素血症仍然是最常见的需要治疗干预的临床疾病之一。然而，仍然缺乏可靠的胆红素毒性指标。这促使我们研究（a）胆红素浓度增加所产生的细胞毒性事件的进展； （b）膜脂质包装与胆红素与红细胞结合的相关性； （c）氯仿提取与白蛋白提取的可靠性，以评估与红细胞结合的胆红素和细胞毒性。
材料与方法：在pH值下孵育4小时后，在4摄氏度或37摄氏度下测定了人类红细胞的形态学改变，游离胆红素，红细胞结合胆红素（白蛋白和氯仿提取物），溶血和膜释放的脂质7.4，随着胆红素与白蛋白的摩尔比增加（0.5-5）。通过形态分析评估白蛋白洗涤引起的细胞毒性的可逆性。
结果：在摩尔比下，观察到37℃vs. 4℃观察到游离胆红素降低，白蛋白提取（表面/非聚集胆红素）降低的红细胞结合胆红素浓度和氯仿提取（深层/聚集胆红素）的较高值。 > 1.胆红素浓度和温度升高时，胞浆菌增多，白蛋白洗涤不能完全逆转。摩尔比为1时，溶血作用已经很明显，摩尔比为3和5时，溶血作用因温度而增强（P <0.01）。摩尔比>或= 0.5时，膜脂的损失在4摄氏度和37摄氏度时均显着（P <0.01），尽管与胆红素浓度的相关性仅在37摄氏度时才显着（r = 0.971； P <0.01 ）。
结论：这些结果表明增加脂质流动性和高胆红素浓度促进膜胆红素易位和毒性。他们还显示白蛋白不能取代位于膜内深处或聚集的胆红素，而胆红素又被氯仿去除。因此，在严重高胆红素血症期间，用氯仿提取的胆红素而不是白蛋白提取的胆红素是评估红细胞结合胆红素的更准确的参数。

BACKGROUND & AIMS: :Surfactant protein (SP)-A and SP-D, immunoglobulins, and complement all modulate inflammation within the lung by regulating pathogen clearance. For example, SP-A binds to and opsonizes a variety of bacteria and viruses, thereby enhancing their phagocytosis by innate immune cells such as alveolar macrophages. Immunoglobulins, which bind to antigen and facilitate Fc receptor-mediated phagocytosis, can also activate complement, a family of soluble proteins with multiple host defense functions. Previous studies showed that SP-A and complement protein C1q interact. Since complement protein C1q binds to IgG and IgM immune complexes, the hypothesis tested in this study was that SP-A, which is structurally homologous to C1q, also binds to IgG and affects its functions. SP-A binds to the Fc, rather than the Fab, region of IgG. Binding is calcium dependent but not inhibited by saccharides known to bind to SP-A's carbohydrate recognition domain. The binding of SP-A does not inhibit the formation of immune complexes or the binding of IgG to C1q. In contrast, SP-A enhances the uptake of IgG-coated erythrocytes, suggesting that SP-A might be influencing Fc receptor-mediated uptake. In summary, this study shows a novel interaction between SP-A and IgG and a functional consequence of the binding.

背景与目标： ：表面活性蛋白（SP）-A和SP-D，免疫球蛋白和补体均通过调节病原体清除率来调节肺内炎症。例如，SP-A结合并调理多种细菌和病毒，从而通过先天性免疫细胞（如肺泡巨噬细胞）增强其吞噬作用。结合抗原并促进Fc受体介导的吞噬作用的免疫球蛋白还可以激活补体，补体是具有多种宿主防御功能的可溶性蛋白家族。先前的研究表明SP-A和补体蛋白C1q相互作用。由于补体蛋白C1q与IgG和IgM免疫复合物结合，因此在这项研究中测试的假设是与C1q结构同源的SP-A也与IgG结合并影响其功能。 SP-A与IgG的Fc区结合而不是与Fab区结合。结合是钙依赖性的，但不受已知结合SP-A的碳水化合物识别结构域的糖类的抑制。 SP-A的结合不会抑制免疫复合物的形成或IgG与C1q的结合。相反，SP-A增强了IgG包被的红细胞的摄取，表明SP-A可能影响Fc受体介导的摄取。总而言之，这项研究显示了SP-A与IgG之间的新型相互作用以及结合的功能性结果。

BACKGROUND & AIMS: :The in vitro effects of xipamide in a concentration range of 10(-8) to 10(-2) M were investigated on various Na+ and K+ transport systems in human red blood cells. Xipamide inhibited the anion carrier or DIDS-sensitive LiCO3- -influx starting from a concentration of 10(-5) M. However, a decrease in the Na+, K+-pump and the Na+, K+-cotransport activity and a rise in the passive permeability of the cell membrane was only observed starting from a concentration of 10(-4) M xipamide.

背景与目标： ：对人红细胞中各种Na和K转运系统，研究了Xipamide在10（-8）至10（-2）M浓度范围内的体外作用。 Xipamide从10（-5）M浓度开始抑制阴离子载体或DIDS敏感的LiCO3--流入。但是，Na，K泵和Na，K-共转运活性降低，而被动吸收升高仅从浓度为10（-4）M的Xipamide开始观察到细胞膜的通透性。

BACKGROUND & AIMS: :Deformability of erythrocytes from four patients with different types of glucosephosphate isomerase (D-glucose-6-phosphate ketoisomerase, GPI) deficiency has been determined by cell filtration. Young as well as whole erythrocyte populations had a markedly increased rigidity and an abnormally strong attachment of haemoglobin to the inner surface of isolated membranes. Acidic environment may enhance membran rigidity in vitro and also during passage of the erythrocytes through the spleen. The decrease of deformability at a pH of 6.8 was most pronounced in the splenectomized patients, and likewise in erythrocytes from the splenic artery, which were obtained from one patient during splenectomy. It is suggested that the metabolic environment of the spleen, with its low pH, impairs the deformability of GPI-deficient erythrocytes and predisposes them to splenic sequestration. The clinical improvement of all patients following splenectomy which is accompanied by an increase of the erythrocyte survival time and by unchanged reticulocyte counts, is in accordance with this view.

背景与目标： ：已通过细胞过滤确定了四名患有不同类型葡萄糖磷酸异构酶（D-葡萄糖-6-磷酸酮异构酶，GPI）缺乏症患者的红细胞变形能力。年轻的和整个红细胞群体的刚度显着增加，并且血红蛋白异常强烈地附着在离体膜的内表面上。酸性环境可以在体外以及在红细胞通过脾脏的过程中增强膜的刚性。在脾切除的患者中，pH值为6.8时变形能力的降低最为明显，同样，在脾切除术中从一名患者获得的脾动脉的红细胞中也是如此。提示脾脏的代谢环境具有较低的pH值，会损害GPI缺失型红细胞的变形能力，并使它们易于进行脾隔离。该观点认为，脾切除术后所有患者的临床改善均伴随着红细胞存活时间的增加和网织红细胞计数的不变。

BACKGROUND & AIMS: :The passage of nucleosides across the plasma membrane of erythrocytes is a membrane-mediated process which is strongly inhibited by derivatives of 9-beta-D-ribofuranosylpurine (1) with S, O, or N atoms at the purine 6 position bearing variously substituted arylalkyl groups. In this structure-activity study, nucleoside derivatives were compared in respect to their ability to inhibit a transport-dependent aspect of nucleoside metabolism in erythrocytes, the synthesis of inosine from external guanosine and hypoxanthine. 6-Benzylthio, 6-benzylamino, and 6-benzyloxy derivatives of 1 were inhibitory at 10(-5)-10(-6) M and the similarity of their activities suggested that alkylation of the transporter as the mechanism of transport inhibition was unlikely. The hydrophobicity of the 6-position substituents appeared to contribute importantly to inhibitory activity. Although replacement of the ribofuranose moiety by other sugars reduced inhibitory activity, compounds with 9-butyl groups were inhibitory. 6-[(2-Hydroxy-5-nitrobenzyl)thio] derivatives of 1 were the most potent of the inhibitors tested, being active at about 10(-7) M.

背景与目标： ：核苷通过红细胞的质膜是一种膜介导的过程，被9-β-D-呋喃呋喃糖基嘌呤（1）的S，O或N原子在嘌呤6位具有不同取代的衍生物强烈抑制芳基烷基。在这项结构活性研究中，比较了核苷衍生物抑制红细胞中核苷代谢的运输依赖性方面，由外部鸟苷和次黄嘌呤合成肌苷的能力。 1的6-苄硫基，6-苄氨基和6-苄氧基衍生物在10（-5）-10（-6）M时具有抑制作用，并且它们的活性相似，表明转运蛋白的烷基化不能抑制转运。 6位取代基的疏水性似乎对抑制活性起重要作用。尽管用其他糖替代呋喃核糖部分降低了抑制活性，但具有9-丁基的化合物具有抑制作用。 1的6-[[（2-羟基-5-硝基苄基）硫基]衍生物是最有效的抑制剂，在约10（-7）M时有活性。

BACKGROUND & AIMS: :Nitric oxide (NO) activates soluble guanylyl cyclase in smooth muscle cells to induce vasodilation in the vasculature. However, as hemoglobin (Hb) is an effective scavenger of NO and is present in high concentrations inside the red blood cell (RBC), the bioavailability of NO would be too low to elicit soluble guanylyl cyclase activation in the presence of blood. Therefore, NO bioactivity must be preserved. Here we present evidence suggesting that the RBC participates in the preservation of NO bioactivity by reducing NO influx. The NO uptake by RBCs was increased and decreased by altering the degree of band 3 binding to the cytoskeleton. Methemoglobin and denatured hemoglobin binding to the RBC membrane or cytoskeleton also were shown to contribute to reducing the NO uptake rate of the RBC. These alterations in NO uptake by the RBC, hence the NO bioavailability, were determined to correlate with the vasodilation of isolated blood vessels. Our observations suggest that RBC membrane and cytoskeleton associated NO-inert proteins provide a barrier for NO diffusion and thus account for the reduction in the NO uptake rate of RBCs.

背景与目标： 一氧化氮（NO）激活平滑肌细胞中的可溶性鸟苷酸环化酶，从而诱导脉管系统中的血管舒张。但是，由于血红蛋白（Hb）是有效的NO清除剂，并且以高浓度存在于红血球（RBC）内部，因此NO的生物利用度将太低，以至于在存在血液的情况下不会引起可溶性鸟苷酰环化酶的活化。因此，必须保留任何生物活性。在这里，我们提出的证据表明，RBC通过减少NO流入而参与了NO生物活性的保存。通过改变带3与细胞骨架的结合程度，红细胞对NO的吸收增加和减少。高铁血红蛋白和变性的血红蛋白与RBC膜或细胞骨架的结合也显示出有助于降低RBC的NO吸收率。 RBC吸收NO的这些变化（因此没有NO的生物利用度）被确定与孤立血管的血管舒张相关。我们的观察结果表明，RBC膜和细胞骨架相关的NO惰性蛋白为NO扩散提供了障碍，因此可以解释RBC的NO吸收率降低。

BACKGROUND & AIMS: :We evaluated the usefulness of enumeration of fecal leukocytes and erythrocytes in making an early diagnosis of Shigella infection, where Shigella is a leading cause of invasive diarrhea. Stool specimens from 561 invasive diarrhea patients were submitted for microscopic examination. A presumptive diagnosis of shigellosis based on microscopic examination was made in 389 of them; 227 had stool cultures positive for Shigella spp (Shigella patients). One hundred sixty-two patients with no detectable Shigella infection (non-Shigella invasive diarrhea cases) served as a comparison group. Two hundred twenty-seven randomly selected Shigella patients and 227 non-Shigella infectious diarrhea cases from the surveillance system database of the hospital constituted another group for comparative evaluation. The stool specimens of the patients were examined under the microscope, and isolation, biochemical characterization and serotyping of Shigella were performed. In comparison with non-Shigella invasive diarrhea cases, the presence of >50 WBC/hpf in association with any number of RBC in the fecal sample had a modest sensitivity of 67%, specificity of 59%, positive predictive value of 70%, negative predictive value of 56%, accuracy of 64%, and positive likelihood ratio of 1.6 in predicting shigellosis. Comparison between Shigella and non-Shigella infectious diarrhea patients revealed the presence of >20 WBC/hpf was a less accurate predictor of shigellosis (sensitivity 51%, specificity 88%, positive predictive value 81%, negative predictive value 64%, accuracy 69%, and positive likelihood ratio 4.1). Direct microscopical examination of stool specimens for the presence of WBC and RBC may facilitate the early diagnosis of shigellosis, and may be a cheap alternative to stool culture in this setting.

背景与目标： ：我们评估了粪便白细胞和红细胞计数在早期诊断志贺氏菌感染中的作用，其中志贺氏菌是侵袭性腹泻的主要原因。将561例浸润性腹泻患者的粪便标本进行了显微镜检查。他们中有389例根据显微镜检查推定为志贺菌病； 227人的粪便培养物对志贺氏菌属呈阳性（志贺氏菌患者）。将没有发现志贺氏菌感染的162例患者（非志贺氏菌性腹泻病例）作为比较组。从医院监视系统数据库中随机选择的277例志贺氏菌患者和227例非志贺氏菌感染性腹泻患者构成了另一组进行比较评估。在显微镜下检查患者的粪便标本，并进行志贺氏菌的分离，生化鉴定和血清分型。与非志贺氏菌性腹泻病例相比，粪便样品中> 50 WBC / hpf与任意数量的RBC联合存在的中度敏感性为67％，特异性为59％，阳性预测值为70％，阴性预测志贺菌病的预测值为56％，准确性为64％，正似然比为1.6。志贺氏菌和非志贺氏菌感染性腹泻患者的比较显示，> 20 WBC / hpf的存在是志贺菌病的较不准确的预测指标（敏感性51％，特异性88％，阳性预测值81％，阴性预测值64％，准确性69％ ，以及正似然比4.1）。对粪便标本进行直接显微镜检查是否存在WBC和RBC可能有助于志贺菌病的早期诊断，并且在这种情况下可能是粪便培养的廉价替代品。

BACKGROUND & AIMS: :Glutathione export from uninfected human erythrocytes was compared with that from cells infected with the malaria parasite Plasmodium falciparum using two separate methods that distinguish between oxidized (GSSG) and reduced (GSH) glutathione. One involved enzymatic recycling with or without thiol-masking; the other involved rapid derivatization followed by HPLC. Glutathione efflux from uninfected erythrocytes under physiological conditions occurred predominantly as GSH. On exposure of the cells to oxidative challenge, efflux of GSSG exceeded that of GSH. Efflux of both species was blocked by MK571, an inhibitor of mammalian multidrug-resistance proteins. Glutathione efflux from parasitized erythrocytes was substantially greater than that from uninfected erythrocytes. Under physiological conditions, the exported species was GSH, whereas under energy-depleted conditions, GSSG efflux occurred. Glutathione export from parasitized cells was inhibited partially by MK571 and more so by furosemide, an inhibitor of the 'new permeability pathways' induced by the parasite in the host erythrocyte membrane. Efflux from isolated parasites occurred as GSH. On exposure to oxidative challenge, this GSH efflux decreased, but no GSSG export was detected. These results are consistent with the view that the parasite supplies its host erythrocyte with GSH, much of which is exported from the infected cell via parasite-induced pathways.

背景与目标： ：使用两种分别区分氧化型（GSSG）和还原型（GSH）谷胱甘肽的方法，将未感染的人类红细胞的谷胱甘肽输出与感染疟原虫恶性疟原虫的细胞的谷胱甘肽输出进行了比较。一种涉及有或没有硫醇掩蔽的酶循环。另一个涉及快速衍生化，然后进行HPLC。在生理条件下，未感染红细胞的谷胱甘肽外排主要以GSH形式发生。当细胞暴露于氧化性刺激后，GSSG的流出超过了GSH。两种物种的外流都被MK571阻断，MK571是一种哺乳动物多药抗性蛋白的抑制剂。寄生的红细胞的谷胱甘肽外排显着大于未感染的红细胞的谷胱甘肽外排。在生理条件下，出口物种为GSH，而在能量消耗条件下，发生了GSSG外排。谷胱甘肽从寄生虫细胞中的输出受到MK571的部分抑制，而速尿则更受抑制，速尿是由宿主红细胞膜中的寄生虫诱导的“新通透性途径”的抑制剂。来自分离的寄生虫的外排以GSH发生。暴露于氧化刺激后，该GSH外排量减少，但未检测到GSSG出口。这些结果与以下观点一致：寄生虫为其宿主红细胞提供了GSH，其中GSH大部分是通过寄生虫诱导的途径从被感染的细胞中输出的。

BACKGROUND & AIMS: BACKGROUND:Hip resurfacing arthroplasty is known to increase the metal ion concentration in the serum, urine and whole blood, with potentially adverse effects on the organism. However, only few data are available about the metal ion concentrations in erythrocytes, although they are directly exposed to the higher concentrations of the serum. METHODS:The ion levels of chromium, cobalt, nickel and molybdenum in erythrocytes of 25 patients with a hip resurfacing implant were analysed with high resolution ICP-sf-MS (inductively-coupled-plasma-sector-field-mass-spectrometry). The results were compared to the ion levels in the serum and urine of the patients, and also to the ion levels of 27 control persons without an implant. RESULTS:Compared to the control group, ion levels in the erythrocytes of the hip resurfacing group were markedly increased for cobalt (0.10 vs. 3.26 microg/kg) and slightly for chromium (6.04 vs. 7.38 microg/kg). In contrast, ion levels in the serum of the hip resurfacing group were increased for cobalt (0.21 vs. 1.92 microg/l), chromium (1.48 vs. 5.64 microg/l), nickel (1.53 vs. 4.25 microg/l) and molybdenum (2.17 vs. 3.78 microg/l). CONCLUSION:Ion concentrations of cobalt and chromium are also increased in erythrocytes after hip resurfacing arthroplasty. Further research is required to evaluate the impact of the elevated ion levels on the erythrocytes, and to evaluate if metal ions also accumulate in other tissues of the body.

背景与目标： 背景：髋关节置换术会增加血清，尿液和全血中的金属离子浓度，对生物体可能产生不利影响。但是，关于红细胞中金属离子浓度的数据很少，尽管它们直接暴露于较高浓度的血清中。
方法：采用高分辨率ICP-sf-MS（电感耦合等离子体场场质谱法）分析了25例髋关节表面置换植入物患者红细胞中铬，钴，镍和钼的离子水平。将结果与患者血清和尿液中的离子水平进行了比较，还与27名没有植入物的对照组的离子水平进行了比较。
结果：与对照组相比，髋关节表面置换组的钴中红细胞的离子水平显着增加（0.10 vs. 3.26 microg / kg），而铬略有升高（6.04 vs. 7.38 microg / kg）。相比之下，钴（0.21比1.92微克/升），铬（1.48比5.64微克/升），镍（1.53比4.25微克/升），钼（钼）增加了髋关节置换组血清中的离子水平。 （2.17对3.78微克/升）。
结论：髋关节置换术后红细胞中钴和铬的离子浓度也增加。需要进一步的研究来评估升高的离子水平对红细胞的影响，并评估金属离子是否还会在人体其他组织中蓄积。

BACKGROUND & AIMS: :Porphyromonas gingivalis, an anaerobic gram-negative bacterium associated with chronic periodontitis, can agglutinate human erythrocytes. In general, hemagglutination can be considered the ability to adhere to host cells; however, P. gingivalis-mediated hemagglutination has special significance because heme markedly accelerates growth of this bacterium. Although a number of studies have indicated that a major hemagglutinin of P. gingivalis is intragenically encoded by rgpA, kgp, and hagA, direct evidence has not been obtained. We demonstrated in this study that recombinant HGP44(720-1081), a fully processed HGP44 domain protein, had hemagglutinating activity but that an unprocessed form, HGP44(720-1138), did not. A peptide corresponding to residues 1083 to 1102, which was included in HGP44(720-1138) but not in HGP44(720-1081), could bind HGP44(720-1081) in a dose-dependent manner and effectively inhibited HGP44(720-1081)-mediated hemagglutination, indicating that the interdomain regional amino acid sequence may function as an intramolecular suppressor of hemagglutinating activity. Analyses by solid-phase binding and chemical cross-linking suggested that HGP44 interacted with glycophorin A on the erythrocyte membrane. Glycophorin A and, more effectively, asialoglycophorin, which were added exogenously, inhibited HGP44(720-1081)-mediated hemagglutination. Treatment of erythrocytes with RgpB proteinase resulted in degradation of glycophorin A on the membrane and a decrease in HGP44(720-1081)-mediated hemagglutination. Surface plasmon resonance detection analysis revealed that HGP44(720-1081) could bind to asialoglycophorin with a dissociation constant of 3.0 x 10(-7) M. These results indicate that the target of HGP44 on the erythrocyte membrane appears to be glycophorin A.

背景与目标： 牙龈卟啉单胞菌（Porphyromonas gingivalis）是一种与慢性牙周炎有关的厌氧革兰氏阴性细菌，可凝集人的红细胞。通常，血凝可以被认为具有粘附宿主细胞的能力。然而，牙龈卟啉单胞菌介导的血细胞凝集具有特殊意义，因为血红素显着加速了该细菌的生长。尽管许多研究表明，牙龈卟啉单胞菌的主要血凝素由rgpA，kgp和hagA内源编码，但尚未获得直接证据。在这项研究中，我们证明了重组HGP44（720-1081）（一种完全加工的HGP44域蛋白）具有血凝活性，而未经加工的形式HGP44（720-1138）则没有。 HGP44（720-1138）中包含但不包含在HGP44（720-1081）中的对应于残基1083至1102的肽可以剂量依赖的方式结合HGP44（720-1081），并有效抑制HGP44（720- 1081）介导的血凝反应，表明域间区域氨基酸序列可能起血凝活性分子内抑制剂的作用。通过固相结合和化学交联的分析表明HGP44与红细胞膜上的糖蛋白A相互作用。外源添加的糖精蛋白A和更有效的去唾液酸糖蛋白抑制了HGP44（720-1081）介导的血凝反应。用RgpB蛋白酶处理红细胞导致膜上的糖蛋白A降解，并减少了HGP44（720-1081）介导的血凝反应。表面等离振子共振检测分析表明，HGP44（720-1081）可以以3.0 x 10（-7）M的解离常数结合去唾液酸糖皮质激素。这些结果表明，HGP44在红细胞膜上的靶标似乎是糖蛋白A。

BACKGROUND & AIMS: :Simple and efficient transfection methods for genetic manipulation of Plasmodium falciparum are desirable to identify, characterize and validate the genes with therapeutic potential and better understand parasite biology. Among the available transfection techniques for P. falciparum, electroporation-based methods, particularly electroporation of ring-infected RBCs is routinely used. Nonetheless, transfection of P. falciparum remains a resource-intensive procedure. Here, we report a simple and economic transfection method for P. falciparum, which is termed as the lyse-reseal erythrocytes for transfection (LyRET). It involved lysis of erythrocytes with a hypotonic RBC lysis buffer containing the desired plasmid DNA, followed by resealing by adding a high salt buffer. These DNA-encapsulated lyse-reseal erythrocytes were mixed with P. falciparum trophozoite/schizont stages and subjected to selection for the plasmid-encoded drug resistance. In parallel, transfections were also done by the methods utilizing electroporation of DNA into uninfected RBCs and parasite-infected RBCs. The LyRET method successfully transfected 3D7 and D10 strains with different plasmids in 63 of the 65 attempts, with success rate similar to transfection by electroporation of DNA into infected RBCs. The cost effectiveness and comparable efficiency of LyRET method makes it an alternative to the existing transfection methods for P. falciparum, particularly in resource-limited settings.

背景与目标： ：对于恶性疟原虫的基因操作，简单有效的转染方法对于鉴定，表征和验证具有治疗潜力的基因以及更好地了解寄生虫生物学是理想的。在恶性疟原虫的可用转染技术中，常规使用基于电穿孔的方法，特别是环感染的红细胞的电穿孔。尽管如此，恶性疟原虫的转染仍然是资源密集的过程。在这里，我们报告了一种简单而经济的恶性疟原虫转染方法，被称为用于转染的裂解-重新密封红细胞（LyRET）。它涉及用含有所需质粒DNA的低渗RBC裂解缓冲液裂解红细胞，然后通过添加高盐缓冲液进行重新密封。将这些DNA包被的裂解-重新密封的红细胞与恶性疟原虫滋养体/裂殖体阶段混合，并进行质粒编码的耐药性的选择。同时，转染也通过利用将DNA电穿孔到未感染的RBC和寄生虫感染的RBC中的方法进行。 LyRET方法在65次尝试中有63次成功用不同的质粒成功转染了3D7和D10菌株，成功率与通过将DNA电穿孔到感染的RBC中的转染相似。 LyRET方法的成本效益和可比的效率使其成为恶性疟原虫现有转染方法的替代方法，尤其是在资源有限的环境中。

BACKGROUND & AIMS: :Previous studies of erythrocyte ion (potassium and sodium) transport during marasmus and kwashiorkor have indicated increased passive permeation to both ions in both syndromes, and increased Na,K pump activity in kwashiorkor and reduced activity in marasmus. Children with severe cerebral palsy (CP) frequently suffer secondary protein energy malnutrition (PEM). Unlike marasmus and kwashiorkor, this PEM is uncomplicated by micronutrient deficiency, parasitism and infections. Because of deformities classification of PEM cannot be performed in these children by stature-based anthropometry, therefore we used triceps skinfold thicknesses less than the fifth percentile and absence of weight gain in the previous year as criteria for malnutrition. K influx data from well- and malnourished CP children, and from well-nourished controls reveal that ouabain-sensitive K influx is highest in malnourished CP, followed by well-nourished CP (P = 0.02), and lowest in controls (P less than 0.001, vs. malnourished). Determinations of ouabain-sensitive Na efflux, though less precise and therefore more variable, were consistent with this finding of no decrease of Na,K pump activity occurring during the development of this malnutrition. There were no statistically significant differences in ouabain-insensitive fluxes of either Na or K. Ion transport in undernourished CP children thus resembles that found in kwashiorkor rather than in marasmus; but oedema is rarely seen in this form of secondary PEM.

背景与目标： ：先前对马拉索斯和克氏针期间红细胞离子（钾和钠）转运的研究表明，两种综合征中两种离子的被动渗透增加，而克氏针中Na，K泵的活性增加而马拉索斯活性降低。患有严重脑瘫（CP）的儿童经常患有继发性蛋白质能量营养不良（PEM）。与Marasmus和Kwashiorkor不同，该PEM不会因微量营养素缺乏，寄生和感染而变得复杂。由于基于身高的人体测量学无法对这些儿童进行PEM畸形分类，因此我们将肱三头肌的皮褶厚度小于5％且上一年没有体重增加作为营养不良的标准。来自营养良好和营养不良的CP儿童的K流入数据以及营养良好的对照组的数据表明，对哇巴因敏感的K流入量在营养不良的CP中最高，其次是营养良好的CP（P = 0.02），而在对照组中最低（P小于0.001，而营养不良）。对哇巴因敏感的钠流出的测定虽然精确度较低，因此变化较大，但与在营养不良发生期间没有发生Na，K泵活性降低的发现相一致。营养不良的CP儿童中，对哇巴因不敏感的Na或K通量没有统计学上的显着差异。但这种继发性PEM形式很少见到水肿。

BACKGROUND & AIMS: :An inherited and complete deficiency of diphosphoglycerate mutase was discovered in the erythrocytes of a 42-yr-old man of French origin whose blood hemoglobin concentration was 19.0 g/dl. Upon physical examination he was normal with the exception of a ruddy cyanosis. The morphology of his erythrocytes was also normal and there was no evidence of hemolysis. The erythrocyte 2,3-diphosphoglycerate level was below 3% of normal values and, as a consequence, the affinity of the cells for oxygen was increased. Diphosphoglycerate mutase activity was undetectable in erythrocytes as was that of diphosphoglycerate phosphatase. The activities of all the other erythrocyte enzymes that were tested were normal except for nomophosphoglycerate mutase which was diminished to 50% of the normal value. The levels of reduced glutathione, ATP, fructose 1,6-diphosphate, and of triose phosphates were elevated, whereas those of glucose 6-phosphate and fructose 6-phosphate were decreased. This report sheds new light on the role of diphosphoglycerate mutase in the metabolism of erythrocytes.

背景与目标： ：在一名法国血统血红蛋白浓度为19.0 g / dl的42岁男性的红细胞中发现了遗传性和完全缺乏的二磷酸甘油酸突变酶。经身体检查，除红病外，他一切正常。他的红细胞形态也正常，没有溶血的迹象。红细胞2,3-二磷酸甘油酸水平低于正常值的3％，因此，细胞对氧气的亲和力增加。与磷酸二甘油酯磷酸酶一样，在红细胞中不能检测到磷酸二甘油酯突变酶的活性。测试的所有其他红细胞酶的活性都是正常的，除了磷酸磷酸甘油酯变位酶降低到正常值的50％。还原型谷胱甘肽，ATP，果糖1,6-二磷酸和磷酸三糖的水平升高，而葡萄糖6-磷酸和果糖6-磷酸的水平降低。该报告为二磷酸甘油酸突变酶在红细胞代谢中的作用提供了新的启示。