Band 3 and glycophorin are progressively aggregated in density-fractionated sickle and normal red blood cells. Evidence from rotational and lateral mobility studies.

Band 3 aggregation in the plane of the red blood cell (RBC) membrane is postulated to be important in the pathophysiology of hemolysis of dense sickle and normal RBCs. We used the fluorescence photobleaching recovery and polarized fluorescence depletion techniques to measure the lateral and rotational mobility of band 3, glycophorins, and phospholipid analogues in membranes of density-separated intact RBCs from seven patients with sickle cell disease and eight normal controls. The fractions of laterally mobile band 3 and glycophorin decreased progressively as sickle RBC density increased. Normal RBCs also showed a progressive decrease in band 3 fractional mobility with increasing buoyant density. Rapidly rotating, slowly rotating, and rotationally immobile forms of band 3 were observed in both sickle and normal RBC membranes. The fraction of rapidly rotating band 3 progressively decreased and the fraction of rotationally immobile band 3 progressively increased with increasing sickle RBC density. Changes in the fraction of rotationally immobile band 3 were not reversible upon hypotonic swelling of dense sickle RBCs, and normal RBCs osmotically shrunken in sucrose buffers failed to manifest band 3 immobilization at median cell hemoglobin concentration values characteristic of dense sickle RBCs. We conclude that dense sickle and normal RBCs acquire irreversible membrane abnormalities that cause transmembrane protein immobilization and band 3 aggregation. Band 3 aggregates could serve as cell surface sites of autologous antibody binding and thereby lead to removal of dense sickle and normal (senescent) RBCs from the circulation.     

Strain sensitivity of band structure and electron mobility in perovskite BaSnO3: first-principles calculation

A first-principles electronic structure calculation is utilized to contrastively investigate the crystal structure, band structure, electron effective mass and mobility of perovskite BaSnO₃ under hydrostatic and biaxial strain. Strain-induced changes in relative properties are remarkable and more sensitive to hydrostatic strain than biaxial strain. The structure of BaSnO₃ remains cubic under hydrostatic strain, while it becomes tetragonal under biaxial strain. Originating from the strain sensitivity of the Sn 5s orbitals in the conduction band minimum, the band gaps of BaSnO₃ decrease for both types of strain from −3% to 3%. BaSnO₃ under tensile hydrostatic strain exhibits higher electron mobility than it does under tensile biaxial strain because of the smaller electron effective mass in the corresponding strain. In contrast, the opposite phenomenon exists in compressive strain. Our results demonstrate that strain could be an alternative way to modify the band gap and electron mobility of BaSnO₃.

A first-principles electronic structure calculation is utilized to contrastively investigate the crystal structure, band structure, electron effective mass and mobility of perovskite BaSnO₃ under hydrostatic and biaxial strain.     

IgG red blood cell autoantibodies in autoimmune hemolytic anemia bind to epitopes on red blood cell membrane band 3 glycoprotein.

Red blood cell (RBC) autoantibodies from patients with IgG warm-type autoimmune hemolytic anemia were labeled with iodine 125 and their RBC binding behavior characterized. Epitope-bearing RBC membrane polypeptides were identified after autoantibody immunoprecipitation of labeled membranes and immunoblotting. Immunoaffinity isolation of labeled membrane proteins with 12 different IgG hemolytic autoantibodies with protein A-agarose revealed a major polypeptide at Mr 95 to 110 kd, which coelectrophoresed on sodium dodecylsulfate-polyacrylamide gel electrophoresis with a membrane component isolated with sheep IgG anti-band 3. Immunoprecipitation studies with chymotrypsinized RBCs resulted in the recovery of two labeled membrane polypeptides with molecular weights characteristically resulting from the chymotryptic fragmentation of band 3. Immunoblotting with sheep IgG anti-band 3 of the immunoprecipitated polypeptides confirmed that hemolytic autoantibody binding led to recovery of band 3 or its fragments. Two 125I-labeled IgG hemolytic autoantibodies showed binding behavior consistent with epitope localization on band 3. The labeled RBC autoantibodies bound immunospecifically to all types of human RBC tested, including those of rare Rh type (Rh-null, D--) at a site density of approximately 10(6) per RBC. The 125I-IgG in two labeled autoantibodies was 84% and 92% adsorbable by human and higher nonhuman primate RBCs. Antigen-negative animal RBC bound less than 10% (dog, 2.6%; rhesus monkey, 7.4%), consistent with immunospecific RBC binding. IgG-1 was the major subclass in five autoantibodies tested; one of six fixed complement; and autoantibody IgG appeared polyclonal by isoelectric focusing. We conclude that IgG eluted from RBCs of patients with autoimmune hemolytic anemia consists predominantly of a single totally RBC-adsorbable antibody population that binds to antigenic determinants on band 3. Unlike RBC autoantibodies from antiglobulin-positive normal blood donors, autoantibodies from patients with autoimmune hemolytic anemia did not show Rh-dependent properties as judged by antigen site density, absence of differential binding to RBC of different Rh phenotypes, failure to immunoprecipitate 30 kd Rh-epitope bearing membrane polypeptides, absence of multiple antibody populations, and the lack of a significant nonadsorbable IgG population that has been associated with the presence of antiidiotypic IgG. The absence of antiidiotypic IgG in hemolytic autoantibodies may be a contributory factor in the development of autoimmune hemolytic anemia.     

Transbilayer mobility and distribution of red cell phospholipids during storage.

We studied phospholipid topology and transbilayer mobility in red cells during blood storage. The distribution of phospholipids was determined by measuring the reactivity of phosphatidylethanolamine with fluorescamine and the degradation of phospholipids by phospholipase A2 and sphingomyelinase C. Phospholipid mobility was measured by determining transbilayer movements of spin-labeled phospholipids. We were unable to detect a change in the distribution of endogenous membrane phospholipids in stored red cells even after 2-mo storage. The rate of inward movement of spin-labeled phosphatidylethanolamine and phosphatidylserine was progressively reduced, whereas that for phosphatidylcholine was increased. These changes in phospholipid translocation correlated with a fall in cellular ATP. However, following restoration of ATP, neither the rate of aminophospholipid translocation nor the transbilayer movement of phosphatidylcholine were completely corrected. Taken together, our findings demonstrate that red cell storage alters the kinetics of transbilayer mobility of phosphatidylserine, phosphatidylethanolamine, and phosphatidylcholine, the activity of the aminophospholipid translocase, but not the asymmetric distribution of endogenous membrane phospholipids, at least at a level detectable with phospholipases. Thus, if phosphatidylserine appearance on the outer monolayer is a signal for red cell elimination, the amount that triggers macrophage recognition is below the level of detection upon using the phospholipase technique.     

The influence of rotational movement exercise on the abdominal muscle thickness and trunk mobility – Randomized control trial

BackgroundTrunk rotations are important functional movements which form the foundations of human motion pattern, especially in the functions of walking and running. They prevent the functional impairments and structural lesions resulting from axial overloading in static positions such as sitting.ObjectivesThe aim of the study was to assess the influence of rotational movement training exercises on the abdominal muscle thickness and spinal mobility range. Study design: Randomized controlled trial.MethodsThe study involved 73 individuals aged 18–45. The subjects were randomly divided into two groups. The study group (TG) comprised 40 people who performed rotational movement exercises over the period of 4 weeks (16 training sessions). In the control group (CG) the training was not applied. Changes in the thickness of selected abdominal muscles on ultrasound imaging were evaluated, as well as trunk mobility, based on the trunk lateral flexion test.ResultsThe analysis of the obtained data has demonstrated a statistically significant increase in the thickness of the abdominal internal (IO) (p < 0.05) and external oblique muscles (EO) (p < 0.001) in the study group (TG) between measurements I and II, and measurements I and III. A similar increase in the thickness was found in the summation measurement of TrA + IO + EO. Bilateral increase in the trunk lateral flexion range in the frontal plane has also been noted.ConclusionsRotational movement training of the trunk leads to an increase in the thickness of the abdominal oblique muscles. Rotational movement exercise training increases trunk mobility in the frontal plane.     

Electrophoretic mobility and agglutinability of red blood cells: a "new" polymorphism in mice

A quantitative method has been developed to determine agglutinability of mouse red blood cells. Tests with different inbred strains of mice revealed only two phenotypes. The same inbred strains were tested with the cytopherometer to determine the electrophoretic mobility of the corresponding red cells. Again, two phenotypes were uncovered, and faster mobility was found in the red cells that had higher agglutinability. The genetic control of this character is autosomal and codominant, and segregates independently of H-2 and coat color.     

Inhibition of protein synthesis in intact HeLa cells.

Polysome analysis has proved to be a sensitive probe for the mode of action of inhibitors of protein synthesis in intact HeLa cells. To classify the active compounds as inhibitors of initiation, elongation, or termination, their effects on the cellular polyribosome pattern were compared under three conditions. These conditions tested (i) their direct effect on the polyribosome profile; (ii) their effect on ribosome run-off produced by hypertonicity; and (iii) their effects on recovery from hypertonicity. Using this technique, diacetoxyscirpenol, 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide, and three alkaloids, harringtonine, isoharringtonine, and homoharringtonine, were found to be inhibitors of initiation. Polysome analysis indicated that in HeLa cells 7.8 x 10(-7) M pactamycin, which inhibited protein synthesis 94%, interfered with elongation as well as initiation under these conditions. Emetine, anisomycin, cycloheximide, and trichodermin each gave polysome patterns consistent with inhibition of elongation. Fusidic acid and aurintricarboxylic acid inhibited incorporation of [(14)C]leucine into intact HeLa cells, but polysome analysis did not localize any specific inhibitory effects to the initiation, elongation, or termination steps of protein synthesis. The use of specific inhibitors of initiation of protein synthesis has indicated that most, if not all, mammalian messenger ribonucleic acids contain a single initiation site.     

Detection, characterization, and bioavailability of membrane-associated iron in the intact sickle red cell.

It is hypothesized that membrane-associated iron in the sickle red cell is of pathophysiologic importance, but the actual existence of such iron in the intact cell has been questioned. Using a strategy whereby membrane iron can be detected through its bioavailability for catalyzing peroxidation, we used phospholipid exchange protein to load membranes of intact erythrocytes (RBC) with approximately 2% phosphatidylethanolamine hydroperoxide (PEOOH) and monitored the development of peroxidation by-products during subsequent incubation. Normal RBC loaded with PEOOH developed very little peroxidation, but vitamin E-replete sickle RBC showed an exuberant peroxidation response that was not seen in cells loaded with control nonoxidized phosphatidylethanolamine. Ancillary studies of sickle RBC revealed that the catalytic iron included both heme iron and free iron located at the bilayer inner leaflet. Significantly, these studies also revealed that peroxidation after PEOOH loading is promoted by cellular dehydration and inhibited by hydration, thus identifying a dynamic interaction between hemoglobin (sickle > normal) and membrane lipid. High-reticulocyte control RBC and sickle trait RBC behaved exactly like normal RBC, while HbCC RBC and RBC having membranes gilded with hemoglobin iron because of prior exposure to acetylphenylhydrazine showed an abnormal peroxidation response like that of sickle RBC. Indeed, the peroxidation response of RBC loaded with PEOOH paralleled amounts of iron measured on inside-out membranes prepared from them (r = 0.783, P < 0.01). These studies corroborate existence of membrane-associated heme and free iron in the intact sickle cell, and they document its bioavailability for participation in injurious peroxidative processes. That association of cytosolic sickle hemoglobin with membrane lipid is modulated by cell hydration status provides a mechanism that may help explain increased development of oxidative membrane lesions in abnormally dehydrated sickle RBC regardless of the mechanism underlying their formation.     

Differential antiviral activities and intracellular metabolism of 3'-azido-3'-deoxythymidine and 2',3'-dideoxyinosine in human cells.

Dideoxynucleosides such as 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (ddI) can effectively inhibit the replication of human immunodeficiency virus (HIV) in T lymphoid cells. There is evidence that HIV can infect and replicate in other cells including monocytoid cells and macrophages. The present study compared the antiretroviral activities of ddI and AZT in three lineages of human cells, i.e., MOLT4 (T lymphocytoid, CD4+), U937 (monocytoid, CD4+), and HT1080 (fibroblastoid, CD4-) cells. Feline leukemia virus, a retrovirus that causes immunodeficiency in cats, was used to infect the cells. The drug concentrations needed to reduce the viral p27 antigen titers in cell lysates by 50% (IC50s) were determined. The data show that AZT and ddI inhibited viral replication in all three cell lines. The IC50s of AZT were 0.02, 1.75, and 2.31 microM in MOLT4, HT1080, and U937 cells, respectively. For ddI, the IC50s were 4.31, 9.52, and 43.5 microM, respectively. These data indicate differential antiviral activities of ddI and AZT in the different cells with the following rank order of drug sensitivity: MOLT4 > HT1080 > U937. A study of the intracellular metabolism of [3H]AZT and [3H]ddI shows that the antiretroviral activities of AZT and ddI in the three cell lines correlated with the levels of their intracellular triphosphate metabolites.     

Whole body, long-axis rotational training improves lower extremity neuromuscular control during single leg lateral drop landing and stabilization

Background

Poor neuromuscular control during sports activities is associated with non-contact lower extremity injuries. This study evaluated the efficacy of progressive resistance, whole body, long-axis rotational training to improve lower extremity neuromuscular control during a single leg lateral drop landing and stabilization.

Methods

Thirty-six healthy subjects were randomly assigned to either Training or Control groups. Electromyographic, ground reaction force, and kinematic data were collected from three pre-test, post-test trials. Independent sample t-tests with Bonferroni corrections for multiple comparisons were used to compare group mean change differences (P ≤ 0.05/21 ≤ 0.0023).

Findings

Training group gluteus maximus and gluteus medius neuromuscular efficiency improved 35.7% and 31.7%, respectively. Training group composite vertical–anteroposterior–mediolateral ground reaction force stabilization timing occurred 1.35 s earlier. Training group knee flexion angle at landing increased by 3.5°. Training group time period between the initial two peak frontal plane knee displacements following landing increased by 0.17 s. Training group peak hip and knee flexion velocity were 21.2°/s and 20.1°/s slower, respectively. Time period between the initial two peak frontal plane knee displacements following landing and peak hip flexion velocity mean change differences displayed a strong relationship in the Training group (r² = 0.77, P = 0.0001) suggesting improved dynamic frontal plane knee control as peak hip flexion velocity decreased.

Interpretation

This study identified electromyographic, kinematic, and ground reaction force evidence that device training improved lower extremity neuromuscular control during single leg lateral drop landing and stabilization. Further studies with other populations are indicated.     

地中海贫血血细胞质控品的研制和评价

摘要：目的：研制适用于地中海贫血的RBC计数、Hb、红细胞比容（HCT）、红细胞平均体积（MCV）、红细胞平均血红蛋白量（MCH）等指标实验室室间质量评价（EQA）的血细胞质控品,为广西地区地中海贫血的三级防控提供质量保证。 方法：用ACD保存液抗凝的献血者新鲜静脉血做基质,添加适量稳定剂,制备高、中、低3个浓度水平的质控品。对质控品均匀性、开瓶稳定性、长期稳定性、运输稳定性进行评价,在常见品牌血液分析仪如Sysmex XE2100、ABX Pentra120、Coulter LH 780、Mindray 6800和Celltac F MEK 8222K上作适用性评价。 结果：质控品随机抽检结果显示组内、组间红细胞各指标差异均无统计学意义（P均＞0.05）;质控品2~8 ℃保存,开瓶后可稳定6 d,未开瓶质控品可稳定35 d;运输稳定性评价显示,质控品在室温与2~8 ℃放置5 d内结果一致（P＞0.05）。红细胞指标中较稳定的是RBC、Hb、MCH;较不稳定的是HCT、MCV,开瓶后会随时间延长而升高。不同品牌血液分析仪检测RBC、Hb、MCH的变异系数（CV）均小于2.5%,检测小红细胞质控品时MCV差异稍大（总CV最大为3.89%）。 结论：自制全血质控品有良好的均匀性、稳定性、适用性,能满足广西地中海贫血MCV/MCH筛查的实验室EQA计划需求。     

Differential profiling of human red blood cells during storage for 52 selected microRNAs

BACKGROUND: MicroRNAs (miRNAs), the negative regulators of cellular mRNAs, are present in mature red blood cells (RBCs) in abundance relative to other blood cells. So far, there are no studies aimed at identifying large‐scale miRNA profiles during storage of RBCs. STUDY DESIGN AND METHODS: RNA samples from each RBC bag stored at 4°C were collected on Days 0, 20, and 40 and subjected to miRNA profiling by using a membrane‐based array. Fifty‐two selected miRNAs of cellular apoptotic pathway represent the array. Through bioinformatics analyses, we identified potential target genes for selected miRNAs. RESULTS: Differential profiling of RBCs for 52 miRNAs revealed two distinguishable patterns during storage: Forty‐eight miRNAs demonstrated no trend at all, while four miRNAs, miR‐96, miR‐150, miR‐196a, and miR‐197, demonstrated an increase up to Day 20 and subsequently decreased during storage. We selected miR‐96 and subjected it to standard bioinformatics analyses for target gene predictions, which identified several mRNAs including the RBC proapoptotic calpain small subunit‐1 (CAPNS1) as potential targets of miR‐96. To validate these predictions, we selected CAPNS1 mRNA as an example and confirmed its presence in the RBCs. Future experimental verification would help define miR‐96–CAPNS1 interaction, if any, in the stored RBCs. CONCLUSIONS: This study for the first time provided a differential profile of stored RBCs for selected miRNAs related to cellular apoptotic pathway and opened new avenues toward identification of novel in vitro RBC biomarkers of storage lesions. Future studies focusing on target gene‐miRNA interactions in stored RBCs would also unravel underlying mechanisms of storage lesions.     

Functional mobility and postural control in essential tremor

OBJECTIVE: To evaluate functional mobility and postural control in participants with essential tremor (ET). DESIGN: Cross-sectional cohort study. SETTING: Motor performance research laboratory. PARTICIPANTS: Sixteen participants with ET including head tremor (age, 59.4+/-12.0 y), 14 participants with ET and no head tremor (age, 57.1+/-15.9 y), and 28 healthy controls (age, 58.4+/-12.4 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: We assessed the Timed Up & Go, time to ascend and descend stairs, Dynamic Gait Index, and Berg Balance Scale (BBS). Participants completed the Activities-specific Balance Confidence Scale and the Human Activity Profile. We assessed postural control using center-of-pressure measures from force platform recordings of quiet standing in 5 conditions. RESULTS: Participants with ET including head tremor performed worse than controls on all functional mobility performance and self-report measures (P<.05) except the BBS and stair descent time. Mean performance of ET participants without head tremor was intermediate between the other 2 groups. Sway speed measures of postural control showed similar patterns, but no significant group differences in post hoc analysis. There were no statistically significant or clinically important correlations between measures of tremor status and functional mobility status. CONCLUSIONS: Participants with ET show reduced functional mobility, especially those with head tremor.     

Freezing red blood cells prepared for quality control of antiglobulin sera

A small-aliquot freezing technique was employed to store at −80 C red blood cells (RBC) prepared for quality control of antiglobulin sera. These RBC were used to test the specificity and potency of both polyspecific and monospecific antiglobulin sera on the day of use. Following deglycerolization, about 83 per cent of test RBC were recovered. They were then stored as 5 per cent suspensions in 0.9% NaCl at 4 C for up to two weeks and tested for specific agglutination. EIg (RBC sensitized with immunoglobulins) and EC4 (RBC sensitized with the fourth component of human complement) remained reactive for the two- week period. EC43 (RBC sensitized with both the fourth and third components of human complement) tended to lose reactivity only with anti-C3c antibodies following deglycerolization and storage. EC3d (RBC sensitized with the C3d fragment of the third component of human complement), produced in vivo as a result of Mycoplasma pneumoniae infection, remained reactive for the two-week period, whereas EC3d prepared by the alternative pathway of complement activation was useful only for one week. Use of deglycerolized test RBC improved quality- control procedures by saving materials and technician time as well as by providing a constant supply of uniformly prepared test RBC.     

Removing IgG antibodies from intact red cells: comparison of acid and EDTA, heat, and chloroquine elution methods

BACKGROUND: To accurately phenotype red cell from patients with a positive direct antiglobulin test (DAT), nonlytic elution procedures were assessed for their ability to dissociate IgG from antibody-coated red cells without altering red cell antigen expression. STUDY DESIGN AND METHODS: Antibodies coating red cells that were sensitized in vivo (warm-reactive autoantibodies: 8 patients) or in vitro (42 alloantibodies) were eluted by using glycine-HCl and EDTA (acid/ EDTA), heat (56 degrees C, 10 min), or chloroquine method. RESULTS: Acid/EDTA elution gave the best results, reducing DAT positivity to microscopic levels or rendering the DAT negative in 48 of 50 instances, whereas 4 samples remained resistant to heat elution and 24 to chloroquine. Standard DAT agglutination scores demonstrated that both acid/EDTA and heat elution were superior to the chloroquine method (p < 0.0001). With the gel low-ionic-strength saline indirect antiglobulin test, acid/ EDTA was superior to heat (p < 0.001). Overall, acid/ EDTA elution dissociated more antibodies than heat (p < 0.0001), especially for Kell system (K, k, Kpa, Kpb) alloantibodies. Common red cell antigens, other than Kell system antigens, were unaffected by acid/EDTA elution. In contrast, the expression of most blood group antigens was diminished after heat elution. However, it was possible to type red cell antigens by using gel low-ionic-strength saline indirect antiglobulin tests or tube agglutination methods. CONCLUSION: Although heat elution may be used on a limited basis, the acid/EDTA method appears to be the procedure of choice for typing red cell coated with warm-reactive IgG alloantibodies or autoantibodies.