Whole Blood Erythrocyte Research Articles

Influence of Mirabegron, Adrenaline, and Ascorbic Acid on the Intensity of Iron-Induced Biochemiluminescence of Erythrocytes in Whole Blood in Men

Influence of Mirabegron, Adrenaline, and Ascorbic Acid on the Intensity of Iron-Induced Biochemiluminescence of Erythrocytes in Whole Blood in Men

Comparative evaluation for the globin gene depletion methods for mRNA sequencing using the whole blood-derived total RNAs

BackgroundThere are challenges in generating mRNA-Seq data from whole-blood derived RNA as globin gene and rRNA are frequent contaminants. Given the abundance of erythrocytes in whole blood, globin genes comprise some 80% or more of the total RNA. Therefore, depletion of globin gene RNA and rRNA are critical steps required to have adequate coverage of reads mapping to the reference transcripts and thus reduce the total cost of sequencing. In this study, we directly compared the performance of probe hybridization (GLOBINClear Kit and Globin-Zero Gold rRNA Removal Kit) and RNAse-H enzymatic depletion (NEBNext® Globin & rRNA Depletion Kit and Ribo-Zero Plus rRNA Depletion Kit) methods from 1 μg of whole blood-derived RNA on mRNA-Seq profiling. All RNA samples were treated with DNaseI for additional cleanup before the depletion step and were processed for poly-A selection for library generation.ResultsProbe hybridization revealed a better overall performance than the RNAse-H enzymatic depletion method, detecting a higher number of genes and transcripts without 3′ region bias. After depletion, samples treated with probe hybridization showed globin genes at 0.5% (±0.6%) of the total mapped reads; the RNAse-H enzymatic depletion had 3.2% (±3.8%). Probe hybridization showed more junction reads and transcripts compared with RNAse-H enzymatic depletion and also had a higher correlation (R > 0.9) than RNAse-H enzymatic depletion (R > 0.85).ConclusionIn this study, our results showed that 1 μg of high-quality RNA from whole blood could be routinely used for transcriptional profiling analysis studies with globin gene and rRNA depletion pre-processing. We also demonstrated that the probe hybridization depletion method is better suited to mRNA sequencing analysis with minimal effect on RNA quality during depletion procedures.

Passive Coagulability Assay Based on Coherence-Gated Light Scattering

Coagulation monitoring relies on in vitro tests where the clot formation is induced using external stimuli. We report an optical method capable of revealing the propensity of coagulation based solely on the natural dynamics of erythrocytes in whole blood. In contrast to traditional techniques, our approach provides means to assess the blood coagulability without the need to chemically trigger the coagulation. Results of correlations with standard clinical methods suggest that this optical assay could be used for continuous management of blood coagulation during clinical procedures.

Dry Fasting Physiology: Responses to Hypovolemia and Hypertonicity

Objective: The aim of this study was to provide a deeper insight into dry fasting (DF) physiology. Design: Ten participants performed DF for 5 consecutive days. Methods: The following parameters were monitored daily: cortisol, aldosterone, high-sensitivity C-reactive protein (CRP), erythropoietin, albumin, uric acid, and vitamin C in serum; vasopressin (ADH), adrenocorticotropic hormone (ACTH), renin, angiotensin II, and total antioxidant capacity (TAC) in plasma; hematocrit and erythrocytes in whole blood; osmolality, noradrenaline, dopamine, adrenaline, Na⁺, and K⁺ in 24-h urine; waist circumference and body, urine, and stool weight. Results: The following parameters increased: ADH (60 ± 11%), ACTH (176 ± 34%), cortisol (495 ± 75%), urine osmolality (20 ± 4%), CRP (167 ± 77%), renin (315 ± 63%), angiotensin II (74 ± 21%), aldosterone (61 ± 21%), TAC (80.4 ± 17%), uric acid (103 ± 19%), albumin (18.4 ± 2.4%), erythrocytes (13.4 ± 2.2%), hematocrit (11 ± 1.8%), and the excretion of noradrenaline (40.3 ± 10%) and dopamine (17 ± 5%). The following parameters decreased: waist circumference (8.20 ± 0.61 cm), body weight (7.010 ± 0.3 kg), erythropoietin (65 ± 18%), and the excretion of adrenaline (38 ± 4%) and Na⁺ (60 ± 16%). The excretion of K⁺ remained unchanged. Vitamin C decreased, showing a half-life of 4.8 ± 0.7 days. The percent ratios of lost weight components were: urine (52.2 ± 3.7%), insensible water loss (32.2 ± 1.4%), stool (5 ± 0.3%), and respiratory gases, i.e., expired CO₂ – incorporated O₂ (10.6 ± 5.4%). Conclusion: The mechanisms underlying the hypertonicity and hypovolemia compensation and the ratio analysis of lost weight components were presented. DF demonstrated short-term antioxidant, anti-ischemic, immune-stimulating, anti-edematous, and anti-inflammatory effects. The results may have an impact on developing new concepts for the treatment of edema, obesity, and inflammatory and ischemic diseases.

Identification of a Hemolysis Threshold That Increases Plasma and Serum Zinc Concentration

Background: Plasma or serum zinc concentration (PZC or SZC) is the primary measure of zinc status, but accurate sampling requires controlling for hemolysis to prevent leakage of zinc from erythrocytes. It is not established how much hemolysis can occur without changing PZC/SZC concentrations.Objective: This study determines a guideline for the level of hemolysis that can significantly elevate PZC/SZC.Methods: The effect of hemolysis on PZC/SZC was estimated by using standard hematologic variables and mineral content. The calculated hemolysis threshold was then compared with results from an in vitro study and a population survey. Hemolysis was assessed by hemoglobin and iron concentrations, direct spectrophotometry, and visual assessment of the plasma or serum. Zinc and iron concentrations were determined by inductively coupled plasma spectrometry.Results: A 5% increase in PZC/SZC was calculated to result from the lysis of 1.15% of the erythrocytes in whole blood, corresponding to ∼1 g hemoglobin/L added into the plasma or serum. Similarly, the addition of simulated hemolysate to control plasma in vitro caused a 5% increase in PZC when hemoglobin concentrations reached 1.18 ± 0.10 g/L. In addition, serum samples from a population nutritional survey were scored for hemolysis and analyzed for changes in SZC; samples with hemolysis in the range of 1–2.5 g hemoglobin/L showed an estimated increase in SZC of 6% compared with nonhemolyzed samples. Each approach indicated that a 5% increase in PZC/SZC occurs at ∼1 g hemoglobin/L in plasma or serum. This concentration of hemoglobin can be readily identified directly by chemical hemoglobin assays or indirectly by direct spectrophotometry or matching to a color scale.Conclusions: A threshold of 1 g hemoglobin/L is recommended for PZC/SZC measurements to avoid increases in zinc caused by hemolysis. The use of this threshold may improve zinc assessment for monitoring zinc status and nutritional interventions.

Ultrastructural changes of erythrocytes in whole blood after exposure to prospective in silico-designed anticancer agents: a qualitative case study

BackgroundNovel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16). These compounds were designed to have improved bioavailability when compared to their source compound, 2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II, present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for the handling of the whole blood sample.The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female’s erythrocytes was examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds. Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by determining morphological changes after a 24 hour storage period at 22°C and 37°C.ResultsNo significant morphological changes were observed in the erythrocyte morphology after exposure to the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural deformity or distress.ConclusionsFrom this research the optimal temperature for ex vivo exposure of whole blood samples to ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal incubation conditions up to 24 hours for whole blood were established as a secondary outcome.

Diffraction Phase Cytometry: blood on a CD-ROM

We demonstrate Diffraction Phase Cytometry (DPC) as a single shot, full-field, high throughput quantitative phase imaging modality, dedicated to analyzing whole blood smears. Utilizing a commercial CD as a sample substrate, along with dynamic spatial filtering via a liquid crystal spatial light modulator, we have developed a compact instrument capable of making quantitative, physiologically relevant measurements. To illustrate the ability of the system to function as a highly sensitive cytometer we imaged a large number (N=1,537) of live human erythrocytes in whole blood without preparation. We retrieved a comprehensive set of geometrical parameters including cell volume and surface area, which are not directly available using existing cytometers. Furthermore, we retrieved the minimum cylindrical diameter, through which red blood cells can pass, and deliver oxygen. These initial results prove the concept for an inexpensive lab-on-a-chip blood screening device.

Evaluation of a Protein Microarray Method for Immuno‐Typing Erythrocytes in Whole Blood

All donor blood samples must be tested pre‐transfusion to determine the blood type of donor erythrocytes, based on the ABO typing system. Current methods of testing are well characterised, but require a number of processing steps prior to analysis. In addition, standard testing protocols require additional assays such as hepatitis C and HIV testing be performed separately. We describe and evaluate a protein microarray platform for ABO blood typing that has the potential to be a simple reliable high throughput method, with the added capability for the integration of other important pre‐transfusion tests. Sixty seven donor blood samples were incubated on microarrays printed with multiple spotted replicates of blood type antigen specific antibodies. We utilised a hold‐out cross validation approach, combined with Receiver Operator Characteristic (ROC) curves to define thresholds within which a sample could be defined as being of a particular blood type. The threshold values from the ROC curve analysis demonstrated an excellent ability to accurately separate samples based on ABO blood type. The results obtained when the thresholds from the training sets were applied to test sets were also very encouraging, with misclassified samples being present in only 2 of the training sets and a mean classification error of 4.28%. When the mean thresholds were applied to the 67 donor samples, 95.5% were correctly blood typed (64 of 67 samples). We have demonstrated the ability of our protein microarray platform to successfully and accurately type human whole blood samples. We believe that this flexible platform provides a strong basis for an integrated approach for combined blood typing and pathogen testing in human whole blood.

Comparison of sample fixation and the use of LDS-751 or anti-CD45 for leukocyte identification in mouse whole blood for flow cytometry

Flow cytometry methods used to measure leukocyte function often entail sample preparation procedures that cause artifactual cell activation. To avoid leukocyte activation by isolation techniques, some preparation methods use fluorescent markers to discriminate leukocytes from erythrocytes in whole blood. One of these markers, laser dye styryl-751(LDS-751), has been used to distinguish leukocytes by staining nucleic acid, but has been found to stain other blood cells and dead cells indiscriminately. Thus, LDS-751 may not be an appropriate reagent for leukocyte identification in whole blood. Fixing samples with formaldehydes increases cell permeability and causes surface protein cross-linking that may alter staining of both intra- and extracellular markers. The degree of this sample alteration by formaldehyde fixation, however, remains in question. In addition, little is known about flow cytometry and sample preparation methods in mouse whole blood. The purpose of this study was to determine if labeling leukocytes with a monoclonal antibody specific to leukocyte common antigen (CD45) was superior to labeling with LDS-751 and to determine the effect of sample fixation on a mouse whole blood preparation for flow cytometry. Samples were incubated with CD16/CD32 Fc receptor blocker, and either 10 μg/ml LDS-751 or phosphate buffered saline (PBS). The samples were then fixed with paraformaldehyde or diluted with PBS followed by incubation with 5 μg/ml PerCP-conjugated anti-CD45, 5 μg/ml FITC-conjugated anti-CD11b, or 80 μM dichlorofluorescein diacetate. We found that samples labeled with LDS-751 demonstrated decreased fluorescence intensity for granulocyte CD11b expression and ROS production compared to samples labeled with anti-CD45. In addition, sample fixation decreased mean fluorescence intensity in samples labeled with either LDS-751 or anti-CD45. We conclude that labeling leukocytes with monoclonal antibody CD45 in a mouse whole blood preparation is preferable, as it provides improved measurement of leukocyte indices compared to LDS-751. Also, while sample fixation prior to antibody staining caused a decrease in overall fluorescence; it can be used to successfully identify extra-cellular markers.

Separation of lipids from blood utilizing ultrasonic standing waves in microfluidic channels.

A method to continuously separate different particle types in a suspension is reported. Acoustic forces in a standing wave field were utilized to discriminate lipid particles from erythrocytes in whole blood. The presented technology proposes a new method of cleaning, i.e. removing lipid emboli from, shed blood recovered during cardiac surgery. Blood contaminated with lipid particles enter a laminar flow micro channel. Erythrocytes and lipid particles suspended in blood plasma are exposed to a half wavelength standing wave field orthogonal to the direction of flow as they pass through the channel. Because of differences in compressibility and density the two particle types move in different directions, the erythrocytes towards the centre of the channel and the lipid particles towards the side walls. The end of the channel is split into three outlet channels conducting the erythrocytes to the centre outlet and the lipid particles to the side outlets due to the laminar flow profile. The separation channel was evaluated in vitro using polyamide spheres suspended in water, showing separation efficiencies approaching 100%. The system was also evaluated on whole blood using tritium labelled lipid particles added to bovine blood. More than 80% of the lipid particles could be removed while approximately 70% of the erythrocytes were collected in one third of the original fluid volume. The study showed that the further reduced micro channel dimensions provided improved performance with respect to; (i) separation efficiency, (ii) actuation voltage, and (iii) volumetric throughput as compared to earlier work.

Low power laser protects human erythrocytes In an In vitro model of artificial heart-lung machines.

The protective effect of the low power helium-neon (He-Ne) laser against the damage of human erythrocytes in whole blood was examined in a perfusion model using an artificial heart-lung machine. Preserved human whole blood was diluted and perfused in 2 closed circuits with a double roller pump. The laser irradiated one of the circuits (laser group), and none the other (control group). In the laser group, erythrocyte deformability and erythrocyte adenosine triphosphate (ATP) levels were significantly higher, and free hemoglobin levels were significantly lower than those in the control group. Subsequent morphological findings by means of scanning electron microscope were consistent with these results. Low power He-Ne laser protected human erythrocytes in the preserved diluted whole blood from the damage caused by experimental artificial heart-lung machines. The clinical application of low power laser treatment for extracorporeal circulation is suggested.

Aggregation and Disaggregation of Erythrocytes in Whole Blood: Study by Backscattering Technique

The aggregation phenomenon is of great importance for the evaluation of performance of the microcirculation system because of its influence on the blood viscosity at low shear stresses. Some important features and consequences of this phenomenon in vivo can be predicted in the in vitro experiments using optical methods. These methods are considered to be the most informative and applicable not only for the basic study of the aggregation phenomenon, but also for the diagnosis of a number of diseases and for the monitoring of therapeutic treatment in clinics. Results presented in this paper prove that the backscattering technique allows one to detect different changes of aggregational ability and deformability of erythrocytes and to get reliable and reproducible results distinguishing normal blood and blood with different pathologies. © 1999 Society of Photo-Optical Instrumentation Engineers.

Photosensitized inactivation of Plasmodium falciparum- and Babesia divergens-infected erythrocytes in whole blood by lipophilic pheophorbide derivatives.

Blood transfusions can transmit parasitic infections, such as those caused by Plasmodium (malaria), Trypanosoma cruzi (Chagas' disease), and Babesia (babesiosis). A higher degree of blood transfusion safety would be reached if methods were available for inactivating such parasites. We evaluated the effectiveness of photosensitization using lipophilic pheophorbide and red light illumination to eradicate red blood cells infected with Plasmodium falciparum, and with Babesia divergens, in whole blood. Fluorescence microscopy and conventional fluorometry showed the specific accumulation of pheophorbide derivatives in the RBC infected with either parasite, compared with uninfected RBC. The effectiveness of different derivatives in eradicating infected RBC was first estimated in parasite cultures. The best photosensitizer was the N-(4-butanol) pheophorbide derivative (Ph4-OH) at 0.2 microM concentration and 5-min illumination. In whole blood, the eradication of RBC infected with B. divergens and P. falciparum was obtained with 2 microM Ph4-OH and 10 and 20 min illumination, respectively. Under these conditions of photosensitization, low levels of RBC hemolysis were noted even after 2 weeks of storage at 4 degrees C and a subsequent 48-hour incubation at 37 degrees C. No reduction of negative charges on treated RBC was noted and no increase in methemoglobin content. In plasma, Ph4-OH is mainly transported by high-density lipoproteins (HDL). This high affinity for HDL may explain the selective accumulation of lipophilic pheophorbide derivatives in the intracellular parasites. Photosensitization with pheophorbide derivatives may be a promising approach to inactivation of transfusion-transmissible parasites and viruses in blood bank units.

Photosensitized Inactivation of Plasmodium falciparum- and Babesia divergeas-Infected Erythrocytes in Whole Blood by Lipophilic Pheophorbide Derivatives

Background and objectives: Blood transfusions can transmit parasitic infections, such as those caused by Plasmodium (malaria), Trypanosoma cruzi (Chagas’ disease), and Babesia (babesiosis). A higher degree of blood transfusion safety would be reached if methods were available for inactivating such parasites. Materials and methods: We evaluated the effectiveness of photosensitization using lipophilic pheophorbide and red light illumination to eradicate red blood cells infected with Plasmodium falciparum, and with Babesia divergens, in whole blood. Fluorescence microscopy and conventional fluorometry showed the specific accumulation of pheophorbide derivatives in the RBC infected with either parasite, compared with uninfected RBC. The effectiveness of different derivatives in eradicating infected RBC was first estimated in parasite cultures. Results: The best photosensitizer was the N-(4-butanol) phrophorbide derivative (Ph4-OFI) at 0.2 μM concentration and 5-min illumination. In whole blood, the eradication of RBC infected with B. divergens and P. falciparum was obtained with 2 pM Ph4-OH and 10 and 20 min illumination, respectively. Under these conditions of photosensitization, low levels of RBC hemolysis were noted even after 2 weeks of storage at 4°C and a subsequent 48-hour incubation at 37°C. No reduction of negative charges on treated RBC was noted and no increase in methemoglobin content. Conclusion: In plasma, Ph4-OH is mainly transported by high-density lipoproteins (HDL). This high affinity for HDL may explain the selective accumulation of lipophilic pheophorbide derivatives in the intracellular parasites. Photosensitization with pheophorbide derivatives may be a promising approach to inactivation of transfusion-transmissible parasites and viruses in blood bank units.

Aspirin improves the enhanced erythrocyte aggregability in patients with cerebral infarction

We examined the effect of aspirin on the enhanced erythrocyte aggregability in 19 patients with cerebral infarction during the chronic phase. Aspirin (81 mg/day) was administered for 8 weeks. The rate of erythrocyte aggregation (aggregability) was measured using the whole-blood erythrocyte aggregometer developed by us (Am. J. Physiol. 251, H1205–H1210, 1986) before, and at 4 and 8 weeks after initiation of the aspirin administration. Concomitant measurements were made of certain blood factors such as the hematocrit, albumin-globulin ( A G ) ratio and fibrinogen concentration. The erythrocyte aggregation rates before, and at 4 and 8 weeks were 0.154 ± 0.019/s, 0.144 ± 0.020/s, and 0.143 ± 0.020/s, respectively. The erythrocyte aggregation rates at 4 and 8 weeks were significantly ( P < 0.05, Bonferroni's modified t-test) lower than that before aspirin administration. Although the hematocrit value and A G ratio were not changed at 4 and 8 weeks after the initiation of aspirin, the concentration of fibrinogen were significantly ( P < 0.05) reduced at 4 weeks. The above results suggest that aspirin can improve the enhanced erythrocyte aggregability in patients with cerebral infarction during the chronic phase.

Aspirin improves the enhanced erythrocyte aggregability in patients with cerebral infarction

We examined the effect of aspirin on the enhanced erythrocyte aggregability in 19 patients with cerebral infarction during the chronic phase. Aspirin (81 mg/day) was administered for 8 weeks. The rate of erythrocyte aggregation (aggregability) was measured using the whole-blood erythrocyte aggregometer developed by us (Am. J. Physiol. 251, H1205–H1210, 1986) before, and at 4 and 8 weeks after initiation of the aspirin administration. Concomitant measurements were made of certain blood factors such as the hematocrit, albumin-globulin () ratio and fibrinogen concentration. The erythrocyte aggregation rates before, and at 4 and 8 weeks were 0.154 ± 0.019/s, 0.144 ± 0.020/s, and 0.143 ± 0.020/s, respectively. The erythrocyte aggregation rates at 4 and 8 weeks were significantly (P < 0.05, Bonferroni's modified t-test) lower than that before aspirin administration. Although the hematocrit value and ratio were not changed at 4 and 8 weeks after the initiation of aspirin, the concentration of fibrinogen were significantly (P < 0.05) reduced at 4 weeks. The above results suggest that aspirin can improve the enhanced erythrocyte aggregability in patients with cerebral infarction during the chronic phase.

Effect of single intravenous administration of batroxobin on erythrocyte aggregability in patients with acute-stage cerebral infarction

(Received 27.09.1994; accepted 24.11.1994) We examined the effect of defibrinogenation with batroxobin on the erythrocyte aggregability (RBC-A) in 16 patients with cerebral infarction during the acute phase (less than 72 hours after onset). Eight patients received a single intravenous administration of 10 units of batroxobin (BU), while the other 8 patients received 5 BU. The RBC-A was examined using the whole-blood erythrocyte aggregometer developed by us (Am. J. Physiol. 251, H1205H1210, 1986) with concomitant measurement of the hematocrit, albumin:globulin ratio and concentration of fibrinogen. The RBC-A values before, and at 1,2, and 7 days after 10 BU administration were 0.154±0.017/s, 0.117 ±0.029/s (76% of the value before the administration, P<O.Ol), 0.105±0.029/s (68%, P<O.Ol), and 0.124±0.015/s (81 %, P<O.Ol), respectively. The fibrinogen concentrations before, and at 1,2, and 7 days after the batroxobin were 364±81 mg/dl, 150±35 mg/dl (41 %, P<O.Ol), 129±35 mg/dl (35%, P<O.01), and 211±42 mg/dl (58%, P<O.Ol), respectively; On the other hand, the RBC-A values before, and at 1, 2, and 7 days after 5 BU administration were 0.143±0.021/s, 0.131±0.036/s, 0.130±0.028/s, and 0.141±0.029/s, respectively. The fibrinogen concentrations before, and at 1,2, and 7 days after the batroxobin were 318±57 mg/dl, 255±60 mg/dl (80%, P<O.Ol), 259±68 mg/dl (81 %, P<O.Ol), and 319±87 mg/dl, respectively. We conClude that a single intravenous administration of batroxobin at 10 BU reduced the RBC-A in patients with cerebral infarction during the acute phase in accordance with a decrease in the fibrinogen level. The effect appeared to last for more than 7 days.

Effect of lipopolysaccharide on the physical conformation of the erythrocyte cytoskeletal proteins

Red blood cell deformability is important for effective circulation in the capillaries. It is known that red cell deformability is significantly reduced during septic shock. Surface to volume ratio, physical effects of the cytoskeletal proteins and the fluidity of lipid bilayer are some of the important intrinsic factors that regulate this mechanical function. Alterations in the physical conformation of cytoskeletal proteins in septic conditions could significantly alter their function. In this study, erythrocytes in whole blood were treated with lipopolysaccharide, the outer covering of Gram-negative bacteria released during Gram-negative sepsis. Electron paramagnetic resonance spectroscopy in conjunction with a protein-specific maleimide nitroxide spin label covalently bound to cytoskeletal proteins was used to investigate the resulting changes occurring in the physical state of cytoskeletal proteins in isolated membranes. Treatment of red blood cells with a lipopolysaccharide concentration as low as 40 μg/mL of blood solution for 90 minutes showed a significant decrease in the relevant EPR parameter (p0̌.01) of the spin label bound to subsequently isolated membranes, suggestive of a decreased segmental motion of the spin label and an increase in cytoskeletal protein-protein interactions. These results suggest a marked conformational alteration in the cytoskeletal proteins induced by the lipopolysaccharide and may explain, in part, the marked reduction in red blood cell deformability during septic shock. Bacterial lipopolysaccharide does not exert most of its effects on the host directly, but rather elicits the production of host factors that leads to complex septic shock. Leukocytes, endothelial tissue and many other cells release these mediators. Leukocytes are thought to be a particularly important source of such mediators, including cytokines (tumor necrosis factor, interleukins, etc.), oxygen free radicals, proteases, and hydrolyses. In order to characterize the possible mechanism by which the lipopolysaccharide acts on the physical state of the erythrocyte cytoskeleton, erythrocytes void of leukocytes and plasma were treated with lipopolysaccharide. The relevant EPR parameter showed no significant change over the control value. These results indicate that the leukocytes and their factors are responsible for the rearrangements seen in the cytoskeletal proteins of the erythrocyte membrane.

Ticlopidine improves the enhanced erythrocyte aggregability in patients with cerebral infarction.

We examined the effect of ticlopidine hydrochloride on the enhanced erythrocyte aggregability in 14 patients with cerebral infarction during the chronic phase (over 1 month after onset). Ticlopidine (100 mg BID) was administered for 8 weeks. We measured the rate of erythrocyte aggregation (aggregability), using the whole-blood erythrocyte aggregometer that we developed, before and at 4 and 8 weeks after the initiation of ticlopidine administration. Concomitant measurements were made of such blood factors as the hematocrit, albumin-globulin ratio, and fibrinogen concentration. The erythrocyte aggregation rates before and at 4 and 8 weeks after were 0.147 +/- 0.017/s, 0.138 +/- 0.019/s, and 0.133 +/- 0.017/s, respectively. The erythrocyte aggregation rates at 4 and 8 weeks were significantly lower (P < .05 by Bonferroni's modified t test) than those before ticlopidine administration. At 4 and 8 weeks after the initiation of ticlopidine treatment, the hematocrit value and concentration of fibrinogen were also significantly (P < .05) reduced. Our results suggest that ticlopidine can improve the enhanced erythrocyte aggregability in patients with cerebral infarction during the chronic phase.

Factors affecting whole blood O2 transfer kinetics: implications for theta(O2)

Half times (t0.5) of O2 uptake (on) and release (off) were measured on thin layers of whole blood (WB). Increasing film thickness (L = 1-6 microns) increased t0.5(on) from 10 to 20 ms and t0.5(off) from 20 to 45 ms. At all L, for changes in saturation (S) 0----1, t0.5(off) was about twofold greater than t0.5(on). Doubling initial onloading PO2 gradient decreased t0.5(on) 36%; t0.5(off) was unchanged. A 10-Torr change in WB O2 affinity (P50) had no significant effect on t0.5. Specific O2 conductance [theta(O2), ml O2.(min.Torr.ml WB)-1] was also computed as a function of S from these data. A bell-shaped theta(O2) vs. S relationship was always obtained; maximum values of theta(O2) reached 25. Measured values for theta(O2) at any S were highly sensitive to choice of experimental conditions, i.e., L, affinity, flux direction, initial PO2 gradient, and initial level of S. We show experimentally and theoretically that no unique relationship exists for theta(O2) as a function of S for erythrocytes in WB.