A microcomputer oximeter for whole blood

A system is described for measuring the percent of oxyhemoglobin in flowing whole blood. The system consists of an Apple II+ computer, a commercially available analog-to-digital converter, a simple electronic circuit to illuminate blood with light at two appropriate wavelengths, and a short program in BASIC. This oximeter is shown to yield a linear measurement of oxyhemoglobin saturation when compared with standard oxygen analyses. Data can be sampled at a sufficiently rapid rate to yield an essentially continuous measurement in typical cardiovascular studies. The system can be implemented by the average investigator from standard electronic components. The oximeter program can be combined into a general-purpose data acquisition program, and 15 analog-to-digital channels remain available for gathering other data.     

Resting whole blood viscosity of elite rowers is related to performance

This study investigated the relationships between resting whole blood viscosity (WBV), haemoglobin concentration (HGB), haematocrit (HCT), and performance in 25 highly-trained national squad rowers (11 women and 14 men). The WBV and HGB were measured at rest prior to a 2500 m simulated race on a Concept rowing ergometer when performance (P) was measured by average velocity. A group of 12 rowers were measured on just one occasion, another 11 were measured twice with an intervening 5 weeks of continued training and 2 were measured three times, the third test after another 4 weeks. Regression analyses making simultaneous use of both intra- and interindividual data indicated a significant inverse relationship between P and WBV (at both high and low shear rates), a relationship which was strengthened after statistically controlling for the effects of HGB, this effect being slightly more significant than HCT. A significant positive regression also emerged between P and HGB, but only after statistically controlling for the influence of WBV at high shear rate. Overall, stronger relationships were demonstrated in the male rowers compared with the female. These data, in the light of previous evidence that fitter people tend to have lower WBV, would indicate that blood rheology unrelated to HGB (or HCT) is related to performance in relatively homogeneous and already highly-trained athletes.     

Silicon-based microfilters for whole blood cell separation

This paper reports on the comparison analysis of four main types of silicon-based microfilter for isolation of white blood cells (WBCs) from red blood cells (RBCs) in a given whole blood. The microfilter designs, namely, weir, pillar, crossflow, and membrane, all impose the same cut-off size of 3.5 μm to selectively trap WBCs. Using human whole blood, the microfilters have been characterized and compared for their blood handling capacity, WBCs trapping efficiency and RBCs passing efficiency. Based on the experimental results, the crossflow microfilter is superior and can be integrated with downstream components for on-chip genomic analysis.     

Serum is more suitable than whole blood for diagnosis of systemic candidiasis by nested PCR

PCR assays for the diagnosis of systemic candidiasis can be performed either on serum or on whole blood, but results obtained with the two kinds of samples have never been formally compared. Thus we designed a nested PCR assay in which five specific inner pairs of primers were used to amplify specific targets on the rRNA genes of Candida albicans, C. tropicalis, C. parapsilosis, C. krusei, and C. glabrata. In vitro, the lower limit of detection of each nested PCR assay was 1 fg of purified DNA from the corresponding Candida species. In rabbits with candidemia of 120 minutes' duration following intravenous (i.v.) injection of 10(8) CFU of C. albicans, the sensitivities of the PCR in serum and whole blood were not significantly different (93 versus 86%). In other rabbits, injected with only 10(5) CFU of C. albicans, detection of candidemia by culture was possible for only 1 min, whereas DNA could be detected by PCR in whole blood and in serum for 15 and 150 min, respectively. PCR was more often positive in serum than in whole blood in 40 culture-negative samples (27 versus 7%; P < 0.05%). Lastly, experiments with rabbits injected i.v. with 20 or 200 microgram of purified C. albicans DNA showed that PCRs were positive in serum from 30 to at least 120 min after injection, suggesting that the clearance of free DNA is slow. These results suggest that serum is the sample of choice, which should be used preferentially over whole blood for the diagnosis of systemic candidiasis by PCR.     

Cryopreservation of human whole blood for pyrogenicity testing

Human whole blood assays are increasingly employed to test immune function or detect pyrogenic contamination, since they offer advantages, such as ease of performance, few preparation artifacts and a physiological cell environment. However, the approach is often limited by the availability of freshly drawn blood, putative safety concerns in the case of infected donors and interindividual donor differences. To overcome these limitations, a method was developed and optimized to produce batches of cryopreserved blood that can be used directly after thawing without any washing steps. Mononuclear cells remained intact as shown by FACS analysis. Cytokine release could be induced by a variety of immunological stimuli. The cell preparation released higher amounts of interleukin-1beta (IL-1beta) and IL-6 compared to fresh blood, but no TNF. These differences could be attributed to the presence of the cryoprotectant dimethylsulfoxide (DMSO) alone by addition of DMSO to fresh blood. Large batches of cryopreserved blood could be produced by mixing blood donations of up to 10 donors, independent of differing blood groups. The detection limit for the World Health Organization (WHO) lipopolysaccharides (endotoxin, LPS) reference preparation (EC-6) with regard to the induction of IL-1beta release was at least 0.5 endotoxin equivalent units (EU)/ml. Endotoxin spikes at the limit concentrations prescribed in the European Pharmacopoeia could be detected in a series of drugs, showing that the In vitro Pyrogen Test (IPT) can also be run with cryopreserved blood. Further possible applications include high-throughput screening for immunomodulators or toxins as well as preservation of patient samples for later analysis of cell functions.     

Fragile X expression in short-term whole blood cultures is affected by cell density

The effect of cell density on expression of the fragile site at (X)(q27.3) in short-term whole-blood cultures from patients with fragile X [fra(X)] or Martin-Bell syndrome was studied. A significant increase in fra(X) frequency was observed in 7 of 8 samples when cell density was decreased. Higher fra(X) frequency was not always noted at below-routine density, but in some cases fra(X) expression was depressed at above-routine density. We conclude that decay of the FUdR effect explains the fact that fra(X) expression is affected by culture density. It is significant that a relationship exists between the two; it suggests that in order to maximize fra(X) expression in cases with low-percentage fra(X) with standard methods, cell density may have to be adjusted. It is possible that in individuals who are normally nonexpressing, such as some obligate female carriers and nonpenetrant males, fra(X) expression may be sensitive to cell density effects.     

Comparison of haemolytic activity of tentacle-only extract from jellyfish Cyanea capillata in diluted whole blood and erythrocyte suspension: Diluted whole blood is a valid test system for haemolysis study

In this paper, we utilized two different test systems to compare the haemolysis of tentacle-only extract (TOE) devoid of nematocysts from jellyfish Cyanea capillata, the 1% whole blood and 0.45% erythrocyte suspension approximately with the same erythrocyte concentration from the blood samples of sheep, rabbit, mouse, rat and human, respectively. Without exception, the haemolytic activity of TOE was dose-dependent in both test systems from all the five kinds of blood samples, while it was generally stronger in erythrocyte suspension than that in diluted whole blood at the relatively high concentration of TOE. When various aliquots of plasma were added into the erythrocyte suspension test system, the haemolytic activity of TOE was declined with the plasma quantity increasing, and dropped to about 20% at the presence of two aliquots of plasma. If serum albumin of 0.5 mg/ml, approximately the same albumin content in 1% whole blood, was added into the erythrocyte suspension test system instead, the haemolysis of TOE was similarly inhibited. The effects of GSH, ascorbic acid and protease inhibitor on the haemolytic activity of TOE were detected in the erythrocyte suspension and diluted whole blood simultaneously, and the test results were coincident between the two systems. These results suggested that the inconsistency of TOE haemolysis between the erythrocyte suspension and diluted whole blood is a universal occurrence in the mammals, and blood plasma plays a dose-dependent protective role against haemolysis which may be due to serum albumin. Diluted whole blood is a valid and convenient test system for haemolysis study in vitro.     

Nuclear magnetic relaxation of aqueous solutions of proteins,blood plasma,erythrocytes, and whole blood

V. I. Ul'yanov-Lenin Kazan' University. (Presented by Academician of the Academy of Medical Sciences of the USSR A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 112, No. 10, pp. 378–381, October, 1991.