Integral viscosity: a useful parameter in evaluating whole blood and plasma viscosities in health and disease

An integral viscosity determination (Q) is proposed for whole blood (QB) and plasma (Qp). The integral viscosity determination was carried out at four shear rates in a Brookfield LTV Viscometer and calculation were obtained by means of a modification of Casson's mathematical model. Polycythemic and paraproteinemic patients resulted to be well differentiated from normals. The ratio QB/Qp represent a continuous sum of relative viscosities and may also supply information on erythrocytes deformability. Integral viscosity determinations are indicated for research purposes and for examining special cases.     

A novel approach to blood plasma viscosity measurement using fluorescent molecular rotors

Molecular rotors, a group of fluorescent molecules with viscosity-dependent quantum yield, were tested for their suitability to act as fluorescence-based plasma viscometers. The viscosity of samples of human plasma was modified by the addition of pentastarch (molecular mass 260 kDa, 10% solution in saline) and measured with a Brookfield viscometer. Plasma viscosity was 1.6 mPa x s, and the mixtures ranged up to 4.5 mPa x s (21 degrees C). The stimulated light emission of the molecular rotors mixed in the plasma samples yielded light intensity that was nonoverlapping and of significantly different intensity for viscosity steps down to 0.3 mPa x s (n = 5, P < 0.0001). The mathematical relationship between intensity (I) and viscosity (eta) was found to be eta = (kappaI)(nu). After calibration and scaling the fluorescence based measurement had an average deviation versus the conventional viscometric measurements that was <1.8%. These results show the suitability of molecular rotors for fast, low-volume biofluid viscosity measurements achieving accuracy and precision comparable to mechanical viscometers.     

Extracellular circulating nucleic acids in human plasma in health and disease

The concentration of extracellular DNA and RNA in blood plasma of healthy donors, trauma patients, patients with breast and lung cancer, nonmalignant breast tumors and nonmalignant lung diseases were estimated. Significant amounts of extracellular RNA were found in plasma of trauma patients. The concentration of DNA and RNA in plasma of trauma patients correlates with the extent of posttraumatic organ failure. Extracellular RNA was not found in the plasma of breast cancer patients and patients with nonmalignant breast tumors, whereas a very high concentration of extracellular RNA was found in patients with malignant and nonmalignant diseases of lung.     

The role of plasma noradrenaline in health and disease

This paper proposes that plasma noradrenaline plays a central role in the physiology and pathophysiology of the macrocirculation, the heart, veins and arteries. The proposal stems, in the final analysis, from the finding that noradrenaline dilates the canine lateral saphenous vein when it is released from its microcirculation, the vasa venarum. The concentration threshold of the effect is estimated to be at least eight times lower than the threshold of the constrictor effect of intralumenal noradrenaline. Combined with other evidence, the finding indicates that, contrary to opinion, plasma noradrenaline has an effective β₁-agonist hormonal effect on the macrocirculation, at normal concentrations. It also indicates that noradrenaline has a bi-polar effect. The reason it has that effect is that noradrenaline is a primary biological stimulus and like all primary stimuli investigated to date, it can be expected to have two cross-inhibitory components, commonly referred to as excitor and lateral inhibitory. When examined, neuronal noradrenaline shows the features characteristic of excitor components in general and plasma noradrenaline shows those characteristic of inhibitory components. The most significant of the latter is that inhibitory components are the most potent physiological modulators of excitor components. A striking example of that modulation effect occurs in smooth muscle contraction where muscles contracted by neuronal noradrenaline stimulation appear to be incapable of relaxing without stimulation by plasma noradrenaline-hence the proposition that vascular stenosis and cardiospasm are caused by a pathological loss of plasma noradrenaline stimulation. By triggering turbulence, cholesterol plaques increase the arterial microcirculatory flow and so increase the β₁-agonist effect of plasma noradrenaline. This paper proposes that this cholesterol driven increase in the microcirculatory effect of plasma noradrenaline is the cause of the arteriosclerotic syndrome, a proposition that is consistent with the success of beta-blockade in treating manifestations of the syndrome. The paper concludes by examining a variety of conditions, including dissecting aneurysms, dilator cardiac failure, angina, myocardial infarction, hypertension, eclampsia and cirrhosis and pointing out that all of them, like varicose veins, show evidence consistent with having been caused by a turbulence induced increase in a β₁-agonist stimulatory effect of microcirculatory plasma noradrenaline.     

Effect of plasma exchange on blood viscosity and cerebral blood flow.

The effects of plasma exchange using a low viscosity plasma substitute on blood viscosity and cerebral blood flow were investigated in eight subjects with normal cerebral vasculature. Plasma exchange resulted in significant reductions in plasma viscosity, whole blood viscosity, globulin and fibrinogen concentration without affecting packed cell volume. The reduction in whole blood viscosity was more pronounced at low shear rates suggesting an additional effect on red cell aggregation. Despite the fall in viscosity there was no significant change in cerebral blood flow. The results support the metabolic theory of autoregulation. Although changes in blood viscosity appear not to alter the level of cerebral blood flow under these circumstances, plasma exchange could still be of benefit in the management of acute cerebrovascular disease.     

The human GLUD2 glutamate dehydrogenase and its regulation in health and disease

Whereas glutamate dehydrogenase in most mammals (hGDH1 in the human) is encoded by a single functional GLUD1 gene expressed widely, humans and other primates have acquired through retroposition an X-linked GLUD2 gene that encodes a highly homologous isoenzyme (hGDH2) expressed in testis and brain. Using an antibody specific for hGDH2, we showed that hGDH2 is expressed in testicular Sertoli cells and in cerebral cortical astrocytes. Although hGDH1 and hGDH2 have similar catalytic properties, they differ markedly in their regulatory profile. While hGDH1 is potently inhibited by GTP and may be controlled by the need of the cell for ATP, hGDH2 has dissociated its function from GTP and may metabolize glutamate even when the Krebs cycle generates GTP amounts sufficient to inactivate hGDH1. As astrocytes are known to provide neurons with lactate that largely derives from the Krebs cycle via conversion of glutamate to α-ketoglutarate, the selective expression of hGDH2 may facilitate metabolic recycling processes essential for glutamatergic transmission. As there is evidence for deregulation of glutamate metabolism in degenerative neurologic disorders, we sequenced GLUD1 and GLUD2 genes in neurologic patients and found that a rare T1492G variation in GLUD2 that results in substitution of Ala for Ser445 in the regulatory domain of hGDH2 interacted significantly with Parkinson's disease (PD) onset. Thus, in two independent Greek and one North American PD cohorts, Ser445Ala hemizygous males, but not heterozygous females, developed PD 6-13 years earlier than subjects with other genotypes. The Ala445-hGDH2 variant shows enhanced catalytic activity that is resistant to modulation by GTP, but sensitive to inhibition by estrogens. These observations are thought to suggest that enhanced glutamate oxidation by the Ala445-hGDH2 variant accelerates nigral cell degeneration in hemizygous males and that inhibition of the overactive enzyme by estrogens protects heterozygous females. We then evaluated the interaction of estrogens and neuroleptic agents (haloperidol and perphenazine) with the wild-type hGDH1 and hGDH2 and found that both inhibited hGDH2 more potently than hGDH1 and that the evolutionary Arg443Ser substitution was largely responsible for this sensitivity. Hence, the properties acquired by hGDH2 during its evolution have made the enzyme a selective target for neuroactive steroids and drugs, providing new means for therapeutic interventions in disorders linked to deregulation of this enzyme.     

Simultaneous measurement of dothiepin and its major metabolites in plasma and whole blood by gas chromatography—mass fragmentography

A method for the simultaneous measurement of dothiepin and two of its major metabolites, northiaden and dothiepin S-oxide in both plasma and whole blood is described. The method involves the use of gas chromatography—mass fragmentography. It is selective, sensitive (1 μg/l) and reproducible.It has been used to analyse both plasma and blood samples following single oral doses of 75 mg dothiepin in seven volunteers.