Simultaneous temporal analysis of whole human or rat blood luminescence and erythrocytes sedimentation rate (ESR) in same blood using special computerized optoelectronic devices for single photon counting and for high temporal resolution of the rate of sedimentation of red blood/plasma boundary revealed correlation between both time series. Correlation was observed in vitro in normal blood, after action of physical (height of blood column) and of chemical (hydrogen peroxide) factors, and in experimental cerebral ischemia. An ischemia was invoked in rats by occlusion of both common carotid arteries. ESR was studied with the device "ESR-scan" and the dynamics of respiratory burst (RB) by a luminol-dependent luminescence method on the same blood samples. There was a noticeable increase of intensity of RB in whole rat blood and significant acceleration of ESR in blood diluted on 50% in 90 minutes after applying a ligature on carotid arteries. The individual differences between animals attesting to different degree of RB and ESR
activation in blood both in intact animals and after operational intervention was obtained. Revealed correlation points to
considerable relation between blood energy and its mechanical properties.
An automatic device for high-temporal resolution of the process of red blood sedimentation was designed. The position of the boundary between red blood and plasma may be registered each 30 sec in several pipettes simultaneously with +/- 10 mkm precision. Fractional rates of the boundary movement are deduced with high accuracy. Data are processed by a PC and presented as velocity-time curves and the curves describing time evolution of the boundary position. The results obtained with this equipment demonstrate non- monotonous character of the rate of red blood sedimentation and revealed several peculiarities of macroscopic stages and microkinetic of the process. Changes of biophysical-chemical state of blood related to changes in physiological state of a donor or under the action of different substances may result either in improving, or in worsening of studied parameters of blood sedimentation dynamics. A slight dilution of blood with saline in vitro leads as a rule to decreasing of sedimentation rate and improving of microkinetic parameters of the process of the red blood sedimentation. Adding of highly diluted hydrogen peroxide to blood samples of patients provides for the oxygenation of the system, and result in the improving of sedimentation dynamics: decreasing of average sedimentation rate and prolonging of macrokinetic states. The parameters of this reaction depend upon the metabolic potential of blood, the functional state of all its cellular and plasma elements, and in complex they reflect the physiological state of the individual Any changes in parameters of macro- and microkinetics may reflect the pattern of oxygen release- delivery-consumption in blood. ESR-graphy may give much more information in comparison to the routine ESR-test and allows to consider blood sedimentation not from the viewpoint of separate physico-chemical parameters, but treating it as a metabolically active system.
An automatic device for high-temporal resolution of the process of erythrocytes sedimentation in blood was designed. The position of the boundary between red blood and plasma is registered each 30 sec in several pipettes simultaneously with +/- 10 mkm precision. Data are processed by a PC and presented as velocity-time curves (ESR-grams) and the curves describing time evolution of the boundary position. ESR-grams demonstrate non-monotonous character of erythrocytes sedimentation in blood. Blood of particular donor being in a stable physiological state taken on different days is characterized by similar ESR-grams. Pathological deviations from a normal physiological state are reflected in the shortening of duration of each process stage and increasing of average sedimentation rate. Intravenous infusion of some medical preparations may lead either to improving (prolonging of macrokinetic stages, decreasing of sedimentation rate), or to worsening of studied parameters depending on an individual. The low extent of blood dilution with saline in vitro lead as a rule to decreasing of sedimentation rate and improving of microkinetic parameters of the process. Adding of highly diluted hydrogen peroxide to blood samples of patients resulted in the improving of sedimentation kinetics. ESR-graphy may widen opportunities of practical medicine in diagnostics, prognostics and drug therapy.
The sedimentation properties of blood of 13 ischemic heart disease patients and 2 healthy volunteers have been analyzed using a special computerized optical device for high temporal resolution tracing of red blood/plasma boundary movement rate (ESR-graphy). The kinetic curves of red blood sedimentation are substantially nonmonotonic and exhibit multiple accelerations, decelerations and even backwards movement of the red blood/plasma boundary. The intensity of blood sedimentation rate oscillations is significantly higher in the blood of patients and voluteers on days of enhanced geomagnetic activity than on quiet days. In healthy donors, blood oscillations were also observed on active geomagnetic days, however, their intensity was lower, the sedimentation rate started to oscillate after a longer time upon pipette installation, and the oscillation frequency was lower than in the patients' blood. Thus, blood is highly responsive to changes in geomagnetic field activity. Possibly oscillatory behavior mechanism of blood sedimentation rate and the diagnostic and prognostic merits of the ESR graphs are discussed.
An automatic device for high-temporal resolution of the process of red blood sedimentation was designed. The position of the boundary between red blood and plasma may be registered each 30 sec in several pipettes simultaneously with +/- 10 mkm precision. Fractional rates of the boundary movement are deduced with high accuracy. Data are processed by a PC and presented as velocity-time curves (ESR-grams) and the curves describing time evolution of the boundary position. Several unexpected phenomena in the process of red blood sedimentation have been revealed. Upward fast movements of the boundary up to 1 mm were noted. In patients' blood sets of 5 - 10 milliHz oscillations of sedimentation rate were usually developing at early stages of blood sedimentation. In non-diluted healthy donors' blood high amplitude periodic oscillations were either absent, or were emerging only after blood resided in pipettes for several hours. When blood was diluted to a certain degree with physiological saline or with own plasma long-term low frequency (1 - 3 milliHz) rate oscillations regularly appeared. Non-trivial dependence of patterns of ESR-grams on diluting of blood with own plasma or saline was observed. Thus, non-linear dynamic behavior of living blood has been revealed due to application of the principles of the system of technical vision for the detailed analysis of red blood sedimentation kinetics.
We have demonstrated recently that luminol- or lucigenin- dependent chemiluminescence (LM-CL and LC-CL, respectively) registered from whole non-diluted blood may reach rather high intensities. Here we demonstrate, that parameters of both LM-CL and LC-CL in non-diluted human blood during RB development is influenced by a reflective screen of aluminum foil surrounding a test tube with a blood sample. Photons reflected back to a sample accelerate slowly developing and retard rapidly development RB. As demonstrated by histochemical NBT test, wrapping samples with blood in aluminum foil effected also the reductive neutrophil activity in. These results indicate that weak light fluxes generated in the course of RB may influence physiological processes in blood. They also provide the basis for an explanation for the phenomenon of the interaction two blood samples in one of which RB in the presence of luminol was stimulated through the optical channel previously reported by us.
Nonenzymatic glycation of free or peptide bound amino acids (Maillard reaction, MR) plays an important role in aging, diabetic complications and atherosclerosis. MR taking place at high temperatures is accompanied by chemiluminescence (CL). Here kinetics of CL development in MR proceeding in model systems at room temperature has been analyzed for the first time. Brief heating of glycine and D-glucose solutions to t greater than 93 degrees Celsius results in their browning and appearance of fluorescencent properties. Developed In solutions rapidly cooled down to 20 degrees Celsius a wave of CL. It reached maximum intensity around 40 min after the reaction mixture heating and cooling it down. CL intensity elevation was accompanied by certain decoloration of the solution. Appearance of light absorbing substances and development of CL depended critically upon the temperature of preincubation (greater than or equal to 93 degrees Celsius), initial pH (greater than or equal to 11,2), sample volume (greater than or equal to 0.5 ml) and reagents concentrations. Dependence of total counts accumulation on a system volume over the critical volume was non-monotonous. After reaching maximum values CL began to decline, though only small part of glucose and glycin had been consumed. Brief heating of such solutions to the critical temperature resulted in emergence of a new CL wave. This procedure could be repeated in one and the same reaction system for several times. Whole CL kinetic curve best fitted to lognormal distribution. Macrokinetic properties of the process are characteristic of chain reactions with delayed branching. Results imply also, that self-organization occurs in this system, and that the course of the process strongly depends upon boundary conditions and periodic interference in its course.
Measurement of erythrocyte sedimentation rate (ESR) is one of the most widely used diagnosis techniques. It is believed that high ESR testifies to inflammatory conditions or decay in body tissues. However, standard ESR test data do not allow definitive determination of such conditions. ESR is currently determined manually. The ambiguity of standard ESR tests that have found general use in medicine raises the necessity of increasing their information merit without serious modification to the test method. The present device and method allow completely automatic hourly ESR measurement and determine several parameters of blood sedimentation dynamics (BSD). Analysis of BSD introduces a number of parameters that allow pathology to be detected in case of normal standard ESR test data and allow more efficient planning of medical treatment than is available with hourly ESR data. The results were used to elaborate and produce a pilot device for automatic clinical ESR testing to obtain BSD data. The device and BSD method are being clinically tested at the Intensive Care Unit of the Moscow Central Railway Hospital. All the technical solutions have been patented. BSD can be applied as a new powerful diagnostic tool, once large statistics has been obtained.
Comparison of lucigenin- and luminol-dependent chemiluminescence (LC-CL and LM-CL, respectively) in nondiluted healthy donors' blood revealed significant differences in their patterns. LM-CL was low in fresh blood and disappeared after it storage for 3 hours. LC-CL was already high in fresh blood and was steadily increasing with blood storage. Serial dilution of blood with saline after addition of chemiluminescence indicators resulted in elevation of LM-CL, but decrease in LC-CL. LM-CL elevation after the initiation of respiratory burst (RB) in blood with zymosan was observed only in aerated samples and immediately dropped down when air supply to a blood sample was ceased. On the contrary, LM-CL did not depend on air supply to a blood sample for about 30 min. after RB initiation. The results suggest that there are at least two mechanisms for reactive oxygen species production in nondiluted blood. The first one is reflected predominantly by LM-CL. It is activated during RB and uses prevalently oxygen dissolved in cell medium. Another one is reflected predominantly by LC- LM. It does not depend upon initiation of RB in neutrophils, operates in blood constantly, and uses oxygen supplied by erythrocytes. It needs blood integrity for its manifestation.
Addition of Luminol to nondiluted blood of healthy donors results in a short and weak increase of chemiluminescence (CL) from it. Contrary to that in 25 cases of stable angina pectoris the intensity of CL from blood of patients sharply increased upon addition of luminol exceeding that form healthy donors' blood 10-100-fold. 24 hours after the 3D intravenous low-level treatment CL burst in patients' blood in the presence of Luminol was in general significantly lower than before the beginning of the treatment. After the 7th treatment the pattern of CL kinetics was in most cases similar to that of healthy donors' blood. However, after the 10th treatment intensity of Luminol-enhanced CL usually increased and for blood of some patients even exceeded its values obtained before the treatment. Some correlation CL from nondiluted blood with neutrophil activity studied by NTB-test and plasma viscosity of same blood was noted. Using highly sensitive single photon counters it is possible to reveal abnormal levels of CL from no more than 0.1-0.2 ml of blood within 3-5 min.
To study a possibility of interaction of two optically, but not chemically coupled samples of whole human blood the following experimental setup was used. A quartz cuvette with either nondiluted blood or saline was placed inside a glass vial. Saline diluted whole blood was poured into the vial and respiratory burst (RB) was initiated in it with phorbol ether or zymosan. Luminol-dependent chemiluminescence (LCL) was registered using liquid scintillation counter (coincident circuit off). Effect of blood placed in the cuvette upon photon emission from blood placed in the vial was evaluated. It was shown that blood of some donors consistently attenuated photon emission from the sample in which RB was induced. Blood of another group of donors enhanced photon emission from the `partner' sample. Some properties of blood taken from the cuvette after being in the contact with the sample in which RB was induced changed in comparison with the same blood that was contacting with the non-stimulated sample. Exposed blood has lost the ability to attenuate light emission from the fresh portion of blood in which RB was induced. Its own LCL in response to addition of zymoscan was different from that of the parallel sample of same blood not exposed to sample undergoing RB. These results suggest that two chemically separated but optically coupled samples of blood can interact.
Parameters of chemiluminescence (CL) from nondiluted human blood were studied. Kinetics and intensity of CL depended upon donor's state of health, time after blood extravasion, conditions of its storage, conditions of its counting. Peculiar dependence of Luminol-enhanced CL on sample volume changes during respiratory burst (RB) was revealed. When 0.5 ml aliquots were consecutively taken from blood and transferred into another vial of the same configuration, each subtraction of blood was followed by an acceleration of CL intensity growth. Summation of portions of blood in the second vial resulted in deceleration of CL intensity increase from it. At equal volumes of blood CL intensity from the first sample was manifold higher than from the second one and this difference was increasing on with further transfers. When blood was transferred back to the first sample, CL intensity from the 'donor' sample began to increase at a faster rate, while CL intensity from the 'recipient' sample stabilized. Such behavior was characteristic of nondiluted healthy donors' blood. Diluted blood or blood of sick people demonstrated different behavior. It is suggested that CL parameters of nondiluted blood may be informative of integrative properties of blood tissue.
Possibility of interaction between the two optically coupled samples of whole human blood as studied. A quartz cuvette was placed inside a vial for a liquid scintillation counter. Saline diluted whole blood was poured into the vial and either undiluted blood or saline was poured into the cuvette. Respiratory burst (RB) was initiated in blood placed into the vial, and luminol-dependent chemiluminescence (LCL) was registered. Blood placed in the cuvette affected photon emission from blood placed in the vial. Blood of another group of donors enhanced photon emission from the 'partner' sample. Some properties of blood taken from the cuvette after being in the contact with the sample in which RB was induced changed in comparison with the same blood that was contacting with the non-stimulated sample. Exposed blood has lost the ability to attenuate light emission from the fresh portion of blood in which RB was induced. Besides its own LCL in response to addition of zymosan was different from that of the control These results suggest that two chemically separated but optically coupled samples of blood can interact.
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