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.
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.
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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