Experimental setup of diffraction phase microscope (DPM) with double low-coherence lighting system is presented in the paper. Algorithm of interference picture processing and optical thickness, height, volume and mean cells volume (MCV) of RBC calculating is shown. We demonstrate results of experiments with blood smears and ability of the method to calculate 3D model of the biological cells shape. Investigation change dynamics of RBC morphology after injection glucose for diabetes by DPM is shown in the paper.
In our research we brought out the age difference in the group of long-livers and the continuous character of the
biochemical basal metabolism indexes changing. The results allowed us to carry out the polynominal high-powered
approximation to study the dynamics of laboratory indexes. We revealed the progressive reduction of the cholesterol,
triglycerides, glucose and creatinine levels starting from 90 years of age, and this reduction showed the non-linear
character with interchange of local minimums and maximums. During the speed characteristics analysis we revealed the
cooccurrence of the speed maximums of all the examined biochemical indexes, except the speed of changing the
concentration of cholesterol, which maximum took the lead over the other indexes by four years. The phase-plane
portrait analysis of the regulatory systems on the plane "time - speed" showed the unfulfilled attempt of system
stabilization by all the searched parameters nearby the special spot - "stable focus". The standard deviation values
analysis of the researched parameters showed their progressive reduction in the long-livers. That fact can be considered
as the regulatory systems physiological "backlash" reduction among the centenarians.
Blood flow structure was studied by PC integrated video camera with following slide by slide analysis. Volumetric blood
flow velocity was supporting on constant level (1 ml/h). Silicone tube of diameter comparable with coronary arteries
diameter was used as vessel model. Cell-cell interactions were studied under glucose and anticoagulants influence.
Increased adhesiveness of blood cells to tube walls was revealed in patient with coronary heart disease (CHD) compare
to practically healthy persons (PHP). In patients with stable angina pectoris of high functional class and patients with
AMI shear stress resistant erythrocyte aggregates were predominating in blood flow structure up to microclots formation.
Clotting and erythrocytes aggregation increase as response to glucose solution injection, sharply defined in patients with
CHD. Heparin injection (10 000 ED) increased linear blood flow velocity both in patients with CHD and PHP.
After compare our results with other author's data we can consider that method used in our study is sensible enough to
investigate blood flow structure violations in patients with CHD and PHP. Several differences of cell-cell interaction in
flow under glucose and anticoagulant influence were found out in patients with CHD and PHP.
Blood plasma pattern formation is a process sensitive to environment and carrier properties, and plasma biochemical
content. 96 patients with coronary heart disease (CHD) were involved in the study. Control group include 12 practically
health persons (PHP). Platelets poor plasma and serum were used to study functional morphology. Plasma and serum
samples of equal volume were placed on degreased glass carrier with after going wedge dehydration. The result of wedge
dehydration is a formation of a special structure called facia. To the samples of compare albumin solution was added.
Morphology of prepared facies was studied by means of light microscopy ("Lomo Biolam P2-1") with 10 times
magnification.
All received facies were of the same principle structure with central, intermediate and edge zones. Zone index was
increasing in samples with albumin adding. Special structures obligatory to atherosclerosis, vessels stiffness increase and
hypoxia were found in facies of plasma and serum of patients with CHD. Quantity of these structures correlated to
protein concentration (p = 0.021). Samples' drying period was also increasing in samples of compare, and differed
significantly in patients with CHD and PHP. In our study gross proteins concentration increase modified plasma and
serum morphology. Albumin solution can be proposed as a probe to elucidate differences of facies of patients with CHD
and PHP.
Immune state monitoring is an expensive, invasive and sometimes difficult necessity in patients with different disorders.
Immune reaction dynamics study in patients with coronary atherosclerosis provides one of the leading components to
complication development, clinical course prognosis and treatment and rehabilitation tactics.
We've chosen intravenous glucose injection as metabolic irritant in the following four groups of patients: men with
proved coronary atherosclerosis (CA), non insulin dependent diabetes mellitus (NIDDM), men hereditary burden by CA
and NIDDM and practically healthy persons with longlivers in generation. Immune state parameters such as quantity of
leukocytes and lymphocytes, circulating immune complexes levels, serum immunoglobulin levels, HLA antigen
markers were studied at 0, 30 and 60 minutes during glucose loading. To obtain continues time function of studied
parameters received data were approximated by polynomials of high degree with after going first derivatives.
Time functions analyze elucidate principally different dynamics studied parameters in all chosen groups of patients,
which couldn't be obtained from discontinuous data compare. Leukocyte and lymphocyte levels dynamics correlated
HLA antigen markers in all studied groups. Analytical estimation of immune state in patients with coronary
atherosclerosis shows the functional "margin of safety" of immune system state under glucose disturbance. Proposed
method of analytical estimation also can be used in immune system monitoring in other groups of patients.
Dynamics of glucose concentration in human organism is an important diagnostic characteristic for it's parameters
correlate significantly with the severity of metabolic, vessel and perfusion disorders.
36 patients with stable angina pectoris of II and III functional classes were involved in this study. All of them were men
in age range of 45-59 years old. 7 patients hospitalized with acute myocardial infarction (aged from 49 to 59 years old)
form the group of compare. Control group (n = 5) was of practically healthy men in comparable age. To all patients
intravenous glucose solution (40%) in standard loading dose was injected. Capillary and vein blood samples were
withdrawn before, and 5, 60, 120, 180 and 240 minutes after glucose load. At these time points blood pressure and
glucose concentration were measured. In prepared blood smears shape, deformability and sizes of erythrocytes, quantity
and degree of shear stress resistant erythrocyte aggregates were studied. Received data were approximated by
polynomial of high degree to receive concentration function of studied parameters, which first derivative elucidate
velocity characteristics of morphofunctional erythrocyte properties during intravenous glucose injection in patients with
coronary heart disease and practically healthy persons.
Received data show principle differences in dynamics of morphofunctional erythrocyte properties during intravenous
glucose injection in patients with coronary heart disease as a possible mechanism of coronary blood flow
destabilization.
Blood cell-cell and cell-vessel wall interactions are one of the key patterns in blood and vascular pathophysiology. We
have chosen the method of reconstruction of pulsative blood flow in vitro in the experimental set. Blood flow structure
was studied by PC integrated video camera with following slide by slide analysis. Studied flow was of constant
volumetric blood flow velocity (1 ml/h). Diameter of tube in use was comparable with coronary arteries diameter.
Glucose solution and unfractured heparin were used as the nonspecial irritants of studied flow.
Erythrocytes space structure in flow differs in all groups of patients in our study (men with stable angina pectoris (SAP),
myocardial infarction (MI) and practically healthy men (PHM). Intensity of erythrocytes aggregate formation was
maximal in patients with SAP, but time of their "construction/deconstruction" at glucose injection was minimal.
Phenomena of primary clotting formation in patients with SAP of high function class was reconstructed under
experimental conditions. Heparin injection (10 000 ED) increased linear blood flow velocity both in patients with SAP,
MI and PHP but modulated the cell profile in the flow.
Received data correspond with results of animal model studies and noninvasive blood flow studies in human. Results of
our study reveal differences in blood flow structure in patients with coronary heart disease and PHP under irritating
conditions as the possible framework of metabolic model of coronary blood flow destabilization.
Erythrocytes system study can provide a framework for detailed exploration of blood cell-cell and cell-vessel wall
interactions, one of the key patterns in blood and vascular pathophysiology. Our objective was to explore erythrocytes
system in patients with stable angina pectoris II f.c. (Canadian classification). The participants (N = 56, age 40 - 55 years) without obesity, glucose tolerance violations, lipid lowering drugs treating,
heart failure of II and more functional classes (NYHA), coronary episode at least 6 months before study were involved in
the study. Blood samples were incubated with glucose solutions of increasing concentrations (from 2.5% to 20% with 2.5% step) during 60 mm (36° C). In prepared blood smears erythrocyte's sizes were studied. Plasma total cholesterol, triglyceride and glucose levels were also measured. Received data were approximated by polynomials of high degree, with after going first and second derivations. Erythrocytes system "behavior" was studied by means of phase pattern
constructing. By lipids levels all the patient were divided into five groups: 1) patients with normal lipids levels, 2) patients with borderline
total cholesterol level, 3) patients with isolated hypercholesterolemia, 4) patients with isolated hypertriglyceridemia and
5) patients with combined hyperlipidemia. Erythrocytes size lowering process was of set of "stages", which characteristics
differ significantly (p > 0.05) in all five groups. Their rate and acceleration characteristics allow us to detect type of lipid
profile in patients. Erythrocyte system disturbing by glucose concentration increase show to be most resistant in group of
patients with isolated hypercholesterolemia.
Blood plasma can be considered as a special water system with self-organization possibilities. Plasma slides as the results of wedge dehydration reflect its stereochemical interaction and their study can be used in diagnostic
processes.
46 patients with coronary heart disease were studied. The main group was formed of men in age ranged from 54 to
72 years old with stable angina pectoris of II and III functional class (by Canadian classification) (n=25). The group
of compare was of those who was hospitalized with diagnosis of acute coronary syndrome, men in age range 40-82.
Clinical examination, basic biochemical tests and functional plasma morphology characteristics were studied.
A number of qualitative and quantitative differences of blood plasma morphology of patients with chronic and acute
coronary disease forms was revealed.
The protocol of our study includes men with acute myocardial infarction, stable angina pectoris of II and III functional classes and unstable angina pectoris. Patients with arterial hypertension, disorders in carbohydrate metabolism were excluded from the study. Blood samples taken under standardized conditions, were stabilized with citrate sodium 3,8% (1:9). Erythrocytes and platelets aggregation activity under glucose influence (in vitro) was studied by means of computer aided microphotometer -- a visual analyzer. Erythrocyte and platelets were united in special subsystem of whole blood. Temporal and functional characteristics of their aggregation were analyzed by creation of phase patterns fragments. The received data testify to interrelation of erythrocytes and platelets processes of aggregation under conditions of increasing of glucose concentration of the incubatory environment, which temporal and functional characteristics may be used for diagnostics and the prognosis of destabilization coronary blood flow at an acute coronary syndrome.
The aggregation ability of erythrocytes of patients with coronary heart disease comparing to practically healthy persons and patients with coronary heart disease combined with non insulin dependent diabetes mellitus depending on different glucose concentration in unguentums of blood incubates with the help of computer microphotometer - visual analyzer was studied. Two-phase behavior of erythrocytes size changing of practically healthy persons depending on glucose concentration in an incubation medium and instability erythrocyte systems of a whole blood to the influence of high glucose concentration were revealed. Influence of high glucose concentration on aggregation ability of erythrocytes of patients with coronary heart disease and its combination with non insulin dependent diabetes mellitus was revealed.
The human blood sedimentation peculiarities caused by chemical agents adding or coronary heart disease of a patient was studied by two coherent-domain optical techniques. These are OCT and spatially-modulated laser beam transillumination technique. The OCT method due to its high sensitivity was used to study the sedimentation of a whole and less diluted blood, i.e. the sedimentation of the aggregated blood. The spatially-modulated method was used to study a highly diluted blood, i.e. sedimentation of individual and weakly-interacting erythrocytes and was tested in clinical research.
The application of laser measuring system with spatially- modulated probing laser beam for study of properties of thin and bulk scattering media is considered. The diluted human blood of the man was used as the scattering medium. Time- dependent dynamics of scattering properties of blood suspensions is investigated in the course of spontaneous sedimentation and aggregation of erythrocytes.
The intravenous glucose tolerance test was performed to estimate the kinetics of blood glucose and insulin levels. Glucose was injected in individual standardized dose (0.5 g. per 1 kg of body weight). Three groups of patients were checked up: 1) patients with coronary heart disease verified by cicatricial alterations in myocardium found by electrocardiographic and echocardiographic methods; 2) children of patients with transmural myocardial infarction practically healthy at the moment of study; 3) persons practically healthy at the moment of study without any indications on cardiovascular diseases and non-insulin dependent diabetes mellitus among all ancestors and relatives who frequently were long-livers. Last groups didn't differ by age and sex. Peripheral blood glucose level, immunoreactive and free insulin (tested by muscular tissue) were studied just before glucose injection (on an empty stomach) and 4 times after it. The received discrete data were approximated by high degree polynomials, the estimation of blood glucose and insulin time functions symmetric was performed. The deceleration of degradation of insulin circulating in peripheral blood and the time decrease of second phase of insulin secretion were analytically established. This fact proves the complicated mechanism of insulin alterations in atherosclerosis, consisting not only of insulin resistance of peripheral tissues but of decrease of plastic processes in insulin- generating cells.
On the basis of experimental data obtained at clinical examination of 5 specially fitted groups of the patients the analytical system of early diagnostic of metabolic condition appropriate for an atherosclerosis and non-insulin dependent diabetes mellitus was designed.
The diagnostic algorithm of glucose-insulinic violations for the patients with a clinically obvious atherosclerosis of coronary arteries, non-insulin dependent diabetes mellitus and persons with the heritable predisposition to these forms of pathology was designed. The realization of intravenous glucose tolerance test in specially fitted groups of patients served as basis of the algorithm.
The time functions of plasma glucose and insulin obtained during intravenous glucose tolerance test were approximated by sections of Fourier series. The convincing quantitative and quality distinctions of amplitudes both phases of the first and second harmonics of decomposition of the indicated time functions are obtained. These distinctions were used as a basis of diagnostic algorithm of metabolic violations appropriate for atherosclerosis and non-insulin dependent diabetes mellitus in clinically obvious and preclinical stages.
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