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2. The objects and methods of investigation
3. The exemination of uv-spectra of the dogs’& rats’ blood plasma absorption in the process of the peritonitis progressing
5. The investigation of the absorption spectra of the dogs’ whole blood in the process of the peritonitis progressing in the vis
The Optical Method of the Investigation of the Peritonitis Progressing Process.
S. H. Guminetskiyа, O. G. Ushenkoа, I. P. Polyanskiyв, A. V. Motrychа,
F. V. Grynchukв
Chernivtsi National University named after Yu. Fed’kovychа
Bukovinian State Medical Universityв
There have been given the results of the spectrophotometric examination of the dogs’ and rats’ venous and whole blood plasma taken in the process of the peritonitis progressing within the spectral interval nm (for plasma) and nm (for the whole blood). It has been defined that D-optical density values in the field of the long-waved maximum of plasma absorption intensity of the venous blood at nm depend upon the intensity of the inflammatory process and also upon the circumstances against the background of which it started to progress. It was found out that the dynamics of D- values changes for (or 570) nm in the process of the peritonitis progressing in case of the whole blood taken from a portal vein is a mirror symmetrical if to compare to the same dynamics for the blood from cava inferior. The defined conformities with regularities may have a diagnostic meaning. It was also found out that the biggest influence upon the dynamics of D-values at = 280nm of the venous blood plasma has the content of the circulating immune complexes, necrosis factor of α –tumors and interleukin – 2, the changes of which explain for almost on 100% the distribution of the optical density parameters and what proves a possible immunologic explanation of its changes.
^ : absorption spectrum, blood plasma, whole blood, peritonitis, optical density.
The laboratory clinical diagnostics of the acute surgical diseases of the abdominal cavity organs has more been improved for the last years. Herewith both the traditional biochemical and the new spectrophotometric methods are being used with the purpose to find simple and objective criteria, which would give an opportunity to evaluate the effectiveness of the medical treatment process. Mainly it has been defined in  , that D-optical density values of a human venous blood plasma at nm could be used as an optical testing for the express-diagnostics of the acute inflammatory destructive diseases of the abdominal cavity organs such as appendicitis, cholecystitis, ileus, pancreatitis, perforating gastroduodenal ulcer. In  we have the results of the dynamics investigation of the optical properties changes of the venous blood plasma in the UV-spectrum field taken from patients with the same pathology of the abdominal cavity organs within the postoperative period of the medical treatment process which gave an opportunity to determine the objective criteria of the evaluation of the medical treatment process effectiveness in the postoperative period. However, in the mentioned papers the blood plasma has not been examined in case of such a critical disease as peritonitis. Besides, the background of the defined changes of the absorption spectra has not been examined yet.
That’s why the purpose of writing this paper is in the investigating the dynamics of the optical properties changes of the venous and whole blood plasma in the process of the peritonitis progressing and also in those cases of its appearance against the background of by-diseases such as diabetes mellitus as well as the acute renal and hepatic insufficiency, which up to the data [3, 4] might change essentially the clinical and biochemical factors and decrease their diagnostic value, also in searching the possible factors which cause the changing of the optical properties.
10 inbred dogs and 75 white non-linear rats of both sexes became an object of the investigation. They were affected up to the method  by the stomach perforation what caused the appearance of the peritonitis. The acute renal and hepatic insufficiency was obtained up to the method  and the diabetes mellitus up to . The venous blood samples for the testing were taken before the peritonitis modeling start up and in some periods of time after its initiating mainly within 6 hours for the rats and in 1 hour for the dogs. The blood samples were taken from the rats’peripheric vein and from dogs’ portal vein and the cava inferior. The rats were separated into 3 groups, by 25 in each: the intact ones with a modeled in advance renal and hepatic insufficiency and diabetes mellitus which affected in such a way that it caused the appearance of the peritonitis.
UV-spectra of blood plasma absorption were examined as well as the absorption spectra of the whole blood in the visible area, also the content of the circulating immune complexes and cytokins: the interleukins 2, 6, 10 and the necrosis factor of α – tumors.
The experimental research has been done at the Surgery Department & Ophthalmic Diseases of Chernivtsi State Medical University.The blood and blood plasma patterns have been examined within 3 days, right after they were taken-not later. They were kept at the t -70° С in the refrigerator before the experimental testing started.
The studying of UV-spectra of the blood plasma and the whole blood absorption in the visible area was carried out with the help of the sphere photometer in accordance with the methods descried in  at the Optics and Spectroscopy Section of the Engineering Department at Chernivtsi National University. The content of the cytokins and the circulating immune complexes were examined on the base of the automatic immunofermental analyzer by means of “Biosuorce” Belgian reagents in accordance with the current instructions.
The research data was obtained on the grounds of the experiments done with the dogs and rats because such kind of testing on humans in case of a critical inflammatory disease is not possible to be carried out.
The blood plasma absorption spectra were measured within a spectral interval nm, but as the area adjoining nm  is an informative one, then we have to analyze the results received within the rangenm. Thus, for example, in fig.1 we have the spectra of one of the dog’s venous blood plasma absorption for the cava inferior in dependence upon the peritonitis passing time, but we see the analogous spectra for the rat’s blood plasma in the fig. 2. The experimental plasma under the testing was diluted with a distilled water in the relation of 1:100.
It is vivid from the above figures that the range of changes of the plasma D-optical density in the process of the peritonitis progressing in case of the experiment with a dog is bigger within the whole spectrum than in case with a rat. Besides, we also see that the biggest D-changes cover nm, i.e. it’s in fact the most informative point. That’s why in the table 1 for the dogs and in the table 2 for the rats the mean values of D-optical density are given in dependence upon the peritonitis progressing time for all tested dogs and all tested rats at nm. Moreover, the diagrams of D-changes at nm for the blood plasma of three groups of the tested rats are given in the fig. 3.
From the given in the tables data and from the fig.3 we may come to a conclusion that the dogs’ blood plasma in case of the peritonitis progressing D-mean values based on 10 tested objects in the first three hours decrease both for the portal vein and the cava inferior. Starting from the forth hour this regularity has being essentially changed and is a different one for the portal vein and cava inferior. This is proved by a different content of the definite substances which cause the changing of the optical density in the blood of the tested veins especially after the sixth hour from the peritonitis initiating point, when the difference in D-values for the portal vein and cava inferior blood plasma becomes a real big one. Herewith, D-values for the portal vein in all cases is, as usual, bigger than the latter for the cava inferior.
Fig.1. Spectra of the dog’s blood plasma absorption from the cava inferior in dependence upon the peritonitis passing time: 1 – before start up (t = 0), 2 – after two hours, 3 – after four hours, 4 – after six hours.
Fig.2. Spectra of the rat’s blood plasma absorption in dependence upon the peritonitis passing time: 1 – before start up (t = 0), 2 – after six hours, 3 – after twelve hours, 4 – after twenty four hours.
Table 1. D-mean values of the dogs’ venous blood plasma.
Table 2. D-mean values of the rats’ venous blood plasma.
Fig.3 Diagrams of D-mean values changes at nm of the rats’ blood plasma in the process of the peritonitis progressing (1); and of the peritonitis on the background of a toxic affection (2), and of the peritonitis on the background of the diabetes mellitus (3).
- For the rats’ blood plasma in case of the peritonitis progressing being modeled on the base of the renal and hepatic insufficiency D-mean values on five tested objects are usually smaller than for the simple form of the peritonitis. And on the contrary, in case of the peritonitis initiated on the base of the diabetes mellitus D-values are considerably bigger. Herewith in all cases after the sixth hour from the peritonitis initiating in D-values the minimum has being observed.
- For the rats’ blood plasma D-values in all cases are considerably bigger than for the dogs, and they are similar to as for the humans ; the regularities of their changes in the process of the peritonitis progressing are also different .
The content of the examined cytokins and the immune complexes reflects the functional activities of the different components of the immune system . It has been defined while comparing the dynamics of the optical density to the examined indexes that the biggest connection that took place covered the changing of the interleukin-2 level (fig.4). After using the factor analysis by the method of the predominant components we found out that the biggest influence upon the optical density had the content of the circulating immune complexes, necrosis factor of α-tumors and interleukin–2, the changes of which interpret for almost on 100% the changing of the optical density parameters. Taking into consideration that the interleukin’s-2 producer is mainly represented by the activated T-helpers and one of its main functions is the stimulation of B-immunoglobulins secretion by the lymphocytes , what leads towards the enlargement of the circulating immune complexes content. And such case is possible that the variation of the optical density indexes depicts the process of the antibodies formation, which are globulins by their structure  in response to the abdominal peritoneum destruction caused by the peritonitis. This is also proved by the following:
- The defined indexes of the optical density of the blood plasma have specific distinctions that proves that there are definite species of the substrate structure which causes the changing.
- The optical density of the dogs’ blood plasma taken from the portal vein in 3 hours from the point of the peritonitis initiating overbalances the blood plasma taken from its cava inferior. It means that the substrate is concentrated predominantly in the portal vein, which picks blood from the abdominal peritoneum organs  and from the abdominal cavity itself.
- The patients with the acute surgical pathology of the abdominal cavity organs with the absence of the destructive changes of the affected organ and, consequently, the abdominal peritoneum, the optical density indexes do not differ from those defined of healthy donors .
Fig.4 The diagrams of the mean values 1 – of D-optical density at λ = 280 nm, 2- interleukin content (pkg/ml) in the blood plasma of the intact rats during the process of the peritonitis progressing.
There have been examined the absorption spectra of ten dogs’ whole blood diluted by a distilled water in the relation of 1:150 in the spectral interval of λ=350-620nm. It is known  that the absorption spectra of the whole blood in the visible area are determined by the appearance of the absorption spectra of its forming components: erythrocytes, thrombocytes and leukocytes, which in the area of nm do not practically absorb , and the blood plasma, as it is seen from the point 3, does not absorb in the area of nm. This caused us to choose such a spectral range for our investigation. For example, in the fig.5 the indicated spectra are given in dependence upon the time of the peritonitis progressing for the cava inferior of one of the tested dogs. They are analogous for the portal vein. It is seen that in the absorption spectra there are two narrow enough maximums in the range of nm and nm and a very strong strip of absorption in the range ofnm, the so called Soret strip . In order to obtain the chart of the absorption spectrum in this strip it’s necessary to have the whole blood to be diluted up to the relation of 1:500 which is not convenient while at testing. That’s why in order to evaluate the influence of the peritonitis over the absorption spectra of the whole blood it is sufficiently enough to analyze their changes in the field of the definite wave lengths nm and nm. They are peculiar as they are conditioned mainly by the hemoglobin absorption in erythrocytes [8, 11, 12].
Fig.5. Absorption spectra of the whole blood from the cava inferior: 1 – checking, 2 – after two hours. 3 – after four hours from the peritonitis initiating.
The results of ten tested dogs’ blood analysis in the way of D- mean values are shown on the diagram in fig.6 for nm. It is seen that on the peritonitis progressing periods including even that one before its starting (t=0), D-value for the cava inferior is considerably lower than for the portal vein, which is analogously as to UV-spectra of the dogs’ blood plasma (tabl.1).
Fig.6. Dynamics of D-values changing of the optical density for nm at peritonitis: 1 – cava inferior; 2 – portal vein.
This phenomenon could be explained by the differences in the blood components with various substances content of the cava inferior and the portal vein that are formed in the tissues of the abdominal cavity  and might serve as a substrate that causes the changing of the optical characteristics of blood. The dynamics of their changes in the process of the peritonitis progressing for both veins is a mirror opposite relation (fig. 6), what indicates on the regular differences of the blood components conditioned by the developing of the inflammatory and destructive process in the abdominal cavity. The results for = 570 nm are totally analogous.
It appears from the above investigations done that:
1. It has been proved for the first time that the dynamics of changing of the optical density of the venous blood plasma in the range of = 280 nm and for the whole blood at = 540nm (or = 570nm) has specific and location differences.
2. The dynamics of D-values changing at = 280nm in the process of the peritonitis progressing and also for the peritonitis that was initiated against the background of the initiated in advance diseases is practically identical.
3. The content of the circulating immune complexes, necrosis factor of α-tumors and interleukin–2 has the strongest influence upon the dynamics of D-values at = 280nm of the venous blood plasma, the changes of which explain for almost on 100% the changing of the optical density parameters.
1. Pat.77081, Ukraine, І.Yu. Polyanskiy, B.М. Nitsovych, S.G. Guminetskiy, F.V. Grynchuk, S. P. Brodovskiy, Method of Spectrophotometric Diagnostics of the Inflammatory Destructive Diseases in the Abdominal Peritoneum, Bulletin №4, 2004.
2. S.G. Guminestskiy, I.J. Polianskiy, A.V. Motrich, F.W. Grіnchuk, Spectrophotometeric properties of vein blood plasma in UV- zone patients with urgent surgical pathology of abdominal region organs, SPIE. - Vol. 6254, P. 485-489, 2006.
3. L. Andriescu, E. Patrascanu, R. Danila, Acute surgical pathology in elderly patients, Rev. Med. Chir. Soc. Med. Nat. Iasi.Vol.109, №2.– P.286–289, 2005.
4. H. Doran, T. Patrascu, C. Radu, Acute abdomen in diabetic patients – diagnostical questions, Chirurgia (Bucur).– Vol.98, №2.– P.119–125, 2003.
5. Pat. 4766, Ukraine, F. V. Grynchuk, І.Yu. Polyanskiy, Method of Modeling of the Acute Peritonitis, Bulletin № 2, 2005.
6. Pat. 31164А, Ukraine, О.S. Fedoruk, Method of Modelingof the Acute Renal Insufficiency, Bul. № 3, 2001.
7. J.C. Henquin, Metabolic control of potassium per meability in pancreatic islet cells, Biocem. J.- Vol. 186, № 3.- Р. 541 – 550, 1980.
8. S.G. Guminetskiy, About the Method of Measuring the Index of Reflection of the Objects on the Device in the Way of the Coupled Photometric Integrators, Optics and Spectroscopy. Vol.78, p. 496-501, 1995.
9. S.G. Guminetskiy, А.V. Моtrych, Spectrophotometric Research of the Biological Objects, Naukovyy Visnyk Chern. State Univer., Issue 261, Physics, Electronics - Chernivtsi: Chern. State Univer, p. 37-43, 2005.
10. Yu.М. Gain, S.I. Leonovich, N.V. Zavada, The Immune Status in Peritonitis and the Ways of its Pathogenic Correction, ООО “Unipress”,p. 256, 2001.
11. O.V. Pishak, S. G. Guminietskiy, V.P. Pishak, P. M. Grigorishin, The investigations of absorptive properties of blood plasma and urin, Bukovinskij Medychnyj Visnik, 2, №1, p. 137-144, 1998.
12. V.Е. Kholmogorov, V.А. Krylyenko, М.А. Оsmanov, Primary Photoprocesses in Blood and its Components Under the Optical Radiation Effect, Molecular Mechanism of the Biological Action of the Optical Emission. Moscow.: Nauka, P.284, 1988.
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