Objective: to provide a rationale for the use of perfluorane to correct erythrocyte morphological changes and blood biochemical indicators in acute blood loss.

Material and methods. Experiments were carried out in outbred male rats weighing 250—300 g under nembutal anesthesia. Blood loss was induced by arterial blood exfusion until blood pressure (BP) reached 40 mm Hg, following which this BP level was maintained for 60 minutes. Then the amount of blood withdrawn was reinfused. Perfluorane was injected intraarterially at a dose of 3 ml/kg 45 min before the start of blood loss. On the following day arterial and venous blood samples were harvested. The values of metabolic parameters (acid-base balance (ABB), hemoglobin (Hb), packed cell volume (PCV), and blood biochemistry), the morphological characteristics of erythrocytes (the size of discocytes and the percentage of stomatocytes and echinocytes), and the levels of reticulocytes in peripheral blood were studied. Results. Perfluorane exerted a normalizing effect on the values of blood ABB. Twenty-four hours after blood loss and reinfusion, Group 1 (a non-perfluorane group; n=8) showed reductions in Hb, PCV, and serum iron as compared to Group 3 (intact animals; n=12). In Group 2 (a perfluorane group; n=10), these indicators were decreased to a lesser extent. In these time periods, hypercholesterolemia was less marked in Group 2 than in Group 1. In Group 2, the mean diameter of a discocyte was larger than that in the animals of the two other groups. Group 1 displayed as high as 32% increases in the count of echinocytes with their virtually complete absence in the intact animals. At the same time, the levels of echinocytes in Group 2 animals were similar to those in the intact animals. As compared to Group 3, Group 1 exhibited a 4.5-fold drop in the percentage of reticulocytes (р≤0.05) while Group 2 showed a 2.2-fold rise in this indicator (р≤0.05).

Conclusion. The injection of perfluorane at a dose of 3 ml/kg body weight 45 min prior to blood loss causes an increase in the values of Hb, PCV, and serum iron and a reduction in total cholesterol. The administration of perfluorane

Objective: to reveal the patterns of a change in heart rhythm and breathing in patients with acute systemic injury due to cold in hypothermic and early posthypothermic periods.

Subjects and methods. Thirty patients aged 18 to 60 years (3 groups of 10 patients with mild, moderate, and severe cold injury) were examined in hypothermic and posthypothermic periods. The patient groups did not differ in gender, age, and weight. Within the first 24 hours after admission, all the patients underwent high-resolution Holter electrocardiographic monitoring that recorded cardiac arrhythmias and breathing disorders.

Results. During the therapy performed, as the degree of acute systemic cold injury increased, the patients were found to have a heart rate reduction (from 102 [90; 122] beats/min in Group 1 to 49 [38; 58] beats/min in Group 3) and a circadian index increase (from 105 [88; 125]% in Group 1 to 210 [185; 223]% in Group 3). With increased hypothermia, the victims were detected to have progressive cardiac rhythm and cardiac electrical conduction disturbances, such as supraventricular pacemaker migration, single and paired supraventricular premature beats, paroxysmal atrial tachycardia, atrial fibrillations, and ventricular premature beats. There was decreased heart rhythm variability in all the study groups, to the greatest extent in the patents with severe systemic cold injury. Late ventricular potentials were found in 2 and 7 patients with moderate and severe cold injury, respectively. Breathing disorders were recorded in all the study groups, the greatest increase in the frequency and duration of apnea/hypopnea episodes was noted in the patients with severe hypothermia. A fatal outcome occurred in 4 of the 10 patients with critical hypothermia due to the occurrence of idioventricular rhythm with transition to asystole.

Conclusion. Systemic hypothermia is accompanied by cardiac rhythm and cardiac electrical conduction disturbances and respiratory depression, which progress with the higher degree of acute systemic cold injury and, in case of critical hypothermia, may lead to a fatal outcome.

Objective: to develop a procedure to image native cells in a physiological medium and to measure the local stiffness of fixed red blood cell membranes in liquid in vitro experiments.

Material and methods. Blood was taken from 4 healthy donors during prophylactic examinations and placed in EDTA-containing Microvette tubes. The red blood cells were fixed in buffer solution onto a polylysine-coated substrate. Cell images were obtained using an ACM NTEGRA Prima microscope (NT-MDT, the Russian Federation). Membrane stiffness was estimated by atomic force spectroscopy.

Results. The values of effective Young's modulus were obtained for a statistical sample of fixed red blood cell membranes. Under the action of verapamil on blood, Young's modulus was equal to 528±21 kP. Under the action of heavy metal ions on blood, the mean local stiffness of the dried membrane was increased up to 942±56 at a depth of 35 nm.

Conclusion. The measurements of the force characteristics of red blood cells in liquid can yield reproducible results of examining the elastic properties of the membranes. Atomic force microscopy and atomic force spectroscopy can be employed analyze the properties of a live cell in the physiological medium.

A search for substances that are able to protect brain cells from the damaging effect of hypoxia remains one of the most relevant issues in modern neurobiology and medicine. Whether neurotrophic factors, brain-derived neurotrophic factor (BDNF) protein in particular, can be used to treat neurological diseases is the subject of wide speculation in the literature now. However, how the expression of this protein in the brain neurons changes after systemic circulatory arrest in the postresuscitation period remains uncertain.

Objective: to estimate the level of BDNF expression in the highly ischemia-sensitive neuronal population of cerebellar Purkinje cells and the value of BDNF in the resistance of neurons to ischemia-reperfusion.

Materials and methods. In mature outbred male albino rats (n=11), the heart was stopped under ether anesthesia at 12 minutes via intrathoracic ligation of the vascular fascicle, followed by revivification. A control group included pseudo-operated animals (n=11). On days 7 after revivification, a morphometric analysis of Nissl-stained paraffin sections 5—6 μm thick was used to determine the total number of Purkinje cells per 1 mm of their layer length. The expression of BDNF protein in the Purkinje cells was immunohistochemically examined by an indirect peroxidase-antiperoxidase test using primary polyclonal antibodies against BDNF. The count of Purkinje cells with different immune responses to BDNF protein was calculated. The intensity of BDNF expression was estimated from the mean optical density. Results. 12-minute systemic circulatory arrest in the rats resulted in a 12.5% reduction in the number of Purkinje cells. The immunohistochemical examination revealed a lower numbers of BDNF– neurons in the resuscitated rats. In this case, the count of BDNF+ and BDNF++ neurons corresponded to their reference level. Consequently, only BDNF-negative neurons, i.e. those that failed to express BDNF protein, died. Analysis of the mean optical density indicated that the remaining neurons had a higher BDNF protein expression than those in the controls. The found facts suggest that this protein has a neuroprotective effect in the postresuscitation period.

Conclusion. The capability for BDNF expression is an important factor that enhances neuronal resistance to death in the postresuscitation period. This offers promise for BDNF use to elaborate novel approaches to protecting the brain in ischemia-reperfusion.

Objective: to study the impact of goal-oriented infusion therapy using balanced salt solution, 4% succinylated gelatin, and 6% hydroxyethyl starch 130/0.42 solution on pulmonary extravascular water levels in coronary heart disease patients operated on under extracorporeal circulation.

Materials and methods. A prospective, randomized, single-blind clinical study investigating the impact of goal-oriented infusion therapy using balanced salt solution, 4% succinylated gelatin, and 6% hydroxyethyl starch 130/0.42 solution on pulmonary extravascular water levels was conducted in coronary heart disease patents operated on under extracorporeal circulation. The hemodynamic target was the global end-diastolic volume index which was maintained in the range from 680 to 850 ml/m2. Pulmonary extravascular water was measured by a transpulmonary thermodilution technique; hydrobalance, lactate, mixed venous oxygen saturation, hemodynamic parameters, and troponin I concentrations were also investigated.

Results. Analysis of the values of the pulmonary extravascular water index revealed no differences between the groups in pulmonary extravascular water levels throughout the study. Large volumes of balanced crystalloid solution were required to maintain intraoperative and postoperative hemodynamics. Balance after surgery was considerably higher and amounted to 2000 [1415—2200] ml in the crystalloid group, 1020 [650—1563] ml in the gelatin group, and 1070 [550—1675] ml in the hydroxyethyl starch solution (p=0.001). Mixed venous oxygen saturation, lactate, and troponin I concentrations were comparable between the groups at the study stages.

Conclusion. It was found the infusion therapy using balanced salt solution failed to result in an increase in pulmonary extravascular water levels in coronary heart disease patients operated on under extracorporeal circulation as did colloid solution-based infusion therapy. Achieving hemodynamic targets requires a large volume of crystalloid solution than the use of colloid infusion solution.

Genetic predisposition partially accounts for the clinical variability of the course of an infectious process. A total of 750 people, including 419 (81.1%) male patients aged 42.9±0.9 years, admitted to the clinics of the V. A. Negovsky Research Institute of General Reanimatology (Moscow, Russia), were genotyped to establish the influence of genetic factors on their susceptibility to critical conditions.

Materials and methods. Tetra-primer allele-specific polymerase chain reaction was used to investigate single-nucleotide polymorphisms (SNP) in the xenobiotic detoxification and oxidative stress genes (CYP1A1 (three sites), AhR, ABCB1, SOD2, GCLC and CAT) and in the vascular homeostasis genes (ACE, AGT, AGTR1, NOS3, VEGFα and MTHFR).

Results. A total of 268 nosocomial pneumonia (NP) cases were registered in a patient group. Individual SNP analysis has shown that among the patients with NP the risk of acute respiratory distress syndrome (ARDS) is associated with the carriage of the following genotypes: CYP1A1 rs2606345-Т/Т (p=0.0027, OR=2.38; 95% CI: 1.35—4.17) and AhRrs2066853-G/A-A/A (p=0.0012, OR=2.94; 95% CI: 1.54—5.60). The frequency of the C allele of the AGTR1 gene (re5186) was much higher among the survivors (in the NP group). The assessment of a multiplicative genetic model of genes that had demonstrated the highest single-locus effects because of a ARDS risk, as well as in-hospital mortality, could establish the complex genotype including a combination of risky alleles of the detoxi- fication and vascular homeostasis genes (CYP1A1 rs2606345-T — AhR rs2066853-A and ACE rs4340-D — AGT rs699-C — AGTR1 rs5186-C), which was associated with the increased risk of both NP and ARDS, as well as with the likelihood of a fatal outcome.

Conclusion. An understanding of the risk factors of NP and ARDS will aid in predicting the outcome of the underlying disease and in developing possible preventive measures. 

On the 75-th Anniversary

The rise in the incidence of acute necrotic pancreatitis in the past decade and high mortality rates of as high as 20—45% all suggest that the disease is of high social, medical, and economic importance.

Objective: to study the efficiency of extracorporeal detoxification methods in the combination treatment of patients with severe destructive pancreatitis.

Subjects and methods. The investigation enrolled 20 patients aged 40 to 53 years with destructive pancreatitis treated in the intensive care unit, Almaty City Clinical Hospital Four.

Results and discussion. Analysis of the findings over time could reveal that early incorporation of extracorporeal detoxification methods in the combination treatment of pancreatic necrosis provides a reduction in the clinical signs of endotoxicosis, which could prevent anuric acute renal failure in 61% of the cases and completely safeguard against multiple organ dysfunction in 22%. The investigation showed the necessity and efficiency the early use of albumin dialysis, hemodiafiltration in the combination treatment of sepsis and septic shock in destructive pancreatitis, leading to a significant reduction in mortality rates. The maximum detoxification potential of albumin dialysis in sepsis complicating the course of destructive pancreatitis is achieved by a combination of different mechanisms for elimination of toxic substances, such as convection, sorption, and diffusion, in the extracorporeal circuit.

Conclusion. Early use of extracorporeal detoxification methods in patients with severe destructive diseases of the pancreas results in a significant reduction in mortality rates.