Macro- and microstructure of erythrocyte membranes under acute massive hemorrhage and subsequent blood reinfusion.

The authors studied changes in erythrocyte membrane nanostructure using a rodent model of hemorrhagic hypotension and resuscitation. Both macro- and microstructural elements were examined using atomic force microscopy. Membrane "roughness" was characterized using spatial Fourier transformation and was stratified according to the periodicity of the membrane. Acute hemorrhage resulted in an increase in the diameter and height of erythrocytes, which returned to baseline levels by the end of the hemorrhagic hypotensive period. The effect of hypotension on the erythrocyte surface was nonuniform. In those regions where damage was considerable, the rate of restoration of the membrane microstructure to baseline levels was prolonged. The less damaged surfaces were restored more rapidly to control values after reperfusion. More detailed use of atomic force microscopy in the definition of the erythrocyte membrane microstructure may further define the mechanisms of cellular functional restoration after hemorrhage.

[Conditioning in resuscitated rats with originally different behavioral patterns].

Male white rats divided in three groups on the basis of their behavior in the elevated plus-maze were subjected to 10-min cardiac arrest. Analysis of the results of their subsequent food and active avoidance conditioning revealed the effects of the following factors: (1) a factor of initial typological features of the higher nervous activity, (2) a factor of general brain mechanisms of postresuscitation, and (3) a factor of postresuscitation features in rats with different behaviors, which represented changes in different mechanisms of conditioned reflex performance in these animals.

[Postresuscitation cerebral structural and functional changes associated with the initial type of behavior]. / Postreamanitsionnye strukturno-funktsional'nye izmeneniia mozga, sopriazhennye s iskhodnym tipom povedeniia.

The elevated cross-shaped labyrinth test carried out on highly, moderately, and low active experimental animals revealed significant differences in the baseline density and composition of neuroglial populations of numerous formations of the brain, as well as in the early structural and functional sequels of experienced clinical death. The most pronounced postresuscitation abnormal changes were observed in highly active animals and the least marked ones were seen in moderately active animals.

Water status and LPO in rat tissues during massive blood loss and irradiation with He-Ne laser.

We revealed a relationship between water balance and LPO in the myocardium, liver, and blood plasma during massive blood loss and irradiation with He-Ne laser. Low-intensity laser irradiation of the plasma inhibits LPO and normalizes water balance in rat tissues during massive blood loss.

[Various forms of water in the rat myocardium and liver during blood loss and the early postresuscitation period]. / Zakonomernosti izmeneniia razlichnykh form vody v miokarde i pecheni krys pri krovopotere i v rannem postreanimatsionnom periode.

Different forms of water content were studied in myocardium and liver of male rats at 2-h hypotension and during the early postreanimation period. The phenomenon of redistribution of free and bound water was found. Comparison of water balance during the postreanimation period in the myocardium and liver revealed that normalization of different forms of water occured in myocardium. In hepatocytes oedema remained over the whole period studied.

[Acute and massive hemorrhage: mechanisms of compensation and damage]. / Ostraia massivnaia krovopoteria: mekhanizmy kompensatsii i povrezhdeniia.

To evaluate the body's compensatory reserves, experiments were made on anesthetized (with heparine) mongrel dogs of both sexes weighing 8-25 kg. The experiments have indicated that early monitoring of physiological parameters (external respiration, cardiovascular performance, hemostasis, red blood cell morphometry) in the first 5-10 minutes of acute massive hemorrhage is of high informative value. The generalization of vasoconstriction resulting in higher overall blood volume, the maintenance of venous return, tachypnea, active spontaneous hemodilution, and increased heart rate are the leading mechanisms of perfusion pressure maintenance in early acute hemorrhage. The severity of acute damage to the membranes of red blood cells and endotheliocytes and hence changes occurring in the suspension structure of blood, which drastically impairs its rheological parameters and fluidity, can play the key role in the pathogenesis of decompensatory cardiovascular and visceral functions in prolonged arterial hypotension. The findings open up new possibilities for early evaluation and prediction of the ensurina course of delayed massive hemorrhage.

Morphologic changes of red blood cells during hemorrhagic shock replicate changes of aging.

Blood loss leads to the reduction in vitality of red blood cells (RBCs). However, the changes in morphology at different stages of hemorrhagic shock have not been studied. Thus, the aim of this study was to identify and quantitate the sequence of morphological changes in RBCs during hemorrhage. This study was performed on 15 adult inbred dogs. Blood samples were taken before hemorrhage, when the mean arterial pressure reached 40 mm Hg (initial stage of shock), and at a mean arterial pressure level of 20 mm Hg (decompensated stage of shock). The volume of blood removed averaged 33.6+/-8.9 and 55.1+/-6.9 mL/kg, respectively. Evaluation of RBC morphology was performed by computerized light microscopic morphometry and scanning electron microscopy. At the early stage of hemorrhage the number of "young-appearing" RBCs with large visible surface areas (41-50 microm2) increased from 17.7%+/-3.1% to 26.6%+/-3.5% (P < 0.05). Concomitantly, the number of "old-appearing" RBCs with small visible surface area (20-30 microm2) significantly decreased from 5.3%+/-2.7% to 2.7%+/-2.3% (P < 0.01). At the stage of decompensated blood loss, the opposite phenomenon was observed. The number of "old-appearing" RBCs increased to 8.2%+/-1.1% (P < 0.01), whereas the number of "young-appearing" RBCs decreased to 12.3%+/-4.2% (P< 0.01). The changes in visible surface area of RBCs was accompanied by significant alterations in their shape. The percentage of abnormal shaped RBCs increased from 8.9%+/-1.1% before the hemorrhage to 36.4%+/-5.8% at the stage of decompensated hemorragic shock (P < 0.01). Thus, during the late decompensated stage of hemorrhagic shock, RBCs assume shape and surface area changes that are similar to those seen in aging. These changes in RBC size and shape may be due to the effects of shock-induced oxidative stress.

[Effect of laser exposure on hemostatic parameters in the preagonal and post-resuscitation period]. / Deistvie lazernogo izlucheniia na parmetry gemostaza v preagonal'nom i postreanimatsionnom periode.

Effects of intravascular low-intensive laser exposure of the blood on the hemostasis during acute blood loss and the early postresuscitation period after 4-min clinical death were studied on narcotized dogs (8-17 kg) of both sexes with different initial levels of heparin. During the preagonal period laser exposure caused hypercoagulation in animals with initial heparin content below 60 micrograms/ml. This acceleration of blood clotting prevented a drop in the activity of antithrombin III and hypercoagulation by the third hour of postresuscitation period. In animals with initial heparin content more than 60 micrograms/ml laser exposure caused hypocoagulation in the presence of reduced activity of antithrombin III by the third hour of the resuscitation period.

[Hemocoagulation while dying of blood loss and restoration to life after clinical death]. / Gemokoaguliatsionnye parametry pri umiranii ot korovopoteri i ozhivlenii posle klinicheskio smerti.

Hemostatic system function was studied in dogs dying of acute blood loss and restoring to life after 4-min clinical death. Phasic changes in hemostatic system of two and three types occurred in the blood loss and reanimation, respectively. Dogs with favorable postresuscitation period exhibited hypercoagulation when dying, hypocoagulation 1 hour after reanimation and normal coagulation 3-6 hours after clinical death.

[Low-energy laser irradiation of blood as a method of prevention of blood circulation decompensation in hemorrhagic shock]. / Nizkoénergeticheskoe lazernoe obluchenie krovi kak sposob profilaktiki dekompensatsii krovoobrashcheniia pri gemorragicheskom shoke.

Effect of low-intensity laser exposure of the blood on the central hemodynamics, oxygen transporting function of the blood, oxygen balance of the organism, and surface configuration of erythrocyte membranes was studied in dogs exposed to 2-h arterial hypotension (arterial pressure 40 mm Hg). Blood exposure was started from the tenth min of hypotension and went on for 45 min (group 2) or 120 min (group 3); group 1 was control. The best results were attained after 45-min laser exposure. A longer exposure creates prerequisites for complications, such as dysadaptation of the vascular tone and delayed decrease of hemoglobin concentration.

[The effect of low-intensity laser radiation on the parameters of the blood anticoagulation system in the early postresuscitation period]. / Vliianie nizkointensivnogo lazernogo izlucheniia na parametry protivosvertyvaiushchei sistemy krovi v rannem postreanimatsionnom periode.

The effect of low-intensive laser exposure at a wavelength of 634 nm and power 1 mWt at the tip of the light-guide on the blood anticoagulative system was studied in mongrel narcotized dogs of both sexes weighing 8 to 16 kg during the postresuscitation period after 4-min clinical death from massive blood loss. The anticoagulative system in the blood plasma was depleted during the reanimation period (the activities of plasmin and fibrinogen-heparin complexes and the level of heparin dropped). Intravascular laser exposure of the blood (for 30 min during blood loss after drop of the mean arterial pressure to 40 mm Hg and at the beginning of the second hour of the postreanimation period) boosted the activities of plasmin and fibrinogen-heparin complexes but failed to increase the level of heparin in the postreanimation period.

[Changes in the functional status of the left-ventricular myocardium during dying and in the early postresuscitation period]. / Izmeneniia funktsional'nogo sostoianiia levogo zheludochka pri umiranii i v rannii postreanimatsionnyi period.

Experiments on dogs have shown that left-ventricular contractility reduces and the processes of contraction and relaxation are discoordinated during the first minutes of dying from acute blood loss with the mean arterial pressure of 40 mm Hg. During the postreanimation period following a ten-min clinical death spontaneous respiration is resumed late (after 15.1 +/- 3.2 min), as are the corneal reflexes (after 46.1 +/- 8.5 min). If there are no changes in left-ventricular contractility in the course of dying, vital functions of the organism and neurological reflexes recover sooner (respiration after 6.1 +/- 2.0 min and corneal reflexes after 32.6 +/- 4.1 min). A relationship has been revealed between the severity of the postreanimation period and the degree of reduction of left-ventricular myocardial contractility during hypoperfusion (1 to 6 hrs after reanimation): the contractility index in animals with late neurologic recovery was lower than in those with a relatively early neurologic recovery.

[Changes in the oxygen balance of the body in the acute period of hemorrhagic shock and after resuscitation under the effect of intravascular helium-neon laser irradiation of the blood]. / Izmeneniia kislorodnogo balansa organizma v ostrom periode gemorragicheskogo shoka i posle reanimatsii pod vliianiem vnutrisosudistogo gelii-neonovogo lazernogo oblucheniia krovi.

The effect of intravascular laser irradiation of the blood (ILIB) on the oxygen budget of the body and blood acid-base balance in the acute period of hemorrhagic shock and after resuscitation was studied in experiments on dogs. Duration of hypotension was 2 h. Laser therapy was started from the 5th-8th min of hypotension and lasted for 45 min; it was performed with a He-Ne laser (ALOK-1, 1 mWt power at the tip of the light guide, wavelength 633 nm) introduced in the right jugular vein. Exposure to ILIB in the acute period of hemorrhagic shock was conducive to creation of favorable conditions for realization of the compensatory mechanisms aimed at improvement of circulation and gas exchange in the lungs, at provision of delivery of oxygen to tissues and its utilization, and, hence, promoted the survival of animals by triggering the regulatory mechanisms of redistribution of the regional bloodflow, improvement of tissue perfusion, and possibly, of oxyhemoglobin dissociation processes in red cells.

[The effect of intravascular helium-neon laser blood irradiation on the state of the compensatory processes in the acute period of hemorrhagic shock and after resuscitation]. / Vliianie vnutrisosudistogo geliineonovogo lazernogo oblucheniia krovi na sostoianie kompensatornykh protsessov v ostrom periode gemorragicheskogo shoka i posle reanimatsii.

The effect of low-intensity intravascular laser blood irradiation on morphofunctional characteristics of erythrocytes and circulation parameters has been studied experimentally on 22 adult inbred anesthetized dogs of both sexes 9 to 23 kg of weight during a 2-hour hemorrhagic shock and in the first hours after resuscitation. It has been established that the use of intravascular laser blood irradiation during 45 min of hemorrhagic shock stabilizes erythrocyte membranes and improves myocardial function. Hyperdynamic and hyperperfusion syndromes were significantly more pronounced in the postresuscitation period in animals treated by intravascular laser blood irradiation than in the control. Thus, the efficacy of intravascular laser blood irradiation was associated both with the recovery of erythrocyte cytoarchitectonics and the enhancement of myocardial contractility.

[Early prognostic signs of disorders of the regional pulmonary circulation in the dynamics of hemorrhagic shock and in the post-resuscitation period]. / Rannie prognosticheskie priznaki narushenii regionarnogo krovoobrashcheniia legkikh v dinamike gemorragicheskogo shoka i v postreanimatsionnom periode.

The earliest significant unfavourable signs of shock are as follows: an increase in total pulmonary resistance, reduced pulmonary blood flow and tachypnea. Ventilation-perfusion disturbances in the lungs precede the onset of circulation decompensation, which in the pulmonary flow manifests as enhanced pulmonary blood content and elevated pressure in the pulmonary artery. Pulmonary flow damage in the early postresuscitation period is to a great extent predetermined by the degree of circulation disturbances in the lungs, developing during hypotension period, the leading among them being congestive processes in the lungs (increased blood content, hypertension) which progressed in case of unfavourable outcome of resuscitation and reduction of the microvascular bed.

[The effect of the functional status of the CNS in the postresuscitation period on restoration of the neurologic status in dogs following systemic circulatory arrest]. / Vliianie funktsional'nogo sostoianiia TsNS v postreanimatsionnom periode na vosstanovlenie nevrologicheskogo statusa u sobak posle sistemnoi ostanovki krovoobrashcheniia.

The use of caffeine or diazepam in experiments on dogs recovered after a 12-min circulatory arrest has no significant effect on the end points of resuscitation, as compared to the control. Diazepam has a more favourable effect on the recovery of central nervous system functions than caffeine. Combined use of caffeine and diazepam or lidocaine at certain stages of the early postresuscitation period accelerates the recovery of the neurological status and improves the outcome of resuscitation.

[Characteristics of the activation of lysosomal enzymes in the rat liver after death and during resuscitation]. / Osobennosti aktivatsii fermentov lizosom pecheni krys pri umiranii i posle ozhivleniia organizma.

Alterations in activity of lysosomal acid phosphatase and cathepsin D, as well as of hepatocyte ultrastructure were studied in rat liver tissue after experimental heart arrest within 10 and 30 min and during the early post-resuscitation period. Cathepsin D free activity in a supernatant fraction as well as both enzymatic activities in lysosomal fraction were increased after 30 min heart arrest. Activity of acid phosphatase and cathepsin D in lysosomes was decreased, while the activity of the free enzymes was increased within 1 and 4 hrs after resuscitation. Triton X-100 (0.025%) caused labilization of lysosomal membranes. Alterations in ultrastructure of hepatocytes were observed within 30 min of the heart arrest and within first hour of the post-resuscitation period. The lysosomal membranes tended to normalization within 24 hrs after the post-resuscitation period, whereas the enzymatic activity remained elevated. Role of lysosomes in regulation of intracellular metabolism is discussed.