Evaluation of the Effectiveness of Systemic Therapy of Spinal Cord Injury of Moderate Severity with Human Umbilical Cord Placental Blood Mononuclear Cells Using Indicators of Dispersion of Articular Angles in the Swimming Test.

Female Sprague-Dawley rats were used as models of moderate contusion spinal cord injury to evaluate the efficiency of single systemic (intravenous) infusion of human mononuclear cord blood cells for restoration of the motor function of hind limbs. The dynamics of recovery of hind limb motor function was assessed using a specially designed method based on calculation of selective dispersion and amplitude-dependent dispersion of hind limbs joint angles measured in the swimming test. The obtained data suggest that systemic application of human mononuclear cord blood cells significantly (p<0.05) promoted recovery of hind limb motor function in the animal models of contusion spinal cord injury of moderate severity in comparison with control animals (without cell therapy).

A Method of Assessment of the Motor Function of Hind Limbs by Swim Test in Spinal Cord Injury Models.

In most studies, various load tests are used to assess the recovery of functions after spinal cord injury in animals. However, the existing methods of assessing the movement in animals are not sufficiently accurate and objective. We developed a new method for assessing motor activity of laboratory animals that allows objective and highly accurate evaluation of movements in animals with serious neurological disorders caused by spinal cord injury. The swimming test was used as the main load test. Motor activity of swimming animals was assessed by measuring angles relative to the axis of motion, and the degree of angle spread for each joint and limb was estimated using the dispersion parameters depending on the values of the angles of the joints and the dispersion depending on the amplitudes of the angles. In Sprague-Dawley rats, contusion of the spinal cord at the Th9 level was modeled. In the swimming test, healthy control animals showed stability of both variance indicators over 6 weeks. In rats with spinal cord injury, motor activity of the hind limbs tended to increase from the first to the third weeks and remained at this level from the third to sixth weeks. The results suggest that the proposed method can become a good analogue of modern methods for assessing motor activity.

Comparison of the Efficiency of Systemic and Local Cell Therapy with Human Umbilical Cord Blood Mononuclear Cells in Rats with Severe Spinal Cord Injury.

Rat model of severe contusion spinal cord injury was used to study the effect of single intravenous and intraspinal injection of human umbilical cord blood mononuclear cells on the restoration of motor function of the hind limbs. Recovery of the motor function of the hind limbs was assessed using load tests and open-field test according to BBB scale. Cell injection via both routes significantly improved (p≤0.05) the recovery of the motor function of the hind limbs by 35-40% relative to the level of "self-recovery"; the effects of intravenous andintraspinal administration did not differ significantly.

Collagen Implant and Mononuclear Cells of Umbilical Blood Allow the Restore of Movements of Hind Limbs after Removing the Site of Spinal Cord.

Replacement of the removal site of the spinal cord on a collagen implant restores the motor function of the hind limbs in rats to the level of movements in the two joints for 8 weeks. After intravenous administration of mononuclear cells of human umbilical blood, recovery accelerated, significantly improved to the level of motion in the three joints, and there is a tendency to improve further recovery of movements.

Thiamine Induces Long-Term Changes in Amino Acid Profiles and Activities of 2-Oxoglutarate and 2-Oxoadipate Dehydrogenases in Rat Brain.

Molecular mechanisms of long-term changes in brain metabolism after thiamine administration (single i.p. injection, 400 mg/kg) were investigated. Protocols for discrimination of the activities of the thiamine diphosphate (ThDP)-dependent 2-oxoglutarate and 2-oxoadipate dehydrogenases were developed to characterize specific regulation of the multienzyme complexes of the 2-oxoglutarate (OGDHC) and 2-oxoadipate (OADHC) dehydrogenases by thiamine. The thiamine-induced changes depended on the brain-region-specific expression of the ThDP-dependent dehydrogenases. In the cerebral cortex, the original levels of OGDHC and OADHC were relatively high and not increased by thiamine, whereas in the cerebellum thiamine upregulated the OGDHC and OADHC activities, whose original levels were relatively low. The effects of thiamine on each of the complexes were different and associated with metabolic rearrangements, which included (i) the brain-region-specific alterations of glutamine synthase and/or glutamate dehydrogenase and NADP+-dependent malic enzyme, (ii) the brain-region-specific changes of the amino acid profiles, and (iii) decreased levels of a number of amino acids in blood plasma. Along with the assays of enzymatic activities and average levels of amino acids in the blood and brain, the thiamine-induced metabolic rearrangements were assessed by analysis of correlations between the levels of amino acids. The set and parameters of the correlations were tissue-specific, and their responses to the thiamine treatment provided additional information on metabolic changes, compared to that gained from the average levels of amino acids. Taken together, the data suggest that thiamine decreases catabolism of amino acids by means of a complex and long-term regulation of metabolic flux through the tricarboxylic acid cycle, which includes coupled changes in activities of the ThDP-dependent dehydrogenases of 2-oxoglutarate and 2-oxoadipate and adjacent enzymes.

Efficiency of transplantation of human placental/umbilical blood cells to rats with severe spinal cord injury.

The effects of a single intravenous injection of human umbilical blood were studied on the model of severe spinal cord contusion injury in rats. Rats receiving no umbilical blood (spontaneous recovery) served as the control. All rats exhibited pronounced hind limb paraplegia and autonomic dysfunction of pelvic organs after the injury. Recovery of the hind limb function was evaluated by loading tests and locomotor activity testing in the open field using BBB score for open-field testing. Testing was carried out weekly for 8 weeks after the injury. Open-field testing showed a significant (p < 0.05) increase of the rate and volume of the hind limb motor activity recovery in the groups receiving umbilical blood infusions.

Modification of injection mapping of vascular-capillary bed in cerebral nervous tissue.

A simple original method of simultaneous staining of the cell elements on cerebral cortex and its vascular bed is developed. It is based on perfusion of polyvinyl acetate emulsion prepared on 2% paraformaldehyde solution with phosphate buffer at pH 7.4.

Changes in the volume of the intercellular space of the cerebral cortex in conditions of peripheral stimulation in rats.

Changes in the volume of the intercellular space of the rat cerebral cortex in response to peripheral repetitive stimulation were studied. The volume of the intercellular space and its changes were assessed by a modification of the four-electrode impedance method. The results suggest that evoked electrical activity in the cerebral cortex was accompanied by 3-5% decreases in the volume of the intercellular space.

Long-term prognosis of chronic glomerulonephritis.

The dependence of actuarial survival rates on morphological and clinical manifestation of chronic glomerulonephritis (CGN) has been studied in 520 patients followed up for 2-42 years. In grouping the patients along two signs--histological lesion and relapse incidence--the survival in different morphological forms of CGN was found to be similarly dependent on the illness activity. It was high in rare and the lowest in frequent and persisting relapses. Hypertension, high-grade tubulo-interstitial changes and sclerosis over 50 per cent of glomeruli indicate a poor prognosis as signs of severe renal damage under which each relapse may hasten the lethal outcome. Identification of the histological appearance is of high importance in CGN prognosis because of their different manifestations and tendency to relapse.

Haemoperfusion in the treatment of chronic renal failure in patients on haemodialysis.

The paper is concerned with materials characterizing physicochemical properties of haemosorbents of varying grades. Adsorption of different uraemic toxins was studied. The possibility of microembolization of internal organs and tissues with haemosorbent particles was explored in the course of prolonged treatment with the use of haemosorption. Different approaches to connecting the sorption column to the dialyzer were examined to disclose an optimal variant. The authors report the results of applying haemosorption in patients who had received haemodialysis therapy to prevent complications. The blood response to the prolonged haemosorption treatment was analyzed. It is recommended that long-term (6-7 hours) sessions of haemosorption might be performed using carbons having high sorption capacity.

Epidemiology of chronic renal diseases.

Autopsies of all uraemic patients in Leningrad for three years, and materials of the City Nephrological Service have demonstrated that the structures of nephrological diseases in their early and terminal stages were different. Chronic glomerulonephritis has been noted in patients with normal renal function just as often as chronic pyelonephritis but the former prevails considerably among the causes of uraemia. The proportion of polycystic kidney disease, amyloidosis, and diabetic nephropathy increases in patients with chronic renal failure. Due to these changes and the difference in the death age of patients with various diseases the majority of patients suitable for treatment with long-term dialysis suffer from chronic glomerulonephritis and only 14.89-20.5% from chronic pyelonephritis.

Peculiarities of erythropoiesis in patients with chronic renal failure.

In 600 patients suffering from chronic renal insufficiency the cellularity of bone marrow, erythroid cells proliferative activity, erythroid cells destruction and iron incorporation rate, data of ferrokinetics, intracellular iron distribution, porphyrin synthesis rate were examined. On the basis of the obtained data the suggestion is put forward that metabolic disturbances are important in anemia development in uremic patients. One of the aspects of this problem is the role of disorders in the protein turnover causing changes in the synthesis of globin and porphyrin which are the primary components for heme synthesis. Special importance is attached to the changes in iron turnover, i.e. to its redistribution between stromal and heme pools.

Carbohydrate metabolism in glomerulonephritis.

Exogenous insulin sensitivity as well as the dynamics of blood serum levels of immunoreactive insulin and growth hormone (GH) after oral glucose loading were studied in patients with glomerulonephritis of different clinical manifestations and varying renal functions. A correlation between disturbances in carbohydrate metabolism on the one hand, and the degree of renal failure and protein depletion on the other, was established. Disturbances in carbohydrate metabolism were detected as early as in the preazotemic phase of the disease. The clinical significance of the disorders and methods for evaluation of insulin response to oral glucose are discussed.

Present-day concepts on gastric pathology in patients with chronic renal failure.

Patients with chronic renal failure often reveal a peculiar "uremic gastropathy" characterized by a decreased secretory activity of the gastric glands. Most of such patients should not be referred to the group of high risk of ulceration. This point is extremely important, when cases are screened for kidney transplantation. Immunomorphological changes occurring in the gastric mucosa provide just one of the possible etiological factors, which also include uremic intoxication and, probably, the intragastric effect of nitrous metabolic products. A long-term (months) therapy by programmed hemodialysis is not, as a rule, accompanied by the development of gastric hyperchlorhydria and hypersecretion.

Androgen therapy for anemia in renal failure.

Forty-six patients marked renal failure and anemia underwent androgen therapy (up to 500 mg/week), as well as intravenous injections of iron preparations (600-800 mg/month) and hemotransfusions (2500 ml/month) for 4 month. Ferrokinetic indices, total marrow cellularity and erythrocyte and hemoglobin levels were determined at the end of therapy. Androgens were responsible for the increase in Hb level by 1.2 g%. High total bone marrow cellularity was observed in these patients (27.4 +/- 3.2 X 10(9) cells/kg, compared with 14.1 +/- 1.4 X 10(9) cells/kg in normal individuals). Hemotransfusions resulted in a decrease of Hb level by 1.3g% and total bone marrow cellularity to 8.3 +/- 2.2 X 10(9) cells/kg. Ferrokinetic indices became poor. The data obtained are discussed.

Possible role of the proximal convoluted tubules of human kidney in chronic glomerulonephritis. A quantitative electron-microscopic study.

The amount of forming pinocytic (coated) microvesicles on the apical plasma membrane of kidney proximal tubule cells was assessed in kidney biopsies of 10 patients suffering from chronic glomerulonephritis. A significant correlation was found between the amount of these vesicles and diurnal proteinuria levels (r = 0.889; p less than 0.01). The possible mechanisms of protein reabsorption via pinocytosis in both normal and pathological conditions are considered.

Iron absorption in gastric and duodenal pathology in patients with iron deficiency anaemias.

Absorption of radioactive iron was studied in 87 patients with different types of iron deficiency anaemias and in 23 healthy subjects. The subjects were given 1...2muci of radiactive iron in the form of FeSO4 together with 5 mg of nonradioactive iron as a carrier and 100 to 150 g of white bread, radioactivity on the whole body being studied with a big liquid scintillation counter 4 pi (BLSC-2). In clinical observations and in single experiments on volunteers there was no conformity of the values of absorption with the levels of acid-formation. But in the same time the gastric juice from an anaemic horse almost doubled iron absorption in healthy individuals. Marked morphological changes in the gastric mucosa inhibited the absorption in the intestine and the degree of increase of absorption in patients with anaemia depended to some extent on the morphological conditions of the gastric mucosa. When healthy subjects and patients with iron deficiency anaemia were given bread "enriched" with iron before baking instead of common bread with "external" mark there was observed similar correlation between the values of absorption but the figures were somewhat lower.

Proliferative activity of erythrone and intramarrow hemolysis in iron deficiency anemias.

3H-thymidine incorporation into normoblasts, proliferation rate of erythroid precursors and degree of intramarrow hemolysis have been studied in vitro on the bone marrow. The normal proliferation rate of normoblasts is 26 +/- 2% i.e. during 24 hours about a quarter of dividable elements of erythropoiesis is renewed. Acute blood loss increases the proliferation rate up to 57 +/- 9% but the value of 3H-thymidine incorporation into cells is not changed as compared to normal. In chronic blood loss both 3H-thymidine incorporation into dividing erythroid precursors at different stages of maturity and the rate of erythroid production are 2 to 3 times lower than normal. In healthy persons the degree of intramarrow hemolysis is 7 +/- 2% of erythroid precursors incubated for 24 hours. In iron deficiency anemia intramarrow destruction sharply increases, presenting at an average 30% of incubated nucleated elements of erythropoiesis. A type of chronic iron deficiency, which is not associated with blood loss, is described. In this type of anemia the proliferation rate of normoblasts and the degree of intramarrow hemolysis do not differ from normal values.