Restoration of gastrocnemius muscle in MDX mice of different age after injury and implantation of xenogenic muscle tissue.

The intensity of regeneration of crossed gastrocnemius muscle was evaluated in two groups of mdx mice of different age 2 weeks after implantation of crushed muscle tissue from newborn rats into the wound defect area. The effect of xenoplasty manifested in increased weight of the damaged muscle. The effect was observed in mice aging 12-16 weeks but not in those aged 40-48-weeks. Structural changes in the skeletal muscle tissue intrinsic of mdx mice and augmenting with age were detected in intact mice before the experiment. Activity of muscle fiber regeneration in intact and injured muscle of 40-48-week-old mice was significantly lower than in 12-16-week-old ones. Myoblasts of the xenogenic transplant retained viability in recipient muscles for at least 2 weeks. posttraumatic regeneration was stimulated in only 12-16-week animals. Xenoplasty was ineffective in older animals and even somewhat enhanced the destructive processes in the muscle. It seems that age-specific regeneration activity of the recipient skeletal muscle tissue should be taken into consideration in the development of effective strategy of cell therapy for progressive muscular dystrophy.

Cardiotropic effect of extracardiac transplantation of embryonic human myoblasts to mice with bradycardia: various effects of cell material.

Animals with bradycardia were detected in reproductive colony of mdx mice. Low pulse rate was associated with poor survival and predisposition to sudden death, but did not directly depend on the presence of dystrophin mutant gene or animal age. Heart rate increased in old mice with bradycardia after extracardial, intramuscular, and intravenous injection of human embryonic myoblasts. Stable normalization of the pulse was observed 2 weeks after transplantation, but early peak of heart rate was observed as early as 24 h after cell transplantation. Cell suspensions, which could contain stem cells (blood mononuclears and CD34+ lymphocytes), also corrected heart rhythm. Unlike the effect of myoblasts, cardiotropic effect of mononuclears was preceded by a period of tachycardia, while the effect of CD34+ lymphocytes was very unstable. The cardiotropic effect of myoblasts was combined with life span prolongation and certain rejuvenation in some animals. Erythrocytes and supernatant obtained during blood cell fractionation did not modify the heart rhythm in mice with bradycardia. After injection of myoblasts to mice with rare and normal pulses serum creatine kinase activity decreased with different rates. These data attest to a variety of biological effects of stem cells and/or their derivatives and to ambiguous mechanisms of these effects.

Hereditary muscular dystrophy in MDX mice as a homologous model for introduction of cell technologies in the treatment of progressive muscular dystrophies in humans.

Life-time monitoring of the main clinical and laboratory manifestations of hereditary muscular dystrophy in mdx mice confirmed the presence of mutation in exon 23 of dystrophin gene and the absence of this protein in skeletal muscles of mutant animals. Muscular dystrophy in mice was similar to human progressive muscle disorder, which allows the use of this model for the development of cell technologies for the treatment of hereditary muscular diseases in humans.

Xenotransplantation of embryonic precursors of human myogenesis for the correction of dystrophinopathy in mice with hereditary muscular dystrophy.

Human embryonic myogenic precursors were transplanted into muscles of mdx mice with hereditary dystrophin-deficient muscular dystrophy. Transplantation induced the synthesis of human dystrophin. The number of dystrophin-positive fibers progressively decreased, however, some of them were preserved even 5 months after transplantation. Our results indicate that xenogeneic transplantation of embryonic myogenic precursors compensates the genetic defect in dystrophin-deficient mice.

[Hereditary diseases of nervous system in the population of Vladimir province]. / Nasledstvennye bolezni nervnoi sistemy v populiatsii Vladimirskoi oblasti.

The first population genetic study of hereditary disorders of the nervous system (HDNS) in Vladimir oblast was carried out. A total of 1,622,900 subjects, including 1,306,200 from the urban and 316,700 from the rural population, were tested. The population examined was characterized by virtually homogenous ethnic structure, with Russians constituting 95.76%. Pooled prevalence of HDNS in Vladimir oblast corresponded to the average prevalence for other Russian populations. Substantial differences between urban and rural populations in respect of the population load of HDNS and its nosological structure were not observed. A total of 22 nosological forms of HDNS were revealed, including thirteen autosomal dominant (193 families with 272 affected individuals), seven autosomal recessive (59 families with 66 affected individuals), and two X-linked (15 families with 17 affected individuals) diseases. The composition of the HDNS spectrum "nucleus" in Vladimir oblast displayed a number of differences from that in the majority of other populations examined. The HDNS in different regions of the area tested were characterized by different prevalence and spectrum. The data obtained may constitute a basis for regional registration of HDNS in Vladimir oblast.

[Pathogenetic role of intestinal microbiocenosis in the pathogenesis of hereditary myodystrophy]. / Patogeneticheskaia rol' kishechnogo mikrobiotsenoza v klinike nasledstvennykh miodistrofii.

The effect of the inclusion of probiotic preparations for the correction of disturbances in normal intestinal microflora into the complex therapy of patients wish Duchenne's childhood muscular dystrophy and Becker's myopathy was analyzed. Probiotic therapy made it possible to improve the clinical state of patients, manifested by an increase in muscular strength and accompanied by positive shifts in electromyographic, immunological, biochemical, hormonal characteristics. Intestinal microbiocenosis plays seemingly a certain role in the formation of hereditary pathology.

A new variant of Charcot-Marie-Tooth disease type 2 is probably the result of a mutation in the neurofilament-light gene.

Charcot-Marie-Tooth (CMT) disease is the most common inherited motor and sensory neuropathy. The axonal form of the disease is designated as "CMT type 2" (CMT2). Although four loci known to be implicated in autosomal dominant CMT2 have been mapped thus far (on 1p35-p36, 3q13. 1, 3q13-q22, and 7p14), no one causative gene is yet known. A large Russian family with CMT2 was found in the Mordovian Republic (Russia). Affected members had the typical CMT2 phenotype. Additionally, several patients suffered from hyperkeratosis, although the association, if any, between the two disorders is not clear. Linkage with the CMT loci already known (CMT1A, CMT1B, CMT2A, CMT2B, CMT2D, and a number of other CMT-related loci) was excluded. Genomewide screening pinpointed the disease locus in this family to chromosome 8p21, within a 16-cM interval between markers D8S136 and D8S1769. A maximum two-point LOD score of 5.93 was yielded by a microsatellite from the 5' region of the neurofilament-light gene (NF-L). Neurofilament proteins play an important role in axonal structure and are implicated in several neuronal disorders. Screening of affected family members for mutations in the NF-L gene and in the tightly linked neurofilament-medium gene (NF-M) revealed the only DNA alteration linked with the disease: a A998C transversion in the first exon of NF-L, which converts a conserved Gln333 amino acid to proline. This alteration was not found in 180 normal chromosomes. Twenty unrelated CMT2 patients, as well as 26 others with an undetermined form of CMT, also were screened for mutations in NF-L, but no additional mutations were found. It is suggested that Gln333Pro represents a rare disease-causing mutation, which results in the CMT2 phenotype.

[Hereditary nervous system diseases in Mordovia]. / Nasledstvennye bolezni nervnoi sistemy v Mordovii.

A medical genetic study on hereditary neural diseases was performed in 21 districts of the Mordovian Republic. The total number of persons examined was 936,800. The population load of autosomal dominant, autosomal recessive, and X-linked recessive diseases was 0.1696 +/- 0.0129, 0.1075 +/- 0.0107, and 0.0341 +/- 0.0010, respectively. Twenty-eight disease entities were revealed, including 10 autosomal recessive (AR), 15 autosomal dominant (AD), 2 X-linked recessive (XR) diseases, and 1 genetically heterogenous disease. These diseases were nonuniformly distributed among different populations of Mordovia. The incidence of AR diseases was highest in the Mordovian and Tatar populations; that of AD diseases, in the Russian population; and that of XR diseases, in the Mordovian population.

[Screening for deletion in patients with Duchenne's myodystrophy by multiplex amplification]. / Skrining deletsii u bol'nykh miodistrofiei Diushenna metodom mul'tipleksnoi amplifikatsii.

Patients with Duchenne muscular dystrophy were analyzed using the method of polymerase chain reaction in order to reveal deletions in the dystrophin gene. Deletions of different lengths and locations were detected in 28 of 78 ill boys. The highest number of deletions was detected in the 3'-end of the gene (the 45-50th exons).

[Status of the feto-placental system in pregnant women with a history of habitual abortion during complex therapy including metabolic correction]. / Sostoianie feto-platsentarnoi sistemy u beremennykh, stradaiushchikh privychnym nevynashivaniem, na fone kompleksnoi terapii s primeneniem metabolicheskoi korrektsii.

Fifty-two women suffering from habitual abortions, referred to high-risk group in respect of placental insufficiency development, were followed up over the course of pregnancy. Metabolic correction was added to the complex of therapy of some of these patients, the rest were administered basic therapy alone. The examinations revealed placental insufficiency signs in 78.8% of these women. Placental insufficiency in the examined group was characterized by enlarged intervillous space and a longer period necessary for filling this space, reduced linear rate of the blood stream in this space, as well as by reduced levels of the fetoplacental hormones and activation of lipid peroxidation. Addition of metabolic therapy to the complex of therapeutic measures helped reduce the incidence of placental insufficiency by 17.4% and reduce more than twofold the incidence of fetal hypoxia and hypotrophy .

[Increase of the negative charge of erythrocyte membrane proteins in hereditary neuromuscular diseases]. / Uvelichenie otritsatel'nogo zariada belkov membrany éritrotsitov pri nasledstvennykh nervno-myshechnykh zabolevaniiakh.

The parameters of erythrocyte ghost protein's fluorophores by nitrate's anions were studied in patients with various hereditary myodystrophy. In all the groups under examination the share of fluorophores accessible to a quencher was close to 1. In erythrocyte membranes of healthy donors the relevant constant quenchering was about 17.3 +/- 1.9 M-1 while those of patients were decreased by 3.1 (Duchenne's myodystrophy) and by about 2.0 (other forms of primary and secondary progressive muscular dystrophies). The most probable reason for the decreasing constant of quenchering is the increase of negative charges on the erythrocyte membrane proteins.

[Central hemodynamics in pregnant women with pre-eclampsia]. / Sustoianie na tsentralnata khemodinamika pri bremenni zheni s preeklampsiia.

Radionuclidic method is used in women with preeclampsia to evaluate the state of the central hemodynamics. The examined women are divided into 6 groups. The results are presented on 2 tables with 23 investigated parameters as intergroup correlation is made. The obtained results show the presence of hypoglycemia, increased peripheral resistance, etc., which are in direct dependence on the degree, gravity and duration of the disease. It is possible to discover occurring changes quite early by this method and thus to begin timely proper therapy, which reduces perinatal morbidity and mortality. It is very important that data are obtained by this method, which could not be determined by other methods. However its usage should be performed electively by very strict indications after the possibilities of other methods are tried (ultrasound, rheography, ect.).

[Signs of developmental dysplasia and function of the genetic apparatus of children and adolescents]. / Priznaki displazii razvitiia i sostoianie geneticheskogo apparata u detei i podrostkov.

Using scanning cytospectrophotometry and radiometry of beta-particles of 3H-thymidine incorporated into DNA the authors studied chromatin in interphase nuclei of lymphocytes in children and adolescents with Down's syndrome, childhood cerebral paralysis, and undifferentiated oligophrenia, as well as in healthy subjects. The findings obtained are compared with the results of cytogenetic examination of metaphase chromosomes and the number of stigmas of dysembryogenesis. Aberrations of metaphase chromosomes were found in 4.4% of patients with childhood cerebral paralysis and oligophrenia while the pattern of changes in interphase chromatin corresponding to the chromosomal imbalance was detected in 36% of these individuals. In all cases they had from 15 to 30 stigmas of dysembryogenesis.

[Discriminant analysis of methods for detecting the heterozygote carrier state for the Duchenne muscular dystrophy gene]. / Diskriminantnyi analiz metodov vyiavleniia geterozigotnogo nositel'stva gena myshechnoi distrofii Diushenna.

The authors used the procedure of a step-wise discriminant analysis for comparing the informative value of different methods for revealing heterozygotic carriers of the gene of Duchenne's myodystrophy by means of an analysis of the blood serum and physicochemical properties of erythrocytes in 11 mothers suffering from Duchenne's muscular dystrophy who were obligate (by the findings of a genealogical analysis) carriers of the gene of Duchenne's myodystrophy. Employment of a complex of four methods (determination of the constant of the rate of chloride-bicarbonate metabolism through the erythrocyte membrane, erythrocytic deformability, hemolytic stability of erythrocytes upon their heating at 55 degrees C, and analysis of the activity of serum creatine kinase) has increased 1.8-fold the rate of detecting heterozygotic carriers of the gene of Duchenne's myodystrophy as compared to the creatine kinase test.

[Study of the kinetics of anion metabolism in the erythrocytes of patients with various forms of hereditary neuromuscular diseases]. / Izuchenie kinetiki anionnogo obmena v éritrotsitakh bol'nykh s razlichnymi formami nasledstvennykh nervno-myshechnykh zabolevanii.

The authors have shown an increase in the rate of the anion transport in erythrocytic membranes in patients with hereditary neuromuscular diseases, as well as in probable heterozygotic carriers of Duchenne's progressive muscular dystrophy. It is suggested that the kinetic parameters of chlorine-bicarbonate exchange be used for the early diagnosis and identification of the heterozygotic carriership of Duchenne's myodystrophy gene.

[Chemiluminescence of blood plasma and functional status of the erythrocytes of patients with various hereditary neuromuscular diseases]. / Khemiliuminestsentsiia plazmy krovi i funktsional'noe sostoianie éritrotsitov u bol'nykh s nekotorymi nasledstvennymi nervno-myshechnymi zabolevaniiami.

The authors studied the parameters of osmotic resistance and deformability of erythrocytes in patients with Duchenne's, Bekker's, Erb-Roth's and Landouzy-Déjérine's progressive muscular dystrophies, with Charcot-Marie's spinal amyotrophy, Kugelberg-Welander neural amyotrophy and with Thomsen's myotonia. Along with an increase in the erythrocytic osmotic resistance and deformability patients with different forms of hereditary neuromuscular diseases showed decreased plasma chemiluminescence (PCL) as well as an elevated ability of the blood plasma to inhibit PCL in the model chemiluminescence system in patients with Duchenne's progressive muscular dystrophy and in their close relatives.

[Destruction of muscle tissue and purine compound metabolism in hereditary muscular dystrophy]. / Destruktsiia myshechnoi tkani i obmen purinovykh soedinenii pri nasledstvennykh myshechnykh distrofiiakh.

The distribution of purine compounds in the skeletal muscles of the anterior and posterior limbs of 129/Re mice with hereditary muscular dystrophy (HMD) was investigated in a comparative study. The results revealed unidirectional metabolic disorders in both groups of muscles which was manifested in a quantitative redistribution of phosphorus-containing purine components and a decrease in the pool of adenylates in muscle tissue. Elevated concentrations of purine metabolites (inosine, hypoxanthine, xanthine, and uric acid) indicated an augmented metabolism of purines in this abnormality. Impairment of metabolic transformations of purines in the muscle tissue in HMD may be one of the factors responsible for the development of the pathological process in the muscle in the given disease.

[Metabolism of purine compounds in skeletal muscle and blood and physical properties of the erythrocytes of mice with hereditary muscular dystrophy]. / Obmen purinovykh soedinenii v skeletnoi myshtse i krovi i fizicheskie svoistva éritrotsitov u myshei s nasledstvennoi myshechnoi distrofieí.

The authors studied the metabolism of purine compounds in the skeletal muscle of 129 Re mice with hereditary muscular dystrophy (MD). The study showed impairment of purine metabolism which was expressed in a sharp decrease in ATP levels and an increase in the content of AMP, IMP and uric acid. No changes were revealed in the pool of purine nucleotides in murine red blood cells. A study of some physical properties of the red blood cells in mice with myopathy showed no alterations in the osmotic resistance of erythrocytes, yet there was a reduction in their pliability as compared to control. Examination of the temperature resistance revealed anomalies of red blood cells in myodystrophic mice at 50 degrees C. The detected changes of some physical properties of erythrocytes seem to be related to abnormalities of the sumbembranous contractile apparatus of these cells.