[The First Russian Consensus on the Multilevel Abobotulinumtoxin A Injections in Spastic Forms of Cerebral Palsy]. / Pervyi Rossiiskii konsensus po primeneniyu mnogourovnevykh in''ektsii abobotulinumtoxin A pri lechenii spasticheskikh form detskogo tserebral'nogo paralicha.

Spasticity treatment is one of the key aspects of the contemporary cerebral palsy (CP) rehabilitation that influences on the effectiveness of other methods. The paper presents the first Russian document that unites the recommendations for the BTA treatment of CP and could be used as the guideline for the multilevel injections. The Russian consensus on the multilevel botulinum toxin A (BTA) treatment of spastic CP is based on the international data and the results of national studies. The authors describe typical CP spasticity patterns in the upper and lower extremities, give recommended intervals for the BTA (Abobotulinum toxin A) dosages for the whole injection procedure and for the separate muscles. The method of dosage calculation for functional segments is also described. Attention is paid to the frequency, optimal intervals between the repeated injections and the whole duration of BTA treatment. The authors discuss effectiveness and safety of BTA, factors that potentially influence the results of the injections, including ultrasound and electromyography control, and indications for the continuation and termination of treatment.

[The results of single-event multilevel orthopedic surgeries and the early rehabilitation used in complex with botulinum toxin treatment in patients with spastic forms of cerebral palsy].

OBJECTIVE: To evaluate motor possibilities of patients with children spastic palsy (CSP) one year after single-event multilevel orthopedic low extremity surgeries in combination with early rehabilitation treatment including botulinum toxin treatment. MATERIAL AND METHODS: Authors studied the results of operative orthopedic treatment in 55 patients with CSP, aged from 5 to 17 years (mean 11.9 ± 2.5 years), who underwent multilevel surgeries with early functional rehabilitation using 1.5 treatment courses with 6-8 week treatment-free periods during 9-12 months in combination with a single injection of disport in the post-operative period. We performed 74 surgeries 140 episodes of botulinum toxin treatment using average doses of Botulinum toxin 10 U per kg of body mass injected into low extremity muscles. RESULTS: In the post-operative period, authors recorded a significant reduction in pain syndrome, assessed with a pain intensity scale, from 8.6 ± 1.2 to 4.3 ± 1.1 scores (p < 0.001). An analysis of gait demonstrated an improvement of gait patterns in all patients able to move independently. An increase in motor abilities, measured with the Gillette Functional Assessment Questionnaire, by 1 level was identified in 28 (50.9%) patients, by 2 levels in 2 (3.6%) patients, no changes were observed in 25 (45.5%) patients. CONCLUSION: Single-event multilevel orthopedic surgeries in children with CSP reduce a number of repeated surgeries. The effective control over pain syndrome in patients with CSP using multilevel botulinum toxin treatment in the post-operative period promotes the functional rehabilitation, increases rates of loading during training sessions and the motivation of CSP patients to restore the lost activity.

Specific regions of the extracellular domain of dlk, an EGF-like homeotic protein involved in differentiation, participate in intramolecular interactions.

The level of expression of dlk, an EGF-like protein possessing six EGF-like repeats in its extracellular region, is critical for 3T3-L1 fibroblasts to differentiate into adipocytes in response to IGF1. The mechanism of action of dlk is not well understood, but its localization on the cell membrane suggests that dlk may function as a receptor, as a ligand or as a regulatory protein modulating the binding, the signaling, or the expression of other molecules involved in cell differentiation and growth. In this work, we demonstrate, by using the Yeast Two-Hybrid system, that dlk interacts with itself through specific regions of its extracellular domain. The strongest interactions were observed between specific EGF-like repeats and between a non EFG-like region where unknown proteases act to generate soluble forms of dlk. These observations suggest that the interaction between two membrane dlk molecules belonging to the same or to different cells, or the interaction between soluble and membrane dlk variants, may be important to regulate dlk function.

betac cytokine receptor-induced stimulation of cAMP response element binding protein phosphorylation requires protein kinase C in myeloid cells: a novel cytokine signal transduction cascade.

We have recently shown that IL-3R occupancy activates a phosphatidylcholine-specific phospholipase C, and the sustained diacylglycerol accumulation subsequently activates protein kinase C (PKC). In human IL-3-dependent myeloid cells (TF-1), the novel PKCepsilon isoform regulates bcl-2 expression and cell survival. The report of a PKC activatable cAMP response element (CRE) in the bcl-2 promoter and a role for PKC in bcl-2 expression in B cells led us to the hypothesis that PKC phosphorylation activates transcription factor CREB after IL-3R engagement. We found that IL-3 and GM-CSF induced phosphorylation of CREB on Ser(133) in TF-1 cells, and this phosphorylation was blocked by two structurally unrelated classes of PKC inhibitors. An inhibitor of cyclic nucleotide-dependent kinases did not block this phosphorylation. IL-4, which is biologically active in these cells but does not use the beta common subunit, did not phosphorylate CREB on Ser(133). Inhibition of mitogen-activated protein kinase kinase activity also inhibited IL3-induced CREB phosphorylation. The PKC inhibitors, but not a cyclic nucleotide-dependent kinase inhibitor, blocked IL-3 activation of CRE-dependent transcription from an egr-1 promoter/chloramphenicol acetyltransferase (CAT) reporter construction transiently transfected into TF-1 cells. Finally, TF-1 cells stably overexpressing PKCepsilon, but not the delta isoform of PKC, enhanced CRE-dependent CAT expression from the promoter/reporter construction. Therefore, it is likely that a PKCepsilon kinase cascade resulting in CREB phosphorylation represents a novel signal transduction cascade for regulating cellular gene expression through the beta common cytokine receptor.

Kinetics and thermodynamics of beta 2-microglobulin binding to the alpha 3 domain of major histocompatibility complex class I heavy chain.

The major histocompatibility complex (MHC) class I molecule plays a crucial role in cytotoxic lymphocyte function. Functional class I MHC exists as a heterotrimer consisting of the MHC class I heavy chain, an antigenic peptide fragment, and beta2-microglobulin (beta2m). beta2m has been previously shown to play an important role in the folding of the MHC heavy chain without continued beta2m association with the MHC complex. Therefore, beta2m is both a structural component of the MHC complex and a chaperone-like molecule for MHC folding. In this study we provide data supporting a model in which the chaperone-like role of beta2m is dependent on initial binding to only one of the two beta2m interfaces with class 1 heavy chain. beta2-Microglobulin binding to an isolated alpha3 domain of the class I MHC heavy chain accurately models the biochemistry and thermodynamics of beta2m-driven refolding. Our results explain a 1000-fold discrepancy between beta2m binding and refolding of MHC1. The biochemical study of the individual domains of complex molecules is an important strategy for understanding their dynamic structure and multiple functions.

ICAM-1 enhances MHC-peptide activation of CD8(+) T cells without an organized immunological synapse.

In addition to the TCR-ligand interaction, other receptor-ligand pairs, such as LFA-1 and ICAM-1, play a major role in the activation of T cells. Recent studies of T cell activation suggest a coordinated movement of LFA-1 and ICAM-1 in forming a defined zone in the immunological synapse. It is unclear from these studies whether the organized molecular geometry of the immunological synapse is necessary for ICAM-1 enhancement of T cell activation. In this report, we demonstrate that ICAM-1 can enhance the activation of CD8(+) T cells by MHC-peptide in the absence of an organized immunologic synapse. Therefore, although the molecular organization of the immunologic synapse may amplify stimuli, it is not an absolute requirement for either CD8(+) T cell activation or the ICAM-1 enhancement of TCR activation.

[Metabolic effects of berlipril-5 in patients with non-insulin dependent diabetes mellitus and arterial hypertension]. / Issledovanie metabolicheskikh éffektov berliprila-5 u bol'nykh insulinonezavisimym sakharnym diabetom i arterial'noi gipertoniei.

AIM: To study metabolic effects of berlipril-5 (enalapril) in patients with non-insulin-dependent diabetes mellitus (NIDDM) and arterial hypertension (AH). MATERIALS AND METHODS: 24 patients with NIDDM and AH were divided into three groups by the level of basal C-peptide: > 2 ng/ml (group 1), 2-4 ng/ml (group 2) and < 4 ng/ml (group 3). RESULTS: A correlation was found between the level of basal C-peptide and duration of AH (r = 0.7) and NIDDM (r = -0.47), between the level of triglycerides (TG) and glycolized hemoglobin Hb A1c (r = 0.48). Berlipril treatment reduced basal C-peptide level in groups 2 and 3 by 20.65 +/- 1.95% and elevated it in group 1 by 16.4 +/- 1.5%. Fasting glucose levels lowered by 9.2 +/- 1.95% indicating better sensitivity of the liver to insulin. Blood glucose levels 2 hours after meal fell by 8.3 +/- 0.95% (p < 0.05) and Hb A1c by 8.14 +/- 1.25% showing indirectly diminishing insulin-resistance at the level of peripheral tissues. TG and VLDLP significantly declined. CONCLUSION: Inhibitors of angiotensin converting enzyme (enalapril, in particular) produce a positive effect on carbohydrate and lipid metabolism in patients with NIDDM and AH.

[Effect of brerlipril in patients with non-insulin dependent diabetes mellitus and mild or moderate arterial hypertension as indicated by 24-h blood pressure monitoring]. / Effektivnost' berliprila u bol'nykh insulinonezavisimym sakharnym diabetom v sochetanii s miagkoi i umerennoi arterial'noi gipertoniei po dannym sutochnogo monitorirovaniia arterial'nogo davleniia.

AIM: To study a hypotensive effect of berlipril, an ACE inhibitor, using 24-h BP monitoring in patients with non-insulin-dependent diabetes mellitus (NIDDM) combined with stable mild or moderate arterial hypertension (AH). MATERIALS AND METHODS: 22 NIDDM patients with mild or moderate AH were treated with berlipril. 24-h monitoring of BP was made in all the patients before and 3 months after the treatment. RESULTS: A stable hypotensive effect of berlipril was achieved at its therapeutic dose 1.5 tablets a day once a day or divided into two doses a day. On treatment week 3-4 a hypotensive effect of berlipril enhanced in 5 patients. This may be due to ACE inhibitors action on the tissue component of the reninangiotensin system. CONCLUSION: Berlipril produces a high hypotensive effect and brings about a positive response of insulin-resistance in NIDDM associated with mild and moderate AH.

A phosphatidylcholine phospholipase C inhibitor, D609, blocks interleukin-3 (IL-3)-induced bcl-2 expression but not c-myc expression in human IL-3-dependent cells.

Activation of the interleukin-3 (IL-3) receptor is required for the induction of cell proliferation and suppression of apoptosis in primitive hematopoietic progenitor cells. A rapid activation of tyrosine kinases and a phosphatidylcholine-specific phospholipase C has been observed in these cells in response to IL-3. The signal transduction cascades regulating cell proliferation and the suppression of apoptosis are poorly understood. Using human IL-3-dependent TF-1 cells, we have found that the tyrosine kinase inhibitor genistein blocks both the IL-3 suppression of apoptosis and the expression of the cell survival gene bcl-2. In addition, we have found that D609, a specific inhibitor of phosphatidylcholine-specific phospholipase C, also inhibits IL-3-induced expression of the bcl-2 gene without affecting IL-3-induced tyrosine phosphorylation. D609 also drove these cells into apoptosis even in the presence of IL-3. Significantly, genistein inhibited the IL-3 induction of both bcl-2 and c-myc gene. The latter gene is related to the induction of cell proliferation. D609, however, blocked the induction only of the cell survival gene bcl-2. Thus, phosphatidylcholine hydrolysis appears linked to the induction of genes related to cell survival. These data fit with the hypothesis that there is a bifurcation in the signaling pathways downstream of IL-3 receptor-induced tyrosine phosphorylation.

Overexpression of protein kinase C isoform epsilon but not delta in human interleukin-3-dependent cells suppresses apoptosis and induces bcl-2 expression.

Hematopoietic progenitor cells die by apoptosis after removal of the appropriate colony-stimulating factor (CSF). Recent pharmacologic data have implicated protein kinase C (PKC) in the suppression of apoptosis in interleukin-3 (IL-3) and granulocyte-macrophage (GM)-CSF-dependent human myeloid cells. Because IL-3 and GM-CSF induce increases in diacylglycerol without mobilizing intracellular Ca++, it seemed that one of the novel Ca++ independent isoforms of PKC was involved. We report here that overexpression of PKC epsilon in factor-dependent human TF-1 cells extends cell survival in the absence of cytokine. Overexpression of PKC delta does not have this effect. By 72 to 96 hours after cytokine withdrawal, the PKC epsilon transfectants remain distributed in all phases of the cell cycle, as shown by fluorescence-activated cell sorting (FACS) analysis, while little intact cellular DNA is detectable in vector or PKC delta transfectants. PKC epsilon induces bcl-2 protein expression fivefold to sixfold over the levels in empty vector transfectants, whereas the levels in PKC delta transfectants are similar to those in vector controls.

[Elaboration of a rapid method of determining the sensitivity of Brucella to antibiotics]. / Razrabotka ékspressnogo metoda opredeleniia antibiotiko- chuvstvitel'nosti brutsell.

Optimal conditions were developed for determination of antibiotic sensitivity in Brucella by using enzyme immunoassay directly in the primary cultures of the material tested. The Brucella concentration in the material tested should be not lower than 1.10(6) microbial cells/ml and the time of culture incubation be 24 hours at 37 degrees C. The obligatory condition is to use a liquid medium, i.e. the Albimi broth with 1% glucose. To inhibit the foreign microflora it is recommended to use polymyxin B and amphoglucamine in a concentration of 3 microgram/ml. The use of enzyme immunoassay was shown that it was possible to determine the antibiotic sensitivity of Brucella in practice.

[The possibility of creating a vaccinal strain of Brucella abortus 19-BA with multiple antibiotic resistance]. / O vozmozhnosti sozdaniia vaktsinnogo shtamma Brucella abortus 19-BA s mnozhestvennoi ustoichivost'iu k antibiotikam.

The possibility of preparing B. abortus vaccine strain 19-BA with multiresistance to antibiotics was shown. The strain was obtained by the spontaneous induction of resistance to rifampicin with the subsequent transformation of nonconjugative hybrid plasmid pOVI which, in addition to rifampicin resistance, governed the resistance of brucellae to tetracycline, doxycycline, ampicillin, and streptomycin. Experiments on guinea pigs demonstrated the immunization with both multiresistant vaccine strain GSA1 and B. abortus initial vaccine strain 19-BA.

[A rapid immunofluorescent method of determining the antibiotic sensitivity of brucella]. / Immunofliuorestsentnyi ékspress-metod opredeleniia antibiotikochuvstvitel'nosti brutsell.

Optimal conditions for rapid assay of Brucella antibiotic sensitivity with the immunofluorescent method were developed. With this method high sensitivity of the main Brucella species to tetracycline, doxycycline and rifampicin was confirmed. It was found actually possible to use the immunofluorescent method for rapid assay of Brucella antibiotic sensitivity in practice.

[Trial of different nutrient media in studying the antibiotic sensitivity of brucellae]. / Ispytanie razlichnykh pitatel'nykh sred pri izuchenii antibiotikochuvstvitel'nosti brutsell.

The possible use of the Unimicon-s nutrient medium for determination of antibiotic sensitivity of Brucella by the methods of serial dilutions in dishes and agar diffusion with paper disks was studied. Previously this method was not used in manipulations with Brucella. The results indicated that the dry Unimicon-s nutrient medium was applicable for determination of Brucella antibiotic sensitivity. It was shown that the agar diffusion method with the use of paper disks may be recommended for determination of antibiotic sensitivity of Brucella. The optimal time for recording the results of the laboratory assays is 48 hours.

[Use of acidic rose bengal antigen in the plate agglutination test for brucellosis in humans]. / Isopol'zovanie kislogo antigen roz-bengal v plastinchatoi reakstii aggliutinatsii pri brutselleze u liudei.

The results obtained in the study of the activity of an antigen stained with rose bengale in the plate agglutination test are presented. The analysis of 1309 serum samples from healthy persons and brucellosis patients has shown the high specificity and diagnostic sensitivity of the rose bengale test. The evaluation of the positive result of the rose bengale test should be differentiated in accordance with the degree of agglutination observed in the test. The correlation between the degree of agglutination with the rose bengale antigen and the titer value in Wright's test has been established. The new accelerated test is recommended for large-scale approbation in clinical conditions, epidemiological practice and in the examination of donors at blood donor centers.