Morphological Changes in Tissue When Using Polypropylene Implants with Adsorbed Multipotent Stromal Cells in Experiment.

The subcutaneous tissue of rats after implantation of polypropylene materials with adsorbed bone marrow-derived mesenchymal multipotent stromal cells (MMSCs) was studied using light microscopy. Inflammation in response to implantation was mild, and the foreign material was encapsulated into a thin strip of dense fibrous connective tissue with multinucleated macrophages. By 1 year after introduction of the monofilament and 6 and 12 months after implantation of the mesh product, some threads were deformed, broken, and had sharp edges. Small fragments of foreign material appeared in the adjacent tissues surrounded by their own relatively thick acellular capsule. As a result of preliminary adsorption of MMSCs on polypropylene, the thickness of the connective tissue capsule decreased, its vascularization increased, and the severity of inflammatory infiltration decreased. However, all effects of MMSCs adsorption in rats were transient and disappeared within 1 week.

VALUE OF AMBULATORY BLOOD PRESSURE MONITORING IN THE VERIFICATION OF ARTERIAL HYPERTENSION IN SCHOOL AGE CHILDREN.

Arterial hypertension is a common pathology in children of different ages. The introduction of daily monitoring of blood pressure into the practice of pediatric cardiologists makes it possible to more accurately establish a diagnosis, determine the prognosis of the course of the disease and monitor treatment of hypertension. Objective - to assess the daily fluctuations of blood pressure in schoolchildren with arterial hypertension. 70 children of school age were examined. The main group (38 people) included children with high blood pressure, the control group included 32 clinically healthy children. All children underwent tonometry. The results for each child evaluated by percentile nomograms regarding age, gender and height. Verification of the diagnosis of arterial hypertension performed according to the recommendations of the American Academy of Pediatrics (AAP). In addition, children underwent ambulatory blood pressure monitoring. In 79% of children of the main group, the level of blood pressure assessed as arterial hypertension of the first stage, in 21% of children - arterial hypertension of the second stage. When conducting daily monitoring of blood pressure in 35 children (92.1%) of the main group, 2 peaks of systolic blood pressure observed: the first peak between 23:00 and 01:00 at night (from 5.5 to 18.8 mm Hg.), the second peak - in 28 children (73.7%) between 6.30 and 8.00 (from 6.8 to 10.1 mm Hg). At the same time, peaks in the level of diastolic blood pressure appeared in fewer children and were not so pronounced. In schoolchildren with stage 1 hypertension, a night peak observed in 60% of children, and a morning peak was in 22% of children. Among children with second stage of arterial hypertension a night peak observed in 100% persons and a morning peak observed in 72% of children. This suggests that the nocturnal peak of blood pressure may be a marker of the severity of arterial hypertension. In healthy children, there were no peaks in the rise in blood pressure. The presence of a non-dipper circadian profile in a school-age child in combination with the morning and/or night peak of systolic blood pressure can serve as a marker for the development of arterial hypertension. Therefore, such children must be attributed to the risk group for the development of this pathology.

[Pattern of excitation in isolated heart of hibernator ground squirrel Citellus undulatus].

Aim of our study was to measure conduction velocity and pattern of excitation during hypothermia in hearts of ground squirrels Citellus undulatus, known to be most resilient hibernators. We imaged electrical conduction in intact isolated hearts of summer active and winter hibernating ground squirrels at temperatures varying from +37 degrees C to +3 degrees C. Electrical activity was mapped using CCD camera (500 frames/sec) and voltage-sensitive dye di-4-ANEPPS during normal sinus rhythm and ventricular pacing. No spontaneous tachyarrhythmia was observed in all hearts at any temperature. Hearts were able to maintain spontaneous sinus rhythm and normal pattern of epicardial excitation throughout the whole range of studied temperatures. Despite responsiveness to pacing in all hearts ventricular conduction velocity was significantly reduced (about 10-fold) at low temperatures +3 degrees C. Our data provides the first direct demonstration that isolated heart of the summer active and winter hibernating ground squirrel Citellus undulatus is able to maintain normal excitation pattern in a range of temperatures from +37 degrees C to +3 degrees C.

[Clinical effectiveness and safety of citalopram in patients with depression after myocardial infarction]. / Klinicheskaia éffektivnost' i bezopasnost' tsitaloprama u bol'nykh s depressiei posle perenesennogo infarkta miokarda.

Citalopram (20 mg/day for 42 days) was given to 20 patients (mean age 54.4+/-2.5 years) with depression after myocardial infarction. This was associated with substantial antidepressive effect (50% and more lowering of the total score of the Beck depression questionnaire) in 89% of patients, reduction of number and severity of somatic complaints, and improvement of parameters of quality of life. Citalopram did not affect blood pressure and according to Holter ECG monitoring data produced no arrhythmogenic or proischemic effects. Overall tolerability of citalopram was good however 21% of patients experienced slight drowsiness, dizziness or sweating.

Expression in plants and immunogenicity of plant virus-based experimental rabies vaccine.

A new approach to the production and delivery of vaccine antigens is the use of engineered amino virus-based vectors. A chimeric peptide containing antigenic determinants from rabies virus glycoprotein (G protein) (amino acids 253-275) and nucleoprotein (N protein) (amino acids 404-418) was PCR-amplified and cloned as a translational fusion product with the alfalfa mosaic virus (AlMV) coat protein (CP). This recombinant CP was expressed in two plant virus-based expression systems. The first one utilized transgenic Nicotiana tabacum cv. Samsun NN plants providing replicative functions in trans for full-length infectious RNA3 of AlMV (NF1-g24). The second one utilized Nicotiana benthamiana and spinach (Spinacia oleracea) plants using autonomously replicating tobacco mosaic virus (TMV) lacking native CP (Av/A4-g24). Recombinant virus containing the chimeric rabies virus epitope was isolated from infected transgenic N. tabacum cv. Samsun NN plants and used for parenteral immunization of mice. Mice immunized with recombinant virus were protected against challenge infection. Based on the previously demonstrated efficacy of this plant virus-based experimental rabies vaccine when orally administered to mice in virus-infected unprocessed raw spinach leaves, we assessed its efficacy in human volunteers. Three of five volunteers who had previously been immunized against rabies virus with a conventional vaccine specifically responded against the peptide antigen after ingesting spinach leaves infected with the recombinant virus. When rabies virus non-immune individuals were fed the same material, 5/9 demonstrated significant antibody responses to either rabies virus or AlMV. Following a single dose of conventional rabies virus vaccine, three of these individuals showed detectable levels of rabies virus-neutralizing antibodies, whereas none of five controls revealed these antibodies. These findings provide clear indication of the potential of the plant virus-based expression systems as supplementary oral booster for rabies vaccinations.

Inactivation of peroxynitrite in multiple sclerosis patients after oral administration of inosine may suggest possible approaches to therapy of the disease.

Peroxynitrite has been implicated in the pathogenesis of multiple sclerosis (MS) and its animal model experimental allergic encephalomyelitis (EAE). Previously, we have shown that administration of uric acid (UA), a peroxynitrite scavenger, is therapeutic in EAE We have also shown that MS patients have lower levels of serum uric acid than healthy individuals or those with other neurological diseases. The aim of this investigation was therefore to raise serum UA levels in MS patients. Oral administration of UA failed to increase low serum UA levels, evidently due to its degradation by gastrointestinal bacteria. However, serum UA could be raised and maintained at elevated levels for a year and more without reported side-effects by oral administration of its precursor inosine. Three of 11 patients given inosine showed some evidence of clinical improvement and there was no sign of disease progression in the remaining patients. Gadolinium-enhanced lesions, observed in two patients before receiving inosine, could not be detected after either 10 or IS months inosine treatment These data provide evidence that serum UA levels can be readily manipulated and that the benefit of higher levels to individuals with MS should be studied further in greater number of patients.

The central nervous system inflammatory response to neurotropic virus infection is peroxynitrite dependent.

We have recently demonstrated that increased blood-CNS barrier permeability and CNS inflammation in a conventional mouse model of experimental allergic encephalomyelitis are dependent upon the production of peroxynitrite (ONOO(-)), a product of the free radicals NO* and superoxide (O2*(-)). To determine whether this is a reflection of the physiological contribution of ONOO(-) to an immune response against a neurotropic pathogen, we have assessed the effects on adult rats acutely infected with Borna disease virus (BDV) of administration of uric acid (UA), an inhibitor of select chemical reactions associated with ONOO(-). The pathogenesis of acute Borna disease in immunocompetent adult rats results from the immune response to the neurotropic BDV, rather than the direct effects of BDV infection of neurons. An important stage in the BDV-specific neuroimmune response is the invasion of inflammatory cells into the CNS. UA treatment inhibited the onset of clinical disease, and prevented the elevated blood-brain barrier permeability as well as CNS inflammation seen in control-treated BDV-infected rats. The replication and spread of BDV in the CNS were unchanged by the administration of UA, and only minimal effects on the immune response to BDV Ags were observed. These results indicate that the CNS inflammatory response to neurotropic virus infection is likely to be dependent upon the activity of ONOO(-) or its products on the blood-brain barrier.

Uric acid levels in patients with multiple sclerosis: analysis in mono- and dizygotic twins.

Presence of nitrotyrosine in cells surrounding plaques indicates that peroxynitrite may be the cause of brain lesions in multiple sclerosis. Low levels of uric acid, a natural scavenger of peroxynitrite, were demonstrated in blood of patients with multiple sclerosis in comparison with control individuals. These observations were now extended to 132 sets of twins with one sibling affected by multiple sclerosis. In blood of both mono- and dizygotic twins the uric acid levels were lower in the twin with the disease than in the healthy twin.

The peroxynitrite scavenger uric acid prevents inflammatory cell invasion into the central nervous system in experimental allergic encephalomyelitis through maintenance of blood-central nervous system barrier integrity.

Uric acid (UA), a product of purine metabolism, is a known scavenger of peroxynitrite (ONOO(-)), which has been implicated in the pathogenesis of multiple sclerosis and experimental allergic encephalomyelitis (EAE). To determine whether the known therapeutic action of UA in EAE is mediated through its capacity to inactivate ONOO(-) or some other immunoregulatory phenomenon, the effects of UA on Ag presentation, T cell reactivity, Ab production, and evidence of CNS inflammation were assessed. The inclusion of physiological levels of UA in culture effectively inhibited ONOO(-)-mediated oxidation as well as tyrosine nitration, which has been associated with damage in EAE and multiple sclerosis, but had no inhibitory effect on the T cell-proliferative response to myelin basic protein (MBP) or on APC function. In addition, UA treatment was found to have no notable effect on the development of the immune response to MBP in vivo, as measured by the production of MBP-specific Ab and the induction of MBP-specific T cells. The appearance of cells expressing mRNA for inducible NO synthase in the circulation of MBP-immunized mice was also unaffected by UA treatment. However, in UA-treated animals, the blood-CNS barrier breakdown normally associated with EAE did not occur, and inducible NO synthase-positive cells most often failed to reach CNS tissue. These findings are consistent with the notion that UA is therapeutic in EAE by inactivating ONOO(-), or a related molecule, which is produced by activated monocytes and contributes to both enhanced blood-CNS barrier permeability as well as CNS tissue pathology.

[The effects of combined electrolytic brain stem-orbitofrontal and brain stem-hippocampal damages to the rat brain]. / Effekty sochetannogo élektroliyicheskogo stvolovo-orbitofrontal'nogo i stvolovo-gippokampal'nogo povrezhdeniia mozga krys.

Morphofunctional studies of animals with associated electrolytic orbitofrontal and hippocampal brainstem lesions as compared to variants of isolated brainstem coagulation showed participation and specific role of orbitofrontal cortex and hippocampus in adaptive-compensatory brain reactions of rats with brainstem lesions. Associated brainstem-orbitofrontal damages result in aggravation of the animals' condition and highly probable lethality within the first two weeks after surgery due to blood circulation dysregulation of hemorrhagic type and probably due to secondary hypothalamus dysfunction. Associated brainstem-hippocampal coagulation intensifies primarily brainstem neurologic symptoms and prolongs time of their reverse development, i.e. supports the brainstem centre of stable pathological activity.

Protection of myelin basic protein immunized mice from free-radical mediated inflammatory cell invasion of the central nervous system by the natural peroxynitrite scavenger uric acid.

Peroxynitrite (ONOO(-)), the product of nitric oxide (NO(radical)) and superoxide (O(2)(-radical)), is believed to be a major contributor to immunotoxicity when produced by activated cells expressing inducible nitric oxide synthase (iNOS). Uric acid (UA) is a natural scavenger of ONOO(-) that is present at high levels in the sera of humans and other higher order primates relative to most lower mammals. We have previously shown that UA treatment is therapeutic in experimental allergic encephalomyelitis (EAE), a rodent model of multiple sclerosis (MS). In this study we have examined the effect of UA therapy on the dynamics of the appearance of iNOS-positive cells in central nervous system (CNS) tissue of mice subjected to the stimuli that cause EAE. The results indicate that UA prevents activated monocytes from entering CNS tissue where they may contribute to the pathogenesis of MS and other CNS diseases.

Uric acid, a peroxynitrite scavenger, inhibits CNS inflammation, blood-CNS barrier permeability changes, and tissue damage in a mouse model of multiple sclerosis.

Peroxynitrite (ONOO(-)), a toxic product of the free radicals nitric oxide and superoxide, has been implicated in the pathogenesis of CNS inflammatory diseases, including multiple sclerosis and its animal correlate experimental autoimmune encephalomyelitis (EAE). In this study we have assessed the mode of action of uric acid (UA), a purine metabolite and ONOO(-) scavenger, in the treatment of EAE. We show that if administered to mice before the onset of clinical EAE, UA interferes with the invasion of inflammatory cells into the CNS and prevents development of the disease. In mice with active EAE, exogenously administered UA penetrates the already compromised blood-CNS barrier, blocks ONOO(-)-mediated tyrosine nitration and apoptotic cell death in areas of inflammation in spinal cord tissues and promotes recovery of the animals. Moreover, UA treatment suppresses the enhanced blood-CNS barrier permeability characteristic of EAE. We postulate that UA acts at two levels in EAE: 1) by protecting the integrity of the blood-CNS barrier from ONOO(-)-induced permeability changes such that cell invasion and the resulting pathology is minimized; and 2) through a compromised blood-CNS barrier, by scavenging the ONOO(-) directly responsible for CNS tissue damage and death.

Expression of alfalfa mosaic virus coat protein in tobacco mosaic virus (TMV) deficient in the production of its native coat protein supports long-distance movement of a chimeric TMV.

Alfalfa mosaic virus (AlMV) coat protein is involved in systemic infection of host plants, and a specific mutation in this gene prevents the virus from moving into the upper uninoculated leaves. The coat protein also is required for different viral functions during early and late infection. To study the role of the coat protein in long-distance movement of AlMV independent of other vital functions during virus infection, we cloned the gene encoding the coat protein of AlMV into a tobacco mosaic virus (TMV)-based vector Av. This vector is deficient in long-distance movement and is limited to locally inoculated leaves because of the lack of native TMV coat protein. Expression of AlMV coat protein, directed by the subgenomic promoter of TMV coat protein in Av, supported systemic infection with the chimeric virus in Nicotiana benthamiana, Nicotiana tabacum MD609, and Spinacia oleracea. The host range of TMV was extended to include spinach as a permissive host. Here we report the alteration of a host range by incorporating genetic determinants from another virus.

Definition of a natural killer NKR-P1A+/CD56-/CD16- functionally immature human NK cell subset that differentiates in vitro in the presence of interleukin 12.

Human natural killer (NK) cell differentiation from immature lineage negative (Lin-) umbilical cord blood cells was examined in vitro. Cells expressing differentiation antigens of mature NK cells (CD56, CD16, CD2, CD8, NKR-P1A) were generated from Lin- cells cultured with interleukin (IL)-2 and a murine bone marrow stromal cell line expressing the human membrane-bound form of stem cell factor. Two subsets of NK cells were identified in these cultures: one expressed both NKR-P1A and CD56 and, in variable proportions, all other NK cell differentiation antigens; the second subset expressed only NKR-P1A and, unlike the former, was not cytotoxic. Neither subset expressed interferon (IFN)-gamma mRNA even after stimulation with phorbol di-ester and Ca2+ ionophore, but both expressed tumor necrosis factor alpha mRNA and the cytotoxic granule-associated proteins TIA-1, perforin, and serine esterase-1. After 10-d culture with IL-2, IL-12, and irradiated B lymphoblastoid cells, approximately 45% of the NKR-P1A+/ CD56- cells became CD56+, and the same cultures contained cells capable of cytotoxicity and of IFN-gamma production. These results indicate that NKR-P1A expression in the absence of other NK cell markers defines an intermediate, functionally immature stage of NK cell differentiation, and that effector functions develop in these cells, concomitantly with CD56 expression, in the presence of IL-12. These cells likely represent the counterpart of a CD3-/NKR-P1A+/ CD56-/CD16- cell subset that, as shown here, is present both in adult and neonatal circulating lymphocytes.

[Local destruction of Deiters' lateral vestibular nucleus (an experimental model of a focal brain stem lesion)]. / Lokal'noe razrushenie lateral'nogo vestibuliarnogo iadra Deistersa (éksperimental'naia model' ochagovogo porazheniia stvola mozga).

The experimental rat model of the local brainstem destruction by isolated electrical coagulation of the lateral vestibular nucleus Deiters was offered for studying the adaptive-compensatory CNS reactions after acute brainstem lesions. Morphological changes and functional derangements were correlated in 22 animals. Neurological disorders and behavioural reactions were reproducible and stable in rats with exact localization of coagulated foci. The results demonstrate the adequacy of the proposed model.

[The effect of maximal and submaximal physical loads on alimentary dyslipidemia]. / Vliianie maksimal'noi i submaksimal'noi fizicheskikh nagruzok na alimentarnuiu dislipidemiiu.

Short-term maximal and long-term submaximal regimens of physical exercise had been assessed for effect on serum lipoproteins in healthy subjects, in preclinical coronary heart disease (CHD) sufferers and in manifest CHD patients having food fat-induced lipemia. Fat loading caused hypertriglyceridemia in all the examinees. In healthy subjects it was associated with a rise in apo AI, while in CHD patients with relevant fall. Maximal exercise in preexisting alimentary hyperlipidemia provoked elevation of total cholesterol, triglycerides, LDL cholesterol, being atherogenic. Apo AI grew with a decrease in apo B/apo AI ratio in healthy subjects, in coronary patients apolipoprotein changes were of atherogenic origin. Prolonged submaximal exercise at the height of food lipemia resulted in lowering of total cholesterol, triglycerides and LDL cholesterol along with elevation of HDL cholesterol and apo AI both in healthy and coronary subjects evidencing anti-atherogenicity. It is noted that the response of lipids and apolipoproteins to fat and physical exercise loads was similar in subclinical and manifest CHD patients.

[Monoclonal antibodies to insulin influencing its biological activity]. / Monoklonal'nye antitela k insulinu, vliiaiushchie na ego biologicheskie aktivnost'.

Somatic hybridization was employed for obtaining 335 hybridomas producing monoclonal antibodies to insulin. Twelve hybridomas were cloned by a method of maximum dilutions, and specific immunoglobulins, secreted by them, were characterized by solid phase immunoenzyme assay. Monoclonal antibodies possessed different activity with relation to human, porcine and cattle insulins, indicating their specificity to different epitopes on the insulin molecule. Antibodies of IN-2 and IN-3 clones were capable of inhibiting insulin biological activity in vivo. In experiments on rabbits antibodies of IN-2 clone decreased a 3-fold insulin hypoglycemic action, and antibodies of IN-3 clone caused an increase in the life span of mice after administration of insulin at high doses to them.

[Prognosis of the outcome of myocardial infarction by using early dynamic exercise tests]. / Prognozirovanie iskhoda infarkta miokarda s ispol'zovaniem rannikh prob s dinamicheskoi nagruzkoi.

A total of 158 males with a history of myocardial infarction were examined. The examination involved collection of information on their histories, course of the disease and symptoms in the hospital period and early (mean, on day 13) bicycle ergometric exercise tests. A multifactorial analysis made it possible to derive the decision rule to predict the condition of a patient within the first year following the onset of myocardial infarction, which involved prognostically unfavorable history data, such as disability prior to myocardial infarction, alcohol usage, exertional anginal hospital-stage parameters, such as bradycardia, premature contraction, circulatory failure, nodal rhythm, as well as 1 mm or more of ST-segment elevation during the early bicycle ergometric exercise test. The sensitivity of the predictive rule developed was 88.5%, its specificity was 78.5%. The study shows that it is impossible to successfully solve the problem in attempting to predict the outcome from some parameters, including the bicycle ergometric test findings. It is possible to do so only when the complex of data on a patient is taken into account.