The Role of Cancer and Somatic Stem Cells in the Anti-Inflammatory and Antitumor Effects of Aconitum baicalense Extract on Experimental Breast Cancer.

We studied the effects of the extract of the terrestrial part of Aconitum baicalense in BALB/c female mice at the early stages after the injection of N-methyl-N-nitrosourea (MNU). The extract reduced inflammatory activity and tumor growth in the mammary gland. The antitumor and anti-inflammatory effects of the extract are based on the inhibition of cancer stem cells, hematopoietic stem cells, and hematopoietic progenitor cells that promote inflammation. The extract of A. baicalense disrupted the recruitment of epithelial progenitor cells and angiogenesis precursors to the mammary gland preventing neovascularization and transformation of epithelial cells into tumor cells.

Effects of Age and Suntan on the Expression of Second Messenger Signaling Pathways of Necroptosis in Skin Cells during Facelifting Surgery.

We studied the effects of age and suntan on the expression of necroptosis signaling molecules (RIPK1, RIPK3, and MLKL kinases) and first TNF receptor (TNFR1) in isolated skin cells from women undergoing facelift surgery. In women above 50 years, the expression of the TNFR1, kinases RIPK1, RIPK3, and MLKL, the phosphorylated forms of these kinases was significantly (p<0.05) increased in comparison with the corresponding parameters in women under 30 years. The expression of all necroptosis proteins and TNFR1 in women with suntan was significantly (p<0.05) higher than in those without tan. Cells from the surgical material were incubated with TNFα to determine the level of induced necroptosis. In women aged >50 years and women with suntan, the expression of phosphorylated forms of kinases was significantly increased, which attested to necroptosis activation. This study allowed identifying the targets on skin cells for prevention of necrosis and inflammation after facelift surgery.

Age-Related Features of the Early Period of Liver Regeneration after Partial Hepatectomy in Rats.

Regenerative processes in the liver were studied in 3-month-old (young) and 9-month-old (aged) male Wistar rats on day 1 after 30 and 70% hepatectomy. Regardless of the resected liver volume, shifts in the biochemical parameters of the serum in aged rats were more pronounced than in young animals. After 30% hepatectomy, no age differences in the rate of hepatic regeneration were found, while after 70% liver resection this parameter was higher in young rats. Hepatectomy in young rats led to recruitment of MSC, hepatocyte precursors, endothelial and epithelial progenitor cells into the liver parenchyma and increased fluidity of the plasma and mitochondrial membranes of hepatocytes. In aged rats, the recruitment of MSC, hepatocyte precursors, and endothelial progenitor cells into the injured liver was impaired and the rigidity of the mitochondrial membranes of hepatocytes increased.

[Comparative Analysis of MPTP Neurotoxicity in Mice with a Constitutive Knockout of the α-Synuclein Gene].

Aggregated forms of α-synuclein are core components of pathohistological inclusions known as Lewy bodies in substantia nigra (SN) neurons of patients with Parkinson's disease (PD). The role of α-synuclein in selective loss of SN dopaminergic neurons (DNs) in PD is studied in mice knocked out in the α-synuclein gene. The new mouse strain delta flox KO with a constitutive knockout of the α-synuclein gene models the end point of in vivo deletion of the α-synuclein gene in mice with a conditional knockout and has no foreign sequence in the modified genomic locus, thus differing from all other α-synuclein knockout mouse strains. The effect of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which is used to model PD, was compared between delta flox KO mice and mice of the well-known α-synuclein knockout strain AbKO. Subchronic MPTP administration, which models early PD, was found to reduce the dopamine content and to change the ratio of dopamine metabolites in the striatum to the same levels in delta flox KO, АbKO, and wild-type mice. Overt locomotor defects were not observed after MPTP treatment, but gait testing in a CatWalk XT (Noldus) system revealed identical gait deviations in mice of the two strains and control wild-type mice. Based on the findings, a similar mechanism of neurotoxic damage to DNs was assumed for delta flox KO and AbKO mice.

Increased Expression of the Multimerin-1 Gene in α-Synuclein Knokout Mice.

Multimerin-1 (Mmrn-1) is a soluble protein, also known as elastin microfibril interfacer 4 (EMILIN-4), found in platelets and in the endothelium of blood vessels. Its function and role in pathology are still not fully understood. Genetic modifications in alpha-synuclein gene (Snca) locus that mapped 160 Kb apart from Mmrn-1 in mouse genome, could weigh with regulatory elements of Mmrn-1 gene. We have studied the Mmrn-1 expression in brain cortex of three mouse lines with Snca knock-out: B6(Cg)-Sncatm1.2Vlb/J, B6;129-Sncatm1Sud/J, and B6;129X1-Sncatm1Rosl/J. The 35-fold increase for Mmrn-1 mRNA level have been found in B6;129X1-Sncatm1Rosl/J mice that carry in their genome foreign sequences including bacterial gene neo and a strong promoter of a mouse phosphoglycerate kinase (Pgk1) oriented towards Mmrn-1 gene. This effect on regulatory elements of Mmrn-1 gene as a result of modifications in Snca locus should be taken into consideration when using B6;129X1-Sncatm1Rosl/J line, that is widely applied for study of neurodegeneration mechanisms.

Chemokine Expression in Neutrophils and Subcutaneous Adipose Tissue Cells Obtained during Abdominoplasty from Patients with Obesity and Normal Body Weight.

The study was aimed at assessing the role of chemokines in inflammatory changes in tissue following abdominoplasty. The levels of neutrophil-coupled chemokines and their receptors in the serum and blood cells, as well as in cells isolated from the subcutaneous adipose tissue sampled during abdominoplasty were compared in patients with obesity and normal body weight. The levels of chemokines CCL3, CCL3, and CCL5 in blood serum and expression of chemokine receptor CXCR2 and CXCR6 on blood neutrophils were significantly higher (p<0.05) in obese patients in comparison with patients with normal body weight. Elevated expression of chemokines CCL2, CCL3, CCL4, CCL5, CCL18, and CCL20 (p<0.05) was detected in subcutaneous adipose tissue cells isolated obese patients in comparisons with persons with normal body weight. These findings attest to favorable conditions for enhanced neutrophil migration to the adipose tissue in patients with obesity, which can promote leukocyte infiltration of the suture site after abdominoplasty and serves as additional risk factor for the development of postoperative complications associated with activity of neutrophil-derived proteolytic enzymes.

Effects of Release-Active Antibodies to CD4 Receptor on the Level of lck-Kinase in Cultured Mononuclear Cells from Human Peripheral Blood.

For evaluation of effects of release-active antibodies to CD4 on cultured lymphocytes from human peripheral blood, we measured intracellular content of lck-kinase cell-based ELISA. In cells treated with release-active antibodies to CD4, the content of intracellular lck-kinase significantly (p<0.01) decreased in comparison with the control (purified water processed in a similar way). Phytohemagglutinin had no effect on the concentration of lck-kinase in cells. The decrease in the content of CD4-associated lck protein suggests that the preparation enhanced intracellular coupling of lck-kinase with T-cell receptor and potentiated T-cell immune response.

[Cellular Model Based on Laser Microsurgery of Cell Spheroids to Study the Repair Process].

In this study, modern techniques of laser microsurgery of cell spheroids have been used to develop a new simple, reproducible model for studying the mechanisms of repair and regeneration in vitro. Nanosecond laser pulses were applied to perform a microdissection of the outer and the inner zones of the spheroids from dermal fibroblasts. To achieve effective dissection and preservation of spheroid viability, the optimal parameters were chosen: 355 nm wavelength, 100 Hz frequency, 2 ns pulse duration, laser pulses in the range of 7­9 µ J. After microdissection, we observed injury of the spheroids : the edges of the wound surface opened and the angular opening reached a value of more than 180°. As early as during the first hour after spheroid microdissection with laser radiation, the surviving cells changed their shape: cells on the spheroid surface and directly in the damaged area became rounded. One day after microdissection, the structure of the spheroids began to partially recover, the cells in the surface layers began to take the original flattened shape; debris of dead damaged cells and their fragments was gradually cleared from the spheroid composition. In the proposed model, the first data on stimulation of structure recovery of injured spheroids from dermal fibroblasts with a P199 synthetic polypeptide, which is used in cosmetology for the initiation of antiaging and regenerative effects in the skin, were received. After microdissection, recovery of the spheroids structure with a few surface layers of flattened imbricated arranged cells and polygonal cells of the inner zone in the presence of P199 peptide was faster than in the control group, and was completed within 7 days, presumably due to the remodeling of the survived cells.

Heat shock protein HSP40 family chaperone DNAJB6/MRJ expression analysis in blood cells obtained from patients with atopic dermatitis in different phases.

Heat shock protein HSP40 family molecular chaperone DNAJB6/MRJ expression has been analyzed in blood cells of patients with atopic dermatitis compared with healthy donors. Severity of disease was estimated according index SCORAD. Methods: Peripheral blood cells were separated using Percoll density gradient. Purified neutrophils and lymphocytes have been stained with antibodies to the heat shock protein DNAJB6/MRJ. Cells were analyzed using flow cytometry. Real time PCR method has been used to verify the bacterial contamination of the skin of patients with atopic dermatitis. Statistical analysis was performed by ANOVA. Results: Expression of DNAJB6/MRJ protein has been found to be elevated in all samples of cells obtained from patients with atopic dermatitis. The highest level of the DNAJB6/MRJ protein expression was shown in neutrophils at the acute phase of severe atopic dermatitis. DNAJB6/MRJ protein expression in lymphocytes of patients with atopic patients was less extensive compared with neutrophil level and was shown to be higher at subacute phase of disease. The DNAJB6/MRJ protein expression was found to be statistically significant higher in lymphocytes from atopic patients compared with healthy donors. The bacterial contamination of skin (verified by PCR) was shown to influence the DNAJB6/MRJ protein level in lymphocytes of atopic dermatitis patients. Conclusion: Expression of the heat shock protein DNAJB6/MRJ was elevated in neutrophils and lymphocytes of patients with atopic dermatitis compared with healthy donors. The highest level of the DNAJB6/MRJ protein was found to be in neutrophils at acute phase of severe atopic dermatitis and gradually decline as continue to the disease.

[Pathogenesis of the vascular consequences caused by cervical instability and their algorithm for pharmacotherapy].

256 patients with radiologic detected cervical instability were examined. This pathology may be the cause of arterial or venous vertebra-basilar disorders. We realized extensive examination the patients with the cervical instability for the pathogenic correction of vascular consequences. We founded normal blood flow only in 19.9% cases without morphology alterations the vessels of both basins. 80.1% patients had different cerebral hemodynamic disorders variations. According to our examination plan we detected the vertebra-basilar insufficiency pathogenesis and proposed medicinal correction of the cerebral instability consequences.

[Cerebral circulation pathophisiology into pneumocephalus craniocerebral dispoportion].

We included 19 patients with a different intensity postoperative pneumocephalus and an inadequate postoperative awakening. Intracranial CT excluded postoperative complications (oedema, haematoma, hidrocephalus) and diagnosed a pneumocephalus (above the frontal and temporal lobes and in the cerebral ventricular system). In two hours after operation we found systolic linear blood flow velocity (BFV syst.) decrease in the extracranial part of internal carotic artery (ICA) (p < 0.001) in patients with pneumocephalus and inadequate postoperative awakening. But in 24-48 hours after operation we diagnosed BFV syst. elevation in the ICA extracranial part (p < 0.001) and preumocephalus diminution in patients with a recovered consciousness.

[Peripheral blood cells luminol-dependent chemiluminescence at the different stages of atopic dermatitis].

UNLABELLED: Aim of this work was to record the luminol-dependent spontaneous and induced chemiluminescence at the different stages of atopic dermatitis. METHODS: Peripheral blood cells were obtained from adult patient with atopic dermatitis followed by the registration of luminol-dependent chemiluminescence on luminograph. Opsonized zymosan as well as yeasts Candida tropicalis have been used to induce the chemiluminescence. RESULTS: Spontaneous and induced chemiluminescence were slightly elevated at the mild atopic dermatitis but were decreased at the severe stage of disease. Statistically significant difference has been found between group with mild and severe atopic dermatitis, Skin contamination by yeasts Candida tropicalis causes the increased level of blood cells chemiluminescence at the first week of atopic relapse when the disease was mild. Severe stage of atopic dermatitis was coupled with statistically significant inhibition of both, spontaneous and induced chemiluminescence. CONCLUSIONS: Luminol-dependent chemiluminescence of peripheral blood cells from adult atopic dermatitis patients may be stimulated at the mild stage and suppressed at severe stage of atopic dermatitis.

Neurodevelopment and phenotype-modulating functions of S100B protein: a pilot study.

The importance of certain neurotrophic proteins found in maternal blood and milk for breastfed infants has remained ambiguous. This study was conducted to present evidence of the impact of an induced deficit of active S100B protein on neonate development. Newborn mice from two groups of mothers, immunized or sham-immunized against S100B, were subjected to various behavioral tests, and the development of their morphological characteristics was recorded from birth until weaning. Morphological problems, including weight gain and fur coating, a delay in the maturation of neurobehavioral systems and a deficit in neuromotor functions, including visual abilities, somato-sensory and posture reactions, muscular strength, locomotion, and fear/orienting processes, were observed in pups of immunized mothers. The S100B protein of external or internal origin in infants may be considered to be a specific factor that determines neuro- and morphological development and a risk-avoidance ('homeward-bent' or fearful) phenotype. The suppression of activity of the S100B protein results in a slower neonatal development and the formation of a risk-tolerant (fearless) phenotype of the offspring. This study thus considers the mechanism of neuroplastic regulation on the extent of sensation-seeking or risk-taking (homeless-like or fearless) and sensation- or risk-avoidance (home-bound or fearful) features in individual phenotypes.

[Leukocyte elastase, alpha 1-proteinase inhibitor and C-reactive protein in patients with delayed negativation of serological reactions after treatment of early syphilis].

It was investigated the activity of leukocyte elastase, alpha 1-proteinase inhibitor and C-reactive protein in patients with delayed negativation of serological reactions after a treatment of syphilis. The patients were divided into three subgroups. The first subgroup consisted of patients who did not have the signs of non-specific immune system response. Second subgroup consisted of patients with clear signs of infection process. The patients this subgroup had elevated values of the contents of inflammatory mediators. The remaining patients with questionable results were attributed in the third subgroup. The obtained results can be of great value to determine further treatment strategy in patients with delayed negativation of serological reactions.

[The effect of the instability of cervical spine on the hemodynamics in the vertebrobasilar system].

256 patients with cephalgia were examined. The main cephalgia cause was a cervical instability. We showed importance of extended examinations for cephalgia pathogenesis. We estimated cerebral circulation by ultrasound (cerebral blood flow registration) methods. We found an intact blood flow in both vascular systems (arterial and venous) in 19.9% cases. Cervical instability more 3 mm vertebra dislocation was not always a cause of vertebral discirculation. Quite often a vertebra dislocation under 3 mm was a result of vertebra-basilar disordes.

[Simulation of repeated local hemorrhagic stroke in rats].

The processes of developed in CNS the complicated stroke and developments of fittings for their pharmaceutical therapy were developed and offering by standardized method of the experimental secondary stroke in rats, suitable for the use in sharp and chronic researches. Variant of repeated hemorrhagic stroke consist of autohemorrhagic right hemisphere stroke by the mechanical damage of brain tissue after 10-daily occlusion of right common carotid artery was studied. A model is comfortable for reproducing of the repeated standardized local damage of brain, is more adequate form of design of transient and chronic cerebrovascular pathology, than the independent use of local hemorrhage of autoblood in the brain of animals. The morphological description of model approaches the clinical variants of development and flow of sharp hemorrhagic stroke after a previous chronic cerebral insufficiency on an ischemic type.

[Influence of high concentration of antibodies to NGF during early embryogenesis on formation of mice behavior in postnatal period].

In this work the influence of high concentration of antibodies to NGF on mouse's progeny has been investigated. During immunization with NGF the highest concentrations of antibodies were created in the first and third days of pregnancy (in different groups of animals). The dependence of abnormalities of mice postnatal development on level of antibodies to NGF at different stages of early embryogenesis has been established. Increasing of abnormalities in the formation of early behavioral acts and more clinically apparent anomalies in the somatic maturation in case of maximum of antibodies on day I of pregnancy has been showed. Immune responses to NGF during early embryogenesis of mice cause lag in the formation of behavioral acts. The latter are characterized by difficulties in sensor-motor coordination of the limbs and more clinically apparent in mice with a maximum of antibodies on day 1 of embryonic development. Infantilism in developing of contacts between progeny and mothers detected in mice with immune reactions may be a sign of serious mental dysontogenesis. The accelerated development of working memory established in mice with immune response to NGF requires further study of the development of cognitive abilities in these animals. The obtained results illustrate the important regulatory role of NGF at the early stages of development of the nervous system.

[The role of innate immunity system in the course of adenomyosis].

The investigation of leukocytic elastase (LE) and alpha1-proteinase inhibitor (alpha1-PI) from patients with different stage adenomyosis and in control group was found activation innate immunity system in all the patients with adenomyosis. The degree of LE activity is a prevalence rate of adenomyosis. The degree of alpha1-PI activity is correlated with antiproteolytic potential that blocks the effects shown by LE. It can lead the prognose of disease and timely treatment.

[Perinatal outcomes in pregnant women with diabetes mellitus and different level of autoantibodies to insulin and its receptors].

The levels of idiotypic (AB1) and antiidiotypic (AB2) antibodies were investigated in pregnants (n = 248) suffer from diabetes. It was proved that AB2 to insulin conditionally can be regards as antibodies to insulin receptor. It was shown that condition of newborns much depends on levels of these antibodies and their proportion. Condition of newborns from women with isolated high levels of AB1 to insulin was much better in comparison with ones from mothers with isolated high levels of AB2 to insulin. Conception about mechanisms of acting of AB1 and AB2 to insulin on fetus was represented.

[The course of the diabetes type I in pregnants with different levels of antibodies to insulin].

The course of the diabetes type I at pregnants (n=120) with different levels of idiotypic (ABI) and antiidiotypic (AB2) antibodies to insulin was investigated. It is known that AB2 to insulin can interact with insulin receptor. It was shown that changes of levels AB1 and AB2 to insulin are often observed at pregnants suffered from diabetes type I. Isolated high levels of AB1 to insulin is relatively good prognostic sign of the course of the diabetes type I at pregnants. On the contrary, isolated high levels of AB2 to insulin lead to decompensation of the diabetes type I and deep glycohemia. High levels of AB1 and AB2 to insulin together lead to unstable course of the diabetes type I. The same situation is observed in case of abnormal low levels AB1 and AB2 to insulin, taking into consideration that serum of health people contains the certain level of autoantibodies (AB1 and AB2) to insulin. The conclusion about significance of detection AB1 and AB2 to insulin during pregnancy of patients with diabetes type I was made.