Chitosan as a Modifying Component of Artificial Scaffold for Human Skin Tissue Engineering.

We compared the structure and mechanical properties of scaffolds based on pure collagen, pure chitosan, and a mixture of these polymers. The role of the composition and structure of scaffolds in the maintenance of cell functions (proliferation, differentiation, and migration) was demonstrated in two experimental models: homogeneous tissue analogues (scaffold populated by fibroblasts) and complex skin equivalents (fibroblasts and keratinocytes). In contrast to collagen scaffolds, pure chitosan inhibited the growth of fibroblasts that did not form contacts with chitosan fibers, but formed specific cellular conglomerates, spheroids, and lose their ability to synthesize natural extracellular matrix. However, the use of chitosan as an additive stimulated proliferative activity of fibroblasts on collagen, which can be associated with improvement of mechanical properties of the collagen scaffolds. The effectiveness of chitosan as an additional cross-linking agent also manifested in its ability to improve significantly the resistance of collagen scaffolds to fibroblast contraction in comparison with glutaraldehyde treatment. Polymer scaffolds (without cells) accelerated complete healing of skin wounds in vivo irrespective of their composition healing, pure chitosan sponge being most effective. We concluded that the use of chitosan as the scaffold for skin equivalents populated with skin cells is impractical, whereas it can be an effective modifier of polymer scaffolds.

[Proteomics as a fundamental tool for subclinical screening, tests verification and assessment of applied therapy].

Proteomics is a science which studies proteins of the body, interactions of proteins and their biological functions. Today, it is an essential partner in establishing preclinical diagnosis protocols. In conjunction with other sciences such as genomics and bioinformatics it will be possible to diagnose diseases on the earliest stages before its clinical onset or to gain the dynamics of pathological processes in the body and response to drug therapy. This article discusses general aspects of proteomics as well as special ones on the basis of models of cardiac diseases and cancer.

[Introduction into PPPM: experience of the past and tomorrow's reality].

Health service today is on the verge of broad changes. The intensive practical application of the achievements of genomics, proteomics, metabolomics and bioinformatics has significantly deepened and continues to deepen our view of the pathological processes taking place at a level of biostructures. In the nearest future this progress will give medical practitioners the opportunity to focus on a subclinical stage of the disease, i.e. on the earliest stages of the pathological process. This will require the prediction of disease development risks, subclinical diagnostics with the precise staging of the pathological process, and, finally, the application as early as possible of targeted pharmacotherapeutic methods in order to prevent the manifestation of the disease or its progression into more severe stages. All these principles create the framework of a fundamentally new "3P" strategy in medicine: predictive, preventive and personalized medicine.

[Development of national and international standards of population age distribution for medical statistics, health-demographic analysis and risk assessment].

The current European standard (CES) and the World population age distribution standard is widely used in medical and demographic studies, performed by international (WHO, etc.) and national organizations. The Russian Federal Service of States Statistics (RosStat) uses CES in demographic yearbooks and other publications. The standard is applied in calculation of the standardized mortality rate (SMR) of the population in different countries and territories. Risk assessment is also used CES. In the basis of the standards there has been laid the idea to assess mortality according to uniform standard, so to get possibility to compare the mortality rate of the population in different countries and regions, different genders and different calendar years. Analysis of the results of test calculations of the values of the SMR for the population of Russia and other countries with the use of current standards has revealed serious shortcomings of the latters and set up the task of improving them. A new concept of the development of standards based on the use of the concept of stable equilibrium of the age distribution of the population and survivorship function is proposed.

[Refutation of traditional views of the disease and formation of the scientific basis of pathology (17-19th centuries)].

This paper focuses on the revision of disease concepts that prevailed during the Galenism era and the formation of the scientific basis of pathology at the time of two scientific revolutions of the 17-19th centuries.

[Autoimmunity and autoimmune syndrome: norm and pathology].

Multicomponent analysis of variations of serum marker antibodies is an efficacious tool for preventive (pre-clinical) diagnosis of immune disorders causing serious diseases. Successful development of this method may lead to revision of the approaches accepted in modern medicine and facilitate practical transition to the prognosis-prevention paradigm. The study of antibody spectra reflecting individual genetic and epigenetic features provides a basis for prognostication of health and pathological conditions. We believe that in most cases early diagnosis of reversible pathologic processes their further development can be arrested which provides a real opportunity to preserve health.

[Diagnostic and pathogenetic implications of the site specificity of antibody proteases in multiple sclerosis].

Disseminated sclerosis is currently regarded as a CNS autoimmune disease. One of the mechanisms behind this pathology is antibody (AB) formation. In this context, recent data on AB with proteolytic activity are of importance because they participate in selective proteolysis of myelin proteins in patients with disseminated sclerosis. This paper focuses on AB-proteases associated with disseminated sclerosis and site-specificity of antibody-mediated proteolysis of myelin basic protein. Protocol of serodiagnostic algorithm to be used in clinical practice is described.

[Quantitative immunohistochemistry: optimization of analyzing the expression of signaling molecules in the myometrium].

The paper analyzes in detail various aspects of semiautomatic quantitative analysis of the results of immunohistochemical reaction using the Morphology 5.0 computer program to analyze microscopic images. Various procedures for assessing the extent of expression of nuclear and cytoplastic markers are compared and a rationale is provided to choose the value of the relative area of a microimage. The authors have developed a new method for calculating the optical density of as an indicator of the intensity of expression of the marker under study, which allows one to effectively separate the study staining from the baseline hematoxylin staining.

[Current protease-antibody-based serodiagnostic tools and their use in the diagnosis of autoimmune thyroid diseases].

The purpose of the investigation was to comparatively study the possible clinicodiagnostic and clinicoprognostic value of thyroglobulin (TG) autoantibodies (anti-TG autoAB) and thyroid peroxidase (TPO) (anti-TPO autoAB) with proteolytic activity (protease-AB) and to develop additional clinicoimmunological criteria for working out a protease-AB-based laboratory serodiagnosis protocol. Sera from 240 patients with autoimmune thyroiditis (AIT), 124 with diffuse toxic goiter (DTG), and 172 with other thyroid diseases were studied. Serum from 40 clinically healthy donors served as a control. Sera were screened for anti-TG and anti-TPO autoAB by enzyme immunoassay and/or radioimmunoassay. Nonspecific proteolytic activity was determined, by incubating a highly purified AB IgG-isotype solution with nonspecific substrate (such as BSA-FITS or thyoredoxine/Trx), followed by the measurement of relative fluorescence shifts at a wavelength of 470 nm. The blood samples taken from patients with AIT or DTG and the highly purified anti-TG or anti-TRO autoAB of IgG isotype were incubated with appropriate human substrates to determine AG-specific proteolytic activity. Proteolysis products were detected by PAAG electrophoresis. An association was found between the rise in the frequency of protease AB, their catalytic activity, a tendency toward autoAB between themselves, and the degree of thyroid tissue degradation in AIT and DTG; some specific features of a serological pattern were noted in diferent dorms of AIT and DTG. For example, the prevailing autoABs have been shown to be TG-specific in AIT and TPO-specific proteases in DTG; also, the detectable catalytic activity of both autoAB in AIT is an order of magnitude higher than that in DTG. No protease ABs have been recorded in other forms of thyroid diseases in which antithyroid autoABs are also frequently detected. The screening procedure for protease AB may be considered as a highly sensitive and specific indicator test in the diagnosis and prediction of AIT and DTG, which allows one to diagnose not only within the framework of major thyroid nosological entities, but much broader, by covering a great variety of syndromal forms of pathology.

[Neuroimmunoendocrine mechanisms of aging].

The review discusses in detail the mechanisms of aging from the position of common signaling molecules produced in three regulatory systems: the nervous, endocrine and immune ones. It is shown that the neuroimmunoendocrine hormonal regulation of homeostasis plays the important role in the complex chain of processes that lead to aging of cells, tissues, organs and body as a whole. Particular attention is paid to morpho-functional involution of nervous, endocrine and immune systems, which is accompanied by a breach of elaborate signaling molecules. Detailed analysis and further development of integrated molecular commonality of views on the regulatory systems of both the central and especially local levels, opens vast new opportunities for deepening knowledge of the mechanisms of aging, as well as for the prevention, diagnosis and treatment of diseases associated with aging, in which pathogenesis the discoordination of neuro-immuno-endocrine signaling mechanisms plays an important role.

[Post-infection clinico-immunologic syndrome: fundamentals of etiopathogenesis and immunogenodiagnostic strategy].

Professor Yu.M. Lopukhin, an ardent advocate of translation of scientific achievements into clinical medicine, is a founder of new scientific disciplines including clinical immunology that evolved from the fusion of new knowledge, original clinical thought, high-tech diagnostic and therapeutic modalities. Highly promising investigations into molecular pathogenetic mechanisms of various diseases, their diagnosis, and prevention make up a most important aspect of his scientific work. The progress of chronic recurring infectious diseases (CRID) is closely related to the associated immune disorders, e.g. post-infection clinico-immunologic syndrome (PIKIS) that reflects the character and severity of disturbances of immune homeostasis. This syndrome associated with CRID is as a rule provoked by infectious agents of different nature, immunopathologic processes inherent in individual patients, specific clinical course ofCRID or inadequate application of antimicrobial medicines. Three forms of PIKIS are described in this paper: (1) post-infection secondary immunodeficiency syndrome:, (2) post-infection autoimmune syndrome, (3) combination of the two.

[From the history of the 19th century pathology. Pathological anatomy and pathology in the first half of the 19th century].

The paper analyzes the first half of the 19th century collective notions on disease as the event that is alien to the human body (which is embodied as a living being), which occurs and develops in accordance with its own laws differing from the laws of human vital functions; on the causes and essence of diseases and the methods of their study. It also shows a place of human pathology in the structure of medical knowledge of that time and its role in the investigation of diseases.

[Antithyroid antibodies of varying specificity in the pathogenesis and diagnosis of autoimmune thyroid diseases].

AIM: To compare the possible pathogenetic and clinicodiagnostic value of antithyroid autoantibodies (autoAB) different in specificity, such as monospecific ones to thyroglobulin and thyroid peroxidase (anti-TG and anti-TPO autoAB) and bispecific ones to thyroglobulin and thyroid peroxidase simultaneously (anti-TGPO autoAB), in patients with autoimmune thyroid diseases. MATERIALS AND METHODS: The sera from 240 patients with autoimmune thyroiditis (AIT) and from 124 with diffuse toxic goiter (DTG) were examined. The sera from 40 healthy donors served as a control. The sera were screened for anti-TG and anti-TPO autoAB, anti-TGPO autoAB, by employing enzyme immunoassay and/or radioimmunoassay. The results were statistically processed using the variation statistics-based programs. RESULTS: The specific features of an autoantigenic component to thyroid tissues were found in the sera of patients with AIT and DTG. An association was established between the progression of disease and the phasic change of autoAB populations or their combinations. CONCLUSION: The procedure for evaluating seropositivity for antithyroid autoAB, which is referred to as non-invasive studies, can be considered as a criterion test in the diagnosis and prediction of the course of AIT and DTG.

[Post-infectious clinical-immunological syndrome and its place in clinical practice].

The progression of chronic-relapsing infectious disease (CRID) depends on a combination of cumulative immune-mediated responses of the human body, which, in turn, are united by a number of the common mechanisms. The mechanisms are called as the post-infectious clinical-immunological syndrome (PICIS) to demonstrate the features and scale of imbalances of immune homeostasis. PICIS usually accompanies most of the known CRID to define the type of the disease, to predict the progression of and risks for the complications to be risen as well. PIFIS is generally provoked by either infectious pathogens of various nature or by the atypical immune responses from the infected patient, or by the onset of the disease itself, or by the inadequate antimicrobial therapy. Three forms of PICIS which depend on two key factors have been described. These included: (i) the spectrum of a microbial colonization landscape; (ii) the antimicrobial immunity itself to generate, for instance, either of three alternative PICIS, namely, (1) postinfectious secondary immunodeficiency syndrome (PISIS); (2) postinfectious autoimmune syndrome (PIAIS), and (3) PISIS combined with PIAIS, i.e. PISIDAS. The dominant monosyndrome-like form of associated immune imbalances in CRID patients is PISIS. PISIS occurs in more than a third of the clinical cases to stress the autoaggression (PIFAS), or combininative form of the immune-mediated imbalances, i.e. PISIDAS. In the process of the development of CRID, PISIS can give a way to either PIAIS or PISIDAS. Besides the immune-mediated imbalances, an essential role in the pathogenesis of CRID and PICIS is also attributed to the infectious factors capable of forming microbial associates in the pathogenesis of PICIS. Therefore, treatment of such patients should be directed not only at the elimination of the infectious pathogen(s), but also at the restoration of the physiological level of the immune homeostasis impaired by PICIS.

[Clinical trials of the new skin test Diaskintest for the diagnosis of tuberculosis].

A new reagent for a skin test given the name Diaskintest has been designed for the screening diagnosis of tuberculosis and preclinical and clinical trials conducted. Preclinical trials were carried out on 315 laboratory animals (guinea-pigs, albino mice). The reagent Diaskintest was ascertained to be nontoxic, to have no sensitizing properties, to be safe and specific, and to induce no positive reactions in BCG-vaccinated animals and healthy guinea-pigs. Its specific activity was comparable with that of the national reference--purified tuberculin PPD-L-2. With progression of tuberculous lesions, the guinea-pigs showed higher responses to Diaskintest dilution and the BCG-vaccinated animals lacked responses to Diaskintest with increased delayed type hypersensitivity. The clinical trial was permitted by the Federal Service for Surveillance in Health Care and Social Development of the Russian Federation. Clinical trials were conducted in 150 persons. The safety, specificity, sensitivity of Diaskintest were first examined in the clinical studies and its action was compared with the results of tuberculin skin test (Mantoux test) with 2 TE of PPD L-2. Diaskintest was ascertained to be highly sensitive when given in a dose of 0.2 microg in 0.1 ml. In patients with active tuberculosis and new cases of Mycobacterium tuberculosis infection, the agent induced a positive skin reaction (a papule of more than 10 mm) in 98-100% of cases (p < 0.05). The agent caused no reaction associated with BCG vaccination. The specificity of the test was 93-100% with 95% significance. The rate of overexuberant reactions (vesicular necrotic changes, lymphangitis, and lymphadenitis) was 4-14% with 95% significance. Tuberculosis patients with significant immunopathological disorders might have no skin sensitivity to Diaskintest, as to PPD L-2 (a negative test). The findings substantiate the use of Diaskintest for mass epidemiological surveys for the differential diagnosis of tuberculosis and BCG vaccination-associated complications. The agent may be also used to evaluate the activity of the process in patients with tuberculosis and the efficiency of treatment in combination with other methods and to make a differential diagnosis of tuberculosis.

[Smallpox is a dormant volcano].

The presence of rodent-associated natural foci containing at least 6 of the known 11 viruses belonging to the genus Orthopoxvirus (Poxviridae, Chordopoxvirinae) within the equatorial, tropical, subtropical, temperate, and subarctic climatic zones; the increasing aggravation of the monkey pox epidemic situation in equatorial Africa with an increase in human mortality by an average of 9.8% with a possibility of 2 to 8 passages in 30-70% of patients; the possible persistence of a virus in the human cadavers buried in the permafrost of Eurasia and America; bioterrorism threat due to the unaccounted viral reserves persisting somewhere or somebody; no postvaccinal human immunity since vaccination and vaccine manufacture stopped 30 years ago as recommended by the WHO, make the risk of the deteriorating epidemic situation with disastrous effects greater now and in the foreseeable future than it was 20-30 years ago. Health care academic circles and bodies do not know methods for rapid diagnosis in the field conditions of species-specific identification smallpox virus or preventive (low-reactogenic, effective vaccines, and those accessible for mass production) and therapeutic (nontoxic drugs, those satisfactory for mass production, inexpedient, effective ones when orally used) agents. Basic studies of biodiversity, functional properties of viral DNA and proteins, pathogenesis, and evolution are required. Live smallpox virus should be used at certain and particularly final stages for these studies that are of scientific and applied significance.

[The current concept of immunological disorders in the case of basal-cell carcinoma].

Imbalance in the interaction of mechanisms of innate and adaptive immunity rather than selective defect of each of the links is a determinant of the development of basal-cell carcinoma of this or that form of immunopathology or that of transformation of one to another form. The lecture gives notions on immunological phenotypes and the parameters of innate and adaptive immunity and on the association of two links of immunity. The major tumor-associated immunological phenotypes are pathogenetically and clinically different, but have signs of two-link immunodeficiency in each case. Tumor-associated secondary immunodeficiency is characterized by defects encompassing both the innate and adaptive links of immunity. Predominant are autoimmune disorders involving mainly the adaptive link of immunological responsiveness in case of tumor-associated autoimmune syndrome concurrent with immunodeficiency.

[Architectonics of cell subpopulations of peripheral blood in patients with autoimmune myocarditis: clinical and pathogenetic aspects].

AIM: To compare the most significant architectonic parameters of peripheral blood cell subpopulations in patients with different variants of an autoimmune myocarditis (AIM) course and their clinical value in therapeutic practice. MATERIAL AND METHODS: Blood cell subpopulations were studied with flow cytometry in 99 blood samples from patients having different AIM variants and myocardiosclerosis as well as in 40 healthy donors. RESULTS: Severe (malignant) AIM was characterized by growing indices of T-/B lymphocyte activation, expression of activation markers on the cells of both differentiation lines, disproportions in composition of subpopulations of the immunoregulatory cells, parallel rise in specific weight of dendritic cells, reduced intensity of apoptosis of autoreactive T-lymphocytes. In benign AIM marked immunopathology was not found. This group can be considered as a separate variant of AIM course necessitating an individual approach to planning pathogenetically sound therapeutic and rehabilitation measures. CONCLUSION: The study of activation markers expression on peripheral blood cells is superior to the study of endomyocardial biopsies providing a non-invasive method of immunodiagnosis.

[Modern technologies in diagnosis and treatment of patients with late stages of hydronephrosis].

To evaluate the role of modern ultrasonic, x-ray and radionuclide methods in staging hydronephrosis; that of stimulators of stem cells of the bone marrow in combined therapy of patients with late hydronephrosis, we examined and treated 18 patients with late-stage hydro- and ureterohydronephrosis. All the patients were operated. Preoperative preparation included drainage of the upper urinary tract and stimulation of bone marrow stem cells. Organ-saving surgery was conducted with a satisfactory functional result in 14 of 18 patients. Modern methods of investigation provide an objective assessment of structural-functional condition of the kidneys and upper urinary tract in patients with late hydro- and ureterohydronephrosis, may evidence for possible reversibility of registered uro- and hemodynamic alterations. Preoperative use of stimulators of bone marrow stem cells promotes activation of repair processes in renal parenchyma which is essential for efficacy of a further reconstructive operation.

[Antibody-mediated proteolysis of myelin-associated proteins as a new mechanism of control over demyelinization processes in multiple sclerosis].

Natural antibodies possessing catalytic activity present a new group of biologically active substances that are found in various autoimmune diseases such as autoimmune thyroiditis, myocarditis, multiple sclerosis, and system lupus erythematosus. Presently, an interconnection between the activity of these antibodies and the extent of organic and tissue lesion in autoaggression have been revealed. Clinical use of catalytic antibodies as a diagnostic criterion to evaluate the severity of disease, a prognostic criterion of the risk of invalidization, and as a pathogenetic basis for medicamentous treatment of autoimmune process is a promising directions of study of the role of catalytic antibodies.