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The objective of the present study was the evaluation of the auxiliary methods for the diagnostics of the intravital formation of the constriction marks; the secondary objective was to determine the pace at which the death and asphyxia occur., Material and Methods: The materials on which the study was based included 17 cases of mechanical strangulation asphyxia involving 13 men and 4 women at the age from 8 to 28 years. All cases of hanging were associated with different blood alcohol levels. Their characteristic feature was the formation of the obliquely ascending constriction marks. The group of comparison was comprised of three cases of death by drowning and one case of manual strangulation. The control group consisted of 10 patients who died from the acute form of coronary heart disease and 5 cases of death from traumatic shock. All the corpses were examined with the use of the traditional methods within the first 24 hours after death. The special laboratory studies were performed by means of the standard histological and immunohistochemical methods with the use of the polyclonal antibodies against total cytokeratin, fibrinogen, immunoglobulin-lambda, fibronectin, and CD-117 antigen. The histological preparations were stained by the method of Spielmeyer and with toluidine blue., Results: The results of the study give evidence of the possibility of diagnostics of mechanical strangulation asphyxia making use of the reaction with anti-fibrinogen antibodies in the stromal tissue of the constriction mark even in the absence of other intravital signs of death. Such diagnostics is also possible with the use of the CD-117 antigen in the pulmonary tissue. The expression of this antigen is characteristic of the cases of alveolar hypoxia. The possibility of application of other markers for the differential diagnostics of mechanical strangulation asphyxia from other causes of death is discussed.

The present review of the literature involves 50 publications concerning various substrates of importance as the biological markers of axonal damages with special reference to the secondary molecular and cellular mechanisms on which to base in vitro and in vivo modeling of the craniocerebral injury. The results of the investigations with the application of mass-spectrometry for the identification of the proteins specifically synthesized in response to the injury are presented; their biological functions are described. The use of the sequential microscopic imaging technique and the immunohistochemical methods made it possible to determine that the majority of the marker proteins are involved in the specific intracellular processes that are triggered in response to the traumatic impact including apoptosis, proliferation, formation of lamellipodia, axon regeneration, actin remodeling, cell migration and inflammation. In addition, a rise in the amount of intracellular actin-associated proteins has been observed. It is concluded that the investigations into the properties and the physiological role of beta-amyloid precursor protein (beta-APP) are of special value for the characteristic of nervous tissue damages and morphological diagnostics of the craniocerebral injury.

Automated image analysis methods are highly important for biotechnology research. The authors developed and tested a program for the morphometric analysis of photomicrographs of the sections processed using the standard immunohistochemical examination protocols. The color deconvolution method used in the algorithm was proven to be effective in mapping the distribution of DAB chromogen in the sample containing multiple dyes. The experiment demonstrated that the level of extracellular matrix proteins could be comparatively quantified in different groups of samples. The effective methods for the quantitative analysis of the Ki-67 labelling index were also tested using the same algorithms. The developed program was published under free GPL 3.0.

The paper presents a method of analysis of cell populations that combines the use of normalized spatial coordinates of the neurons with the morphometric criteria of their evaluation. These algorithms were applied to check the heterogeneity of apopulation of neurons Y-conducting channel in cat at the level of the lateral geniculate body (LGB). As a specific marker of Y-neurons, SMI-32 antibodies were used. Evaluated The dynamics of the distribution of the number of cells and the orientation of their soma within each layer and mediolaterally along the length of LGB dorsal nucleus (LGBDN). Among the SMI-32-positive neurons, the existence of at least two populations was detected differing in number, orientation and distribution of the soma in different layers of LGBDN. The heterogeneity of Y-neuron population in LGBDN detected in this study is consistent with the earlier electrophysiological data. We believe that the described algorithm for neuronal analysis may be successfully applied to study not only LGB, but also other extensive structures of the brain, including those having laminar organization.

The present review of the literature concerns the problem of morphological diagnostics of fat embolism, i.e. mechanical obturation of multiple blood vessels with fat globules, that can be detected by a variety of methods including polarization microscopy, staining of native, frozen, and paraffin-embedded histological sections with the use of immunohistochemical techniques, electron microscopy, etc.

There was performed an assessment of genotoxic effects of rocket fuel component--unsymmetrical dimethylhydrazine (UDMH, heptyl)--on forming germ cells of male mice. Immunocytochemically there was studied the structure of meiotic nuclei at different times after the intraperitoneal administration of UDMH to male mice. There were revealed following types of disturbances of the structure of synaptonemal complexes (SCs) of meiotic chromosomes: single and multiple fragments of SCs associations of autosomes with a sex bivalent, atypical structure of the SCs with a frequency higher than the reference level. In addition, there were found the premature desinapsis of sex bivalents, the disorder offormation of the genital corpuscle and ring SCs. Established disorders in SCs of spermatocytes, analyzed at 38th day after the 10-days intoxication of animal by the component of rocket fuel, attest to the risk of permanent persistence of chromosomal abnormalities occurring in the pool of stem cells.

The objective of the present study was to study the morphological criteria for toxic cardiopathy with the use of histological and immunohistochemical methods. The results of immunohistochemical studies of the sinoatrial node---???---(SAN) and the working myocardium in the patients presenting with alcoholic cardiomyopathy (ACMP) are presented. It was shown that vimentin expression in the SAN structures and the contractile myocardium is slightly increased whereas the expression of sarcomeric actin is decreased and that of fibrinogen is increased too. The authors put forward an assumption about the role of lesions in the membrane apparatus of the pacemaker cells in the development of arrhythmia characteristic of tanatogenesis associated with alcoholic cardiomyopathy.

Human skin structures stained positively for angiomotin or endostatin were studied by indirect immunohistochemical method. Skin specimens from frontal surface of the lower part of the neck (from upper corner of standard autopsy skin incision) from human fetuses died antenatally from 20 to 40 weeks of pregnancy, humans who died from different causes from 1 day to 85 years of life were obtained at autopsy. Positive staining for angiomotin or endostatin in the skin was found in epidermal cells, fibroblasts, sweat and sebaceous glands, blood vessels of the dermis. Blood vessels stained positively for angiomotin were detected in skin samples in all ages. Age-dependent decrease in the content of angiomotin in blood vessels of the dermis was detected. Most prominent decrease in angiomotin content in dermal blood vessels was found in 61-85 years age-group. Endostatin positive blood vessels were also detected in skin samples of all ages. However, the intensity of staining for endostatin in dermal blood vessels was increased during aging. It can be proposed that changes in the content of angiomotin and endostatin yield a negative impact on angiogenesis in human skin during aging.

The objective of the present work was the analysis of the literature concerning the application of the immunohistochemical methods for the improvement of diagnostics of intravitality and prescription of the mechanical injuries in forensic medical practice. Special attention is given to the examples of publication dealing with the methods for addressing this issue. The most promising areas of the application of immunohistochemical methods are considered. They are exemplified by the use of specific antibodies for the establishment of intravitality and prescription of the mechanical injuries. The possibility of using the presence of fibrinogen in the pulmonary alveoli as the marker of prolonged strangulation is illustrated. The results of this literature review provided a basis for the conclusion about good prospects of the application of the immunohistochemical methods with the purpose of establishing intravitality and prescription of the mechanical injuries in forensic medical practice.

The article proposes new (immune cytochemical) mode of evaluation of reaction of blast-cell transformation of human lymphocytes under their stimulation with phytohemagglutinin instead of standard radiometric method using 3H-thymidine as a marker. The reaction was assessed by means of luminescent microscope under indirect immunofluorescence using nuclear antigen Ki-67 as a marker of proliferating cells. The Ki-67 was detected using monoclonal antibodies Ki-67. The functional capacity of lymphocytes was evaluated by amount of visualized (luminescent) Ki-67 positive cells appearing after three day stimulation of proliferation with phytohemagglutinin. The comparison was made between immune cytochemical (with Ki-67 marker) and radiometric (with marking by 3H-thymidine) methods of phytohemagglutinin stimulation of lymphocytes. The article presents substantiation of advantage of immune cytochemical method of evaluation of reaction of blast-cell transformation of human lymphocytes as compared with radiometric method concerning possibility of its broader implementation in laboratory practice.

The paper analyzes the data available in the literature on the morphological and prognostic features of relatively rare high-grade thyroid cancer. It shows the difficulties and unsolved problems of morphological diagnosis and the possibilities of current methods for immunohistochemical and molecular genetic studies to determine the malignant potential of the tumor and prognosis.

It was analyzed the examination results of 156 patients with osteosarcoma. The data show that definition of histogenetic source, diagnostics and prognosis of treatment results are possible and expedient in case of analysis of signs reflecting tumor cells specificity. These signs may be determined by using of clinical parameters, X-ray imaging and light microscopy in case of moderately and highly differentiated sarcomas. Ample opportunities of flow citometry and immunohistochemistry allow to perform histogenetic identification, differential diagnostics and prognosis for low-grade sarcomas.

Immunolabeling method of microsporidium Paranosema locustae proteins on cryosections of locust infected fat body was proposed. In contrast to single parasite cells and artificially infected host cell cultures, this method allows to study molecular mechanisms of host-parasite relationships and in particular the secretory microsporidial proteins either at cellular or tissue level. Immunolocalization of the EPR-specific and cytoplasmic forms of Hsp70 family of molecular chaperones on cryosections showed accumulation of these proteins in the respective compartments of intracellular developmental stages of P. locustae and their absence in host structures. This allows to use them in diagnostics of microsporidiosis lesions in infected tissues as well as in colocalization analysis with P. lociustatre secretory proteins as a marker of parasite. The cytoplasmic chaperone stains cytoplasmic compartment homogeneously, but in the infected host cell during sporogony it disappears partially from the intracellular stages of development which damaged by maturing spores. Thereby study of molecular mechanisms of host-parasite relationships is to be carried out at the earlier stages of infection before active sporogony.

The purpose of the study - to determine the feature of the vascular endothelial growth factor (VEGF) and thyroid-stimulating hormone (TSH) expression in the thyroid gland (TG) in various thyroid diseases. Material - thyroid tissue (operative material) with histologically confirmed diagnosis: 10 - follicular adenoma, 17 - multinodular goiter, 8 - thyroiditis Hashimoto, 8 - papillary carcinoma, 10 - intact (normal) thyroid samples (forensic autopsy). The immunohistochemical study of the material showed the following results: the increase of the Hürtle cells population 40 % or more indicates a hyperthyroidism tendency despite TSH+ receptor status. Under the thyroid pathology TSH and VEGF expression appears in thyrocytes and also in microvascular endothelial cells. VEGF expression is below the norm in the Hashimoto thyroiditis. VEGF is involved not only in angiogenesis, but in pathophysiological shifts in thyroid tissue. Microvessel density (MVD) and TSH positive receptor status under the thyroid pathology testify the absence of the endothelial cells transformation, however, this index can not serve as a biopothential prognostic marker of thyroid disease.

The authors proposed a number of immunohistochemical markers to be used in histological diagnostic studies of the heart and protocols for their identification that extend the possibilities for the estimation of the heart function in case of pathological changes and sudden cardiac death.

The aim of this study was to determine the possibility of immunohistochemical demonstration of various tissue antigens in paraffin sections of zinc-ethanol-formaldehyde-fixed spinal cord, surrounded by the vertebral tissues and adjacent muscles, after decalcification in formic acid, in adult (n = 5) and newborn (n = 2) Wistar rats. In the present work, the antibodies used were against: glial fibrillary acidic protein, neurofilaments, neuronal specific nuclear protein, enzyme tyrosine hydroxylase, protein of synaptic vesicles--synaptophysin and the intermediate filament protein--vimentin. Combination of fixation in zinc-ethanol-formaldehyde and decalcification in 21% formic acid solution was shown to provide an optimal preservation of the antigens studied.

Using immunogold labeling, excitatory serotoninergic synapses of both chemical and mixed types, were found on the ventral dendrite (VD) of goldfish Mauthner neuron (MN).They are characterized by the presence of several mitochondria in the bouton and by an obligatory desmosome-like contact (DLC) besides the active zone (AZ). Their AZs were commonly found to make contact with the unlabeled chemical crested synapses, which, in turn, directly interacted with VD. These synapses were practically devoid of mitochondria and had no DLCs, thus allowing to identify them as the inhibitory ones. This "two-level" organization of excitatory serotoninergic and inhibitory synapses appears to be related to the reciprocal mechanism of the regulation of MN functional activity by visual input.

We studied the immunolocalization of hemeoxygenase-2 in neurons of the medulla oblongata in men (n=8), aged 18-44 years, who died from causes unrelated to the injury of the central nervous system and in people with the lifetime diagnosis of hypertension (n=6). It has been found that neurons with enzyme positive reaction are present in all parts of the medulla oblongata with concentrations ranging from 0.5 to 13.7% of the total number of cells. The high proportion of small neurons with the high or moderate density of deposits was found in the sensory nuclei. Large cells of the motor nuclei often exhibit the negative or low intensity of the enzymatic reaction. In arterial hypertension, a decrease in the proportion AH NO-positive neurons and the average optical density of the reaction product was noted. The reduction was seen in most affected neurons in the rostral part of the solitary tract nucleus and the lateral reticular nucleus. In the motor nuclei and in the dorsal nucleus of the vagus nerve, these parameters decreased as well although the reduction was not as great as observed in the sensory nuclei.

The aim of this investigation was to demonstrate the regulatory elements in the skin epithelium of Enteropneusta which are supposed to be related to the chordate ancestors. Using electron microscopy, it was found that in the skin epithelium of a representative of enteropneusts Saccoglossus mereschkowskii, the basal parts of some epitheliocytes took part in formation of a nerve layer. These cells were considered as receptor ciliated cells. The granular epithelial cells were shown to release secretion according to both exocrine and endocrine mechanism; these cells were characterized as endocrine-like regulatory cells. Fine granular cells possibly represent special receptor-endocrine-like cell type. The immunocytochemical detection of FMRFamid neuropeptide localization in histological sections confirmed the electron microscopic data on the presence of receptor and endocrine-like cells in the epithelium. It is suggested that the skin epithelium of Enteropneusta contains a peculiar neuro-endocrine regulatory system that is represented by receptor cells, receptor-endocrine-like cells of an open type and nerve elements of the nerve layer.

An analysis of modern methods of diagnostics such as morphological, immunohistochemical and spectral, which included the bronchoscopy and spectrometry by using reflectance and autofluorescent regime, was made. The data involved the results of prospective follow-up study of 167 patients (620 biopsies). An obligatory spectrometry of suspicious area was carried out before the forceps biopsy. The microslides, which met the requirements of criteria of one of the carcinogen steps (n=201), were subjected to the in-depth morphological and immunohistochemical investigations. The tendency of angiogenesis (CD31 and CD34), proliferative activity (Ki-67), level of apoptosis (P53), EGFR expression were estimated. The sensitivity of combined endoscopic method was 94,74% by specificity 79,95% and high prognostic value of negative endoscopic diagnosis - 99,4%.

The objective of the present study was the comprehensive evaluation of quantitative and functional metabolic characteristics of human leukocytes following in vitro irradiation of peripheral blood samples at a dose of 0.25, 0.50, 1.0, and 4.0 Gy for the development of a rationale for their application as biological markers of radiation exposure in forensic medical expertise of the subjects affected by supernormal ionizing radiation. The total number of leukocytes, lymphocytes, and their subpopulations as well as the functional and metabolic status of neutrophils were determined. The results of the study indicate the possibility of using a set of functional metabolic characteristics of human leukocytes as the additional forensic medical criteria for diagnostics of radiation impact on living subjects.

A protocol of immunocytochemical demonstration of choline acetyltransferase (ChAT), a key enzyme of acetylcholine synthesis, in paraffin sections of the brain of some laboratory animals, is presented. The method is simple, gives fairly reproducible results and allows for demonstration of ChAT in neurons, nerve fibers, and terminals in preparations of at least three species of laboratory animals including rat, rabbit, and cat. Different kinds of fixation (10% formalin, 4% paraformaldehyde, or zinc-ethanol-formaldehyde) were found suitable for immunocytochemical visualization of ChAT, however, optimal results were obtained with the application of zinc-ethanol-formaldehyde

In this paper we describe the modern immunohistochemical method detecting the neuronal marker PGP 9.5. The application of this method for the study of the innervation of rat and human heart, the detailed examination of the topography of the cardiac nervous apparatus, as well as their changes in pathological states, is demonstrated. Structural organization and the nature of the terminal branches of nerve apparatus suggest that they have afferent function. Protocol of the reaction demonstrating PGP 9.5 on paraffin sections is presented.

The objective of the investigation whose results are given in the paper was to compare a few HER2 testing techniques (immunohistochemistry, fluorescence in situ hybridization, and silver in situ hybridization) for adenocarcinomas of the stomach and esophagogastric junction. Different antibody clones and in situ hybridization kits were comparatively analyzed. Recommendations for the use of different testing methods and for the interpretation of their results are given.

By immunohistochemical method the hydrogen sulfide (H2S)-producing system was studied in digestive tract in fry and adult individuals of the masu salmon Onchorhynchus masou. The total and age-related peculiarities have been revealed in morphology, distribution, and the amount of enteral cystathionine beta-synthase (CBS)-immunopositive neurons. They have been established to be present in fry only in esophagus a stomach, while in adult fish--along the entire length of the digestive tract, the number of marked cells in esophagus, stomach, and distal intestine exceeding that in the proximal and middle intestines. In fry and in adult fish the neurons and nerve fibers are located in branches of vagus, in intermuscular and submucosal nerve plexuses, in serosal and muscle layers. There are detected uni-, bi-, and much seldom the multipolar neurons surrounded by numerous poorly and undifferentiated nerve cells. Besides, the CBS-immunopositive netve fibers were present in the subepithelial area of the duct of swim bladder.

The representatives of herpesviruses family (Herpesviridae--virus of herpes simplex and human cytomegalovirus) are largely widespread in human population. These herpesviruses bring on severe disorders of embryonic development up to fetal death, newborn diseases, neurologic disturbances, deafness, and blindness and in transplantation patients the severe internal organs affections and transplanted organs rejection. Both herpesviruses are able to affect the central nervous system and result in encephalitis with lethal outcome. The particular attention deserve the asymptomatic forms in case of which virus is excretee and can be transmitted both by horizontal line (sexual way included) and by vertical line (in the process of itrauterine development offtetus). The lacking of clear djfefrentiating clinical symtoms frequentyv observed under manifestations of herpesviruses infections brings another danger The comparison is made of fouur methods of detection of herpes simplex in urethra scrapes: the immunocytochemical method of detection of herpes simplex antigen in smears: the rapid cultural method; the immune-enzyme method of detection of herpes simplex antigen; the polymerase chain reaction. The results demonstrated that the rapid cultural method detected herpes simplex in infected samples on all stages of disease and in even more quantity at the stage of exacerbtion as compared with the polymerase chain reaction. The presented data testify the actuality of development of of laboratory diagnostics of herpesviruses injection to relevant diagnosis, determination of form and stage of disease, timely initiation of treatment and monitoring of therapy.

Morphological and clinical characteristics of the urinary bladder cancers (UBC) are important diagnostics and prognostic criteria, however the possibilities of biopsy using/for prognosis of recidivation or efficiency of UBC treatment are limited. The most popular diagnostic and prognostic immunohistochemical markers are the regulators of cell cycle (P53, P21, Ki-67) and cytokeratins. In order to revealed immunohistochemical criteria, objectively reflected the malignancy of UBC, we studied the expression level of P53, proliferative index of Ki-67 and the malignant of UBC according to CK20 in the biopsy of 32 patients with superficial UBC. The immunohistochemical reaction with antibody to P53, CK20 and Ki-67 was carried out at paraffin slices. We detected differences of P53 expression levels at carcinomas with high and low malignancy rate. The expression levels of CK20 and the Ki-67 proliferative index were different between Ta and T1-T2 cancers (p = 0.006). Intensity of nuclear staining with P53 antibodies could be use for estimation of the UBC malignancy rate. Pathological type of CD20 expression and high percentage of Ki-67-positive staining nucleus are evidence of the invasive urothelial tumors. The using of P53, Ki-67 and CD20 could be recommended for pathohistological investigation of biopsy and surgical material of UBC to diagnostic objectification and prognosis of its clinical course.

According to ASCO/CAP guidelines, there are two recommended methods for Her2/neu-testing in breast cancer patients such as immunohistochemistry and fluorescence in situ hybridization. This paper analyses results of alternative immunocytochemical method for Her2/neu detection and its diagnostic pitfalls.

Immune localization of heme oxygenase-2 in neurons of some nuclei of the spinal cord and brain stem in 6 men 18-44 years old who died from causes unrelated to injury of central nervous system was studied. Neurons with positive reaction are determined for all studied regions of the brain where their contents in various nuclei ranging from 0.5 to 16% of the total number of cells detected by methylene blue. In all the sensory nuclei there is a high proportion of small neurons with a high or moderate density of reaction produce deposits. Large cells of motor nuclei often exhibit negative or low intensive enzyme reaction.

We present a case of a rare tumor--melanotic schwannoma of trigeminal nerve root and gasserian ganglion. Differential diagnosis between metastatic melanoma and melanotic schwannoma (MS) is associated with serious difficulties and high responsibility. Metastatic melanoma is a high grade tumor while most MS are benign lesions with good outcome. By the date 105 cases of these tumors are described in the world literature, 3 of them originated from trigeminal nerve root and gasserian ganglion. MS predominantly occur in relatively young patients, they are characterized by presence of Carney's complex and psammomatous bodies and absence of primary focus. MS and metastatic melanoma have similar appearance on MRI due to presence of melanin granules. Indirect signs evident for MS include cystic structure and dumbbell-shaped growth. Metastatic melanoma of cranial nerves is more typical in people older than 40, primary focus in the face in the zone of innervation of affected nerve is common. In case of absence of the listed features differential diagnosis is based on immunohistochemical analysis and electron microscopy of tissue samples.

The objective of the present work was to develop the forensic medical criteria for chronic narcotic intoxication based on the results of morphological studies. The internal organs from 179 cadavers were available for the examination following death from acute poisoning with narcotic substances or as a result of chronic narcotic intoxication. The studies were carried out with the use of routine histological staining techniques and an immunohistological method. The data obtained provided a basis for the development of criteria to be employed in forensic medical diagnostics of acute poisoning with narcotic drugs and chronic narcotic intoxication. These criteria include ischemia of cerebral neurons, pulmonary emphysema with the formation of foreign body-type granulomas and fibrin/erythrocyte thrombi, morphological signs of ventricular fibrillation, the picture of bacterial endocarditis, follicular hyperplasia of the lymphoid organs, chronic portal hepatitis, and nodular degeneration of the adrenal cortex associated with its atrophy.

Phosphorylated H3 histone is of interest for the study of proliferative activity of the cells in various tissues. Its detection is used in histopathological tumor diagnosis. The purpose of this study was to develop immunocytochemical protocols for the demonstration of H3 phosphohistone in the cells of the developing nervous system for classical light microscopy and confocal laser microscopy. The protocols presented allow to identify the dividing cells in the structures of developing brain with high selectivity and good reproducibility. Zinc-ethanol-formaldehyde is recommended as an optimal fixative for H3 phosphohistone.

Forebrain subventricular zone (SVZ)--the putative major source of neural stem cells in the brain of adult mammals--can hardly be visualized using routine histological staining. The present study was focused on the possibility of application of immunocytochemical approach for accurate delineation of the border between SVZ and striatum. It was shown that immunocytochemical reactions demonstrating tyrosine hydroxylase or synaptophysin were optimal for the determination of the border between SVZ and striatum in different mammals.

This paper was designed to illustrate the advantages of immunohistological methods over those traditionally employed in this country for the study of material evidence. A method for the identification of ABO antigens in cellular elements based on the use of Diagnostic BioSystems-Universal HPP kits is described. The reaction is specific and as sensitive as the immunofluorescent techniques. The stained preparations can be stored during a long period without the loss of their properties. The authors recommend to use the method being considered in forensic medical (specifically, in forensic cytological) practice.

The removed pheochromocytomas of 25 patients aged from 25 to 72 years have been studied. Among them 5 tumors have had bilateral character, 8 pheochromocytomas have been diagnosed as malignant, 4 patients have had metastases. The average size of benign pheochromocytomas has been 4.0 cm, malignant - 4.5 cm. The majority of malignant tumors have had areas with diffusing growth patterns. The results of immunohistochemical study of benign and malignant pheochromocytomas have had small differences. The study of tumor ultrastructure hasn't found out their biological potential.

The observation of stromal gastric tumor of 12 years old girl has been investigated. The diagnostics was carried out on tumor biopsy taken by a laparoscopy. An evident edema of stroma caused "pseudo-papillary" organization of epithelioid cell neoplasm prevented the right diagnosis established only by immunohistochemical staining of CD117 and CD34 markers. The absence of mutations in C-kit and PGFRA genes uncovered by molecular-genetic analysis is typical for stromal gastric tumors in child. The next gastric resection allowed to estimate tumor appearance and localization, histological organization, and to repeat immunohistochemical studying. This observation confirmed the correctness of diagnosis and established high level of Ki67 proliferative activity (12-15%) determined prescriptions of target medicamental therapy.

Histological studies of oral cavity mucosa and salivary glands in subjects with chronic alcoholic intoxication revealed changes at the surface of the tongue and in the glandular tissues. Specific features of chronic alcoholic intoxication include acinar and ductal hyperplasia, reduction of the adipose tissue mass in salivary gland stroma, predominance of T-lymphocytes in hard palate minor salivary glands and B-lymphocytes in the stroma of labial minor salivary gland, the absence of plasma cells in the stroma of hard palate minor salivary glands and labial mucosa. Leukoplakia, dysplasia, and hyperplasia of the basal epithelial layer of oral cavity mucosa are considered to be the signs of long-term (over 12 months) alcohol consumption.

Analysis of the results of examining 57 primary and metastatic soft tissues tumors has shown the principal capabilities of immunophenotyping in the cytosmears for differential diagnosis, the determination of histogenesis, and the assessment of their prolferative activity. Its major benefit is short response time and easiness to use. All results were assessed by comparison with the data of immunohistochemical study on paraffin sections.

Serous exudates were studied by immunoperoxidase and immunofluorescence immunocytochemical assays and by laser flow cytofluorimetry. The current high-tech techniques detect sparse tumor complexes in the exudates at the early metastatic stages, which are undetectable by the routine cytological assay, and also reduce the hyperdiagnosis of the tumor process. The use of a set of immunocytochemical markers can specify the source of metastatic spread when the primary tumor focus is not found. Fluorescence immunocytochemistry of the exudates has been shown to be highly effective and rapid during emergency intraoperative studies to clarify the extent of the tumor process.

Terminological and classification aspects are of great importance in the systems approach to managing endocrinological patients, including those with thyroid cancer. The 2007 Bethesda (USA) thyroid cytological study classification system contributes to the improvement of patient selection results for surgery for cancer evidence. Experience with more than 120000 cytological studies and more than 4500 examinations of intraoperative thyroid specimens could reveal some gap between the cytological interpretation of thyroid pathology and the evaluation of individual morphological phenomena that are particularly important in the differential diagnosis of follicular neoplasias, rare variants of papillary carcinoma. Among other things, there was evidence that it was inexpedient to use the term "dysplasia" in relation to the follicular epithelium and in both cytological and histological practice. Analysis of the results of consultations could reveal the most common diagnostic errors during morphological studies.

Unlabelled: Precancerous changes in the epithelium of the cervix uteri include low- and high-grade squamous intraepithelial lesions (LGSIL and HGSIL). Morphologically, these must be differentiated, among other conditions, immature squamous metaplasia (ISM) of the cervix uteri., Materials and Methods: This study involved 119 cases divided into 3 groups: 1) LGSIL (n = 28); 2) HGSIL (n = 56); 3) ISM (n = 35). An immunohistochemical study was conducted using antibodies to p16INK4a, CK17, and Ki-67 according to the standard protocol., Results: Ki-67 expression was mainly low and moderate in 67.9% of cases in the LGSIL group; it was seen in 100% of cases in the HGSIL group, being significant in 73.5%. In the ISM group, Ki-67 was identified in 65.7% of cases, most of which demonstrated a low-to-moderate expression. In the LGSIL group, the expression of p16INK4a was recorded in 46.4% of cases. In the HGSIL group, the reaction was observed in 73.2% of cases; of them 57.2% it was strong. In the ISM group, the reaction was chiefly low in 38.6% of cases. In the LGSIL group, CK17 expression was found in 17.9% of cases. In the HGSIL group, the reaction was recorded in 19.7% of cases, the expression being predominantly low. In the ISM group, the marker was detectable in 88.5% of cases; of them 51.4% showing a high expression. The expressions of p16NK4a and Ki-67 were statistically significantly higher in the HGSIL group (p < 0.05) than those in the LGSIL and ISM groups. On the contrary, the expression of CK1 7 was significantly higher in the ISM group (p < 0.05) than in the HGSIL and LGSIL groups. Thus, evaluation of p16INK4a, Ki-67, and CK17 expressions is a valuable diagnostic tool in the differential diagnosis of HGSIL, LGSIL, and ISM.

The functioning of Jacobson's or vomeronasal organ (VNO) in man is the subject-matter of discussion today. It is generally taken that VNO as an anatomic structure also remains in the adult; however, its receptor apparatus still degenerates in the fetal stage of ontogenesis. Nevertheless, the data available in the literature on the time and specific features of degenerative changes in the human fetal VNO are conflicting and ambiguous. The authors examined the human fetal nasal septum from the 8th week of development to birth, by applying the traditional histological procedures and neuron-specific beta3-tubulin antibodies. An immunohistochemical study could first show the receptor apparatus of the human fetal VNO at weeks 8-26 of development. The immunohistochemical study on a series of sections could reveal the regularities of spatial receptor distribution depending on the time of fetal development. In addition, the developed human fetal vomeronasal nerve and ganglion at weeks 8-26 were described, in human fetuses at weeks 8-26. The neuron-specific marker test has shown the nerve fibers departing directly from the VNO wall, which is inconsistent with the data available in the literature on vomeronasal nerve degeneration in this sign just after the 18th week of development.

The purpose of this study was to investigate the expression of intermediate filament proteins (vimentin and glial fibrillary acidic protein--GFAP) in the cells of embryonic rat neocortex at different time points after its allotransplantation into injured sciatic nerve of adult animals. Using immunohistochemical methods, the differentiation of GFAP-positive astrocytes from vimentin-positive radial glial cells was observed in embryonic rat neocortex, grafted into sciatic nerve. It was shown that the differentiation of the embryonic neocortical astrocytes in the transplants took place a few days earlier than in the rat neocortex during normal ontogenesis. Reactive gliosis was demonstrated in the long-term transplants, as indicated by a large number of intensely stained GFAP-positive cells and vimentin-containing astrocytes. These findings suggest that ectopic neurotarnsplants could serve as a model for fundamental studies of the mechanisms of reactive gliosis development.

The paper deals with the problems of HER2 testing in breast cancer. It describes the optimal immunohistochemical procedure and the principles of assessment of its results.

A panel containing 11 types of monoclonal antibodies (mAb) specific for the recombinant protein pp65 of human herpesvirus type 5 was constructed. Enzyme immunoassay and immunoblotting of the immunochemical properties of mAb indicated that mAb could interact well with both recombinant and native pp65 antigen. Testing the suitability of mAb for an immunocytochemical study established that mAb 5F10 was highly effective in detecting the major tegument protein pp65 in the persistently infected Vero cells. A combination of the immunochemical properties of mAb enables them to be recommended for the immunodiagnosis of human cytomegalovirus infection.

Quantum dots (QD) nanocrystals consisting of CdSe core with ZnS shell are a novel class of fluorophores with tremendous potential in microscopy and cytometry techniques. The unique optical features of Qdots, namely, high photostability and extinction coefficient, wide absorption and narrow emission spectra, and large Stokes shift make them desirable fluorescent tags for diverse biomedical applications. Applications of this novel technology in microscopy and cytometry produce reliable multicolor specimens due to increased photostability, ability for multiplexing and narrow emission spectra of nanocrystals. QD conjugates are available on the market and could be prepared in the laboratory. This paper describes the application of QD-conjugates for immunophenotyping and FISH assessment of cells and tissues, and the requirements for microscope and flow cytometer reengineering for successful use of QD in multiplex fluorescent format. Despite the considerable progress, important methodological issues still need to be solved in terms of QD nanocrystals' size, heterogeneity, functionalization and stability of their conjugates. We discuss practical approaches and challenges that need to be addressed to make QD immunostaining a standard method in biology.

Ezrin (cytovillin or p81 protein) is an actin-binding protein, a member of ERM (ezrin, radixin and moesin) family, which species contribute to stabilization of the plasma membrane-formed structures. The aim of the present study was to demonstrate the ezrin-containing cells in the rat brain and to describe their topography and morphological features. The most pronounced immunohistochemical reaction to ezrin was found in the epithelium of the choroid plexus, cells of the subcommissural organ and ventricular ependyma. Moreover, ezrin staining was also detected in the unidentifiable cells in the subventricular zone, rostral migration pathway and astrocytes in various brain areas. Preferential ezrin localization in the brain cells contributing to formation of barrier structures suggests its involvement in transport processes in the CNS.