[Possibilities of multiphoton microscopy for the diagnosis of the vulvar lichen sclerosus]. / Vozmozhnosti mul'tifotonnoi mikroskopii dlya diagnostiki skleroticheskogo likhena vul'vy.

BACKGROUND: Vulvar lichen sclerosus (VLS) is a chronic and recurrent dermatosis of an inflammatory nature with severe focal atrophy of the skin. Connective tissue changes are polymorphic and are still not taken into account in histological diagnostics due to the difficulty of interpreting routine histological methods. In this work, we use multiphoton microscopy (MPM) as a new imaging technique that provides detailed information about the organization of collagen fibers in the dermis based on a non-linear second harmonic generation (SHG) process. OBJECTIVE: To determine the degree of connective tissue damage in lichen sclerosus using standard histological techniques and to reveal the diagnostic capabilities of multiphoton microscopy. MATERIAL AND METHODS: We studied 42 biopsies with a histopathological diagnosis of VLS and 10 biopsies of normal vulvar skin. Histological, histochemical and immunohistochemical evaluation was used in comparison with MPM data. Quantitative analysis included the determination of the thickness, length of collagen fibers and the average intensity of the SHG signal. RESULTS: A comprehensive study of the skin showed 4 groups of changes that can be regarded as the degree of the dermis damage: initial, mild, moderate, severe. The affected area at the initial and mild degree has subtle changes, however, it is reliably identified by quantitative analysis of the SHG signal. So, the initial degree is characterized by thin (1.3-1.8 µm) long (56-69 µm) collagen fibers, with a moderate degree, the fibers are thickened (3.4-4.3 µm) and fragmented (22-37 µm). The affected area in moderate and severe cases undergoes homogenization, which is associated with the deposition of extremely thin (0.6-0.9 µm) short (16-28 µm) collagen fibers and the expression of type V collagen. CONCLUSION: Multiphoton microscopy in the second harmonic generation mode is a reliable method for identifying collagen fibers in tissues. The study made it possible to identify 4 degrees of the dermis damage in vulvar lichen sclerosus.

Cross-Polarization Optical Coherence Tomography for Clinical Evaluation of Dermal Lesion Degrees in Vulvar Lichen Sclerosus.

The aim of the study was to identify different degrees of dermal lesions in vulvar lichen sclerosus (VLS) using cross-polarization optical coherence tomography (CP OCT) based on attenuation coefficient to detect disease early manifestations and to monitor the effectiveness of treatment. Materials and Methods: The study included 10 patients without pathology and 39 patients with VLS diagnosed histologically. CP OCT was performed in vivo on the inner surface of the labia minora, in the main lesion area. From each scanning point, a 3.4×3.4×1.25-mm3 3D data array was obtained in 26 s. CP OCT examination results were compared with histological examination of specimens stained with Van Gieson's picrofuchsin.Quantitative analysis of OCT images was performed by measuring the attenuation coefficient in co-polarization and cross-polarization. For visual analysis, color-coded charts were developed based on OCT attenuation coefficients. Results: According to histological examination, all patients with VLS were divided into 4 groups as per dermal lesion degree: initial (8 patients); mild (7 patients); moderate (9 patients); severe (15 patients). Typical features of different degrees were interfibrillary edema up to 250 µm deep for initial degree, thickened collagen bundles without edema up to 350 µm deep for mild degree, dermis homogenization up to 700 µm deep for moderate degree, dermis homogenization and total edema up to 1200 µm deep for severe degree.Pathological processes in dermis during VLS like interfibrillary edema and collagen bundles homogenization were visualized using CP OCT method based on values of attenuation coefficient in co- and cross-polarization channels. However, CP OCT method appeared to be less sensitive to changes of collagen bundles thickness not allowing to distinguish thickened collagen bundles from normal ones with enough statistical significance. The CP OCT method was able to differentiate all degrees of dermal lesions among themselves. OCT attenuation coefficients differed from normal condition with statistical significance for all degrees of lesions, except for mild. Conclusion: For the first time, quantitative parameters for each degrees of dermis lesion in VLS, including initial degree, were determined by CP OCT method allowing to detect the disease at an early stage and to monitor the applied clinical treatment effectiveness.

[Heart damage in the combined types of systemic amyloidosis]. / Porazhenie serdtsa pri sochetannykh formakh sistemnogo amiloidoza.

The cardiovascular system is a common target of systemic amyloidosis (SA); amyloid light chain (AL) cardiac amyloidosis (AL-CA), the wild-type transthyretin (ATTRwt-CA), and mutant-type transthyretin (ATTRmt-CA) are the most studied types of SA. The literature describes only single cases of two types of SA in the same patient. OBJECTIVE: To identify and determine the clinical and morphological characteristics of combined types of SA in patients with biventricular chronic heart failure (CHF). MATERIAL AND METHODS: Eighty autopsy protocols for biventricular CHF deaths were retrospectively analyzed. Immunohistochemistry and confocal laser scanning microscopy (CLCM) with antibodies to amyloid A (AA), serum amyloid-P (SAP), prealbumin, and immunoglobulin kappa (κ) and lambda (λ) light chains were performed. RESULTS: The myocardium showed a combination of different types of SA in 6 (7.5%) cases, including Alλ-CA+ATTR-CA, ALκ-CA+ATTR-CA, and AA-CA+ATTR-CA in 4, 1, and 1 cases, respectively. Macroscopically, the heart mass averaged 470±20 g; the thickness of the left and right ventricular myocardium was 1.5±0.2 and 0.4±0.1 cm, respectively; the interventricular septum averaged 1.2±0.2 cm; and the cardiac index was 0.008. The myocardium was dense, dark red with diffuse layers of whitish dense fibrous connective tissue; the heart cavities were enlarged. Microscopically, in 25% of cases, all heart parts had ALλ-CA that was characterized by massive amyloid deposits localized predominantly in the intramyocardial vessel wall, intermuscular connective tissue, and perivascularly. The myocardium also displayed small amyloid deposits of ALλ-CA and ATTR-CA in the intermuscular connective tissue and intramyocardial vessel wall. Amyloid deposits were located in different parts of the myocardium; there were also areas of co-localization of ALλ-CA+ATTR-CA. CONCLUSION: The combined types of SA occurred under the guise of coronary heart disease and the dilated cardiomyopathy phenotype. The combined amyloid AL-CA and ATTR-CA was generally localized in the interstitium and myocardial vessels. There were also small areas of co-localization of amyloid deposits, which were found mainly in the intramyocardial vessels.

[Anatomical pathology of novel coronavirus (COVID-19) infection. First impressions]. / Patologicheskaya anatomiya novoi koronavirusnoi infektsii COVID-19. Pervye vpechatleniya.

To date, descriptive results of a clinical and morphological study of novel coronavirus COVID-19 infection, mainly of the lungs, have appeared. However, in other organs, primarily in the cardiovascular system, there are substantial structural changes that lead to multiple organ dysfunction and contribute to death. OBJECTIVE: To analyze the thanatogenetic significance of novel coronavirus COVID-19 infection in different age and gender groups and to describe the main morphopathological manifestations in various organs. MATERIAL AND METHODS: The investigators carried out a comprehensive analysis of 700 autopsies of people disease from the novel coronavirus COVID-19 infection, which included an examination of gross changes reflected in the autopsy protocols and forensic medical examination reports, as well as that of microscopic changes detected during histological examination of organs. Immunohistochemistry (IHC) using mouse or rabbit antibodies to CD34, CD68, EMA, Ki67, caspase-3, and VEGF was employed in some observations. RESULTS: The novel coronavirus COVID-19 infection was the primary cause of death. Acute respiratory and pulmonary heart failure and multiple organ dysfunction became the leading thanatogenetic mechanisms in COVID-19. Cardiovascular disease, diabetes mellitus, and obesity were the most common diseases in patients with COVID-19. The most pronounced lung changes in COVID-19 were determined with a predominance of multiple total bilateral lesions of the lower lobes of the lungs, which was manifested by virus-induced changes in the parenchyma and stroma, as well as by microcirculation disorders. Acute dyscirculatory and ischemic changes in the parenchymal organs dominated in tissue damage caused by the virus. CONCLUSION: The changes in different organs of those who have died from the new coronavirus COVID-19 infection are stereotyped and include the manifestations of virus-induced action and a systemic inflammatory response with mainly microvasculature alteration, which leads to the development of coagulopathies and, accordingly, to total hypoxia.

THE STUDY OF POLYMORPHISMS OF XPD GENE A751C IN LUNG CANCER PATIENTS WITH DIFFERENT CLINICAL AND MORPHOLOGICAL CHARACTERISTICS OF TUMOR.

There were studied distribution of polymorphic variants of gene of repair of DNA XPD A751C in lung cancer depending on histological type of tumor (small cell / non-small cell lung cancer) and the prevalence of tumor process (with foci / without foci of metastasis). It was found a significant increase in the incidence of minor allele C, CC and AC genotypes of the polymorphic site of gene XPD A751C in patients with lung cancer. We estimated relative risks of lung cancer development in carriers of polymorphic variants of gene XPD A751C. The heterozygous genotype AC polymorphism of gene XPD A751C is characterized by the greatest risk of developing lung cancer with small cell histological type. Homozygous CC genotype of the polymorphic site of gene XPD A751C is associated with non-small cell lung cancer development. Statistically significant differences in the distribution of polymorphic variants of gene A751C XPD depending on spread of cancer were not received.

[The informative value of oral mucosal biopsy for the diagnosis of systemic amyloidosis].

One hundred and thirteen oral mucosal biopsy specimens were analyzed in patients suspected of having systemic amyloidosis. Histological, immunohistochemical, and genetic studies and polarized light microscopy revealed oral amyloid deposits in 72.6% of cases, including 63.5% with metabolic syndrome and 36.5% with another etiology of chronic heart failure (coronary heart disease, cardiomyopathy). Systemic amyloidosis was found in 13.4% of cases (hereditary transthyretin, AL, and senile forms in 1.8, 10.5, and 1.1% cases, respectively). An anterior abdominal wall skin flap was a valid location to reveal the systemic forms of amyloidosis. Patients with metabolic syndrome and periodontal diseases may have local oral amyloidosis.

[Oral status and oral mucosa blood circulation changes in patients with chronic heart failure].

The objective of this study was to characterize dental status and oral mucosa blood flow in patients with chronic heart failure and amyloid deposits in oral mucosa. Histological and immunohistochemical analysis of 80 oral mucosa biopsies taken from patients aged 32-72 years with chronic heart failure I-IV NYHA functional class was carried out. It detected a systemic amyloidosis in 15.7% of cases; a local amyloid deposition in oral mucosa was found in 58.5% of cases. Amyloid deposition in oral mucosa was associated with severe chronic generalized periodontitis in more than a half of cases. Amyloid deposits in oral mucosa were revealed more often in patients with metabolic syndrome (63.5%). The article describes dental status and oral mucosa blood flow in patients with heart failure.

[AL amyloidosis originally considered as systemic vasculitis].

The paper describes a sectional case of the rare unusual manifestation of AL amyloidosis in a 58-year-old woman with multiple small myocardial scars, a lung decay focus, and extensive gastric mucosal necrosis that has led to fatal hemorrhage, which was assessed as the pattern of systemic vasculitis before immunological examination. The pathogenesis of this form of AL amyloidosis is considered as a manifestation of malignant transformation of B lymphocytes.

[Desmin-related cardiomyopathy].

The observation of 26 years old patient with desminopathy declared itself by hypertrophied cardiomyopathy with its transformation into restrictive phenotype is presented. The features of pathologic course at the patient were a dominance and diversity of cardiac manifestations. Endomyocardiac biopsy allowed suspecting the desminopathy confirmed by genetic analysis. Morphological features of desmin-related cardiomyopathy were irregular desmin conglomerates mainly located under sarcolemma and an indirect histological signs of idiopathic cardiomyopathy as well nuclear polymorphism, perinuclear "nimbus", chaotic located myofibrils.

[The implementation of polymerase chain reaction technique: the real time to reveal and differentiate the viruses of human papilloma of high carcinogenic risk].

The polymerase chain reaction technique was applied in "real time" format to evaluate the occurrence rate and infection ratio of various genotypes of human papilloma of high carcinogenic risk in virus-positive women and contact persons. The examination sampling consisted of 738 women aged of 17-50 years. The examination results permitted to establish high percentage of infection of 546 patients (74%) by carcinogenic papilloma viruses. The analysis of detection rate of various genotypes of human papilloma of high carcinogenic risk established that the 56th and 16th types of high carcinogenic risk are revealed more often than others--in 33% and 15.4% correspondingly. In males, first place in occurrence rate is for those types of virus of human papilloma: the 56th n = 10 (33.3%), 16th n = 3 (10%), 45th n = 3 (10%), 51th n = 3 (10%). The rest of genotypes are detected in 3-7% cases.

[Safe drinking water supply to the Vologda Region's population using risk assessment methodology].

To supply the population with qualitative potable water is a priority problem in the provision of sanitary-and-epidemiologic well-being and in the prevention of disease in the Vologda Region. The monitoring of the results of laboratory control over the quality of drinking-water and the assessment of health risk enabled a package of measures to be proposed to optimize the conditions of drinking water supply in the Vologda Region. The risk assessment technology used by a state agency for sanitary-and-epidemiological surveillance makes it possible to substantiate a system of actions to organize household water use and to include scientifically grounded proposals into the developed regional and local programs.

[The regional characteristics of the hygienic evaluation of biological contamination of superficial waters].

The paper gives the data characterizing the specific features of formation of an anthropogenic load on water bodies in north regions; sanitary and hygienic criteria for estimating the biological contamination; and the factors that increase the biological productivity of the algal flora; a classification of eutrophic waters is also proposed.

[Cardiac manifestations of amyloidosis].

Amyloidosis is shown by a group of diseases with a variability of clinical and morphological manifestations determining difficulties in its diagnosis. The cardiovascular system is commonly the site of involvement in different forms of amyloidosis. The variants of cardiac involvement in the pathological process in amyloidosis are outlined depending on various precursor proteins. The specific features of different forms of cardiopathic amyloidosis are described in relation to its type.

[The role of apoptosis in ishemic damage to the myocardium].

36 specimens of the myocardium were obtained during necropsy of patients who had died of acute cardiac failure. Bcl-2, caspase 3 and p53 were studied immunohistochemically. TUNEL method was used to study apoptic DNA fragmentation in cardiomyocytes (Cms). The results suggest that apoptosis contributes to the development of cardiomyocyte injury in ishemia. Apoptotic death of Cms may lead to electric instabilly of the myocardium in early ishemia and cause sudden cardiac death. Apoptosis of the Cms may precede extension of the necrotic area. Further studies of apoptotic death of Cms in ishemic heart disease may help to renew therapeutic strategies aimed at regulation of apoptosis.

DNA-hydrolyzing antibodies from sera of autoimmune-prone MRL/MpJ-lpr mice.

Catalytically active antibodies (abzymes) hydrolyzing proteins, polysaccharides, ATP, DNA, and RNA have been detected in the sera of patients with various autoimmune and some viral diseases, but abzymes from the sera of animals are practically unstudied. The development of lupus-like autoimmune disease of MRL/MpJ-lpr mice is an experimental model for study of autoimmune pathologies and immunopathogenesis. In this work, homogeneous IgG preparations were isolated from the sera of MRL/MpJ-lpr mice. These antibodies (Abs), their Fab-fragments, and isolated light chains were shown to possess catalytic activity in DNA hydrolysis, whereas Abs from the sera of control healthy mice did not hydrolyze DNA. The data demonstrate that DNA hydrolyzing activity is an intrinsic property of Abs from MRL/MpJ-lpr mice. It was shown that various markers of autoimmune pathologies (level of total protein concentration in urea (proteinuria), Abs titers to native and denatured DNA, and DNA-hydrolyzing activity of IgG) increased in animals with aging, but they noticeably increased (2-22 times) only after appearance of obvious indicators of pathology independently of age. The highest increase in proteinuria (25-fold), anti-DNA Abs titers (12-19-fold), and abzyme activity (120-fold) was found in mice after their immunization with DNA-protein complex.

[Damage to the vision organ by non-medical lasers]. / Povrezhdenie organa zreniia lazerami nemeditsinskogo naznacheniia.

The offered study is dedicated to an acute ophthalmology issue, i.e. damage to the vision body by irradiation of non-medical lasers used in every-day life, industry and army. A total of 6 patients with damages to the central eye section were analyzed. The results of ultrasound fluorography as well as of electrophysiological examinations are described and conservative therapy schemes are presented. According to a preliminary conclusion, the outcome of the clinical condition can be accompanied by a low and eccentric vision.

[Conservative therapy in rehabilitating patients with eye injuries]. / Konservativnaia terapiia v reabilitatsii bol'nykh s povrezhdeniiami organa zreniia.

The authors offer recommendations on the choice and use of optimal clinically tried drugs in ophthalmic traumatology with consideration for posttraumatic changes, doses, and mechanism of action. The recommendations are based on the clinical observations of 16,000 patients treated at department of ocular injuries, reconstructive surgery, and ocular prosthetics of Helmholtz Institute of Ocular Diseases in Moscow.

[Clinical and immunomorphological parallels in posttraumatic uveitis]. / Kliniko-immunomorfologicheskie paralleli pri posttravmaticheskikh uveitakh.

Parallels between the clinical diagnosis, immunological parameters of the leukocyte migration inhibition test (LMIT) to eye tissue antigens (uveoretinal, lenticular, and retinal), and morphological picture are studied in patients with posttraumatic uveitis and consequences of grave penetrating injuries to the eye without uveitis symptoms. Cell sensitization to uveoretinal antigen is detected only in posttraumatic uveitis but not in consequences of injuries without uveitis. The authors come to a conclusion that only positive LMIT with uveoretinal or a combination of uveoretinal and lenticular antigens may be considered as an immunological validation of autoimmune posttraumatic uveitis. Further improvement of LMIT with purified fractions of uveoretinal antigens is needed.