Characteristic of Ultrastructure of Mice B16 Melanoma Cells under the Influence of Different Lighting Regimes.

Circadian rhythms of physiological processes, constantly being in a state of dynamic equilibrium and plastically associated with changes in environmental conditions, are the basis of homeostasis of an organism of human and other mammals. Violation of circadian rhythms due to significant disturbances in parameters of main environmental effectors (desynchronosis) leads to the development of pathological conditions and a more severe course of preexisting pathologies. We conducted the study of the ultrastructure of cells of mice transplantable malignant melanoma B16 under the condition of normal (fixed) lighting regime and under the influence of constant lighting. Results of the study show that melanoma B16 under fixed light regime represents a characteristic picture of this tumor-predominantly intact tissue with safe junctions of large, functionally active cells with highly irregular nuclei, developed organelles and a relatively low content of melanin. The picture of the B16 melanoma tissue structure and the ultrastructure of its cells under the action of constant lighting stand in marked contrast to the group with fixed light: under these conditions the tumor exhibits accelerated growth, a significant number of cells in the state of apoptosis and necrosis, ultrastructural signs of degradation of the structure and functions, and signs of embryonization of cells with the background of adaptation to oxygen deficiency.

Structural Changes in the Nervous Fibers of the Colon Mucosa in Experimental Acute Colitis.

Morphological, morphometric, and ultrastructural analysis of the nerve fibers in the colon mucosa was performed in C57BL/6 mice at various terms of development of acute colitis induced by dextran sodium sulphate. The nerve fibers were labeled with antibodies to pan-neuronal marker ßIII-tubulin. The progression of inflammatory and ulcerative processes in the mucosa on days 3-5 was associated with hyperplasia and hypertrophy of nerve fibers that peaked on day 7 after colitis induction. Ultrastructural analysis at all terms of colitis development showed moderate degeneration of axons. Thus, hypertrophy and hyperplasia of the nervous fibers in colon mucosa in experimental acute colitis correlated with aggravation of the ulcerative process in the mucosa. These changes are determined by alteration of histoarchitectonics and regenerative processes in the mucosa.

Quantitative and Qualitative Characterization of Phagocytic Activity of Macrophages of Bone Marrow and Fetal Origin.

We compared phagocytic activity of macrophages of monocyte origin and Kupffer cells under the influence of M1 and M2 inducers and without activation. Cultures of monocyte-derived macrophages and Kupffer cells were characterized by intensive expression of CD68 that was not affected by activation factors. At the same time, these cultures demonstrated different dynamics of phagocytic activity. Monocyte-derived macrophages initially had more pronounced absorption capacity that gradually increased during the experiment. Kupffer cells were characterized by abrupt fluctuations of phagocytic activity: sharp growth and rapid saturation. Despite these differences, the endosomes produced by monocyte-derived macrophages and Kupffer cells had similar degrees of maturity.

Heterodimeric V. nikolskii phospholipases A2 induce aggregation of the lipid bilayer.

We report that the action of the heterodimeric phospholipases A2 (PLA2s) from Vipera nikolskii, which comprises enzymatically active basic subunit and inactive acidic PLA2 homologue, on the lipid bilayer results in the aggregation and stacking of bilayers. These processes are demonstrated using two independent methods (fluorescence spectroscopy and electron microscopy). Aggregation of bilayers is possible because both subunits of the V. nikolskii heterodimer contain a membrane-binding site (also known as IBS). Thus, when the two IBSs bind to the membrane, the heterodimer acts as a connecting agent. Heterodimers induce aggregation of negatively charged bilayers composed of phosphatidylglycerol and do not induce aggregation of neutral bilayers composed of phosphatidylcholine.

Immunohistochemical Analysis and Electron Microscopy of Glial Cells in the Pancreas of Fetuses and Children.

Schwann cells forming peri-insular glial sheath of the pancreatic islets in some mammals can be involved in the pathogenesis of diabetes mellitus. Human pancreatic cells contain small elongated or oval cells of unknown origin with S100-immunopositive processes. We found that cells with processes located in pancreatic islets of human fetuses and children are similar to Schwann cells by their morphological and ultrastructural characteristics, immunopositive reaction for S100, and integration with nerve endings. In the pancreas of fetuses and children, Schwann cells are often seen in forming pancreatic islets and around the pancreatic ducts. The data suggests that Schwann cells can participate in the morphogenesis of human pancreatic islets.

[Pulmonary histological and ultrastructural changes in HIV infection concurrent with tuberculosis].

OBJECTIVE: To analyze cellular and subcellular disorders in the lung in the presence of HIV-associated tuberculosis. MATERIAL AND METHODS: The investigators made a comprehensive morphological examination of 25 autopsy specimens from patients who had died from HIV-associated tuberculosis. Lung pieces from the foci of caseous necrosis at its boundary and adjacent lung segments and from macroscopically intact perifocal pulmonary segments were microscopically explored. Plain stains and Ziehl-Neelsen histobacterioscopy were used for light microscopy. An immunohistochemical study was conducted using tuberculosis mouse monoclonal antibodies (clone 1.1/3/1, Vector). Seven early autopsy specimens (1-2 hours after statement of biological death) were investigated ultrastructurally. Ultrathin sections were contrasted with Reynolds' lead citrate and looked through a Libra-120 transmission electron microscope (Germany). Semithin sections were stained with toluidine blue. RESULTS: HIV-associated tuberculosis is a disease accompanied by dual aggressive infection. Impaired blood-air barrier integrity gives rise to structural and ultrastructural changes primarily in type I alveolocytes that are responsible for physiological gas exchange and detoxification. Type I alveolocytes have been noted to be in close contact with plasmocytes that are most ultrastructurally preserved and at the same time display the phagocytic and detoxifying functions being redistributed, which is necessary to keep cells viable and which is a characteristic sign of this comorbidity. This redistribution is due to severe damage to macrophages that are unable to ensure complete mycobacterial phagocytosis; thereby the plasma cells partly assume this function, as supported by ultrastructural examination. Severe destruction of granulocyte organelles and partial and complete cell necrosis with its preceding apoptotic changes were established. A study of vessels in the microcirculatory bed has indicated that the ultrastructural changes in the endothelial cells are polymorphic, which is characteristic of an exudative-inflammatory response of lung tissue. CONCLUSION: Cellular and subcellular changes, the pattern of intercellular relationships, and necrotic and apoptotic cell rearrangements, which are in one way or another involved in the pathophysiological processes maintaining definite hemostasis, are an important component in the pathogenesis of HIV-associated tuberculosis.

[Early morphogenesis of ciliated cells in human oral cavity].

Ciliated cells were found in the epithelium of the oral cavity of human embryos and fetuses starting from the seventh week of prenatal development. At the early stages of prenatal development (until the 13th week), cells with cilia cover most of the dorsal surface of the tongue and the soft palate, whereas they are found only near the gland ducts in the circumvallate and foliate lingual papillae after 17 weeks of development. The ultrastructure of the axoneme of cilia corresponds to the structure of motile cilia and is represented by nine microtubule doublets that surround the central pair of microtubule singlets. An immunohistochemical study performed on weeks 10-12 of development identified nerve endings associated with the ciliated cells. Until the 14th week of development, the cytoplasm of ciliated cells is immunopositive for NSE. The spatial distribution of ciliated cells in the tongue epithelium until the 13th week of development is not related to the morphogenesis of lingual papillae, and their role in the human oral cavity during the first trimester of pregnancy is unclear and requires further study.

[Role of low-dose acetylsalicylic acid in comprehensive postoperative treatment of patients with lower-limb chronic arterial insufficiency].

The study comprised a total of 107 patients (all men) after endured femoropopliteal bypass grafting above the genicular fissure with a synthetic stent graft manufactured by the Gore Company for stage IIB and III chronic arterial insufficiency of the lower extremities according to the Fontain-Pokrovsky classification. Group One I (control group) was composed of fifty-four patients permanently taking in the postoperative period at the out-patient stage pentoxiphylline (trental 400 mg 1 tablet 3 times daily) and xantinol nicotinate at a dose of 150 mg one tablet thrice daily. Group Two (Study Group) consisted of fifty-three patients taking after reconstructive vascular surgery at the out-patient stage in addition to pentoxiphylline and xantinol nicotinate acetylsalicylic acid (cardiomagnil 75 mg 1 tablet once daily). The Control Group patients within 3 to 6 months of follow up were found to have a considerable progressing improvement of the functional abilities of the microcirculatory bed requiring in 44 (81.5%) cases hospitalization to the Surgical Department for intensive vascular therapy. Despite this fact four (7.4%) patients within the time frame from 6 to 9 months after surgery developed thrombosis of the vascular implant requiring a repeat surgical intervention. In the Study Group patients, the degree of functional capabilities of the microcirculatory bed in the postoperative period was less considerable, reaching the maximum after 10-12 months of follow up, with eighteen (34.0%) patients requiring hospitalization for additional vascular therapy to perform. There were no cases of implants' thrombosis in the Study Group patients. Pathological alterations in the functional state of the peripheral vascular bed correlated with viscosimetric indices and activity of blood platelet aggregation. The addition of antithrombocytic agents to conservative postoperative therapy considerably improved the outcomes of surgical treatment.

Simple method of specimen preparation for scanning electron microscopy.

We compared conventional method for specimen (cell cultures, tissue specimens) preparation for scanning electron microscopy and a method without sputtering and critical-point drying. OTO-method (osmium-thiocarbohydrazide-osmium) with sample impregnation with hexamethyldisilazane followed by air drying was used as an alternative method. Excellent preservation of surface ultrastructures and electrical conductivity was proved. The method is easy to use and does not require additional costs for equipment.

[Morphological and biochemical criteria for cell death].

The state-of-the-art of classifications of and criteria for cell death in the light of the 2009 recommendations of the Nomenclature Committee on Cell Death is presented as a lecture. Motivation is given for the necessity of using the unified criteria in the description of cell death and more than one study in its verification. The major structural and biochemical signs of four typical types of cell death--apoptosis, autophagia, keratinization, and necrosis are compared. Data are given on the major atypical forms of cell death--mitotic catastrophe, anoikis, exitotoxicity, Wallerian degeneration, paraptosis, pyroptosis, pyronecrosis, and entosis.

[Clinical and morphological parallels of various degrees of chronic arterial insufficiency of lower limbs (review of the literature)].

The article deals with the main clinical and instrumental methods of determining severity of chronic arterial insufficiency of the lower extremities, showing possibilities of morphological studies for an accurate diagnosis of the ischaemia degree. Also determined are promising trends of studying the hypoxia-inducible factor (HIF) and vascular endothelial growth factor (VEGF) in assessment of ischaemic impairments of lower-extremities tissue.

[Influence of clinorotation on embryoid bodies morphology].

The compensative effects of gravitation at the first stages of embryonic development stages have been investigated using slow clinorotation of embryoid bodies from mouse R1 stem cell lines. Semithin sections (1-2 microm) analysis and electron microscopy study of embryonic bodies cells allowed to reveal morphological features of cells at different maturation stages. Significant decrease in the number of embryonic stem cells undergoing apoptosis as well as noticeable reduction of "lacunas" relative areas were found in clinorotated embryonic bodies compared to control. We propose that large cyst absence may be caused by initial differentiation and morphogenesis stages delay associated with autophagy processes in embryonic bodies.

[Role of hypoxia-induced transcriptional factor HIF-1 in the regulation of enterocytic metabolism].

The mechanisms responsible for the induction of the desquamation and death of intestinal epithelial cells in the areas of extrusion in normalcy remain unknown so far. The author's ultrastructural data and the data available in the literature suggest that HIF-1 [corrected] may regulate these processes. Its activators may be both lower oxygen concentrations in the upper portions of the intestinal villus ("physiological hypoxia") and some nonhypoxic factors, such as LPS of the enteric microflora. On the one hand, HIF-1 is involved in adaptive processes, by mobilizing specifically cellular and tissue resources, and, on the other hand, it promotes the inclusion of the mechanisms of programmed cell death at a certain stage, by provoking the synthesis of the proapoptotic mitochondrial proteins BNIP3 and BNIP3L, which is morphologically manifested as different types of apoptotic and necrotic cell death.

[Morphological alterations of the rat myocardium in chronic morphine intoxication]. / Morfologicheskie izmeneniia miokarda krys pri khronicheskoi intoksikatsii morfinom.

Morphine was administered intraperitoneally to Wistar rats at increasing doses from 10 to 60 mg pay during 8 days. Cardiomyocyte damage in the form of myocytolysis and wave-like deformations of muscle fibers prevailed at the level of light microscopy. Microcirculatory alterations were characteristic: stasis, sludge-phenomenon, perivascular and interstitial oedema, diapedes hemorrhages, focal proliferation of vascular wall cells. Ultrastructural lesions were as follows: subsarcolemma edema, mitochondrial destruction, formation of giant forms of mitochondria, single contractures, increasing micropinocytosis, intestitial edema.

[Cytological characteristics, elemental composition of alveolar macrophages from bronchoalveolar lavage, lung pathology in liquidators of the consequences of the accident at the Chernobyl power plant]. / Tsitologicheskie osobennosti, élementnyi sostav al'veoliarnykh makrofagov bronkhoal'veoliarnykh smyvov, patomorfologiia legkikh u likvidatorob posledstvii avarii na Chernobyl'skoi AES.

Bronchoalveolar lavage fluid (BALF), broncho- and transbronchial biopsies were examined in Chernobyl wreckers eleven years after the accident. Cytologically and electron-microscopically, there was predominance of alveolar macrophages (AM) with many cytoplasmic foreign bodies containing clear needle-like crystals. Roentgenospectral analysis and laser mass-spectrometry revealed chemical elements which are part of non-active "hot particles" (Al, Si, Ca, Fe) as well as reactor fuel elements and the station constructive materials which are the active part of "hot particles". Programmed BALF allowed to decrease cytosis and the amount of AM containing foreign-body particles as well as concentration of the majority of chemical elements in AM.

[The presence of cesium-137 in the tissue of a lung tumor in someone who cleaned up the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. / Prisutstvie tseziia-137 v tkani opukholi legkikh likvidatora posledstvii avarii na Chernobyl'skoi AES.

A man of 66 participated in the emergency works after the Chernobyl accident. His official dose of radiation comprised 24 roentgens. In March, 1993, he underwent prophylactic examination including computed tomography which revealed peripheral tumor in the lower part of the left lung with penetration in the lymph nodes of the lung root. Biopsy showed it to be adenocarcinoma. Moreover, electron-microscopically, more than 40% of alveolar macrophages contained large optically dense particles in the cytoplasm. After lobectomy in March 1993 x-ray and gamma spectrometry of the removed lung tissue recorded high content of 137Cs (0.1-0.18 Bq). This finding evidences possible long-term deposition of radionuclides in the bronchopulmonary system of radiation-exposed subjects.

[The effect of weightlessness on amphibians. The skeleton and mineral metabolism]. / Vliianie nevesomosti na amfibii. Skelet i mineral'nyi obemn.

The visceral and somatic skeleton of Pleurodeles waltilii was investigated after a series of two-week space flights on the biosatellites. It was shown that under conditions of weightlessness osteoporosis of the skeleton and its demineralisation were stimulated. In weightlessness, calcium, phosphorus and sulfur were lost and potassium accumulated. Trabeculae of endochondral part of the extremity bones were destroyed by osteoclasts. Proliferation of osteoclasts was disrupted, appositional growth stopped, and the cartilage of hyoid system was resorbed. Skeleton readaptation during two months did not result in the complete regeneration of morphological structure of the bones.

[The morphogenesis of the human brain on the 27th-35th day of development with disordered neurulation]. / Morfogenez mozga cheloveka na 27--35-i den' razvitiia pri narushenii neiruliatsii.

Human embryos with anomalies of brain and spinal cord were studied on 27-35th day of the development. It was established that developmental anomalies were associated with the disturbed formation of neural tube. The reconstruction of three-dimensional brain arrangement of embryos has shown the presence of different variants of nonclosed medullar tori. In mild disturbance of neurulation, there is an acceleration of organogenesis and morphological differentiation induced by the shackening of normal mechanical tensions and relaxation of neuroepithelial layers. In presence of substantial parts of open neural tube, the development of brain and ectodermal derivatives becomes disturbed to be a possible cause of anencephaly and hypotelorism. The type of disturbed neurulation inducing developmental anomalies of nervous system is suggested to be a factor which determines forms of secondary craniofacial and cranial pathology.

[An immunohistochemical study of the primordium and primary differentiation of the parafollicular cells in the human thyroid]. / Immunogistokhimicheskoe issledovanie zakladki i pervichnoi differentsirovki parafollikuliarnykh kletok shchitovidnoi zhelezy cheloveka.

Formation and differentiation of calcitonin-secreting cells has been studied in human embryos. The cells form from the entodermal part of the rudiment of the last pharyngeal cavity. They do not secrete calcitonin before incorporation into the thyroid gland. The first calcitonin-secreting cells appear in the thyroid gland as a diffuse net. The properties and origin of this net have been described.