The first imported human infestation with Furuncular myiasis in man in the Slovakia and current knowledge of myiasis.

INTRODUCTION: Furuncular myiasis is caused by the genus of botfly Dermatobia hominis. It belongs to the family Cuterebridae and is indigenous to Central and South America. OBJECTIVE: to present a case report of the first case of this disease in Slovakia. CURRENT STATE OF PROBLEM SOLUTION: The term myiasis refers to infestation of the host (animal, man) by botfly larvae. Its larvae burrow under the skin. They feed on the host's living tissues and fluids. MateriAl and methods: Patient's history analysis, parasitological examination. RESULTS: A 58-year-old woman after returning from Central America found in the skin above her m. gluteus mayor 2 indurations, which contained three botfly larvae. DISCUSSION: Infestation with botfly larvae Dermatobia hominis is for man annoying and from a health point of view dangerous. CONCLUSION: With proper diagnosis, it is possible to remove the larvae safely from furuncles. The authors point to the first case of imported infestation with Furuncular myiasis caused by botfly Dermatobia hominis in man introduced to Slovakia. They note that increasing tourism spread to the countries with the endemic occurrence of Furuncular myiasis will cause its higher prevalence also in Central European countries (Fig. 5, Ref. 45).

Cristobalite and Hematite Particles in Human Brain.

Foreign substances get into the internal environment of living bodies and accumulate in various organs. Cristobalite and hematite particles in the glial cells of pons cerebri of human brain with diagnosis of Behhet disease with scanning electron microscopy (SEM), energy-dispersive microanalysis (EDX), and transmission electron microscopy (TEM) with diffraction were identified. SEM with EDX revealed the matter of irregular micrometer-sized particles sometimes forming polyhedrons with fibrilar or stratified structure. It was found in some particles Ti, Fe, and Zn. Some particles contained Cu. TEM and electron diffraction showed particles of cristobalite and hematite. The presence of the particles can be a result of environmental effect, disruption of normal metabolism, and transformation of physiologically iron-ferrihydrite into more stable form-hematite. From the size of particles can be drawn the long-term accumulation of elements in glial cells.

Ultrastructural changes of kidney in diabetic rats.

OBJECTIVES: To demonstrate histopathological changes in diabetic kidneys of the rat on ultrastructural level with focusing on glomerular cellular environment. METHODS: Male Wistar rats were administered streptozotocin and after 8 weeks, their kidneys were studied using qualitative transmission electron microscope analysis. RESULTS: In diabetic kidneys, the presence of irregularly thickened glomerular membrana basalis (GBM), often up to six times as compared to healthy kidney GBM, mainly around peripheral capillary loops, was striking. We also demonstrated lamination and split appearance of GBM, with presence of delicate fibrillar structure, which was absent within normal GBM. Accumulation of mesangial matrix was found only occasionally. CONCLUSION: Model may be useful to better understand the mechanisms that play role in the progression of DM-induced renal disease and to comprehend the morphological changes, which this process leaves within the tissue. The clear association between ultra-structure and functional markers may not be necessary (Fig. 4, Ref. 27).

Iron and its relation to glycoconjugates in human globus pallidus.

Iron and eosinophilic, argyrophilic spheroid structures of glycoconjugates are observed in the pallido-nigral system of human and nonhuman primates. In the present study, we map the localization and distribution of ferritin and hemosiderin and their relation to neutral and acidic (sulphated and carboxylated) glycoconjugates in human globus pallidus. We investigated tissues of human globus pallidus without any motor abnormalities and psychiatric symptoms. Acidic (Alcian blue, AB pH 2.5) and neutral glycoconjugates (PAS reaction) showed spheroid deposits with the size of 5-15 µm. Staining reaction utilizing AB (pH 1.0) displays sulphated fibers and highly sulphated (AB pH 0.5) glycoconjugates round deposits. AF/AB pH 2.5 for separating sulphated from carboxylated glycoconjugates revealed the presence of both sulphated and carboxylated glycoconjugates. Perls' and Turnbull's positive reaction showed the presence of Fe(III) and Fe(II) (hemosiderin) and Ag-NOR reaction showed ferric phosphate (ferritin) deposits. Scanning electron microscope with microanalysis revealed the iron in glycoconjugates globular deposits. We suppose the presence of glycoconjugates in the samples is the result of elimination and inactivation of iron as inductor of reactive oxygen species. They can be a useful neuroprotective agent in CNS degradation (Tab. 2, Fig. 7, Ref. 44).

Iron-rich complexes in human spleen in hereditary spherocytosis.

OBJECTIVE: Tissue iron plays an important role in the development of certain diseases. Although it is one of biogenic elements, its excess induces the reactive oxygen species (ROS) formation. The aim of the present work is to examine the protection against free or loosely bound iron from the view of morphology and chemical composition of iron-rich complexes in human spleen tissues with hereditary spherocytosis (HS) by scanning and transmission electron microscope with energy-dispersive microanalysis (EDX). RESULTS: The examination of human spleen tissues by scanning and transmission electron microscope showed covering of iron-rich particles. EDX revealed many iron-rich complexes of multi-element composition in HS samples with sulphur and phosphorus as the major elements. Detection was negative in the reference samples. CONCLUSION: The covering of iron-rich particles can be explained by elimination and isolation of ferritin/iron complexes from surrounding environment to prevent the ROS formation. Sulphur, phosphorus and their compounds are probably the most significant elements that influence the ROS formation (Fig. 5, Ref. 16). Full Text in PDF www.elis.sk.

Iron accumulation in human spleen in autoimmune thrombocytopenia and hereditary spherocytosis.

BACKGROUND: Although the iron is an essential element for the physiological functions of cells, tissues and organs, it is also an important inductor of reactive oxygen species (ROS). MATERIAL AND METHODS: Three groups of human spleen with autoimmune thrombocytopenia (AITP), hereditary spherocytosis (HS) and reference samples stained by haematoxylin and eosin, Perls' reaction for nonheme Fe(III) iron and Alcian blue for glycoconjugates detection were studied. RESULTS: Positive Perls' reaction in both AITP and HS groups was seen. Higher positivity in the HS than in AITP group was observed. HS group showed a higher amount of acidic glycoconjugates deposits than AITP group. Iron overload in HS and AITP leads to overproduction of ROS. CONCLUSION: We suggest that acidic glycoconjugates deposits are involved in antioxidant defence by elimination and restriction of iron as a ROS inducer (Fig. 4, Ref. 19).

Endocrine organs and laser scanning confocal microscopy (LSCM) imaging: vascular bed in human spleen.

OBJECTIVE: This work was aimed to utilize the precise method of laser confocal microscopy (LSCM) to depict the image of spatial relationships of the vessel network in the tissue structures of the human spleen. METHODS: With the use of serial paraffin or vibratome sections of more than 20 µm thickness infiltrated with eosin fluorescence dye the images of arterial and venous walls of different calibres, capillaries, and venous sinuses were morphologically revealed. RESULTS: Venous sinuses were frequently found to create mutually communicating branches and their lining projected into the lumen protruding cells with distinct spherically or ovally shaped nuclei, positioned on the brightly fluorescent and fragmented lamina basalis. The presence of lymphocytes was distinct in periarteriolar lymphoid sheath (PALS) and lymphatic follicles. Lining cells of the red pulp veins sporadically contained marked eosinophilic granules. CONCLUSION: The method of LSCM allowed: 1. to reveal two-dimensional and sharp image of the human spleen structures, 2. to investigate the vertical course of venous structures in the tissue, 3. to obtain serial optic sections in z axis to their maximum spatial projections. These data will also serve for the creation of three-dimensional images of vessel network in the human spleen in the future studies.

Perspectives and complexity of an experimental cancer study. the secrets of tumorigenesis. (To the Gupta's, Chaffer's and Weinberg's "perspectives" and to the nurse's "horizons").

We would like to add to the "mysteriousness", our observations from the application of "identical" BP6 cells either intraperitoneally or subcutaneously. In connection with the concept that tumor development is not only a portrayal of cells proliferation, we could presume that different "environment" will result in structurally different tumors. Morphological differences observed are not significant, but they are present (Fig. 2, Ref. 12). Full Text (Free, PDF) www.bmj.sk.

[Changes in membrane enzymes in warm liver ischemia]. / Zmeny membránových enzýmov pri teplej ischémii pecene.

BACKGROUND: The aim of the study is to perform a morphological analysis of certain membrane enzymes and parenchymal alterations during warm ischemia in the pig liver. METHODOLOGY: Standard hepatectomies were performed in a set of 24 pigs. Intraoperative intravenous (portal vein) hydroxyl radical scavenger Stobadine and Pentoxifylline was administered. Tissue specimens were removed from the margo acutum in interval 10, 60, 80 et 100 minutes. Applied methods of morphological analysis were concentrated on studying the activity of enzymes, which are regarded as cell membrane condition markers. First of them was the alkaline phosphatase, the second studied enzyme was the 5'-mononucleotidase. Detected findings in control group were compared to other two groups. We evaluated each sample using Image Forge Version 1.1 software (PROVER LTD., Bratislava). Applying this software we measured percentage of pre-measured activity levels of both enzymes. RESULTS: In normal pig liver, the activity of alkaline phosphatase was regularly distributed in sinusoid walls in the whole lobule. At 10 minutes after the beginning of the warm ischemia, we have found focal losses of enzyme activity, especially in cells of zone 3 of the liver lobuli. One hour after the beginning of warm ischemia were the local losses of enzyme activity even more pronounced. Decreased bile canalicular 5'-mononucleotidase activity reflects cell damage, especially in pericentral areas after 60 min.of ischemia and a further decrease in activity at all surfaces of the membrane. Total amount of enzyme activity is different. CONCLUSIONS: Morhological findings of enzyme activities showed that zone 2 and 3 of the liver lobule are essential for the organ survival. The study showed, that intravenous administration of Pentoxifylline and Stobadine probably protects the liver from warm ischemia injury.

Warm liver ischemia in experiment and lysosomal markers.

BACKGROUND: The aim of the study is to perform a morphological analysis of certain lysosomal enzymes and parenchymal alterations during warm ischemia in the pig liver. METHODS: Standard hepatectomies were performed in a set of 24 pigs. Intra-operative intravenous (portal vein) Pentoxiphylline and hydroxyl radical scavenger Stobadine was administered. Tissue specimens were removed from the margo acutum in 10 minutes interval. RESULT: In normal pig liver, the acid phosphatase (ACP) activity is in not numerous Kupffer cells and on the biliary pole of hepatocytes, diffusely in the whole parenchyma. One hour after the beginning of warm ischemia, there was an increase in ACP activity in the cytoplasm of hepatocytes. The activity in Kupffer cells could not be detected. Lactate dehydrogenase (LDH) is localized exclusively in the cytoplasmic matrix of liver cells, so only cytoplasmic enzymes leak into the blood plasma. LDH activity has remained low in areas around portal and central veins. CONCLUSION: Morphological findings of enzyme activities showed that zone 2 and 3 of the liver lobule are essential for the organ survival and signs of diffusion of lysosomal enzymes into the cytoplasm of hepatocytes indicate one of the possible explanations for the findings after liver reperfusion. The study showed that intravenous administration of Pentoxiphylline and Stobadine protects the liver from warm ischemia injury (Tab. 2, Fig. 2, Ref. 37). Full Text (Free, PDF) www.bmj.sk.

Conformation study of the membrane models by the Maxwell displacement current technique and oxidative stress.

The role of biological membranes as a target in biological radiation damage is still unclear. Recently much attention has been paid to the dynamic behaviour of the cell membrane. Maxwell displacement current technique (MDC) provides new possibility of conformation study of the membrane models. Oxidative stress can impair macromolecules in the cell on a molecular level. MDC technique enables to study the changes in molecular orientations and/or conformations of cell membranes. The combination of different methods in structural biology can clarify membrane chemical and physical properties.

Chondroitin sulphates A, B and C, collagen types I-IV and fibronectin in venous sinus of the red pulp in human spleen.

The morphological relationship of chondroitin sulphates A, B, and C, collagen types I-IV and fibronectin in the wall of venous sinuses of the red pulp in human spleen has not been a focus of interest among morphologists. Regarding the hypothesis that the structure of the spleen lends it the function of a blood filter the substances described in our study might play a significant role in the functional morphology. Of 146 human spleen surgical specimens, groups of 12 specimens each were examined under a light microscope using the method of antibodies against fibronectin, against collagen types I-IV and against chondroitin sulphates A, B, and C. The sections of the red pulp of human spleen stained with hematoxylin and eosin provided limited information about the wall of the sinuses. Chondroitin sulphates A and B were observed on the surface of sinus-lining cells (SLC), and fibronectin was detected on the surface of the annular fibers. Collagen type 11 was observed almost in the same places as chondroitin sulphates A and B. Collagen type IV was present in annular fibers of the wall of the sinus and in the basement membrane, like fibronectin. Chondroitin sulphate was not present in the walls of sinuses. Binding of antibodies against chondroitin sulphate A and against chondroitin sulphate B indicates the presence of chondroitin sulfates on the surface of SLC, where they probably play a role in helping the human organism to recognize alien and self substances. The presence of chondroitin,sulphates A and B probably affects inhibition of binding of cells with collagen type I, but not with fibronectin.

Results and perspectives of the treatment of skeletal neoplasms.

Improvements in the therapy of skeletal neoplasms at the end of the 20th century are the reason for a retrospective view on the development of the therapy of skeletal neoplasms, especially in the Slovak Republic. Up to the sixties years of the 20th century was amputation the imperative in the surgical treatment of malignant tumors, above all tumors of the extremities. Then chemotherapy was introduced into the treatment of these tumors, in Slovak Republic it was later. Bone transplantations were widely used in the 70 and 80 years of 20th century. Autotransplantations of bone grafts were used at the Orthopaedic department in Bratislava already in the year 1953, but low number of appropriate grafts limited their use in clinical practice. In the 70 and 80 years development of bone banks was observed. Results of the treatment of malignant diseases with this technique are considered poor. Only few patients survived more than 5 years after the surgery, mainly due to inappropriate techniques of detection, localization and determination of the size of the tumor. Biopsy played above all at this time an important role in the diagnostics of bone tumors. Apart from basic examinations of the fixed material, processed after it was imbedded in particular medium, perioperative biopsy is recently being performed with the technique of frozen sections from unfixed tissue obtained during the surgery. This type of material processing prolongs the period from obtaining the material till making the diagnosis, but the advantages of this type of examination are undoubted. (Ref. 17.).

[Clinical and histopathologic picture of acute appendicitis in children]. / Klinický a histopatologický obraz detskej akútnej apendicitídy.

A retrospective study of the clinical and histopathological picture of acute appendicitis in children confirms agreement of the evaluation of the finding on the appendix made by the surgeon and pathologist in 72.2% and disagreement in 27.8%. The finding is overrated in 11.3% and underrated in 16.5%. Agreement is closest in phlegmonous appendicitis (88.1%) and lowest in gangrenous appendicitis (54.9%). The most frequently overrated finding is in catarrhal appendicitis (20.7%). The number of missed appendectomies is from the pathologist's point of view significantly lower (21.8%) than from the surgeon's point of view (34.7%). This indicates obviously that it is not possible to make an accurate macroscopic assessment of the progress of appendiceal inflammation. The number of "negative" appendectomies is according to the surgeon 15.8% and according to the pathologist 16.8%, i.e. almost equal.

Endotoxin-induced aggravation of preservation-reperfusion injury of rat liver and its modulation.

BACKGROUND/AIMS: In clinical transplantation, exposure of donors to gut-derived endotoxin occurs frequently and may adversely affect liver transplantation therapy. The aim of this study was to investigate: 1) whether brief exposure of rats to endotoxin before liver procurement aggravates the early phase of reperfusion injury of hepatic explants; and if so 2) whether Kupffer cell activation is a contributing factor to liver injury; and 3) whether heparin and pentoxifylline could minimize this effect. METHODS: Male Wistar rats were injected with 0.2-4.0 mg/kg of Escherichia coli lipopolysaccharide 2 h prior to liver harvest. After preservation in University of Wisconsin cold-storage solution, the livers were reperfused using a blood-free perfusion model. To inactivate Kupffer cells, some rats were pretreated with gadolinium chloride or liposome-encapsulated dichloromethylene-diphosphonate before lipopolysaccharide administration. The other rats received lipopolysaccharide with heparin or pentoxifylline. RESULTS: In a dose-independent fashion, lipopolysaccharide impaired portal flow during graft reperfusion. In a dose-dependent way, lipopolysaccharide increased lactate dehydrogenase release into the perfusate and decreased bile flow and bromosulfophthalein excretion. Gadolinium chloride, liposomal dichloromethylene-diphosphonate, heparin, and pentoxifylline reduced lactate dehydrogenase release by 34%, 43%, 59%, and 64%, respectively, and improved functional parameters of the liver. A 52-fold increased neutrophil infiltration in the liver sinusoids after lipopolysaccharide exposure was not affected significantly by the drugs studied; however, heparin reduced markedly neutrophil activation. CONCLUSIONS: The results of this investigation provide direct evidence that aggravation of preservation-reperfusion injury of rat liver by endotoxin is mediated by Kupffer cell-dependent mechanism(s) and it can be minimized by heparin and pentoxifylline.

[Current knowledge on the functional morphology of the human spleen]. / Súcasný stav poznatkov o funkcnej morfológii l'udskej sleziny.

The knowledge on the functional structure of the spleen is gained from the analysis of diseases, and immunological studies. Significant information is gained from model tests and isolated cells. The latter is advantageous in coincidence with the studies of intercellular relations. The main function of the spleen resides in "purification" of the blood. The vascular system, white pulp, red pulp and the marginal zone participate in this process. T-dependent zones of the white pulp are restricted by the circumference reticulum (CR) of periarterial lymphatic sheaths. CR is a morphologic substrate of retro-directional flow of lymphocytes. In addition to T lymphocytes and a certain amount of B lymphocytes, the T-dependent zones contain interdigitizing cells. The B-dependent zones, in addition to lymphocytes, contain dendrite reticular cells. The cords are constituted by varied cellular and extracellular compartments, especially macrophages, fibroblastic reticular cells and various fibrous components. The latest filtration barrier is formed by the wall of sinuses of the red pulp. The achievement of new functional knowledge resides prospectively in the use of lectins, anti-collagen antibodies, fibronectins, proteoglycanes and other various molecules participating in intercellular relations, or morphometric studies, they can provide information on the newest subpopulations of cells in both red and white pulp, that are so far inaccurately defined.

Metastasizing adenocarcinoma of the female prostate (Skene's paraurethral glands). Histological and immunohistochemical prostate markers studies and first ultrastructural observation.

The case of a 46-year-old women with well-differentiated adenocarcinoma of the female prostate (Skene's paraurethral glands and ducts) with inguinal metastases is reported. Besides adenocarcinomatous structures, also more solid parts of the tumor and anaplastic regions with dark cells were found on histological examination. Clear cancerous cells were typical for glandular and solid tumor parts. The cancerous cells showed distinct immunohistochemical positivity of prostate specific antigen (PSA) and prostate (specific) acid phosphatase [P(S)AcP]. These are the first published results of electron microscopic examination of formalin fixed tissue showing the ultrastructure of female prostate carcinoma, comparable to that of the male prostate carcinoma. In the female, similar to the male, the prostate carcinoma probably originates from the secretory (luminal) cells of the female prostatic glands.

[Present knowledge of human spleen function]. / Súcasný stav poznatkov o funkcii l'udskej sleziny.

The human spleen is an important lymphoid organ participating in immune reactions against all types of circulating antigens. It also removes particles of nonantigenic nature from blood circulation. The structure of the spleen, containing specialized cells types, provides a microenvironment where various intercellular interactions occur and determines the course of development of immune and nonimmune reactions. Though the spleen significantly participates in host defense reactions it is not inevitable for life. However, removing of the spleen increases the risk of overwhelming infections by bacteria with polysaccharide capsules.

[The spleen in hereditary spherocytosis]. / Slezina pri hereditárnej sférocytóze.

Hereditary spherocytosis is the most common inherent, autosomal dominant hemolytic anemia. Mild splenomegaly, venostasis and common decrease of while pulp is characteristic for hereditary spherocytosis. Cords are filled with spherocytes, sinuses can be empty or squeezed. Both sinuses and veins include ghost erythrocytes that lost haemoglobin. They are seen light-microscopically in differential interferent contrast. Macrophages are numerous, sinus lining cells are hypertrophic. Erythrophagocytosis is hardly seen by electron microscope. Not all red blood cells are spherocytes. Spherocytes are seen well in electronmicroscopy. We did not observed transition of erythrocytes through sinus walls. In certain circumstances lots of ferritin is seen both intra and extracytoplasmically. Iron accumulation in cords can result in their fibrosis.

[Fibronectin and the human spleen]. / Fibronektín a l'udská slezina.

Fibronectins are glycoproteins with a function of molecular glue. The aim of the study is to encourage interest in processes in extracellular matrix of the spleen. Undigested cryostat or formol-paraffin sections and commercially available antibodies were used. We were mostly successful in extracellular localization of fibronectin in trabeculae, vessel walls and lamellae of circumferent reticulum of periarterial lymphatic sheaths and in the ring fibres of the sinuses. Changes in the amount of fibronectin occurred, for example, in circulation disturbances. In hereditary spherocytosis the ring fibres were reduced. Increase of fibronectin can suggest the development of fibrosis.