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.

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.).

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.

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.

[Dermoid cyst of the spleen]. / Dermoidná cysta sleziny.

The case report of a 14-year old girl is given. In her spleen of the weight of 1580 g an irregular cyst of glossy inner surface and with striking trabeculation developed. It was filled with clear greenish liquid. The wall was formed by hyalinised collagen tissue which was covered by epidermoid and cuboidal epithelium on the inner surface. Vessel conglomerates were frequent, some bleeding or mononuclear infiltrates, foci of giant cell granulation tissue, lymphoid or fatty tissue were present.

[The significance of prostate markers in the orthology of the female prostate]. / Výnam prostatických markerov v ortológii zenskej prostaty.

In addition to knowledge gained in the first half of the 8th decade, the evidence of cross-antigenicity between male prostate and Skene's glands by means of PSA and PSAcP demonstrations in Skene's glands and ducts justifies utilization of the term prostate in both sexes. The authors compared the results of immunohistochemical examination of prostatic markers by means of the PAP method which was used at the beginning of the 8th decade, with that of BSAP technique. Prostatic tissues of 11 females and children at the age ranging from 5 to 71 years were examined. The results gained by means of the BSAP method were identical with those gained by means of the PAP method. Prostatic markers PSA and PSAcP were expressed on the surface and in apical cytoplasm of cells lining the prostatic ducts, and in prostatic glands. The authors proved the expression also in membranes of the stratified cylindrical epithelial cells of the ducts, and in female prostatic fluid in ducts and glands, especially PSAcP. Even though both immunohistochemical methods brought identical results, the authors recommend to prefer the BSAP method to the PAP method due to optically more contrast expression. (Fig. 8, Ref. 36.)

[The human spleen in idiopathic thrombocytopenic purpura]. / L'udská slezina pri idiopatickej trombocytopenickej purpure.

Idiopathic (autoimmune) thrombocytopenic purpura (ITP, AITP) represents a relatively frequent impairment. It involves a syndrome of various diseases with a shortened thrombocytes survival caused by anti-platelet antibodies. The majority of cases are of secondary character. Spleenectomy often evokes a complete remission of thrombocytopenia. The study describes morphologic findings in spleens of 30 patients with the clinical diagnosis of ITP/AITP. The findings were gained by light microscopy from formol-paraffin blocks and histochemical findings from cryostat sections of non-fixed tissue. The alcaline and acidic phosphatases, nonspecific esterase, chloracetate esterase, and dipeptydilpeptidase IV were investigated enzymohistochemically. Immunoglobulins were examined immunohistochemically and T lymphocytes by means of monoclonal antibodies. The affinity HPA--Helix pomatia agglutinin, PHA--phytohemagglutinin from Phaseolus vulgaris, SBA--soy-bean agglutinin from Glycine max. and PSA--peas bean agglutinin from Pisum sativum were investigated by means of specific antilectin antibodies. The human spleen during idiopathic thrombocytopenic purpura accumulates neutrophilic polymorphonuclear granulocytes; platelets-stagnate and are destroyed. These processes can be identified in histologic sections e.g. also by means of anti-fibrinogen antibodies. The red pulp contains foam cells to various extent. Besides generally known processes, the white pulp also displays alterations in composition of cellular compartment of the periarterial lymphatic sheaths. Human spleen distinguishes modified blood platelets as alien corpuscles, and thus eliminates them from the blood circulation system by its immunologic and other mechanisms, the details of which still remain to be clarified. (Fig. 6, Ref. 44.)

Delayed effects of neonatal exposure to Tween 80 on female reproductive organs in rats.

Neonatal female rats were injected ip (0.1 ml/rat) with Tween 80 in 1, 5 or 10% aqueous solution on days 4-7 after birth. Treatment with Tween 80 accelerated maturation, prolonged the oestrus cycle, and induced persistent vaginal oestrus. The relative weight of the uterus and ovaries was decreased relative to the untreated controls. Squamous cell metaplasia of the epithelial lining of the uterus and cytological changes in the uterus were indicative of chronic oestrogenic stimulation. Ovaries were without corpora lutea, and had degenerative follicles.

Subacute immunotoxicity study of formaldehyde in male rats.

Immune functions were examined in male rats following 28 day oral administration of formaldehyde by gastric tube at dose levels of 0, 20, 40, and 80 mg/kg. Routine parameters examined included hematology, clinical chemistry, and body, thymus, kidney, and liver weights. In addition, cellularity of spleen and lymph nodes, histology of spleen, thymus, lymph nodes, liver kidney, small and large intestines, and histochemistry of spleen and lymph nodes were evaluated. Immune parameters evaluated included serum hemagglutinin antibody response; antibody plaque forming cell response to sheep erythrocytes (lymphocyte-dependent antigen); microbicidal activity of Candida albicans; and phagocytic activity by adhesion of microspheric hydrophilic synthetic particles to leukocyte cell membrane. Body weights were slightly decreased at high dose level (80 mg/kg). The difference was significant when compared to the controls. The lymph node weights were significantly increased in rats receiving formaldehyde. The cellularity of lymphoid organs was not influenced after 28 day exposure to formaldehyde.

[Lectin histochemical analysis of the epithelial lining of the appendix in rabbits]. / Lektínová histochemická analýza epitelovej výstelky králicieho apendixu.

The binding of PSA, PNA, HPA, WGA, Con A, LCL, RCA, SBA, and PHA lectins to epithelial structures of the normal rabbit appendix was studied. Differences were observed in the affinity of some lectins to the epithelium of the intercryptal lining of the rabbit appendix, to the epithelium hemming the glandules and crypts of the domes of Peyer's patches. The obtained results document the difference in the affinity of individual batches of anti-WGA antibodies to this lectin after its binding to the epithelial structures studied. This implies the possibility that differently reacting antibodies may develop to different commercially available batches of WGA. Precipitation of WGA at sites of alkaline phosphatase occurrence demonstrates the relationship of this enzyme to WGA binding sites in tissues.

[The estrogenic effect of the fungicide Novozír MN 80 on the uterus in neonatal rats]. / Fungicíd Novozír MN 80 a jeho estrogénny úcinok na uterus neonatálnych potkanov.

The estrogenic effect of the ethylenebisdithiocarbamate fungicide Novozír MN 80 (Duslo, Sal') was studied by means of the screening method determining the reproductive risk of chemical substances, recommended by the Environmental Protection Agency. Compared to intact controls, a significant increase in relative uterine weight was recorded in the group of animals which received 1/25 of the maximal tolerable concentration of Novozír MN 80. Compared to negative controls (Tween 80), the relative uterine weight was increased significantly after administration of 1/25 and 1/50 of the maximal tolerable concentration of Novozír MN 80. On histologic examination the cuboidal epithelium of the mucous membrane was found to be changed into columnar epithelium, with uneven distribution of enlarged nuclei in the vacuolized cytoplasm. At 1/25 maximal tolerable concentration thickening of the uterine wall was observed.

Combined chronic toxicity/carcinogenicity study of the fungicide trimorphamide in rats.

Trimorphamide, a new fungicide, was fed over 2 years to groups of 50 male and 50 female rats at dietary levels of 0, 67, 330 and 665 mg/kg. No adverse effects on mortality, haematology, urinalysis, organ weights, incidence and severity of tumors were observed. Isolated observations of abnormalities in blood chemistry suggested the possibility of certain renal damage in the highest dosage group; however, the clinical significance of these findings is dubious. According to the results of the present study, 67 mg/kg in the diet can be considered as the "no observed effect level".

Premature delivery and placenta. A morphological study.

A complex morphological examination of the placentas of the prematurely delivered newborns revealed that in a part of them there was a complex of changes detectable either by a naked eye immediately after delivery or by the use of light and electron microscope, respectively. The changes observed were characteristic of premature maturation of placenta, some changes, however, were similar to those present in the placentas of the term delivered intrauterine growth retarded newborns. The most important were the following changes: underweight of placenta, placental infarctions, capillary haemangioma, increased amount of fibrinoid, small and focal circulatory defects, reduction of the microvillous border and of the pinocytic activity of the syncytiotrophoblast, focal to diffuse thickening of the basal membrane with an increased number of collagen fibrils and fibroblasts in the stroma of terminal villi. These changes resulted in deterioration of the intrauterine state of the foetus and in premature delivery of frequently intrauterine growth retarded newborn. Such changes did not occur in all placentas under study. They indicate, however, the possibility of the occurrence of chronic placental insufficiency in a part of the pregnant women treated for the imminent premature delivery. From this follows the necessity of monitoring the placental functions and the state of the foetus with a subsequent differential approach to the therapy of such patients.

Cellular composition of periapical granulomas and its function. Histological, immunohistochemical and electronmicroscopic study.

Periapical granulomas have been investigated histologically, immunohistologically using polyclonal and monoclonal antibodies, as well as electronmicroscopically. Lesions were formed by inflammatory granulation tissue frequently with foci of purulent exudation and fibrosis. Most numerous were plasma cells usually in cellular regions of the granulation tissue where they were tightly pressed. Of other cellular types were numerous lymphocytes, fibroblasts, less frequent were macrophages, scattered granulocytes and mast cells. More than a half of the plasma cells were IgG positive, about 20% IgA positive, up to 10% IgM, rarely IgE and sporadically IgD positive cells. In the vascular walls and their surrounding as well as in the phagocytes fine granular to granular positivities of C3 and C4 components of the complement were present. The majority of lymphocytes beared markers of T lymphocytes of which the T-suppressor markedly prevailed over the T-helper lymphocytes. In electron microscopy the plasma cells were most frequent. They were usually close to each other, sometimes with a disintegrated cytoplasmic membrane and non-damaged organelles being free around the nucleus. Mast cells were numerous and did not show any signs of marked degranulation. Rich production of immunoglobulins as well as the presence of IgG and IgM positive material in phagocytes, and the presence of positivities of the C3 and C4 components of the complement in the surrounding of the vessels and in phagocytes on the other hand supported the presumption that immune complexes participate in the pathogenesis of periapical granulomas. In spite of the presence of the IgE producing cells the morphological picture of mast cells did not suggest the presence of anaphylactic reaction in periapical lesions. Diffuse distribution of T lymphocytes, moreover with the prevalence of T-suppressor/cytotoxic over T-helper lymphocytes and not numerous macrophages in the inflammatory infiltrates did not suggest the participation of a typical cell-mediated immunity reaction in the development of periapical granulomas. Numerous T-suppressor/cytotoxic lymphocytes and low numbers of macrophages can be important factors of the chronicity of periapical inflammatory diseases.

Chronic lymphoproliferative disease of large granular lymphocytes.

An 80-year-old patient has been followed for hepato- and splenomegaly, hemolytic anemia, neutropenia with lymphocytosis with large granular lymphocyte predominance in his peripheral blood, with infiltration of bone marrow, liver and probably also spleen. Determination of surface markers of proliferating lymphocytes in peripheral blood showed a mixed phenotype of T suppressor/cytotoxic and natural killer cells (SIg-, E+, T3+, T8+, EAC+, Leu7-, N901+, NK9+, VIB C5 and VIB E3-, Ia-). An in vitro cytotoxic test showed the functional inactivity of the cells tested also after human leukocyte interferon stimulation. Chromosomal analysis neither of peripheral blood lymphocytes nor of bone marrow cells proved the monoclonality marker. Following long-term prednisone therapy, the improvement of anemia, later also neutropenia accompanied by the decrease of lymphocytes has been achieved. As the disease present in our patient was distinguished only in recent years and in our country has not been reported yet, the details on its clinical, morphologic, hematologic, cytogenetic and mainly immunophenotypic characteristics are given in this paper. The problems concerning classification of the disease and determination of its biological nature are discussed.