PTEN mutations as predictive marker for the high-grade endometrial cancer development in slovak women.

Endometrial carcinoma (ECa) is one of the most common neoplasia of the female genital tract. The phosphatase and tensin (PTEN) homolog is the most frequently mutated tumor suppressor gene in endometrial carcinoma. PTEN encodes a phosphatase, a key regulatory enzyme involved in a signal transduction pathway that regulates cell growth, migration and apoptosis. The study evaluates an association between the morphological appearance of endometrial hyperplasia and ECa, and the presence of PTEN variations, PTEN protein´s level and intracellular localization. A total of 67 archived formalin-fixed and paraffin-embedded human biopsy tissue specimens with normal proliferative and secretory endometrium, endometrial hyperplasia without atypia and endometrial atypical hyperplasia, endometrioid the grade G1 and G3 and serous subtype of ECa were evaluated by sequencing for the presence of mutations in coding regions of PTEN gene of endometrial epithelial cells. The PTEN gene expression and intercellular localization of PTEN protein were evaluated immunohistochemically by immunoreactive score (IRS). PTEN mutation spectrum in endometrial carcinoma was identified for Slovak population. 28 non-silent mutations were identified in PTEN, twelve of them were novel, not annotated in Catalogue of Somatic Mutations in Cancer. Higher frequency of PTEN mutations was observed in serous carcinoma compared to global average. No correlation was observed between samples´ IRS, PTEN cellular localization and identified mutations. PTEN sequencing can be beneficial for patients considering prognosis of disease and sensitivity to treatment.

Pineal gland calcification under hypoxic conditions.

The pineal gland (glandula pinealis) is neuroendocrine gland located at the epithalamus of the brain secreting melatonin. The aim of this study was to explore effects of prenatal hypoxia in rats at the age of 33 weeks on the occurrence of pineal gland calcification. Distribution and chemical composition of calcerous material by light, scanning and transmission electron microscopy was investigated. Melatonin concentrations in blood plasma by direct radioimmunoassay were measured. Rats were exposed to prenatal hypoxia for 12 h at day 20 of development and second group to prenatal hypoxia for 2x8 h at days 19 and 20 of development. Vacuoles of intracellular edema in the pineal samples after 12 h hypoxia were found. Their size ranges up to 30 µm. Some of them were filled with the flocculent and fibrous material. Samples of pineal glands after 2 x 8 h hypoxia revealed the pericellular edema of pinealocytes. The amount of calcium rich particles in 2 x 8 h hypoxia group was lower than in 12 h hypoxia group. Plasma melatonin levels did not differ between control and both hypoxia groups. We concluded that calcification is a process induced by osteoblasts and osteocytes with melatonin as a promotor and it is favored under hypoxic conditions.

Metabolic effects of subchronic peripheral oxytocin administration in lean and obese zucker rats.

Increasing evidence indicates a role of oxytocin in controlling energy metabolism. The aim of his study was to investigate oxytocin effects on obese phenotype in leptin-resistant Zucker fatty rats, focusing on glucose and lipid metabolism. Zucker fatty rats and their lean controls were treated with oxytocin (3.6 µg/100g body weight/day) by osmotic minipumps implanted subcutaneously for 2 weeks. Two-hours intraperitoneal glucose tolerance test was performed in fasting rats. Oxytocin decreased food intake in both phenotypes while body weight gain reduced only in obese animals. In obese rats oxytocin impaired hepatic insulin extraction and enhanced liver triglyceride accumulation. Moreover, in the skeletal muscle of lean rats oxytocin treatment downregulated insulin signal transduction by decreasing of insulin receptor substrate 1 protein level and stimulating of its serine phosphorylation. Concurrently, the gene expression of insulin receptor substrate 1 in the skeletal muscle and adipose tissue was downregulated by oxytocin. In obese rats, oxytocin reduced adipocyte size and normalised mRNA levels of both fatty acid binding protein 4 and fatty acid synthase but attenuated gene expression of glucose transporter 4. The present study in Zucker fatty rats demonstrated ambivalent effects of oxytocin treatment with predominantly negative impact on skeletal muscle insulin pathway in lean animals.

Dominant neurologic symptomatology in intravascular large B-cell lymphoma.

Intravascular large B-cell lymphoma (IVLBCL) is a rare variant of extranodal large B-cell lymphoma and it is characterized by selective intravascular proliferation of malignant cells. Typical features of the disease include aggressive behavior, rapid and frequently fatal course. Clinical picture is non-specific and heterogeneous, depending on the affected organ. It is not uncommon that this unique type of lymphoma is diagnosed post mortem. Herein, we report two cases of IVLBCL with neurologic symptomatology. In our clinical study patient 1 was an 80-year-old male with mixed paraparesis of lower extremities and difficulties with sphincter control. Patient 2 (56-year-old male) had vision malfunction, mental status changes and defect in phatic and motor functions. In both cases definite diagnosis was established by histological examination of necroptic material. We propose to include IVLBCL in differential diagnostic considerations in patients presenting with gradually impairing neurological status and spinal cord damage of unknown etiology (Fig. 2, Ref. 9).

PTEN sequence analysis in endometrial hyperplasia and endometrial carcinoma in Slovak women.

Phosphatase and tensin homolog (PTEN) is a protein that acts as a tumor suppressor by dephosphorylating the lipid second messenger phosphatidylinositol 3,4,5-trisphosphate. Loss of PTEN function has been implicated in the pathogenesis of a number of different tumors, particularly endometrial carcinoma (ECa). ECa is the most common neoplasia of the female genital tract. Our study evaluates an association between the morphological appearance of endometrial hyperplasia and endometrial carcinoma and the degree of PTEN alterations. A total of 45 endometrial biopsies from Slovak women were included in present study. Formalin-fixed and paraffin-embedded tissue samples with simple hyperplasia (3), complex hyperplasia (5), atypical complex hyperplasia (7), endometrioid carcinomas G1 (20) and G3 (5), and serous carcinoma (5) were evaluated for the presence of mutations in coding regions of PTEN gene, the most frequently mutated tumor suppressor gene in endometrial carcinoma. 75% of the detected mutations were clustered in exons 5 and 8. Out of the 39 mutations detected in 24 cases, 20 were frameshifts and 19 were nonsense, missense, or silent mutations. Some specimens harboured more than one mutation. The results of current study on Slovak women were compared to a previous study performed on Polish population. The two sets of results were similar.

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).

[Clinical and microbiological characteristics of severe infections caused by group A streptococci]. / Klinická a mikrobiologická charakteristika závazných infekcii sposobených streptokokmi skupiny A.

AIM OF THE STUDY: Analysis of data on the occurrence of severe infections caused by group A streptococci, emm typing of the isolated strains, and survey of resistance to the selected antibiotics. MATERIAL AND METHODS: In 2012 and 2013, 43 S. pyogenes strains were isolated from severe infections. The isolates were most often recovered from wounds and abscesses (36 patients), followed by blood culture (5 patients), sputum (1 patient), and autopsy samples (1 patient). Antimicrobial susceptibility to selected antibiotics ( penicillin, erythromycin, clindamycin, spiramycin, and tetracycline) was tested by the disk-diffusion method and the minimal inhibitory concentrations (MICs) were determined. RESULTS: In 2012 and 2013, an increase in severe streptococcal infections was observed, with 27 and 16 cases reported, respectively. Resistance to macrolides was detected in 18.5% and 12.5% of isolates, respectively, and resistance to tetracycline in 25.9% and 20% of isolates, respectively. The isolated S. pyogenes strains were most often assigned to emm type 1. Risk factors for streptococcal infection were analyzed, with injury being most often identified as a risk factor- in seven and two patients, respectively. The patients were most frequently treated by beta-lactam antibiotics (penicillin and ampicillin). In 2012, two cases were fatal. CONCLUSION: This study reports cases of invasive group A streptococcal (GAS) infection as seen in clinical practice. An increased occurrence of S. pyogenes was observed in the hospital, with the isolates exhibiting elevated resistance to erythromycin and tetracycline. It is necessary to monitor the trends of antimicrobial resistance and the distribution of emm types among group A streptococcal isolates. Cooperation of the laboratory and clinical professionals in the prevention and therapy of streptococcal infections, invasive in particular, is crucial for risk reduction.

The normal human newborns thymus.

The thymic microenvironment constitutes a unique cell environment composed of thymic epithelial cells, myoid cells, and bone marrow-derived accessory cells for the differentiation, maturation and selection of T lymphocytes. The histological feature of thymus is markedly dependent on the age of individual and on various negative stimuli. Our study group consisted of fourteen newborns whose thymuses were removed during surgery performed for various congenital heart defects. We used a palette of seven monoclonal antibodies for exact localization of different cells creating the thymic microenvironment (cytokeratin AE1/AE3, desmin, actin, S100 protein, CD68, CD20, and CD45RO) as well as three monoclonal antibodies against proteins regulating the process of apoptosis (bcl2 oncoprotein, p53 protein, and survivin). We described and microphotographically illustrated the localization of thymic cytokeratin AE1/AE3-positive epithelial cells (subcapsular part of the cortex and medulla, especially Hassall's corpuscles), dendritic cells (medulla, often inside the Hassall's corpuscles), thymic myoid cells (medulla, often in close contact with Hassall's corpuscles), macrophages (mostly cortex, but also medulla and inside the Hassall's corpuscles), B lymphocytes (thymic medulla) and CD45RO-positive T lymphocytes (mostly thymic cortex). We found p53-positive thymic epithelial cell nuclei in subcapsular part of cortex and in outer epithelial cell layer of Hassall's corpuscles (very similar to the basal layer of epidermis). Bcl2 positive lymphocytes were mostly localized in thymic medulla, especially nearby Hassall's corpuscles. The thymuses were mostly survivin-negative with exception of round cells in border between cortex and connective tissue septa (probably migrating progenitor cells) (Tab. 1, Fig. 14, Ref. 66).

[The viewpoint of an internist on the problem of an aortoenteric fistula]. / Pohlad internistu na problematiku aortoenterálnej fistuly.

Aortoenteric fistula is an uncommon life-threatening disease; it can be divided into primary and secondary one. Primary aortoenteric fistula is the result of ongoing disease in the aorta and the intestine, secondary one is iatrogenic. Typical symptoms are abdominal pain and gastrointestinal bleeding (two-stage process). The most appropriate diagnostic method is CT aortography, treatment is only surgical. 75-year-old patient was admitted to the 5th Department of Internal Medicine in Bratislava due to progression of renal parameters, the patient had undergone an aortofemoral bypass 4 years ago. During the fourth day of his hospitalization the patient had a massive hematochezia with a shock state. After transient stabilization of the patient, 30 minutes after the first hematochezia, a massive hematemesis appeared and then the patient died. An autopsy has confirmed the secondary aortic-enteral fistula between the duodenum and the aneurysm arising from the aortofemoral bypass.

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).

Thymic Hassall's bodies of children with congenital heart defects.

The development of the thymus and heart are closely related while in both, the neural crest cells play an important role. In our preliminary study, the thymic microscopic structures of the infant's thymuses with the congenital heart defects were observed. The study was conducted on 36 specimens of newborn thymuses removed due to surgery for cardiovascular malformations. Standard formalin-fixed paraffin-embedded tissue technique was used. Five-microm-thick sections were stained with hematoxylin and eosin and the microscopic examination was focused on the structure of Hassall's bodies. The Hassall's bodies showed considerable variations in size as well as in quantity. In most cases, the Hassall's bodies were large with the heterogeneous amorphous material enclosed in cystic dilatations. This type of Hassall's bodies is typical for adult thymuses. The most conspicuous changes (huge Hassall's bodies with the cystic dilatation filled up with cell detritus) were observed in patients with ventricular septal defect, atrioventricular septal defect and tetralogy of Fallot. Small-sized Hassall's bodies corresponding with infant age, were observed in cases with pulmonary valve atresia, atrial septal defect and in some cases of transposition of great arteries. We assume that the changes of microenvironment of the thymic medulla are associated with disrupted migration of the neural crest cells which are essential in the normogenesis of both heart and thymus (Tab. 1, Fig. 12, Ref. 33).

Cosmetic perfumes vs. human pheromones (natural chemical scents) of the human female and male in signalling and performing context of their sexual behaviour.

Scent communication in man is undoubtedly of importance, although it is unconscious, rather than active, as compared to subhuman primates. Besides human sexual life it also affects a number of further characteristics of human life and its infrastructure including the mother-child relationship, creation of the odour basis of the family with the possibility to identify the family members solely by their odour as well as other parameters investigated thus far. Pheromones have effect upon the selection of a suitable partner of the opposite sex (or of the same sex in homosexual partners). The formation of specifically significant responses during communication between the two sexes, first of all in sexual life and its manifestations, may also be influenced by pheromone-based perfumes or classical cosmetic perfumes, as far as they are selected and used appropriately. The situation is much easier if the partners are of the olfactory type where for both partners the mutual olfactory parameters are the most attractive for their sexual life and its parameters, which significantly contributes to the quality of their overall coexistence (Ref. 29).