Loss of Expression of Antiangiogenic Protein FKBPL in Endometrioid Endometrial Carcinoma: Implications for Clinical Practice.

Background and Objectives: FK506 binding protein like (FKBPL) is a member of the immunophilin family, with anti-angiogenic effects capable of inhibiting the migration of endothelial cells and blood vessel formation. Its role as an inhibitor of tumor growth and angiogenesis has previously been shown in studies with breast and ovarian cancer. The role of FKBPL in angiogenesis, growth, and carcinogenesis of endometrioid endometrial carcinoma (EEC) is still largely unknown. The aim of this study was to examine the expression of FKBPL in EEC and benign endometrial hyperplasia (BEH) and its correlation with the expression of vascular endothelial factor-A (VEGF-A) and estrogen receptor alpha (ERα). Materials and Methods: Specimens from 89 patients with EEC and 40 patients with BEH, as well as histological, clinical, and demographic data, were obtained from the Clinical Hospital Centre Zemun, Belgrade, Serbia over a 10-year period (2010−2020). Immunohistochemical staining of the tissue was performed for FKBPL, VEGF-A, and ERα. Slides were analyzed blind by two pathologists, who measured the intensity of FKBPL and VEGF-A expression and used the Allred score to determine the level of ERα expression. Results: Immunohistochemical analysis showed moderate to high intensity of FKBPL expression in 97.5% (n = 39) of samples of BEH, and low or no expression in 93.3% (n = 83) of cases of EEC. FKBPL staining showed a high positive predictive value (98.8%) and a high negative predictive value for malignant diagnosis (86.7%). The difference in FKBPL expression between EEC and BEH was statistically significant (p < 0.001), showing a decrease in intensity and loss of expression in malignant tissues of the endometrium. FKBPL expression was positively correlated with ERα expression (intensity, percentage and high Allred score values) and negatively correlated with the expression of VEGF-A (p < 0.05 for all). Conclusions: FKBPL protein expression demonstrated a significant decrease in FKBPL in EEC in comparison to BEH tissue, with a high predictive value for malignancy. FKBPL might be emerging as a significant protein with antiangiogenic and antineoplastic effects, showing great promise for the diagnostic and therapeutic applications of its therapeutic derivatives in gynecological oncology.

Expression of connexin-43 in surgical resections of primary tumor and lymph node metastases of squamous cell carcinoma and adenocarcinoma of the lung: a retrospective study.

Background: Connexins are transmembrane proteins forming gap junctions between the cells, which allow intercellular communication. Significance of gap junctions and connexins in lung carcinoma is not yet understood. The objective of the study was to investigate immunohistochemical expression and the localization of connexin-43 (Cx43) in primary lung carcinoma and its lymphatic metastases. Methods: Surgical specimens of excised tumors from 88 patients (45 men and 43 women, 61.9 ± 7.4 years) with lung carcinoma (52 adenocarcinoma (AC), 36 squamous cell carcinoma (SqCC)) who were operated on at the University Hospital "Bezanijska Kosa" in a five-year period (2012-2016) were used. We conducted immunohistochemical staining for Cx43 and measured the degree of expression (percentage of positive cells and staining intensity) as well as localization of Cx43 in primary tumor and in lymphatic metastases. Results: Immunohistochemical analysis of the primary tumors revealed that SqCC showed significantly higher percentage of tumor cells expressing Cx43 as well as higher staining intensity than AC (p < 0.001). Almost 70% of samples with SqCC showed high Cx43 expression, whereas AC showed no expression in more than 50% of cases. Localization of Cx43 expression was most often cytoplasmic (AC and SqCC) and combined membranous and cytoplasmic (SqCC) with very rare instances of nuclear localization (AC). Almost the same pattern in distribution, intensity, and localization of Cx43 expression was observed in the lymph node metastases; however, almost a third of AC cases changed the pattern of Cx43 expression in the metastasis compared to primary tumor. Conclusion: The results of this study showed that lung carcinomas express Cx43 in more than 65% of cases and that it was aberrantly localized (not membranous localization). We highlighted that SqCC expressed Cx43 more than did AC, both in primary tumor and lymphatic metastases. Further research is needed to establish whether Cx43 could be used as a prognostic biomarker in lung carcinoma.

Teledermoscopy for Skin Cancer Prevention: a Comparative Study of Clinical and Teledermoscopic Diagnosis.

INTRODUCTION: The number of newly diagnosed skin cancers per year is greater than the sum of the four most common cancers: breast, prostate, lung, and colon. The implementation of primary and secondary prevention measures, over the last 2 to 3 decades, has made a major contribution to successful treatment. AIM: Evaluate the accuracy and reliability of teledermoscopic versus clinical diagnosis for skin cancers when diagnostic algorithms are used, and when GPs and surgical specialties are involved in the clinical procedure. METHODS: Digital dermoscope (TS-DD, by Teleskin company) was used for the acquisition of teledermoscopic photographs and specialized teledermoscopic software was used for clinical examination and teledermoscopic consultation. The teledermoscopic procedure itself was performed in two steps. The first step was a clinical examination using the ABCDE rule with digital dermoscopic photography of the suspected lesion. The second step was a 2-step dermoscopic evaluation using the second step ABCD algorithm for the second step. Accuracy and diagnostic reliability were calculated for: teledermoscopic diagnosis versus histopathological diagnosis; clinical diagnosis versus histopathological diagnosis and teledermoscopic diagnosis versus clinical diagnosis. RESULTS: The study included 120 patients with 121 Pigmented Skin Lesions, of which 75 (62%) were benign and 46 (38%) were malignant lesions (6 melanomas and 40 NonMelanoma Skin Cancers). Diagnostic accuracy between teledermoscopic and histopathologic diagnosis was 90.91% and reliability k=0.81; between clinical and histopathological diagnosis the diagnostic accuracy was 82.64% and the reliability k=0.64 and between the clinical and teledermoscopic diagnosis the diagnostic accuracy was 81.82% and the reliability k=0.62. CONCLUSION: The achieved diagnostic accuracy between clinical and teledermoscopic diagnosis, when using diagnostic algorithms, establishes a feasible screening path for skin cancers and indicates that general practitioners and specialized surgeons may equally be involved in prevention.

New software for collecting data from the organized cervical cancer screening program in Serbia. Are we on the threshold of a new screening registry? - A multicentric study.

PURPOSE: The purpose of this study was to present the Screening Registry and the results of organized cervical cancer screening program (OCCSP) in the Republic of Serbia using a database made as an output model, linked with the Screening Registry. METHODS: Data were respectively collected over a onemonth period from 3 state primary health care centers (and related hospitals/clinical center) in central Serbia in which OCCSP was conducted. The sample consisted of women of the target population (25 to 64 years old) who responded the call for Pap test. RESULTS: The most frequent abnormal cytological diagnosis was in the 38-50 years age group, and consisted of atypical squamous cells of undetermined significance - ASCUS (7.5%) and low grade squamous intraepithelial lesions - L-SIL (7.3%). The most frequent abnormal colposcopic finding in the youngest age group of women (25-37 years) was iodine negative epithelium (35.7%) and in the group of women aged 38-50 and 51-64 years acid-white epithelium. The most common histopathological diagnosis was L-SIL. Positive predictive value of colposcopy in relation to the Pap test was 0.64 (95% CI=0.56-0.70). Interrater agreement (between cytotechnicians and supervisors) measured by the Cohen's coefficient was 0.94 (95% CI=0.91 to 0.97), but between cytology (supervisors) and pathology findings it was 0.83 (95% CI = 0.67 to 0.99). CONCLUSION: The existence of a screening registry contributes to a better epidemiological surveillance of a screening program, and to a possibility for development of various epidemiological researches.

Homogeneous static magnetic field of different orientation induces biological changes in subacutely exposed mice.

It has been shown that static magnetic field (SMF) of moderate intensity produces considerable impact on biological systems. SMF can be homogeneous or inhomogeneous. In many studies, inhomogeneous SMF was employed. Aware that inhomogeneous SMF could result in experimental variability, we investigated the influence of a vertical homogeneous SMF of different orientation. Male Swiss-Webster 9- to 10-week-old mice were subacutely exposed to upward- and downward-oriented SMF of 128 mT generated by a cyclotron for 1 h/day during a 5-day period. We found that SMF affected various organs and that these effects were, to some degree, dependent on SMF orientation. Both upward- and downward-oriented SMF caused a reduction in the amount of total white blood cells (WBC) and lymphocytes in serum, a decrease of granulocytes in the spleen, kidney inflammation, and an increase in the amount of high-density lipoprotein (HDL). In addition, upward-oriented SMF caused brain edema and increased spleen cellularity. In contrast, downward-oriented SMF induced liver inflammation and a decrease in the amount of serum granulocytes. These effects might represent a specific redistribution of pro-inflammatory cells in blood and among various organs. It appears that homogeneous SMF of 128 mT affected specific organs in the body, rather than simultaneously and equally influencing the entire body system.

Anxiolytic and antidepressant effect of zinc on rats and its impact on general behavioural parameters.

BACKGROUND/AIM: Zinc is an essential element which has considerable interaction with gamma-aminobutyric acid A type receptors (GABA(A)) and glutamate receptors in the cen tral nervous system (CNS). It is believed that zinc acts as a potent inhibitor of glutamate N-methyl-D-aspartate (NMDA) receptors, and binding to structurally specific site on the GABA(A) receptor leads to inhibition of GABA-dependent Cl-pass. The aim of our research was to test the anxiolytic and antidepressant effects of zinc after single application and its influence on general behavioural parameters after repeated administration. METHODS: Male Wistar rats were treated with increasing doses of zinc histidine dehydrate (10, 20, 30 mg/kg, i.p.). To determine anxiolytic and antidepressant properties of zinc two models were used: elevated plus maze (EPM) and forced swim test (FST). Behavioural parameters (stillness and mobility) were, also, recorded after single and repeated administration of active substance. RESULTS: Testing animals in the EPM showed a statistically significant difference as follows: dose of 20 mg/kg significantly increased the time animals spent in open arms, indicating an acute anxio lytic effect, while doses of 30 mg/kg significantly reduced the time in the open arms, indicating a potentially anxiogenic ef fect. Testing the animals by FST showed a statistically signifi cant difference in immobility time of animals treated with the lowest applied (10 mg/kg) and highest applied (30 mg/kg) doses of zinc, compared to the control group. The first day of testing behavioral parameters showed the tendency to in crease locomotor activity of the animals with the lowest dose of zinc (10 mg/kg), while the following day revealed a reduced activity with the highest dose applied (30 mg/kg). CONCLUSION: Zinc has important effects on the CNS: After single application, in all doses zinc showed antidepressant ef fects. The effects of zinc on anxiety and locomotor activity showed dose-dependent bidirectional effects.

Ultrastructural changes of the peritoneum in a rabbit model of peritoneal dialysis.

BACKGROUND/AIM: The number of patients with end-stage renal diseases treated with chronic dialysis is increasing over the last years. Long-term peritoneal dialysis is associated with progressive development of structural and functional alterations of peritoneal membrane. The aim of the study was to analyze ultrastructural alterations of mesothelial monolayer and submesothelial tissue in a modified nonuremic experimental model of peritoneal dialysis in rabbits. METHODS: The study was performed on 5 healthy Chinchilla rabbits. Surgical procedures of implantation and removal of peritoneal catheter, prevention of catheter clothing, prevention of infection and dialysate instillation were performed according to previously described protocols. Peritoneal tissue samples were collected upon catheter placement and removal after a 5-week follow-up and processed for transmission electron microscopy (TEM) examination. RESULTS: The rabbits tolerated anesthesia, surgical procedure and the applied regimen of dialysate instillations well. The animals recovered completely and no adverse effects were noted. In the animals treated with peritoneal dialysis instillations, TEM revealed alterations of the mesothelial monolayer and submesothelial tissue. The mesothelial cells in direct contact with dialysis fluid were prone to shrinking. They lost the typical cobblestone morphology and assumed a flattened shape. The mesothelial cells were often detached from the basement membrane. These cells showed euchromatic nuclei, higher number of microvilli in their apical part and very numerous vesicles. A higher quantity of collagen fibers was noticed in the peritoneal lamina propria in close relation to the basement membrane of mesothelium. The nuclei of the fibroblasts were also euchromatic. Numerous mitochondria, granules and vesicles were present in their cytoplasm. CONCLUSION: The used rabbit model of peritoneal dialysis is simple, practical to perform, reproducible, not expensive and not requiring advanced devices. It is suitable for obtaining peritoneal tissue samples for histological examination and can be used to analyze the effects of dialysis solutions on the rabbit peritoneal membrane.

DMCM, a benzodiazepine site inverse agonist, improves active avoidance and motivation in the rat.

There are several modulatory sites at GABA(A) receptors, which mediate the actions of many drugs, among them benzodiazepine. Three kinds of allosteric modulators act through the benzodiazepine binding site: positive (agonist), neutral (antagonist), and negative (inverse agonist). The goal of the present study was to examine the influence of the inverse agonist methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) acting on α GABA(A) receptor and compare its dose-response effects on memory and depression-like behavior. We independently studied the effects of DMCM (0.05-1.0 mg/kg) on retention versus acquisition of active avoidance and depression-like behavior in the forced swim test. Throughout the study, drugs were given intraperitoneally, 30 min before testing. ANOVA has showed that treatment with DMCM significantly affected retrieval of avoidance response (p<0.05), exerted promnesic effects in inverted U-shape manner. Dunnett's test indicated that the DMCM avoidance-facilitatory dose was 0.1mg/kg. At the dose facilitating retrieval of avoidance memory, DMCM significantly (p<0.05, comparison of regression coefficients by Student's t-test) and progressively increased acquisition rate during 5 days training, compared to the saline group. In forced swim test, ANOVA indicated statistically significant effects of DMCM (p<0.05). Dunnett's analysis showed that DMCM significantly decreased immobility time at the dose of 0.1mg/kg, exerted acute antidepressant-like effects. Our results experimentally support the findings that under certain circumstances, nonselective benzodiazepine site inverse agonists, produce memory-enhancing and antidepressant-like effects. The molecular and neuronal substrates linking the actions of specific GABA-benzodiazepine receptor complex subunits remains to be further elucidated.

Phyllodes tumors and fibroadenoma common beginning and different ending.

Phyllodes tumors and fibroadenomas are the most common benign breast tumors. They arise from intralobular fibrous tissue as a unique lesion and after a period of time they differentiate in two direction: to fibroadenoma and to phyllodes tumors. Fibroadenomas grow up to 2-3 cm and then stop growing but phyllodes tumors grow continually and sometimes are to 40 cm big. Both these lesions have two components, epithelial and stromal. Clinically fibroadenomas are well circumscibed, hard, oval, movable lesions. They can be solitary, multiple, unilateral and bilateral. They are hormone dependent changes, because they change their own consistency during menstrual cycle and gravidity. The most commonly used histological classification is in two types: pericanalicular and intracanalicular type. Phyllodes tumors make about 1% of all breast tumors. This tumor has many synonyms. It starts as fibroadenoma in intralobular stromal component. It has continuous growth and biologically it can be benign, borderline and malignant. The first description is from Miller (1838). The main goal is to find the divergence point when the developing is direct to fibroadenoma or phyllodes tumor. The second goal is to investigate the fate of epithelial and stromal component in these two lesions. Retrospective analysis is made of all fibroadenomas and phyllodes tumors in Pathology Department of Medical Center "Bezanijska kosa" in the period from 1998 to 2006. In this period, 2919 women were operated for breast changes. 343 fibroadenoma (24, 4%), were diagnosed, benign phyllodes tumor in 95 women (6.7%) and malignant phyllodes in 4 cases or 0.2%. All slides from these patients were analysed for many different histological parameters and immunohistological investigation for steroid receptors was also used, c-erbB2 (Her2/Neu), PCNA (proliferative cellular nuclear antigen) and Ki-67, androgen receptor and p53. All data were statistically investigated (Odds ratio, confidence interval, Fisher exact test, Wilcoxon sum test and Kendall test). It was concluded that fibroadenomas and phyllodes tumors arise from intralobular fibrous tissue, both changes have very close histology in the beginning and divergent growth starts later. Differences are present in stromal component. Phyllodes tumor has two component stroma. Stromal cells in phyllodes tumors are more PCNA positive than in fibroadenomas, also Ki-67 and androgen receptors are more positive in phyllodes tumors. Histologically phyllodes tumors have perforated capsule with finger like projections. These data determine surgical procedure, wide excision in phyllodes and simple excision in fibroadenomas.

Angiofollicular lymphoid hyperplasia--an unusual cause of axillary artery pseudoaneurysm.

BACKGROUND: Angiofollicular lymphoid hyperplasia (Castleman's disease) is a rare inflammatory condition. Its cause is unknown and it can be both localized and general. Eighty-six percent of lesions are found in the mediastinum or hilum, and 91% are of the hyaline vascular type. Although Castleman's disease is primarily involving the chest and retroperitoneum, it may also involve neck and axilla in 2-4% of cases. In this article, we present a very rare case of Castleman's disease causing axillary artery pseudoaneurysm. METHODS AND RESULTS: A 30-year-old woman patient presented with pulsating tumefaction of the left arm which was 3.5 x 10 cm in size, and became evident 15 days before admission. History revealed that she suffered trauma of the left upper arm in childhood; therefore, it was suspected that tumefaction may be due to a post-traumatic aneurysm. Duplex scan and multislice computed tomography examinations were performed and an axillary artery pseudoaneurysm was diagnosed. Since intraoperatively the tumefaction resembled the tumorous formation but not the aneurysmal wall, specimens were sent for pathohistological analysis. Arterial reconstruction was performed using autologous vein graft. Pathohistological findings showed vascular type of angiofollicular lymphoid hyperplasia. CONCLUSION: Although angiofollicular lymphoid hyperplasia is rarely localized in the axillary area, this disease should also be considered when axillary artery pseudoaneurysm is diagnosed.

[Morphometric characteristics of neuropeptide Y immunoreactive neurons of human cortical amygdaloid nucleus].

INTRODUCTION: Cortical amygdaloid nucleus belongs to the corticomedial part of the amygdaloid complex. In this nucleus there are neurons that produce neuropeptide Y. This peptide has important roles in sleeping, learning, memory, gastrointestinal regulation, anxiety, epilepsy, alcoholism and depression. MATERIAL AND METHODS: We investigated morphometric characteristics (numbers of primary dendrites, longer and shorter diameters of cell bodies and maximal radius of dendritic arborization) of NPY immunoreactive neurons of human cortical amygdaloid nucleus on 6 male adult human brains, aged 46 to 77 years, by immunohistochemical avidin-biotin technique. RESULTS: Our investigation has shown that in this nucleus there is a moderate number of NPY immunoreactive neurons. 67% of found neurons were nonpyramidal, while 33% were pyramidal. Among the nonpyramidal neurons the dominant groups were multipolar neurons (41%--of which 25% were multipolar irregular, and 16% multipolar oval). Among the pyramidal neurons the dominant groups were the neurons with triangular shape of cell body (21%). All found NPY immunoreactive neurons (pyramidal and nonpyramidal altogether) had intervals of values of numbers of primary dendrites 2 to 6, longer diameters of cell bodies 13 to 38 microm, shorter diameters of cell bodies 9 to 20 microm and maximal radius of dendritic arborization 50 to 340 em. More than a half of investigated neurons (57%) had 3 primary dendrites. DISCUSSION AND CONCLUSION: The other researchers did not find such percentage of pyramidal immunoreactive neurons in this amygdaloid nucleus. If we compare our results with the results of the ather researchers we can conclude that all pyramidal NPY immunoreactive neurons found in this human amygdaloid nucleus belong to the class I of neurons, and that all nonpyramidal NPY immunoreactive neurons belong to the class II of neurons described by other researchers. We suppose that all found pyramidal neurons were projectional.

[Breast Paget disease: morphologic substrate and Paget cells characteristics]. / Pagetova bolest dojlke: morfoloski substrat i karakteristike Pagetovih celija.

Authors analyzed 143 cases of breast Paget disease with special emphasis on basic tumor process and on immune histochemical characteristics of tumor cells. Breast Paget disease represent special clinical presentation of the breast tumor. The most often tumor in the breast are ductal carcinoma in situ, ductal carcinoma and in rare instance lobular carcinoma and lobular carcinoma in situ. There are several clinical combinations with or without involvement of axillary's lymph nodes. The best prognosis is in cases with in situ as underlying process. All other combinations have worse prognosis in relation to the combination with in situ lesion. The middle ages of patients with Paget disease is 10 years higher than the ages without areola involvement. Paget cells show the same immune histochemical characteristic as tumor cells. The authors propose mastectomy instead central segmentectomy.

[Morphological characteristics of the phyllodes tumours of the breast]. / Morfoloske karakteristike filodnih tumora dojke.

Tumor phyllodes arise from stroma component of the terminal ductulo-lobular unit (TDLU). Stromal and epithelial proliferation are present in the same time. The stromal component grow th more intensive and dominante over the epithelial component. Clinically, macroscopically and microscopically tumor phyllode look like fibroadenoma which is more common lesion than tumor phyllodes. The differentiation between these two lession is important because the fibroadenoma almost never reccur bur the reccurence is ofthen in tumor phyllodes especially in incomplete excision. The fibroadenoma growth is never over 3 cm, but the growth of tumor phyllodes is sometimes over the 20 cm. The clinica differentiation in smaller lesions is very hard, macroscopical differentiation is a litle better, but the final diagnosis is microscopical. Biological behavior is not in the correlation with histological picture. Histologically, we can differentiate benign, borderline and malignant tumor phyllodes.

[Markers of ductal carcinoma in situ]. / Markerske karakteristike duktalnog karcinoma in situ.

The breast cancer has become more frequent with women lately. The reasons for that are longer life, effects of external factors and hormone effects, estrogen in particular either natural or synthetic. Today's trend is to detect these tumors in situ in early stage where two types of lesions are distinguished: ductal carcinoma in situ (DCIS) and lobular carcinoma in situ. It is important to determine by means of histochemistry receptors for estrogen, progesteronee, C-erb B-2, p53 and profilic nucleic antigen (PCNA) in tissue of these tumors not only for prognostic importance, bat for selection of the optimal treatment method. The aim of the study is to analyze the frequency of the said receptors in 22 samples of tissue "ex tempore" where DCIS was found. Out of 50 cases where areas of micro-calcification identified on mammograms, 22 cases were separated with DCIS "ex tempore". Applying the immunohistochemistry method with commercial antibodies produced by the firm "Dako" receptors of estrogen, progesterone, C-erb B-2, p53 and PCNA were identified. Receptors were identified by monoclonal antibodies according to the instruction supplied by the firm. Regarding the type of DCIS the finding was 18% solid, 4.5% comedo, 4.5% parietal, 13.6 cribriform, while mixed types were found in 59.1% cases. Out of 22 women with DCIS, 4 of them or 18.2% had positive P-53; 13 or 59.1% PCNA, 15 or 68.2% positive C-erb B-2, 8 or 36.4% positive estrogen, 4 or 18.2% positive progesterone receptors. When the sample is submitted for patho-histologic testing it is necessary to determine the subtypes of DCIS, number of ducts affected by subtypes, other accompanying profilic changes and it is compulsory to identify receptors for C-erb B-2, estrogen and progesteronee, which can be done in any patho-histologic lab. The benefit of their identification before passing a decision about the therapy is big, i.e. they determine the type of therapy which has a preventive effect on the rest od tissue both affected and sound breast.