Radioimmunoassay of chromogranin A and free metanephrines in diagnosis of pheochromocytoma.

This work discusses the clinical performance of chromogranin A, free metanephrine and normetanephrine determination in plasma using a radioimmunoanalytical methods for the diagnosis of pheochromocytoma and paraganglioma. Blood samples were collected from 55 patients (46 pheochromocytomas, 9 paragangliomas). A sampling of biological materials was performed preoperatively and about one week, six months and one year after adrenal gland surgery. The comparative group without a diagnosis of pheochromocytoma/paraganglioma consisted of 36 pheochromocytoma/paraganglioma patients more than 4 months after adrenal gland surgery, and of 87 patients, 16 of them with multiple endocrine neoplasia, 9 with medullary and 5 with parafolicullar carcinoma of the thyroid gland. The rest were patients with various adrenal gland disorders. Chromogranin A, metanephrine and normetanephrine were determined in the EDTA-plasma using a radioimmunoassay kits Cisbio Bioassays, France and IBL International GmbH, Germany. Clinical sensitivity was 96 % for the combination of metanephrine and normetanephrine, and 93 % for chromogranin A. Clinical specificity was 100 % for the combination metanephrine and normetanephrine, and 96 % for chromogranin A. Falsely elevated levels of chromogranin A were observed in 1 patient with chronic renal insufficiency and 9 analyses were influenced by the administration of proton pump inhibitors. These results were excluded of CGA specificity. Both the combination of plasma free metanephrine, normetanephrine and chromogranin A as determined by radioimmunoassays, which are simple without the necessity of special laboratory material, are effective markers of pheochromocytoma or paraganglioma. Chromogranin A exerts association to malignity and all markers are associated with tumor mass.

Absence of BRAF mutation in pheochromocytoma and paraganglioma.

Pheochromocytomas and Paragangliomas (PHEO/PARA) are rare endocrine tumors originating from the adrenal medulla. More than 20 genes are involved in the tumorigenesis of these tumors, but a substantial part of the causative genetic events remains unexplained. A recent study has reported the presence of the activating BRAF V600E mutation in PCC, suggesting a role for BRAF activation in tumor development. Other studies have not find this mutation. This study investigates the occurrence of the BRAF V600E mutation in these tumors.A cohort of 64 PHEO/PARA were screened for the BRAF V600E mutation using direct Sanger sequencing and QRT-PCR.All cases investigated displayed wild-type without V600E BRAF mutationTaken together with all previously screened tumors up to date, only 1 V600E BRAF mutation has been found among 427 PCCs. These findings imply that the V600E BRAF mutation is a rare event in PHEO/PARA.

Endoscopic ultrasound-guided fine needle aspiration biopsy of pancreatic lesions. An 8-year analysis of single institution material focusing on efficacy and learning progress.

OBJECTIVE: To evaluate the efficacy and the learning curve of the endoscopic ultrasound-guided fine needle aspiration biopsy (EUS-FNAB) diagnostics of pancreatic lesions over 8 years (2007-2014). MATERIAL AND METHODS: EUS-FNAB using a Linear Echoendoscope Olympus was performed in 531 patients, mostly without rapid on-site evaluation (ROSE). Smears plus cytoblock sections were used routinely. Immunocytochemistry was utilised as indicated. RESULTS: The average, satisfactory sampling rate increased from the 61.2% in the first 3-year period to 72.9% in the last 3-year period (P = 0.008). The availability of the material for cytoblocks increased from 36.4% in the first period to 75.3% in the last period (P = 0.017). The efficacy of cytoblocks increased from 39.6% to 46.2% (P = 0.086). Comparing the first and last 3-year periods, the indication for immunocytochemistry did not rise substantially (5.2% and 8.5% respectively), but the predictive value of immunocytochemistry rose from 56.3% to 100.0% (P = 0.001). The most frequent diagnostic result was malignancy confirmation - both primary (41.2%) and metastatic (2.1%). In cases with representative samples and follow-up information, the specificity, sensitivity, positive predictive value and accuracy were high from the beginning. The negative predictive value decreased slightly in the last 3-year period. CONCLUSION: The pancreatic EUS-FNAB without ROSE represents a suboptimal arrangement conditioned with our staff/time/location reality. Nevertheless, within the last 3 years of our activity, nearly three-quarters of patients profit from the diagnostic contribution of this procedure.

[Adrenocortical oncocytoma presenting as Cushing´s syndrome in pregnancy with spontaneous postpartum uterine rupture]. / Adrenokortikální onkocytom jako raritní prícina Cushingova syndromu v gravidite a následná delozní ruptura po porodu.

OBJECTIVE: The description of rare case of the Cushings syndrome in pregnancy resulting in the spontaneous rupture of uterus. DESIGNS: A case report. SETTING: Department of Gynecology and Obstetrics, 1st Medical Faculty of Charles University and Hospital Na Bulovce. CASE REPORT: The authors report the case of a 33 year old woman, who was admitted to the clinic in 30th week of pregnancy for elevated blood pressure and hypokalemia. Arterial hypertension was corrected with the combination of Vasocardin and Dopegyt. In 36th week of pregnancy the patient was admitted to the hospital with premature rupture of membranes. In less than sixteen hours the patient spontaneously gave birth to a healthy girl. Subsequently the patient suffered from abdominal pain. CT scan showed haemoperitoneum and a right adrenal mass and the surgical revision was performed. During the surgery uterine rupture was identified and the patient uderwent abdominal hysterectomy. Based on the endocrinological examination the diagnosis of Cushings syndrome was made. Three months after the delivery she underwent laparoscopic right adrenalectomy. Histological examination revealed adrenocortical oncocytoma. CONCLUSION: Cushings syndrome is rare in pregnancy and misdiagnosis is common. Symptoms mimic pregnancy complications. The disease is often diagnosed with delay even after delivery. The sonography and magnetic resonance are dominant imaging methods. Laboratory diagnosis is difficult due to pregnancy related changes. In spite of rarity of Cushings syndrome in pregnancy we should think about it in terms of the differential diagnosis of hypertension, diabetes, dysbalance of mineralogram and typical cushingoid habitus.

Deconjugated urinary metanephrine, normetanephrine and 3-methoxytyramine in laboratory diagnosis of pheochromocytoma and paraganglioma.

This work discusses the clinical performance of deconjugated metanephrine (MN), normetanephrine (NMN) and 3-methoxytyramine (3MT) determined in the basal first morning urine using a chromatographic method with electrochemical detection for the clinical diagnosis of pheochromocytoma (PHEO) and paraganglioma (PGL). Urine samples were collected from 44 patients (36 with PHEO, 8 with PGL) aged 54+/-17 (20-78) years (22 females, 22 males). A sampling of biological materials was performed preoperatively and about one week, six months and one year after adrenal gland surgery. The control group consisted of 34 PHEO/PGL patients more than 4 months after adrenal gland surgery. All subjects in the control group were without a diagnosis of PHEO or PGL. Clinical sensitivity was 55 % for MN, 64 % for NMN, 80 % for combination of both MN and NMN, and only 23 % for 3TM. Clinical specificity calculated from the control group was 93 % for MN, 95 % for NMN, 95 % for the combination MN and NMN, and 97 % for 3TM. Cut-off values for deconjugated metanephrines in the basal urine were 310 (MN), 690 (NMN) and 250 microg/l (3MT). Chromatographic determination of deconjugated urinary metanephrines, which is simple without the necessity of special laboratory material, can serve for the screening of PHEO or PGL patients. Urine NMN and 3MT exerts an association to malignity, and all markers are associated with tumor mass. However, the principal laboratory diagnosis of PHEO or PGL must be based on plasma-free metanephrines and plasma chromogranin A with better performance in the laboratory diagnosis of PHEO or PGL.

Biochemical Testing After Pheochromocytoma Removal: How Early?

Pheochromocytomas are catecholamine-producing tumors with typical clinical presentation. Tumor resection is considered as an appropriate treatment strategy. Due to its unpredictable clinical behavior, biochemical testing is mandatory to confirm the success of tumor removal after surgery. The aim of the study was to investigate the feasibility of a shorter interval of postoperative testing (earlier than the recommended 2-4 weeks according to recently published Guidelines). We investigated 81 patients with pheochromocytoma before and after surgery. Postoperative examination was performed of stable subjects after their transport from the surgical to the internal ward (7.1±2.2 days after surgery). Plasma metanephrines were used for the diagnosis of pheochromocytoma and confirmation of successful tumor removal. All subjects with pheochromocytoma had markedly elevated plasma metanephrines before surgery. No correlation between postoperative interval (the shortest being 3 days) and plasma metanephrine levels was found. Postoperative plasma metanephrine levels did not differ significantly from those taken at the one-year follow-up. In conclusion, we have shown that early postoperative diagnostic workup of subjects with pheochromocytoma is possible and may thus simplify early postoperative management of this clinical condition.

[Results of the Czech National Cervical Cancer screening programme]. / Výsledky Národního programu screeningu karcinomu delozního hrdla v Ceské republice.

INTRODUCTION: The nationwide Cervical Cancer Screening Programme in the Czech Republic was introduced in 2009. The aim of this article is to describe the results of this programme in the first few years after its transformation into a fully-fledged, organised programme. MATERIAL AND METHODS: During the first few years, a network of 37 accredited laboratories was stabilised, ensuring that examinations would be readily available across the Czech Republic. Although all women aged between 25 and 70 years fall within the target group of the current personalised invitation programme, women not in this age group are reimbursed for preventive screening examinations as well. The programme is equipped with an information support provided by the Institute of Biostatistics and Analyses of the Masaryk University, which runs the Cervical Cancer Screening Registry. RESULTS: Cervical cancer incidence and mortality rates in the Czech Republic have seen a significant decrease over the last decade. By the end of 2013, more than 11 million examinations were reported to the registry by the accredited laboratories, and more than 15,000 serious cervical lesions were detected. Analysis of the data from the registry makes it possible to monitor the quality of individual laboratories and the effectiveness of the entire programme. CONCLUSION: The promising start of the programme provides a strong argument for its continuation. The program--me has the potential to decrease cervical cancer incidence rates in the Czech Republic to values comparable with those reported by countries with advanced health care systems.

Correlation among the BRAF gene mutation status, clinicopathological features of primary tumour, and lymph node metastasizing of papillary thyroid carcinoma.

BACKGROUND: Papillary thyroid carcinoma (PTC) is the most common malignant thyroid tumour. A common mutation of papillary thyroid carcinoma (PTC) is the somatic mutation of the BRAF (V600E) gene. AIM: The aim was to 1) determine the association of lymph node metastases of PTC with the BRAF gene mutation of primary tumour; 2) evaluate association of the BRAF mutation in the -primary tumour with clinicopathological para-meters; 3) examine the extent of genetic heterogeneity by monitoring the BRAF mutation in multicentric tumours. SUBJECTS AND METHODS: Retrospective analysis of the BRAF (V600E) mutation in PTC and PTC neck lymph node metastases in 156 patients operated from 2003 to 2012 in Prague and Zlín, the Czech Republic, using a qPCR assay. The results were correlated with clinicopathological factors. RESULTS: DNA was successfully extracted from 137 samples. The BRAF (V600E) mutation was detected in 78 cases (56.9%). The patients with BRAF p.Val600Glu mutation of primary tumour had only non-significantly higher risk of cervical lymph node metastases [OR=2.39 (95%) CI 1.00-5.75, p=0.052]. In the classic papillary variant, the BRAF (V600E) mutation was found significantly more often than in other PTC subtypes (p=0.022). We did not confirm any significant association between the BRAF (V600E) mutation and other clinicopathological findings. CONCLUSION: Except for the higher prevalence in papillary variant of PTC, BRAF p.Val600Glu mutation was not associated with other prognostic clinicopathological factors of PTC. BRAF mutation cannot be regarded as a reliable marker of node metastases in patients with PTC.

[Evaluation of safe resection margins in tumours of parenchymatous organs]. / Problematika stanovení bezpecných resekcních okraju u nádoru parenchymatózních orgánu

The definition of a safe surgical margin in tumours of parenchymatous organs has been the subject of frequent debates and a number of studies. This topic is not generally unified for parenchymatous organs and has many organ specifics. Moreover, there are still controversies in the diagnostic criteria and the definitions of positive resection margins, in indications of perioperative examinations of resection margins, and in the prognostic significance of margin involvement in different parenchymatous organs. The consensus in terminology and standardization of histopathological examination procedures is also very desirable across the diagnostic areas.

The prognostic value of T1 bladder cancer substaging: a single institution retrospective study.

OBJECTIVE: To evaluate the prognostic value of the depth of lamina propria invasion in patients with T1 bladder cancer. SUBJECTS AND METHODS: 200 patients were treated between the years 2002 and 2009. Tumours with depth of invasion above the muscularis mucosae level were categorised as pT1a and those with depth of invasion up to or beyond the muscularis mucosae as pT1b. RESULTS: Categorisation for pT1a and pT1b was performed in 176 of 200 patients (88%). In 10 patients a muscle-invasive tumour was found in re-transurethral resection samples. 131 (79%) of 166 analysed patients had pT1a tumour and 35 (21%) had pT1b tumour. During the follow-up, in 101 (61%) patients the tumour had recurred and in 27 (16.3%) the tumour had progressed. Of all the investigated parameters, T1 substaging (p < 0.0001), grade (p = 0.0003) and the number of bacillus Calmette-Guérin instillations (p = 0.0490) were significant in predicting progression. The only significant factor for disease-specific survival was T1 substaging in univariable (p = 0.0008) and multivariable (hazard ratio 4.407) analysis. T1 substaging (p = 0.0149) and tumour multiplicity (p = 0.0448) have a statistically significant prognostic value with respect to overall survival. CONCLUSIONS: Deep invasion of the lamina propria is a significant adverse prognostic factor for tumour progression, disease-specific survival and overall survival.

Prediction of recurrence in low and intermediate risk non-muscle invasive bladder cancer by real-time quantitative PCR analysis: cDNA microarray results.

The aim of the study was to define specific genetic profile in Ta and T1 urinary bladder carcinoma patients with and without recurrence by gene expression microarrays. Eleven patients with the time to recurrence shorter than one year (patients with recurrence) and 11 patients with time to recurrence longer than 4 years (patients without recurrence) were enrolled. Data from microarrays were subjected to a panel of statistical analyses to identify bladder cancer recurrence-associated gene signatures. Initial screening using the GeneSpring and Bioconductor software tools revealed a putative set 47 genes differing in gene expression in both groups. After the validation, 33 genes manifested significant differences between both groups. The significant expression was observed in the group of patients without recurrence by 30 genes of which the highest differences were detected by ANXA1, ARHGEF4, FLJ32252, GNE, NINJ1, PRICKLE1, PSAT1, RNASE1, SPTAN1, SYNGR1, TNFSF15, TSPAN1, and WDR34. These genes code for signal transduction, vascular remodeling and vascular endothelial growth inhibition mainly. In the group with recurrence, 3 genes had significant differences, the highest differences were identified by two genes (PLOD2 and WDR72). Loci of genes with significant changes of gene expression were located on characteristic chromosomes for bladder cancer: 7 loci on chromosome 9, 8 loci on chromosomes 1, 2, 3, 12,14,15,16, and 22. We have selected and validated 15 genes that are differentially expressed in superficial bladder cancer. We hope that this cohort of genes will serve as a promising pool of candidate biomarkers for early stage bladder cancer. Our results indicate that it may be possible to identify patients with a low and high risk of disease recurrence at an early stage using a molecular profile.

[What is new in cervical precanceroses cytodiagnostics?]. / Co je nového v cytodiagnostice cervikálních prekanceróz?

Cytopathology investigation of the uterine cervix transformation zone smear (Pap test) has been accepted during the last 80 years worldwide as a potent tool in lowering the incidence of squamous cell cervical cancer; it can reveal a proportion of adenocarcinomas as well and contributes to the diagnostics of cervicovaginal infections. The technique itself and diagnostic criteria have been internationally unified in the systems Bethesda I (1988) and Bethesda II (2002). Nevertheless, the cytodiagnostics of cervical precanceroses continues to develop vividly in the following fields of interest. In processing the cervical sample:Unified polychrome staining has been accepted as compulsory Processing of the sample acquired has split into two branches - conventional preparation - CP and liquid based preparation - LBP. In both types of processing (mainly in LBP) additional tests are employed.In evaluation of the cytology features:Differences of the petrified diagnostic features formulated formerly for CP in the LBP have been described. Differentially-diagnostic pitfalls (look-alikes) are studied. Sensitivity of precanceroses detection in a screening routine with the prevalence of negative findings has been improved with compulsory rescreening of 10-20% random selected negative cases as well as rapid pre- or postscreening of the whole material or involvement of automated pre- or postscreening using image analysis systems.Some cytomorphology findings are followed with additional tests - especially HR HPV detection.Cyto-bioptic correlations are constantly studied.In terms of lowering the incidence of uterine cervix carcinoma: Opportune screening is substituted with nationwide programs aimed at:Involvement of as many women of the target group as possible.Standardized investigation (CP or LBP) in an accredited laboratory with functioning systems of external and internal quality control.Selective additional investigation with non-morphological tests.Appropriate treatment of women with pathology findings.Some newly designed nationwide screening models start with a non-morphological test (HPV) followed by a pap test and colposcopy.Stern control of successful implementation of the achievements listed above is expressed in uterine cervix cancer incidence.

High incidence of cardiovascular complications in pheochromocytoma.

Excess of catecholamines in pheochromocytoma is usually accompanied with classical symptoms and signs. In some cases, severe cardiovascular complications (e. g., heart failure, myocardial infarction) may occur. We performed a retrospective analysis focused on the incidence of cardiovascular complications (classified as follows: arrhythmias, myocardial involvement or ischemia and atherosclerosis, cerebrovascular impairment) before the establishment of diagnosis of pheochromocytoma among 145 subjects treated in our hospital. Cardiovascular complications occurred in 28 subjects, but these subjects did not differ significantly from subjects without complications in age, gender, body mass index, paroxysmal symptoms, symptom duration, tumor dimension, catecholamine secretory phenotype, and incidence of hypertension or diabetes mellitus. Arrhythmias occurred in 15 subjects (2 arrhythmia types in 2 subjects): atrial fibrillation in 9 subjects, supraventricular tachycardia in 3 cases, and ventricular tachycardia in 2 patients. Significant bradycardia was noted in 3 cases. Five subjects presented with heart failure with decreased systolic function (takotsubo-like cardiomyopathy found in 2 cases). One subject suffered from hypertrophic obstructive cardiomyopathy. Seven subjects presented with non-ST-segment elevation myocardial infarction, 2 patients with ST-segment myocardial infarction, and 1 subject underwent coronary artery bypass grafting. Two subjects suffered from significant peripheral atherosclerosis. Among cerebrovascular complications, transient ischemic attack was found in 3 cases, 2 subjects suffered from stroke, and subarachnoidal bleeding occurred in 1 patient. One subject suffered from diffuse neurological impairment due to multiple ischemic white matter lesions. These data show relatively high incidence of cardiovascular complications (19.3%) in subjects with pheochromocytoma. Early diagnosis is mandatory to prevent severe complications in pheochromocytoma.

Paraoxonase-1 (PON1) status in pancreatic cancer: relation to clinical parameters.

Human paraoxonase 1 (PON1) has been shown to decrease the level of systemic oxidative stress, which is thought to contribute to cancer development. The aim of this study was to examine the interrelationships between PON1 status and some clinical characteristics in patients with pancreatic cancer (PC). A group of 73 consecutive patients with PC (stage II-IV) and 73 control subjects were examined. Laboratory studies included five polymorphisms of the PON1 gene (L55M, Q192R, -108C/T, -126C/T, and -162A/G), PON1 arylesterase (PON1-A) and lactonase (PON1-L) activities, as well as some markers of protein metabolism, insulin resistance, and oxidative stress. In comparison with the control group, no difference in the distribution of the PON1 polymorphisms was found in cancer patients, both arylesterase and lactonase activities being significantly lower (-33, -47 %, respectively, both P < 0.001). There was neither statistically significant association of PON1 polymorphisms with tumour stages nor with diabetes mellitus connected with PC. The genotype distribution of L55M and ­108C/T differed only in a subgroup of patients presenting clinically relevant malnutrition (χ² = 6.50, 6.25, respectively, both P < 0.05). In the PC group, PON1-A and PON1-L activities correlated with Nutritional Risk Index (r = 0.351, 0.409, respectively, both P < 0.01), PON1-L with mid-arm muscle circumference (r = 0.328, P < 0.05), and PON1-A and PON1-L with serum albumin (r = 0.352, 0.391 respectively, both < 0.01). Our results suggest that PON1 plays an important role in PC, especially in cancer-associated malnutrition.

Outcomes of pregnant patients with Pap smears classified as atypical glandular cells.

OBJECTIVES: The incidence of cervical cancers increases with age. Due to the trend of increasing age of first pregnancy, abnormal Pap smears including those classified as atypical glandular cells (AGC) are being found more often in early pregnancy. Once invasive cancer is excluded, conservative management of squamous intraepithelial lesions (SIL) in pregnancy is considered safe; however, optimal management of AGC is not well established. The aim of our study was to evaluate the outcome of patients with AGC diagnosed from smears during pregnancy. METHODS: The study included 17 patients referred to us in early pregnancy with Pap smears reported as AGC: 11 not otherwise specified (AGC-NOS), five favour neoplasia (AGC-FN) and one adenocarcinoma in situ (AIS). Thirty-one with high-grade SIL (HSIL) Pap smears confirmed on punch biopsy in early pregnancy comprised a control group. Human papillomavirus (HPV) positivity was found in seven patients with persistent AGC-NOS (including all four who had CIN3 postpartum). All the women were initially examined by expert colposcopy and those with AGC-FN or AIS smears also by transrectal ultrasound to exclude invasive endocervical cancer. Follow-up controls were carried out every 8-12 weeks and, if there were no signs of progression, revaluation was scheduled 6-8 weeks after delivery. RESULTS: The mean age of the women was 31.4 years. Conization in one patient in the study group was performed in the 16th week of pregnancy due to colposcopic signs of microinvasive squamous cell cancer confirmed on histology. Progression to invasive cancer was not found in any of the other 16 patients in the study group or in the control group. Cervical intraepithelial neoplasia or AIS was confirmed postpartum by conization or punch biopsy in 47.1% (8/17) of patients in the study group and, in 77.4% (24/31) of patients in the control group. CONCLUSIONS: Conservative management of women with AGC in pregnancy is safe where invasive cancer is excluded. As histological verification of glandular pre-cancerous lesions by punch biopsy is not reliable and the postpartum regression rate cannot be determined precisely, conization should be performed in all cases with AGC-FN or AIS. Triage of persistent AGC-NOS with HPV testing is useful in distinguishing significant underlying lesions.

[The new system for reporting fine needle aspiration biopsies of the thyroid gland: Bethesda 2010]. / Nový systém pro sjednocené hodnocení tenkojehlových aspiracnich biopsií stitné zlázy--Bethesda 2010.

The newly published Bethesda system for reporting thyroid cytopathology represents an important contribution to the international standardisation of cytodiagnostic reports. The article provides translation of the diagnostic categories into Czech terminology, discusses briefly the explanatory notes in context with the postgraduate teaching and national terminology used. The involvement of the system as a national standard is recommended.

Diagnostic terminology for reporting thyroid fine needle aspiration cytology: European Federation of Cytology Societies thyroid working party symposium, Lisbon 2009.

A European Federation of Cytology Societies (EFCS) working party of 28 members from 14 European countries met at the European Congress of Cytology in Lisbon in September 2009, with two observers from the USA, to discuss the need for standardising thyroid FNA nomenclature in the light of the National Institute of Cancer (NCI) recommendations resulting from the State of the Science conference in Bethesda in 2007. The data were obtained through two questionnaires sent by email and a transcript of the live discussion at the congress, which is presented in full. The surveys and discussion showed that there were currently no national terminologies for reporting thyroid FNA in the different European countries except in Italy and the UK. Personal, 'local', surgical pathology and descriptive terminologies were in use. All but one of the working party members agreed that thyroid FNA reporting should be standardised. Whilst almost a third would adopt the NCI Bethesda terminology, which offers the advantages of a 'risk of cancer' correlation and is linked to clinical recommendations, more than half favoured a translation of local terminology as the first step towards a unified nomenclature, as has been done recently in the UK. There was some disagreement about the use of: a) the six-tiered as opposed to four or five-tiered systems, b) the use of an indeterminate category and c) the 'follicular neoplasm' category, which was felt by some participants not to be different from the 'suspicious of malignancy' category. The conclusions will be passed to the different national societies of cytology for discussion, who will be asked to map their local terminologies to the Bethesda classification, observe its acceptance by clinicians and audit its correlation with outcome.

[False negative PAP test? Cytopathologist as a member of expert group in case of late diagnosis of cervical cancer]. / Falesne negativní PAP test? Cytopatolog v roli clena skupiny znalcu pri pozdní diagóze cervikálního karcinomu.

The Pap test represents a potent tool in reducing the incidence of cervical cancer, though it is unable to eradicate it totally. Under standardized application it has a (very low) failure rate unexpected by the public. In relation to the National screening program of cervical carcinoma an increase in number of litigation cases is to be expected started by women who took part in the screening program and still developed cervical cancer. Together with the international guidelines of Pap test administration and evaluation (Bethesda 2001), widely adopted guidelines for Expert review of histologic slides and Papanicolaou tests in the context of litigation or potential litigation have been formulated (College of American Pathologists 20001). Arguments for adoption of these guidelines by the Czech Society of Pathologists and their implementation into the National Program of Cervical Cancer Screening are presented.

[Efficiency of vitrectomy in diabetic macular edema and morphometry of surgically removed of the internal limiting membrane]. / Význam vitrektomie u diabetického makulárního edému a morfometrie chirurgicky odstranené vnitrní limitující membrány.

AIM: To evaluate the efficiency of vitrectomy with peeling of the internal limiting membrane (ILM) in eyes with diffuse and/or cystoid nontractional diabetic macular edema (ME) refractory to macular laser photocoagulation. Histopathologic and morphometric analysis of the ILM in diabetic eyes was performed. Thickness of the ILM was correlated with the level of glykosylated hemoglobin (HbA1c) and other clinical factors. MATERIAL AND METHODS: The prospective study involved 56 eyes of 52 diabetic patients with a mean age of 63 +/- 7.6 years. Vitrectomy with trypan blue-assisted ILM peeling was performed in standard way. Mean follow-up period was 8.7 months (range 3 - 19 months). The ILM was fixed immediately after peeling in 2.5% glutaraldehyde and submitted for electron microscopic evaluation. The ILM was photographed in standard magnification (x 5000) with the scale of 1 microm in the shot. RESULTS: Statistical analysis of the postoperative visual acuity (VA) in the study group of 56 eyes proved a general improvement with prevalence of the resulted VA in intervals 0.1-0.2 and 0.5-1.0 related to ETDRS chart. The VA was improved by at least 2 lines in 29 eyes of 56 eyes (51.8%), one line in 6 eyes (10.7%) and remained unchanged in 11 eyes (19.6%). The postoperative VA deteriorated by one line in 2 eyes (3.6%) and at least 2 lines in 8 eyes (14.3%). Morphometric analysis demonstrated a significant thickening of the ILM in all eyes with a mean thickness of the ILM 3.61 +/- 1.22 micro m. It was found that a higher thickness of the ILM is related to elevated HbA1c by both types of diabetes mellitus (DM) (p = 0.040). We also found significant dependence of ILM thickness in relation to duration of DM by comparison of men and women (p = 0.026) and significant correlation between ILM thickness and the age of diabetic patients related to their gender (p = 0.029). CONCLUSIONS: Vitrectomy with peeling of the ILM in eyes with chronic diffuse and/or cystoid nontractional diabetic ME mildly improves the VA and extends a hope for its stabilization. We confirmed increased thickness of the surgically peeled ILM and statistically significant correlations to elevated HbA1c by both types of DM and to further clinical characteristics of case series. Morphometric and histopathologic analyses of the ILM contribute to more objective evaluation of ultrastructure of the vitreomacular interface.

BRAFV600E mutation in the pathogenesis of a large series of papillary thyroid carcinoma in Czech Republic.

BACKGROUND: Activating point mutation of the BRAF gene, the most common genetic alteration reported in papillary thyroid carcinomas (PTC), has been associated with poor prognostic characteristics. AIM: Our objective was to determine the frequency of BRAFV600E mutation in PTC tumor tissues from the period 1960-2007 and to correlate it with clinicopathological parameters. SUBJECTS AND METHODS: DNAs were extracted from 242 PTCs, 23 sporadic medullary carcinomas, one anaplastic carcinoma and 6 poorly differentiated carcinomas. The presence of BRAFV600E mutation was determined using single strand conformation polymorphism method and verified by direct sequencing. RESULTS: BRAFV600E mutation was detected in 81 of 242 PTCs (33.5%), in one of 6 poorly differentiated carcinomas (16.7%) and in anaplastic carcinoma. BRAFV600E mutation was much less frequent in the follicular variant compared to classical variant and mixed follicular- classical variant of PTCs (p=0.001). BRAFV600E mutation was significantly associated with presence of nodal metastasis (p=0.029), more advanced TNM stage (p=0.014) and recurrence of disease (p=0.008). The mutation correlated with a higher age at diagnosis (p=0.049) and with a greater tumor size (p=0.041). Multivariate analysis confirmed these findings. The prevalence of BRAFV600E mutation before 1986 was significantly lower than after it (p=0.008). CONCLUSIONS: Our data suggest that BRAFV600E mutation is associated with high-risk clinicopathological characteristics of PTC and worse prognosis of patients. The frequency of the mutation significantly varied during the observed period but rather because of the different age distribution of patients in particular periods than as a consequence of Chernobyl accident.