[The new blood test bio-marker SEPT9 and colorectal carcinoma screening]. / Nový krevní test bio-markeru SEPT9 a screening kolorektálního karcinomu.

Due to its high incidence and mortality rates, the colorectal carcinoma represents a crucial medical issue. However, when it is detected in early stage there is high rate of successful treatment. Thats why, early stage cancer screening programmes were introduced into the clinical practice. They focus on the finding of hidden bleeding, using various laboratory techniques, sigmoidoscopy, and, primarily, colonoscopy. However, screening programmes have not yet reached the effect required. New techniques are therefore being developed, such as the detection of blood bio-markers. This group includes also methylated SEPT9 (mSEPT9) detection in blood. We applied this test on 57 patients; we divided the group into two parts. There were 33 asymptomatic individuals in the first group. In this group, we were got only one positive mSEPT9 result. The consequent colonoscopies were negative. The other group had 24 proven carcinomas. Of them, two had negative mSEPT9 results. The remaining in all 22 patients was tested mSEPT9 positive. After its efficiency is tested by further studies, this test may be used especially for patients with low compliance, as it only requires routine blood drawing.

Mitochondrial DNA phylogeny in Eastern and Western Slavs.

To resolve the phylogeny of certain mitochondrial DNA (mtDNA) haplogroups in eastern Europe and estimate their evolutionary age, a total of 73 samples representing mitochondrial haplogroups U4, HV*, and R1 were selected for complete mitochondrial genome sequencing from a collection of about 2,000 control region sequences sampled in eastern (Russians, Belorussians, and Ukrainians) and western (Poles, Czechs, and Slovaks) Slavs. On the basis of whole-genome resolution, we fully characterized a number of haplogroups (HV3, HV4, U4a1, U4a2, U4a3, U4b, U4c, U4d, and R1a) that were previously described only partially. Our findings demonstrate that haplogroups HV3, HV4, and U4a1 could be traced back to the pre-Neolithic times ( approximately 12,000-19,000 years before present [YBP]) in eastern Europe. In addition, an ancient connection between the Caucasus/Europe and India has been revealed by analysis of haplogroup R1 diversity, with a split between the Indian and Caucasus/European R1a lineages occurring about 16,500 years ago. Meanwhile, some mtDNA subgroups detected in Slavs (such as U4a2a, U4a2*, HV3a, and R1a1) are definitely younger being dated between 6,400 and 8,200 YBP. However, robust age estimations appear to be problematic due to the high ratios of nonsynonymous to synonymous substitutions found in young mtDNA subclusters.

Association of the leukemia inhibitory factor gene mutation and the antiphospholipid antibodies in the peripheral blood of infertile women.

To characterize the impact of the potentially functional mutation--the G to A transition at the position 3400 of the leukemia inhibitory factor (LIF; a pluripotent cytokine that plays a central role in the control of the embryo implantation) gene that leads to the exchange of valine with methionine at codon 64 we evaluated the association of the LIF gene mutation and the levels of antiphospolipid antibodies (aPLs) in the peripheral blood of infertile women (the aPLs examination was part of our routine immunological test during the infertility check-up). Eight infertile mutation-positive women were diagnosed with idiopathic infertility (n=5) and endometriosis (n=3) and their levels of aPLs in serum were compared with 115 infertile women without any LIF gene mutation. Enzyme-linked immunosorbent assay was used for the detection of seven antiphospholipid antibodies; the results were statistically assessed by the Fisher's 2 by 2 exact test to evaluate the association of the LIF gene mutations and aPLs in serum of infertile patients. The presence of aPLs was significantly higher in our study group (100%) than in 30% of aPLs-positives in control infertile patients (p = 0.0035) which indicates that the aPLs are elevated in women with LIF gene mutations.

[The leukemia inhibitory factor gene mutations in the population of infertile women: the heterozygote transition G to A on the position 3400 does not affect the outcome of the infertility treatment]. / Mutace v genu pro leukemický inhibicní faktor v populaci neplodných zen: heterozygotní bodová zámena G za A na pozici 3400 neovlivnuje úspesnost lécby.

OBJECTIVE: The leukemia inhibitory factor (LIF) is one of the most important signaling factors in the embryo-maternal cross talk during the embryo implantation. We investigated the prevalence of the LIF gene mutations in the population of infertile women and their impact on infertility treatment. DESIGN: A cohort study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine and University Hospital of Charles University, Pilsen. SUBJECTS AND METHODS: The population to screen consisted of 399 infertile women. The control population was comprised of 202 healthy fertile subjects. For the mutational analysis, the temperature gradient gel electrophoresis (TGGE) followed by subsequent sequencing of the positive samples, had been used. The groups of fertile controls and infertile patients were compared for statistically significant difference using the Fisher's 2 by 2 Exact test. RESULTS: Twelve potentially functional LIF gene mutations, the G to A transversion at the position 3400 leading to the valin to methionin exchange at codon 64 (V64M) were detected in the group of infertile women. No mutations were identified in the control group, which means that the frequency of functionally relevant mutations of the LIF gene in infertile women is significantly enhanced in comparison with controls (P = 0.01, Fisher's 2 by 2 Exact test ). Seven of these patients were successfully treated by in vitro fertilization (IVF). CONCLUSION: The results suggest that the LIF gene mutation, the heterozygote G to A transition on the position 3400, affects fertility but the infertility treatment can succeed. Even though LIF gene mutations occur infrequently and can be overcome by infertility treatment, their impact on molecular events during early phases of pregnancy should be further elucidated.

Fatal venous thrombembolism complicating imatinib therapy in a patient with metastatic gastrointestinal stromal tumor.

Imatinib, a tyrosine kinase inhibitor, is currently the therapy of choice for gastrointestinal stromal tumor (GIST). The toxic effects of imatinib treatment are usually mild, and serious adverse events are rare. We report here the case of a patient with peritoneal metastases of GIST involving the pelvis treated by imatinib. Abdominal pain deteriorated early in the course of the therapy along with the enlargement of the tumors. The patient died suddenly, and the autopsy revealed pulmonary embolism originating from the deep vein thrombosis caused by the compression of common iliac vein by the tumor. The possibility of deep vein thrombosis caused by the compression of the veins by necrotic tumor should be considered in patients with abdominal or pelvic metastases of GIST, including patients treated with imatinib.

[Frequency view on genome changes testing]. / Frekvencní pohled na vysetrení odchylek genomu.

Laboratories dealing with human genome, both inherited and acquired changes, dispose with similar methods and technology. The spectrum of genetic tests is relatively broad and the number of mutations or variants tested differs substantially. Also the number of examinations carried out in individual laboratories varies. Data presented in the tables come from the year 2004 and indicate the number of examinations requested and number of positive results. Many laboratories mentioned in the registry CZDDNAL (http://www.uhkt.cz/lab_a_vysetreni/nr lab_dna_diag/dna_lab_db) perform the same tests but there is also a great number of tests carried out by only one laboratory. Reasons of the request, cost-effectiveness and clinical utility of genetic testing is being discussed.

Chromophobe renal cell carcinoma with microcystic and adenomatous arrangement and pigmentation--a diagnostic pitfall. Morphological, immunohistochemical, ultrastructural and molecular genetic report of 20 cases.

We present clinical, morphological, immunohistochemical, ultrastructural and molecular genetic features of 20 cases of a peculiar form of chromophobe renal cell carcinoma (CRCC) with morphology differing from that of conventional CRCC. Microscopically, the typical features of the tumors were microcystic arrangement and formation of adenomatous structures. Microcystic areas were composed of smaller eosinophilic and bigger pale cells having cytological appearance typical of conventional CRCC. Cytological features of the adenomatous structures were mostly different from those of conventional CRCC. They had a typical columnar arrangement with nuclei positioned at the base of the glandular structures and a small amount of a deeply eosinophilic cytoplasm often endowed with brush border facing the lumen of the glands. In addition, all the tumors showed a brown pigmentation. The pigmentation was located mostly extracellularly, where it formed pools of heavy deposits. Microscopic calcifications present in all cases formed psammoma bodies or else the calcifications were more extensive and amorphous in shape. Ultrastructurally, the cells showed features characteristic of CRCC: typical cytoplasmic vesicles were 100-700 nm in size and mitochondria had tubulovesicular, lamellar or circular cristae. Some tumor cells contained dark, variously sized electron-dense pigment granules. Neither melanosomes nor membrane-bound neurosecretory granules were seen. Using fluorescence in-situ hybridization probes for chromosomes 1, 2, 6, 10, 13, 17 and 21, the tumors revealed massive loss of tested chromosomes typical for conventional CRCC. Monosomy of chromosomes 1, 2, 6, 10, 13 and 21 was found in 100, 36, 91, 82, 82, 82 and 64% of cases, respectively. None of the cases showed mutation of exons 9, 11, 13 and 17 of the c-kit gene. The important feature of pigmented microcystic chromophobe renal cell carcinoma is a relatively benign biological behavior and the absence of distant metastases and sarcomatoid transformation.

[Tumorous diseases in patients with the testicular feminization syndrome ("androgen insensitivity" syndrome)--description of two cases]. / Náidorová onemocnení pacientu se syndromem testikulární feminizace ("androgen insensitivity" syndrom)--popis dvou prípadú.

OBJECTIVE: To describe tumors occurring in two cases of testicular feminization syndrome. SUBJECT: Case report. SETTING: Dpt. of Special Diagnostics SPAU, University Hospital Plzen. SUBJECT AND METHOD: Two cases of testicular feminization syndrome were selected from four cases in our registry. Patients were 45 and 84-year-old Caucasian "females". Resected material was fixed in formaldehyde, routinely processed and stained with hematoxiline-eosin, inhibin, cytokeratines 20, placentar alkaline phosphatase, CD 99, Melan A, hCG. Sertoli cell adenoma was diagnosed in both patients. Older patient had in addition unclassified sex cord tumor of Leydig cell type. The number of sex chromosomes was examined using FISH analysis in both patients. CONCLUSION: Patients with testicular feminization syndrome are frequently affected by benign or malignant tumors in the cryptorchid testes. We documented two benign Sertoli cell adenomas and one sex cord tumor of uncertain biological behavior in our patients. The testes should be removed after puberty with subsequent estrogen therapy in patients with testicular feminization syndrome.

[Transesophageal echocardiography in adults with congenital heart defects]. / Transezofageální echokardiografie u dospelých s vrozenými srdecními vadami.

The authors give an account of their experience with transesophageal echocardiography (TEE) in 61 adult patients with congenital heart disease. The main indications for examination in this group of patients were: confirmation of an atrial septal defect, unsatisfactory visualization, in particular in complex defects, evaluation of the results of surgical correction, detailed valvular morphology, in particular of the mitral and aortic valve, search for the source of embolization, suspected aneurysm of the aorta. A high diagnostic yield of TEE was obtained in: defects of the atrial septum, or other pathological conditions affecting the atrium, defects of the atrioventricular septum (AV), pathology of the AV valves, in particular the mitral one, pathology of the aortic valve and the efflux tract of the left ventricle, aortic arch and its descendent part, transposition of the large arteries or other complex defects, incl. conditions following surgical correction. A small diagnostic contribution of TEE was obtained when visualizing defects of the ventricular septum, pulmonary valves and the efflux tract of the right ventricle and the distal portions of the ascendant aorta.

[Detection of HER-2/neu in breast carcinoma]. / Prukaz HER-2/neu v karcinomu mlécné zlázy.

BACKGROUND: HER-2/neu protein overexpression has been shown to be an independently adverse prognostic and predictive factor in patients with breast cancer. Recently, HER-2/neu overexpression has gained therapeutic implications: It has been shown that in patients with breast cancer the use of trastuzumab/Herceptin TM, the recombinant humanized monoclonal antibody directed against extracellular domain of HER-2/neu molecule, can block the HER-2/neu protein activation and bring about a clinical remission. Following these developments, demand for pathologists to evaluating properly HER-2/neu in breast cancer specimens has been rapidly increasing. METHODS AND RESULTS: In our series of 449 cases of breast cancer, HER-2/neu protein and gene were examined by means of immunohistochemistry and fluorescence in situ hybridization respectively. Results of HER-2/neu study were compared with the hormonal status, cancer grade and proliferation activity as assessed using immunohistochemistry with MIB1 antibody. All seven cases of breast cancer with strong overexpression of HER-2/neu (score 3+) manifested a gene amplification. In contrast, among 11 cases of breast cancers with mild HER-2/neu overexpression (score 2+), the gene amplification was demonstrated in 5 cases only (45%). CONCLUSIONS: Immunohistochemical assessment and fluorescence in situ hybridization (FISH) are complementary methods for detection of HER-2/neu status in breast cancer. While immunohistochemistry is an excellent screening method, FISH should be used for the confirmation of positive results before the Herceptin treatment.

[Congenital heart defects in adults worldwide and in the Czech Republic]. / Vrozené srdecní vady dospelých ve svete a u nás.

Congenital heart disease of adults is a new area of cardiology with specific diagnostic, therapeutic, social and economic problems of these patients. The First Medical Clinic of the Second Medical Faculty, Charles University assembled so far data on more than 1200 adult patients with congenital heart disease from different districts in Bohemia and Moravia. Atrial septal defects are recorded most frequently (25.7%), followed by ventricular septal defects (25.0%) and coarctation of the aorta (12.4%). 50.3% of the patients were treated by surgery, most frequently patients with a patent ductus arteriosus--in 83.8%, with coarctation of the aorta (82.8%) and an atrial septal defect (60.5%). The authors review reasons why for the care of these patients in other countries specialized centres developed and the foundation of such a centre in the Czech Republic is discussed.

[Role of p27Kip1 protein in the cell cycle and its appearance in lymphoid tissues, particularly non-Hodgkin's B-cell lymphomas. Review]. / Role proteinu p27Kip1 v bunecném cyklu a jeho výskyt v lymfoidních tkáních, zvláste v nehodgkinských B-lymfomech. Prehled problematiky.

Cell cycle progression is governed by cyclin dependent kinases (CDK) that are activated by cyclin binding and inhibited by CDK inhibitors. Protein p27Kip1 functions as a CDK inhibitor, which controls the progression from G1 to S phase. Further, p27Kip1 may have a positive regulative influence. In nonneoplastic tissues and in the majority of tumors investigated so far, the immunohistochemical positivity of p27Kip1 showed an inversely proportional relationship to the proliferation index. Among B-cell non-Hodgkin lymphomas, the exceptions to this rule are represented by mantle cell lymphoma, hairy cell leukemia, and the immunoblastic Epstein-Barr virus latent membrane antigen positive diffuse large B-cell lymphoma in AIDS patients. The loss of p27Kip1 expression is a negative prognostic factor in numerous tumors, including the majority of B-cell lymphomas.

[Amplification and overexpression of HER-2/neu in parotid gland salivary duct carcinoma. Immunohistochemical study and fluorescence in situ hybridization]. / Amplifikace a overexprese HER-2/neu v salivárním duktálním karcinomu príusní zlázy. Imunohistochemická studie a fluorescencní in situ hybridizace.

Salivary duct carcinoma (SDC) is highly malignant salivary gland tumour with aggressive clinical behaviour, characterised by its histological resemblance to invasive ductal carcinoma of the breast. Amplification of gene HER-2/neu and overexpression of its gene product have been shown to have both prognostic and treatment implications in breast cancer. The reports concerning the expression of c-erbB2/HER-2/neu in salivary gland tumours are few and controversial. Thus, eleven cases of SDC were evaluated for HER-2/neu status using immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH). To the best of our knowledge, this is the first molecular genetic analysis of SDCs using FISH. HER-2/neu overexpression, identified as strong membrane staining, was observed in all but one case of SDC in majority of neoplastic cells while only four tumours, of nine cases analysed, revealed HER-2/neu gene amplification by means of FISH analysis. SDCs were associated with poor clinical outcome, 6 patients (55%) died of disseminated carcinoma within 4 to 44 months after therapy. There was no difference in outcome of patients with IHC positive-nonamplified and IHC positive-amplified tumours.

[Pharmacotherapy of diastolic dysfunction in heart failure]. / Farmakoterapie diastolické dysfunkce u srdecního selhání.

Impaired left ventricular diastolic function is an indicator of early stages of heart disease. In cardiac failure it is not necessarily associated with impaired systolic function and correlates more closely with clinical symptoms or load tolerance. Pharmacotherapy leading to improved diastolic parameters improves also the clinical condition of cardiac failure. Based on data in the literature, the authors analyze drugs which exert a favourable effect on left ventricular diastolic function: ACE inhibitors, calcium channel blockers, beta adrenergic agonists, phosphodiesterase inhibitors, diuretic and direct vasodilating agents, from the aspect of clinical administration.

[Emergency states in internal medicine--diagnostic contribution of emergency abdominal ultrasonography]. / Náhlé stavy ve vnitrním lékarství--diagnostická pomoc urgentní abdominální ultrasonografie.

The authors analyzed the activities of the sonographic laboratory during four consecutive months. During this period 896 patients were examined by ultrasonography and 119 of them (13.3%) for urgently indicated ultrasonography. The authors present a detailed evaluation of results in these 119 examined patients, in particular from the aspect of yield and asset of the method, affection of different organs, agreement of clinical and sonographic diagnoses and comparison with some other examination methods. Urgent ultrasonography is recommended as a very useful method, in particular as regards diagnostic yield which makes it possible to establish the diagnosis quickly and thus to start appropriate treatment early and to reduce thus the period spent by the patient in hospital. The method is useful also in serious cases and in some conditions which threaten the patient's life, as it can help early indication of surgery.

[Present views on the diagnosis of cavernous hemangioma of the liver using ultrasound]. / Soucasný pohled na diagnostiku kavernózních hemangiómu jater pomocí ultrazvuku.

The submitted review deals with morphometry, normal sonographic anatomy and the position of ultrasonography in the diagnosis of cavernous haemangiomas of the liver. The authors draw attention to the contemporary equipment used and analyze the position of other diagnostic methods in the diagnosis of cavernous haemangiomas of the liver. They discuss also the problem of percutaneous biopsy of the liver when this benign tumour is suspected. Finally the authors recommend the best procedure in the diagnosis of cavernous haemangiomas of the liver.

[Ultrasonography of the kidneys for the detection of incipient diabetic nephropathy]. / Praktický prínos ultrasonografického vysetrení ledvin pro stanovení zacínající diabetické nefropatie.

At the First Medical Clinic of the Faculty Hospital in Prague 5 Motol a group of 15 men and 14 women treated on account of type I diabetes for a mean period of 12.8 +/- 5.8 years was examined. Their mean age was 26.3 +/- 5.8 years and their BMI 23.91 +/- 2.82. The control group was formed by 18 healthy subjects (10 men and 8 women) mean age 23.8 +/- 8.9 years, BMI 22.44 +/- 2.48. All subjects were subjected to a detailed ultrasonographic examination and, assessment of microalbuminuria. From the assembled sonomorphometric data the volume of the right kidney was related to the BMI and thus the renal volume index (RVI) was obtained. In the group of diabetic subjects a statistically significant correlation was found between RVI and microalbuminuria (r = 0.574, p < 0.01). In the group of patients with RVI higher than 7.44 a close correlation was found between microalbuminuria and RVI (r = 0.993, p < 0.05). In the group of patients with a duration of diabetes under 5 years no correlation between the investigated parameters and the RVI was found, nor a mutual correlation of parameters. In patients with diabetes persisting for 5-20 years correlation was found between microalbuminuria and RVI (r = 0.686, p < 0.01). In the group of patients with diabetes for more than 20 years there was no correlation between RVI and other indicators.

[What are the characteristic signs of cholecystolithiasis in patients with type 2 diabetes?]. / Jaké jsou charakteristické znaky cholecystolitiázy u nemocných s cukrovkou 2. typu?

In a group of 166 patients with type 2 diabetes mellitus, hospitalized at the First Medical Clinic, Faculty Hospital Prague 5-Motol, cholecystolithiasis was found in 91. In these patients sonographic parameters were analyzed. All were hospitalized primarily on account of a non-biliary indications. The group comprised 67 men, mean age 69 years. The control group was formed by 67 patients with a normal glucose tolerance. An enlarged gallbladder was found in 15 patients (16.4%), a gallbladder of reduced size in 8 (8.8%). Stones smaller than 1 cm were in the group of diabetics in 31 subjects (34.3%), stones larger than 1 cm in 60 (65.7%). A wall of the gallbladder wider than 3.5 mm was found in 14 (15.4%) diabetics with cholecystolithiasis. All these results were statistically significant as compared with the control group (p < 0.01). There was no significant difference in the number of stones in diabetics and controls. The authors discuss the causes of these findings. In the conclusion the authors state that it is necessary to seek actively cholecystolithiasis in type 2 diabetics and indicate cholecystectomy in time to prevent numerous, in particular inflammatory, complications. For detection ultrasonography is suited best.

[Cholecystolithiasis in type II diabetics]. / Cholecystolitiáza u diabetiku II. typu.

In a group of 166 type II diabetics hospitalized in a medical department the authors made clinical and ultrasonographic examinations focused on the presence of cholecystolithiasis. The control group was formed by 67 subjects with normal glucose tolerance. None of the patients were hospitalized on account of biliary disease. The purpose of the work was to 1. evaluate the difference in the incidence of cholecystolithiasis in diabetic patients and controls with regard to age and sex, 2. to assess differences in the incidence of obesity, impaired lipid metabolism and a positive biliary family--history in diabetics and controls with lithiasis, 3. to evaluate diabetes and the presence of microalbuminuria. In the authors' group cholecystolithiasis is significantly more frequent in diabetics as compared with controls, in men, women and people above 65 years (p less than 0.01). The group of diabetics and controls with lithiasis does not differ as to the incidence of obesity, hyperlipoproteinaemia and positive family-history of biliary disease. No significant differences in parameters of compensation of diabetes nor differences in the incidence of microalbuminuria were found between diabetics with and without lithiasis. The results suggest that it is useful to screen cholecystolithiasis in diabetic subjects.

[Detection of atrial septal defects using esophageal echocardiography]. / Prukaz defektu sínového septa jícnovou echokardiografií.

By means of transoesophageal (oesophageal) echocardiography (TEE) the authors examined a group of 20 subjects. In 10 there was clinical suspicion (physical finding, ECG, X-ray) of an atrial septal defect. Common precordial echocardiographic examination (TTE) and dilution of indocyan green by means of an acustic sensing unit were negative or only marginally positive and did not provide a final decision. By means of TEE the defect was confirmed in 8 subjects by the finding of anatomical discontinuity of the septum and the presence of coloured Doppler shunt flow. In two subjects the finding was negative, in 10 controls no pathological findings were detected. The authors confirmed that TEE is a reliable method which proves even slight shunts in the area of the atrial septum. It is probably the most sensitive existing method.