Novel biomarkers for prediction of acute kidney injury in acute heart failure.

BJECTIVES: Acute kidney injury (AKI) is a frequent event in patients with an acute heart failure (AHF) and is associated with a poor short and long-term outcome. The aim of this study was to describe diagnostic yield of selected novel biomarkers in prediction of AKI in patients admitted for AHF. METHODS: We performed a prospective cohort study of 72 consecutive patients (46/26 M/F) aged 69±10,3 years admitted for AHF. Renal damage was defined according to KDIGO guidelines. Patients were divided into the two groups: AKI- (without renal injury, n=52) and AKI+ (with renal injury, n=20). Urine samples for AKI biomarkers measurements (NGAL, TIMP2, IGFBP7) were collected at the admission. The ROC and linear logistic regression of new biomarkers and selected clinical variables was performed for the evaluation of the AKI prediction. RESULTS: The patients with AKI+ were older (median age: 75 vs 64 years, p=0.01), had lower BMI (median: 28 vs 29.5 kg/m2, p=0.04), were with a higher proportion of patients with HF with a reduced ejection fraction (55 % vs 23.1 %, p=0.01) and a higher level of serum NTproBNP. Urinary NGAL at admission was significantly higher in the AKI+ compared to the AKI - group (152 vs 19.5 ng/mL, p<0.0001); also median of u-TIMP-2 and u-IGFBP-7 in the AKI+ patients was significantly higher: 194.1 versus 42.5 ng/mL (p<0.0001) and 379 versus 92.4 pg/mL (p<0.0001) resp. Age, u-NGAL, u-TIMP2, u-IGFBP7, s-haemoglobin, NTproBNP and LVEF were associated with the development of AKI. Urine concentration of IGFBP-7 was measured, which is the best marker for the prediction of AKI (AUC 0.94). CONCLUSION: Urine concentrations of NGAL, TIMP2, IGFBP7 at the time of admission for AHF predicted a development of AKI. Age, NTproBNP, LVEF and s-haemoglobin were also associated with AKI in AHF patients (Tab. 3, Fig. 3, Ref. 22). Text in PDF www.elis.sk Keywords: biomarkers, cardiorenal syndrome, acute heart failure.

[Coeliac disease as a possible cause of some gynecological and obstetric abnormalities]. / Celiakia ako mozná prícina niektorých gynekologicko-pôrodníckych ochorení.

OBJECTIVE: To bring a review of available literature sources on the prevalence of coeliac disease and its possible impact on gynecological and obstetric disorders. DESIGN: Review article. SETTING: Gynecology and Obstetrics Clinic, Jessenius Faculty of Medicine, Comenius University in Bratislava, Martin, Slovakia. METHODS: Analysis of literary sources. CONCLUSION: Coeliac disease is an autoimmune enteropathy caused by abnormal immune system response to gluten. Over the last decade when the prevalence of the disease increases rapidly confirming the relationship between coeliac disease and a range of reproductive disorders. Problems in this area are mostly confirmed in untreated women. Among the atypical symptoms of coeliac disease also include infertility such as delayed onset of menstruation, early menopause, secondary amenorrhea, infertility and pregnancy complications, such as recurrent abortions, intrauterine fetal growth restriction, small fetus for gestational age, low birth weight and premature birth.

[Comparison of perinatal mortality in Slovak Republic in the years 2007-2009 and in the years 2010-2012]. / Porovnanie perinatálnej mortality v Slovenskej republike v rokoch 2007-2009 a v rokoch 2010-2012.

OBJECTIVE: Comparison of perinatal mortality in Slovak Republic in the years 2007-2009 and in the years 2010-2012. DESIGN: Epidemiological perinatal nation-wide. SETTING: 1st Department of Gynaecology and Obstetrics School of Medicine, Comenius University and University Hospital, Bratislava, Slovak Republic. METHODS: The analysis of selected perinatal data prospectively collected in the years 2007-2009 and in the years 2010-2012. RESULTS: In the year 2007 there were 63 maternity hospitals, 51,146 deliveries and that of live births 51,650 in Slovak Republic. In the years 2010-2012 decreased the number of maternity hospitals, total number of deliveries and that of live births from 57 to 55, from 55,362 to 54,996 and from 55,901 to 55,643 respectively. Preterm deliveries rate increased from 7.4 to 7.7% and multiple pregnancies rate from 1.4% to 1.5% in the years 2010-2012 compared to years 2007-2009. Perinatal mortality rate decreased from 6.2 in the year 2007 to 5.1 per 1,000 still and live births in the year 2012. During the years 2007-2012 stillbirth participate in perinatal mortality 64%, low birth weight 65% and severe congenital anomalies 19%. Transport in utero to perinatal centres decreased in the years 2007-2012. It was from 64% to 56% for infants with very low birth weight and from 75% to 70% for infants with extremely low birth weight. CONCLUSION: In the year 2012 Slovak perinatology reach the best result in perinatal mortality rate 5.1 (0.51%). Centralisation of high-risk pregnancies, transport in utero very low birth weight infants, prenatal detection of severe congenital abnormalities and obstetric and neonatal intensive care units equipment need still to be improved in Slovak Republic.

[Leiomyomas of the uterine body: Molecular-genetical aspects of formation and development]. / Leiomyómy tela maternice: Molekulárno-genetické aspekty vzniku a rozvoja.

OBJECTIVE: An overview of the molecular-genetical aspects of formation and development of leiomyomas of the uterine body. DESIGN: A review article. SETTING: Department of Gynecology and Obstetrics, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic. METHODS: An analysis of the literature using database search engines PubMed, Blast, Science direct and Web of Knowledge focused on tumorigenesis of leiomyoma. RESULTS: Benign uterine leiomyomas, also known as myoma, fibroids or fibromyomas are the most common tumours located in the pelvic area of women. The prevalence of this disease reaches, on the global scale, values higher than 50%, depending on the ethnicity even up to 80% of women of reproductive age. Despite such a high value, the origin of leiomyomas is still unknown. The main reason is the heterogeneity of the disease, and a number of factors that influence their development. In the case of leiomyomata occurrence, it has so far been observed several genome rearrangements and a number of aberrantly expressed genes. There are several reasons for overexpression or underexpression of a particular gene, from a point mutation in the exon region of the gene, promoter or other regulatory sequences to epigenetic modifications, most commonly the nature of methylation, or more precisely inadequate regulation short molecule miRNA. Many of these genes belong to the group of tumour-suppressor genes, or more precisely to genes, which can affect the cell cycle in a different way and thus can affect even the cell division. The aim of this work is to describe the various factors influencing the formation of leiomyomas and their impact on tumorigenesis.

Determination of malignant potential of cervical intraepithelial neoplasia.

Basic diagnostic procedures in cervical cancer screening are able to set the diagnosis but they do not provide any information about the biological nature and behavior of lesions. The causal link of HPV infection and cervical cancer and discoveries of complex interactions between host and HPV genome opened new possibilities in molecular diagnostics. HPV DNA analysis, determination of viral load, detection of E6 and E7 mRNA transcripts, identifying of methylation profiles, genomic changes, miRNAs, and telomerase activity should be the right choice for exact diagnostics and prediction of behavior of premalignant lesions of the cervix. These findings set a completely new light not only in diagnostic but also in management and treatment of cervical dysplasia and cervical cancer.

[Knowledge about cervical cancer among respondents in Slovakia and the Czech Republic - Aurora Project]. / Vedomosti respondentov na Slovensku a v Cechách o karcinóme cervixu - projekt Aurora.

OBJECTIVE: The Aurora Project, aimed at promotion of cervical cancer prevention, was realised with the support of the European Commission. The project included 14 partners from 11 EU countries. The objective of this contribution was to analyse the level of knowledge on cervical cancer among respondents in the project partner countries and to compare the situations in Slovakia and the Czech Republic. Data were obtained within one of the project outputs: Work Package 2 (WP2) Dissemination of Aurora Project Objectives and Results. METHODS: The questionnaire used included 10 questions (available at the project website www.aurora-project.eu) and has been translated into 11 languages of the project partners. In total, 2111 questionnaires were analysed (91.7% response rate), among them 246 were from Slovakia and 305 from the Czech Republic. Descriptive statistical methods and the χ2 test were used to analyse data. RESULTS: The level of knowledge in Slovak and Czech respondents was comparable in answers to seven questions. Statistically significant differences were observed in answers to questions about anatomy and cervical cancer therapy. Answers to the question, 'What are the symptoms of cervical cancer in the early stages?', should be considered as crucial to understand attitudes of the lay population towards prevention. There were 7% of women in the Czech Republic and 16% in Slovakia with the opinion that there is some clinical manifestation of such a condition. This means that women with such an opinion have no reason to visit a gynaecologist while no signs of a disease are present. The period during which they do not attend a preventive check-up is sufficient for the development of precancerous lesions or even cancer. CONCLUSION: Recommendations of doctors play a key role in primary and secondary prevention of the disease. An important part of interventions includes information campaigns and educational programmes. The internet is another important source of information, especially for younger generations. Together, these can contribute to increased participation in preventive check-ups by education of the general public. Thus, the disease can be detected early and the development of cervical cancer prevented.

[Fetal magnetocardiography: a promising way to diagnose fetal arrhytmia and to study fetal heart rate variability?]. / Fetálna magnetokardiografia: Slubný spôsob diagnostiky fetálnych arytmií a stúdia fetálnej srdcovej variability?

OBJECTIVE: An overwiev of the new diagnostic method of fetal wellbeing - fetal magnetocardiography (fMCG). DESIGN: A review article. SETTING: Department of Gynecology and Obstetrics, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic. METHODS: An analysis of the literature using database search engines PubMed, and SCOPE in field of fMCG. RESULTS: Fetal magnetocardiography is a non-invasive technique able to monitor the spontaneous electrophysiological activity of the fetal heart. Compared to cardiotocography and fetal electrocardiography, this is a more effective method with a higher resolution. The signal obtained from the fetal heart is sufficiently precise and the quality allows an assessment of PQRST complex alterations, and to detect fetal arrhythmia. Thanks to early diagnosis of fetal arrhythmia, there is the possibility for appropriate therapeutic intervention and the reduction of unexplained fetal death in late gestation. fMCG with high temporal resolution also increases the level of clinical trials which record fetal heart rate (FHR) variability. According to the latest theories, FHR variability is a possible indicator of fetal status and enables the study of the fetal autonomic nervous system indirectly. fMCG is an experimental method that requires expensive equipment. It is yet to be shown in the future, if this method will get any application in clinical practice.

[Genetic aspects of pelvic floor defects and stress urinary incontinence in women]. / Genetické aspekty defektov panvového dna a stresovej mocovej inkontinencie u zien.

OBJECTIVE: An overwiev of the genetic aspects of pelvic floor defects and stress urinary incontinence in women. DESIGN: A review article. SETTING: Department of Gynecology and Obstetrics, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Slovak Republic. METHODS: An analysis of the literature using database search engines PubMed, BLAST and Ensembl in field of POP and SUI. RESULTS: Pelvic organ prolapse (POP) and stress urinary incontinence (SUI) affect a large number of women and often requires surgical correction. Moreover, these diseases are not only individual health but also social and economic burden on patients and society. It is possible to prevent or at least timely intervene these pathologies by screening of risk patients. The incidence of these disorders leads to find possible genetic factors. In ethiopathogenetic studies of pelvic floor defects and stress urinary incontinence it is looking for genes associated with these diseases and optimization of molecular biology methods is necessary to determine the possible gene mutations or polymorphism site. Selection of relevant genetic factors tend to clarify the pathomechanism of SUI and POP that can greatly affect their current diagnostics and therapeutics. KEYWORDS: prolapse, stress urinary incontinence, polymorphism, gene expression.

[Correlation between the incidence of PIK3CA mutations in breast cancer and histopathological characteristics of the tumor]. / Korelácia výskytu PIK3CA mutácií v karcinóme prsníka s histologickými vlastnostami nádoru.

OBJECTIVE: To determine the presence of mutations in exon 9 (encoding the helical domain) and exon 20 (encoding the kinase domain) of phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene in DNA obtained from paraffin embedded tissue from patients with carcinoma of the mammary gland and to correlate results with clinicopathological characteristics of cancer. DESIGN: Prospective clinical study. SETTING: Department of Molecular Biology, Department of Obstetrics and Gynecology, Jessenius Faculty of Medicine, Commenius University, Martin, Slovak Republic. METHODS: In set of 95 tissue samples from patients with breast cancer, mutations in exon 9 and 20 were analysed by sequencing. We also observed the associations between mutations and histopathological characteristics of tumor. RESULTS: Overall, mutations were present in 25.3% (24/95) of PIK3CA gene, of this 14.7% (14/95) of mutations were located in exon 9 and 10.5% (10/95) of mutations were in exon 20. We detected three "hotspot" mutations, two were located in exon 9 (E542K, E545K) and the third mutation was found in exon 20 (H1047R). Mutations in exon 9 showed significant correlation with lower grade(p = 0.0074) and pN status without metastases(p = 0.0415). Mutations in exon 20 were associated with higher age of patient (p = 0.0249). The E545K mutation correlated with lower grade (p = 0.0013) and pN status (p = 0.0232) particularly; the H1047R mutation was significantly more frequent in lobular type of breast cancer (p = 0.0354). CONCLUSION: The PI3K signaling pathway plays a critical oncogenic role in the development of human breast cancer and the prevalence of its deregulation advocates its potential as a feasible therapeutic target. In our study we demonstrate a significant correlation between the presence of PIK3CA mutations and some clinicopathological characteristics of tumour. We have shown that the mutations in exon 9 of PIK3CA were associated with favourable prognostic factors. KEYWORDS: "hotspot" mutation, PIK3CA, PI3K pathway, breast cancer.

[Breech presentation - an analysis of results in one perinatal center]. / Poloha plodu koncom panvovým - analýza výsledkov jedného perinatologického centra.

OBJECTIVE: A retrospective analysis of medical records during years 2007-2011 considers maternal and fetal outcome in patients with breech presentation terminated by vaginal delivery versus caesarean section (CS). DESIGN: Retrospective analysis. SETTING: Department of Gynecology and Obstetrics, Jessenius Faculty of Medicine, Comenius University, Martin, Slovak Republic. METHODS: Authors devided patients with breech presentation of fetus (n = 299) to groups of single pregnancies terminated in term (n = 197), before term (n = 67) and to group of multiple pregnancies (n = 35). All groups were devided according to the way of termination of pregnancy by vaginal delivery, by acute CS and by planned CS. Main followed parameters: parity, gestational week, Apgar score, birth weight, birth length, fetal gender, indications for CS, mortality and neonatal morbidity, umbilical artery pH, convulsions, admit to neonatal intensive care unit, intubation of neonate, intracranial bleeding, cervical spine and peripheral nerve injuries. RESULTS: Total cohort of breech deliveries was 299. In the group of single pregnancies in term was 19.8% terminated by vaginal delivery (n = 39), 32.5% deliveries by acute CS (n = 64). The most common indication was fetal hypoxia (43.8%). By planned CS was terminated 47.7% deliveries (n = 94). The most common indication for CS was footling presentation (54.3%). Severe neonatal morbidity was rare and without significant difference according to the type of termination of pregnancy. Neonatal outcome was comparable in the group of preterm deliveries(n = 67) terminated by vaginal delivery or by CS. All neonatal deaths were associated with extreme prematurity and not with type of termination of gravidity. CONCLUSION: Clinical outcomes between vaginal breech deliveries and breech deliveries terminated by CS in term in singleton pregnancies were not significant different. Mortality of neonates delivered by preterm delivery was associated with severe prematurity.

Ethnic disparities in breast cancer between Central Europe Caucasian women of Slavic origin and Middle East Turkish subjects.

The biological, cultural, behavioral and sociodemographic differences across populations modulate breast cancer profile among races or ethnics. Following this, we aimed to identify differences in breast cancer epidemiology, histopathology, and clinical presentation from representatives of central Europe (Slovakia) and Middle-East countries (Turkey) to point on ethnic disparities in cancer biology. The population based cross-sectional study analyzing 414 cases of primary breast carcinomas where 214 represented Caucasian and 200 Turkish subjects. The differences were found for age at the time of diagnosis (<0.0001), education, menopausal status (<0.001), tumor localization (<0.01), size (<0.0001), grade (<0.05) and axillary lymph node status (<0.001) between groups. Although carcinomas in Slovak subjects were of higher grade, negative axillary nodal status was more frequent finding compared to Turkish patients (50.0 vs. 41.0%). The Slovak group showed carcinomas to be more often ER positive (72.4 vs. 54.0%; <0.001), ER/PgR positive (54.6 vs. 49.0%; <0.001), of better Nottingham prognostic index (<0.001), and less frequent Her-2 positive (21.2 vs. 28.5%). Slovak population expressed significantly higher risk of non-sentinel lymph node metastases with increased tumor size, grade, vascular invasion and Her-2 positivity compared to Turkey population. The tumor size >2 cm and high tumor grade (G3) bears a risk of OR=7.62 and OR=3.10 in Slovak compared to OR=3.94 and OR=1.79 in Turkish cases, respectively.There are wide demographic and biological disparities in breast cancer between observed ethnics providing unique information for clinicians working at the level of screening or therapy in these populations.

Arterial peculiarities of the thoracolumbar spinal cord in rabbit.

The aim of this study was to investigate the arterial blood supply of the thoracolumbar spinal cord in rabbit. The study was carried out on twenty adult New Zealand white rabbits. Ten rabbits were used in the corrosion technique and ten rabbits in the dissection technique. After the killing, the vascular network was perfused with saline. Batson's corrosion casting kit no. 17 © was used as a casting medium. After polymerisation of the medium, in ten rabbits the maceration was carried out in KOH solution, and in ten other rabbits, formaldehyde was injected by the dissection technique into the vertebral canal. We found high variability of segmental arteries supplying blood to the spinal cord. There are 12 intercostal arteries and 1 costo-abdominal artery. Dorsal branches arising from the dorsal surface of the aorta thoracica were found as follows: in 70% of the cases, 9 pairs were present; in 20% of the cases 8 pairs; and in 10% of the cases 10 pairs. The paired arteriae lumbales were present in 6 pairs in 90% of the cases and in 5 pairs in 10% of the cases. On the dorsal surface of spinal cord, we found two irregular longitudinal arteries in 70% of the cases, no longitudinal arteries in 20% of the cases and three irregular longitudinal arteries in 10% of the cases receiving dorsal branches of rami spinales. Among the dorsal branches observed in the thoracic region, 60.5% were left-sided, 39.5% right-sided and in the lumbar region, 52.5% were left-sided and 47.5% right-sided.

[Recurrent subareolar non puerperal abscess of breast with fistules of lactiferous ducts (Zuskas disease)]. / Rekurentný subareolárny non- puerperálny absces prsníka s fistuláciami ductus lactiferi (Zuskas disease).

INTRODUCTION: Zuskas disease (ZD) is an illness also known as the recurrent subareolar non-puerperal abscess of breast with fistulas of lactiferous ducts or as a periductal mastitis. ZD is rare, but painful chronic disease of breast characterized by local inflammation and evacuation of viscous content from abscess around the nipple. We present a rare form of the non-puerperal mastitis in patients with recurrence of this disease and with the description of management and treatment. CONCLUSION: The treatment of ZD is often inadequate and it leads to the recurrence of ZD and retraction of the nipple. Definitive treatment of ZD is surgical excision of the fistulation, removing of whole retroareolar fibroglandular tissue, abscess cavity and ductal tissue inside the nipple, including the obstructed ducts. The ratio of patients cured by this method is high, as well as their satisfaction with the final cosmetic effect of the nipple and breast.

Impact of fetal pulse oximetry and ST analysis surveillance withdrawal on rates of obstetric surgery and frequency of low birth umbilical artery pH: a cause of rising caesarean rates?

A retrospective observational study on a sample of 13,413 deliveries analysed the effect of a withdrawal of the CTG additional diagnostic methods of fetal hypoxia (fetal pulse oximetry and ST analysis of the fetal ECG) on operative delivery rates and frequency of the umbilical arterial pH < 7.15. Following the withdrawal, obstetricians are more likely to perform caesarean sections for fetal hypoxia (OR 2.23, 95% CI 1.94-2.55, p < 0.0001) and labour dystocia (OR 1.45, 95% CI 1.18-1.77, p = 0.0003), which increases the overall caesarean rate (OR 1.49, 95% CI 1.38-1.61, p < 0.0001), although decreases the incidence of birth umbilical arterial pH < 7.15 (OR 0.43, 95% CI 0.22-0.85, p = 0.015). This also leads to the significant decline in overall frequency of instrumental vaginal deliveries (OR 0.58, 95% CI 0.48-0.71). In order to decrease the overall caesarean rate, obstetricians need to be supported by more accurate and possibly automated diagnostic tools for intrapartum fetal hypoxia.

[Rapid detection of the most common chromosomal aneuploidies in the second-trimester amniotic fluid using QF-PCR]. / Rýchla detekcia najbeznejsích chromozómových aneuploídií z plodovej vody v druhom trimestri tehotnosti vyuzitím QF-PCR.

OBJECTIVE: To introduce QF-PCR method for detection of the most common chromosomal (trisomy 21, 18 and 13) and gonosomal aneuploidies at our department in the second-trimester amniotic fluid. To test the hypothesis of chromosomal aneuploidies detection using STR markers of Aneufast® kit via analysing free fetal DNA (ffDNA) isolated from plasma of pregnant women with confirmed trisomy 21 in fetus. DESIGN: A prospective clinical study. SETTING: Department of Obstetrics and Gynecology, Jessenius Faculty of Medicine and University Hospital in Martin, Slovak Republic. METHODS: The samples of amniotic fluid were obtained from 67 women (twin pregnancy in 3 cases) in the 2nd trimester (15th to 22nd gestational week (g.w.)). Samples were examined using multiplex QF-PCR via Aneufast kit. In the case of positivity for trisomy 21, they were re-examined using Devyser Resolution 21 kit. All samples were parallelly evaluated by cytogenetic karyotyping. We also analyzed ffDNA from the plasma of 3 high-risk women using Aneufast kit. The plasma samples were obtained in the 2nd trimester(17th to 21st g.w.). Qiaamp DSP Virus kit was used for ffDNA isolation. Trisomy 21 of 3 fetuses was confirmed by karyotyping after 2nd trimester amniocentesis. RESULTS: In the cohort of 70 samples, 7 pathological results (six trisomies 21 and one trisomy 18) were obtained. There was 100% concordance with cytogenetic karyotype in all samples examined by QF-PCR. The amplification of tracked chromosome 21 fragments was not evaluable in the case of ffDNA analysis. CONCLUSION: QF-PCR was approved as reliable, rapid, quite simple and financially bearable method of prenatal diagnostics. Despite the fact of good availability and work implementation of Aneufast® kit, results of ffDNA analysis are insufficient. We did not obtain interpretable results after ffDNA analysis from maternal plasma in trisomy 21 fetuses.

[The p16INK4A mRNA transcripts estimation in cervical smear with different degrees of cervical dysplasia]. / Stanovenie expresie p16INK4A mRNA transkriptu v steroch z krcka maternice s rôznymi stupnami cervikálnej dysplázie.

OBJECTIVE: To determine the presence of HPV infection and expression level of p16INK4A mRNA transcripts in cervical smears as adjunct biomarker in detection of cervical intraepithelial neoplasia or cancer. DESIGN: Prospective pilot clinical study assessing clinical utility and validity of ddCt method for qPCR mRNA expression of p16ink4a in comparison to immunohistochemistry. SETTING: Department of Molecular Biology, Department of Obstetrics and Gynecology, Jessenius Medical Faculty, Commenius University, Martin, Slovak Republic. METHODS: Cervical smears (OC) from patients with different cervical lesions (L-SIL, H-SIL, SCA; n=45) and from healthy controls (n=45) were tested for the presence of HPV infection and p16INK4A mRNA transcripts using relative quantification (RQ). Results were compared to H&E and IHC histological findings from biopsies (conization, hysterectomy). RESULTS: HPV 16 was the most frequent finding (53.3%) in the group of subjects with cervical dysplasia. The p16INK4A mRNA expression analysis revealed the slightly reduced expression in L-SIL group, 4-fold increased expression in H-SIL and 10-fold increase in women with SCA when compared to controls. The p16INK4A mRNA expression in OC was present in 30% of L-SIL, 75% of H-SIL and 85.7% of SCA samples, respectively. The test overall sensitivity was 81.48% (95% CI: 61.92-93.7) and specificity 60% (95% CI: 26.24- 87.84) with PPV of 84.62% and NPV of 54.55%. The likelihood ratio (LR) in case of test positivity was 2.04 and for negativity 0.31. The diagnostic accuracy of p16INK4A expression by RQ method in OC smears for prediction of p16 positivity in cervical dysplasia was 66.7% for the L-SIL lesions, 59.5% for H-SIL lesions, and 100% for SCA (r=0.9897, p<0.0913) when compared to IHC p16 positive findings in surgically treated samples. CONCLUSION: The relative quantification is able to determine the level of p16INK4A mRNA transcripts in cervical smear cells with active carcinogenesis nearly at the same level as IHC staining. The advance of biopsy sparing over IHC is qualifying this diagnostic approach for useful candidate in selective management of women with cervical dysplasia looking for cervix preservation or avoiding the unnecessary overtreatment.

Trastuzumab and lapatinib modulation of HER2 tyrosine/threonine phosphorylation and cell signaling.

Cell surface transmembrane signaling receptors EGFR, HER3, and HER4 are activated by ligand-binding-mediated dimerization and phosphorylation. In contrast, HER2 amplification promotes signaling by increasing homo/heterodimerization and ligand binding. Trastuzumab or lapatinib therapy of HER2 amplicon-positive breast cancer cells induces growth inhibition and intracellular growth pathway signaling modulation. The mechanism(s) by which trastuzumab, an IgG1 humanized antibody, induces modification of cell signaling upon binding to an extracellular determinant on a ligand-less "receptor" membrane protein remains unexplained. Using immune detection methodology comprised of antibodies detecting three distinct domains of HER and five tyrosine/threonine phosphorylation sites, the effects of trastuzumab and lapatinib were defined during steady state growth inhibition. Here, we show that lapatinib markedly reduces HER2 tyrosine phosphorylation, while in contrast, no change in tyrosine phosphate levels is detected during trastuzumab-mediated cell growth inhibition. As trastuzumab treatment does not change either the steady state HER2 protein levels or HER2 mRNA, these findings argue against an antibody-dependent alteration in internalization kinetics. We further show a sequenced relationship between lapatinib-induced blockage of phosphorylation (6-8 h) and induction of delayed cell death (5-6 days), while trastuzumab-treated cells showed no evidence of cell death up to 9 days. Taken together, these results demonstrate that inhibition of HER2 phosphorylation by lapatinib is sufficient to induce apoptosis while trastuzumab binding to the extracellular HER2 domain may function by sterically modulating the detection of phosphate moieties by cytoplasmic signal transducers. This investigation also detected a 20 kD protein, which is down-regulated by lapatinib, further demonstrating the complexity of this signal transduction system.

The effect of transgenesis on rabbit thyroid tissue structure.

This study was aimed to compare structures of the thyroid tissue of transgenic rabbits expressing the human clotting factor VIII under the murine whey acidic protein promoter (mWAP-hFVIII rabbits) with the non-transgenic controls. Thyroid tissue samples were taken from transgenic and non-transgenic New Zealand White rabbits, examined by optical microscopy and analysed morphometrically. The analysis revealed no significant differences (P > 0.05) in the relative volume of basic thyroid structures. Furthermore, no significant differences (P > 0.05) were observed when measuring the epithelial height and nuclear diameter of the follicular cells. Altogether, this study demonstrates no negative effect of the mWAP-hFVIII transgenesis on the rabbit thyroid gland structure.

[Methylation of selected tumor-supressor genes in benign and malignant ovarian tumors]. / Metylácia vybraných tumor-supresorických génov v benígnych a malígnych ovariálnych nádoroch.

OBJECTIVE: To evaluate the usefullness of examination of methylation status of selected tumor-supressor genes in early diagnosis of ovarian cancer. DESIGN: Prospective clinical study. SETTING: Department of Gynecology and Obstetrics, Department of Molecular Biology, Jessenius Medical Faculty, Commenius University, Martin, Slovak Republic. METHODS: In this study we analyzed hypermethylation of 5 genes RASSF1A, GSTP, E-cadherin, p16 and APC in ovarian tumor samples from 34 patients - 13 patients with epithelial ovarian cancer, 2 patients with border-line ovarian tumors, 12 patients with benign lesions of ovaries and 7 patients with healthy ovarian tissue. The methylation status of promoter region of tumor-supressor genes was determined by Methylation Specific Polymerase Chain Reaction (MSP) using a nested two-step approach with bisulfite modified DNA template and specific primers. RESULTS: Gene methylation analysis revealed hypermethylation of gene RASSF1A (46%) and GSTP (8%) only in malignant ovarian tissue samples. Ecad, p16 and APC genes were methylated both in maignant and benign tissue samples. Methylation positivity in observed genes was present independently to all clinical stages of ovarian cancer and to tumor grades. However, there was observed a trend of increased number and selective involvement of methylated genes with increasing disease stages. Furthermore, there was no association between positive methylation status and histological subtypes of ovarian carcinomas. CONCLUSION: RASSF1A and GSTP promoter methylation positivity is associated with ovarian cancer. The revealed gene-selective methylation positivity and the increased number of methylated genes with advancing disease stages could be considered as a useful molecular marker for early detection of ovarian cancer. However, there is need to find diagnostic approach of specifically and frequently methylated genes to determining a methylation phenotype for early detection of ovarian malignancies.

[Biochemical aspects of fetal hypoxia]. / Biochemické aspekty fetálnej hypoxie.

OBJECTIVE: To evaluate validity of biochemical diagnostic methods of fetal hypoxia. DESIGN: A case-control study. SETTING: Department of Gynecology and Obstetrics, Jessenius Faculty of Medicine, Comenius University, Martin, Slovak Republic. METHODS: We included 67 patients, and they were retrospectively divided into group of controls (n=36), and studied group (n=31) according to pH in umbilical artery (UA) <7.15. Acid-base parameters were assessed with Rapidlab 248, Bayer Healthcare LLC, East Walpole, USA. We determined criterion for metabolic acidosis (MAC) as pH UA <7.15, resp. base deficit (BD) UA >12 mmol/l. Postpartal lactate concentration in umbilical vein (UV) and UA was determined with lactatemeter Accutrend Lactate, Roche Diagnostics, Switzerland. Quantitative assessment of fetal human protein S100B was provided with ELISA (Sangtec 100 ELISA, DiaSorin Inc., Stillwater, Minnesota, USA). Fetal erythropoietin concentration in UV was examined with immunoenzymatic assessment Access EPO (Beckman Coulter, Inc., Fullerton, CA, USA). STATISTICS: histograms, Kolmogorov-Smirnov test, Mann-Whitney test, Spearman's rho; statistical significance: p<0.05, Receiver Operating Characteristic curves, Area Under the Curve. RESULTS: The best correlation was between fetal acid-base parameters and lactate in UA (p<0.0005). Significant correlation was between EPO in UV, and protein S100B in UV (p<0.05). EPO in UV significantly correlated with lactate in UA (p<0.05). Correlation between EPO in UV and protein S100B was not significant. According to ROC curves in prediction of fetal hypoxia, we found an excellent accuracy (AUC>0.9) for lactate in UA, good accuracy (AUC>0.7) had EPO in UV. Results for protein S100B were not significant. The highest sensitivity had EPO in UV, while the highest specificity has had lactate in UA. CONCLUSION: An indisputable evidence of labor management quality is the fetal metabolic status. On the basis of our results, the suitable clinical markers are lactate and EPO, in addition to acid-base parameters.