Prevalence and Age Distribution of Human Papillomavirus among Females in Region of Tuzla, Bosnia and Herzegovina

A. Nadarevic, J. Lisicic, A. Saric, M. Omerbasic, M. Brcic, T. Stjepic, J. Lucic

University of Tuzla

Introduction: A human papillomavirus (HPV) is a papillomavirus that induces hyperproliferative lesions in cutaneous and mucosal epithelia. So far more than 100 HPV genotypes have been identified and associated with cancerous and precancerous lesions of the cervix. Certain types, such as HPV 16 and 18 are highly carcinogenic, while others, such as HPV 31 and 33 are moderate. Types, such as HPV 6 and 11 are mildly carcinogenic. Worldwide, right after breast cancer, cervical cancer is the second most common cancer that affects women. The aim of this study was to determinate the prevalence and age distribution of HPV infections among females in the Tuzla Canton between years 2000 and 2008, and to reveal the type of cytological change found in women born in 1971 and later.

Material and Methods: This study has been conducted by a retrospective analysis, using the information from Policlinic for Laboratory Diagnostics-Department of Pathology, University Clinical Center Tuzla. Samples of the cervical smear for the PAP test and HPV analysis were taken during routine gineacological examinations, by using brush and sticks with cotton, taken from the Digene Specimen Collection Kit, from the whole surface of a portion, and by mild rotating moves from the outer cervical entrance. Identification of the presence of HPV was carried out by the Digene Hybride Capture II test in all the cases. The cytomorphological changes found using light-microscope method were classified according to the Bethesda classification of cervical lesion.

Results: From January 2000 until April 2008, a total of 4918 women were recruited into the study and had their Pap smear tested. Positive Pap smear was found in 2091(42,5%) women of all ages. Our results showed that the most tested group was born between 1961 and 1970 (29.28%), followed by women born in 1971 and later (28.54%). The most frequent abnormal PAP test at HRHPV+ women born in 1971 and later was LGSIL (40,09%) followed by CC (30,91%) and HGSIL (28,98%).

Conclusion: HPV is very common among females in the region of Tuzla. Our data indicates an increased number of tested women in all age groups after a conducted media campaign. Our study does not coincide with other studies that were done in USA. The most tested and affected group of the women in our study were those born between years 1961 and 1970.

New Applications for Neonatal and Prenatal Screening / Diagnostics of the Hemoglobinopathies

E. Mantikou1, J.L. Kerkhoffs2, M. Phylipsen1, C.L. Harteveld1, P.C. Giordano1, J. van Beckhoven1

2Sanquin Blood Bank

Introduction: Sickle-cell disease (SCD) and beta-thalassemia major are two common forms of hemoglobinopathy with severe outcome. Children with SCD develop severe painful crises due to multiple infarctions, tissue degeneration and organ damage while children with beta-thalassemia major become transfusion dependent and remain as such life long. In the Netherlands, neonatal screening (NS) for hemoglobinopathies has started in January 1st 2007 using state of the art HPLC technology.

Material and Methods: Screening of 1500 anonymous cord blood samples using the capillary electrophoresis (CE) technology (Sebia, France). Identification of carriers will be confirmed according to published DNA methods. Exploration of the possibility to detect beta-thalassemia carriers (not detected so far) using accurate HbA measurement. Current and new technologies will be tested for their diagnostic application in regular and non-invasive prenatal diagnosis.

Results: We are presenting the validation of an alternative method to identify carriers of SCD by the regular separation of HbS, C, E and D on CE. Moreover, we report sensitivity data on a selective method to identify carriers of beta-thalassemia trait by the levels of hemoglobin A2 present at birth.

Conclusion: Without preventive measures 60 new cases can be expected yearly in the Netherlands. The CE technology is a sensitive alternative to HPLC for the screening of hemoglobinopathies. Identification of neonates-carriers of SCD and beta-thalassemia based on biochemical techniques facilitates prevention of beta-thalassemia major due to combinations of beta-thalassemia and HbS.

Subclinically Increased Troponin T levels in Preeclamptic Patients are Correlated with the Development of Pulmonary Edema

W.H. Tong, W.C.J. Hop, E.A.P. Steegers, J.J. Duvekot

Erasmus MC

Introduction: Pulmonary edema is a serious complication in preeclampsia. Cardiac failure is considered as a possible cause of this complication. Preliminary reports describe subclinically increased Troponin T (TnT) levels, a marker of myocardial damage, in patients with hypertensive disorders of pregnancy. In this present study, the relation of TnT levels and the development of pulmonary edema in preeclamptic patients was examined.

Materials & Methods: A prospective, longitudinal survey was conducted using medical files. From September 2004 until December 2005 all patients diagnosed with preeclampsia (mild or severe) were reviewed. Pulmonary edema was defined using common clinical parameters: clinical dyspnoea, crepitating upon auscultation, oxygen saturation Results: During the study period, 219 patients were admitted with the diagnosis preeclampsia. In 138 patients, 92 patients with mild preeclampsia and 46 patients with severe preeclampsia, at least one TnT measurement was available. Ten patients developed pulmonary edema within two days after delivery. None of the patients developed pulmonary edema before delivery. During these days, at least one TnT measurement was available in 122 patients. Sixteen patients had no TnT measurements, these patients were excluded for further analysis. Combining the data of the 122 patients using Cox regression, preeclamptic women with subclinically increased TnT levels were more likely to have or to develop pulmonary edema (OR = 18.1, 95% CI 3.1-103.2, P=0.001).

Multivariable analysis, adjusted for severity of preeclampsia, revealed that patients with severe preeclampsia were more likely to develop pulmonary edema than those with mild preeclampsia (OR = 6.1, 95% CI: 1.2-61.0, P=0.03). Subsequently, severe preeclamptic patients with elevated TnT levels were more likely to develop pulmonary edema (OR = 16.3, 95% CI: 2.5-106.9, P=0.003).

Conclusion: Subclinically increased TnT levels are strongly correlated with the development of pulmonary edema in patients with predominantly severe preeclampsia. Myocardial failure is a possible cause of pulmonary edema in preeclamptic patients.

Changes in the Endometrial Receptivity in Tubular Infertility: Morphological Study

P. Männik, R.T. Kibur, M. Aunapuu, A. Salumets, A. Arend

University of Tartu

Introduction: Functional salpinxes are one of the most important preconditions for female fertility. Tubular infertility – TFI (Tubular Factor Infertility) is the most significant of salpingeal pathologies, resulting in uni- or bilateral partial or complete salpingeal occlusion. Tubular infertility is one of the leading causes of infertility in the world including Estonia. The most common method treating infertility due to TFI is In Vitro Fertilization (IVF). Occurrence of TFI decreases receptivity of the uterine mucosa, lowering the expression of ανβ3 integrins on the endometrial endothelium and therefore lowers the possibility of implantation. Noticeable changes in cytokine profile (higher levels of TNF-α) of the uterine endothelium in tubular infertility have also been reported, influencing uterine maturation. For the interaction between the endometrium and the embryonic cells special structures called pinopodes are needed. Pinopodes can be studied using electron microscopy. The role of pinopodes in TFI and infertility has not been thoroughly studied yet. Our study centers on finding out the causes of tubular infertility.

Material and Methods: Ten endometrial biopsies of patients with infertility problems were carried out in Nova Vita Clinic. Taken tissue samples were divided into two. Half of them were fixed in 10% buffered formalin solution and the other half in 2.5% glutaraldehyde solution. Sections embedded in paraffin were studied with light microscopy. For electron microscopic assessment sections were embedded in plastic (Epon 812) according to classical methods. Histological methods were used to specify the stage of menstrual cycle. Immunohistochemical staining was used to detect integrin beta3 (CD61) in the endometrium. Scanning electron microscopy (SEM) was used to assess endometrial glands, endometrial epithelial cells and pinopodes.

Results: Light microscopy studies of biopsies revealed no pathologic changes in endometric morphology. The ratio between glandular and stromal components was 1:1 in five samples. In one sample corresponding ratio was 2:1. The sample also showed lots of spiral arteries. Immunohistochemical staining with CD61 showed a very intense reaction in one patient, mild reaction in two patients and low reaction in seven patients. For SEM assessment only 5 samples were suitable. In one patient SEM showed normal endometrial epithelial cells. All the other patients showed morphological changes in ciliated epithelial cells. Pinopodes, essential components for embryonic implantation, were present in only one of the patients samples.

Conclusion: SEM studies have shown that TFI patients have appreciably lower amount of pinopodes needed for the implantation of embryo