Male Human Papillomavirus Infection and Genotyping in Turkey.

BACKGROUND: High-risk (HR) Human Papillomavirus (HPV) has been shown to play an important role in men in various locations in Turkey. This study aims to screen the male persistent infection with the high-risk human papillomavirus (HPV) genotype status in Turkey to provide a reference basis for formulating prevention strategies for the development of genitourinary tract neoplasia. METHODS: The HPV QUANT-21 Quantitative RT-PCR Kit® was used to identify and quantify low-risk HPV (HPV 6, 11, 44) and high-risk (HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82) from male individuals in Turkey. RESULTS: Of the total 1304 samples, 473 were positive for at least one HPV genotype, with an overall frequency of 36.2%. Two-hundred fifty-four patients were positive only for one or more LR   HPV genotypes (54%), and 219 patients were positive for one or more HR HPV genotypes (46%). The LR HPV genotype frequency was 53.7%, while the HR HPV genotype frequency was 46.3%. Our technology had the positive advantage of being able to calculate concentrations for each genotype. Genotype 51 was second in frequency but had the highest average concentration of 5.38 log (copies/sample). CONCLUSION: The presence and genotype of the virus before HPV vaccination are also of increasing importance. The data obtained will serve as a guide for prevention strategies, especially vaccination. Based on our findings there is a need of new estimates of the efficacy of currently available HPV vaccines and to develop a screening program to prevent and reduce the incidence of genitourinary tract neoplasias in Turkey. Further studies are planned to measure and define the high levels of infection that may lead to the development of cervical tumors. Using this technique, it may be possible to make clinical decisions about the extent of cytological alterations.

Analysis of the Prevalence and Quantification of viral load of different human papillomavirus types in Turkish women population.

BACKGROUND: Persistent infection with high-risk (HR) Human Papilloma Virus (HPV) genotypes has been shown to play a significant role in the development of cervical intraepithelial neoplasia (CIN) and CC (cervical cancer). The present study aimed to determine the distribution and quantification of viral load of HPV genotypes in numerous genital samples obtained from women undergoing routine gynaecological care in different regions of Turkey. METHODS: HPV typing was done by HPV QUANT-21 Quantitative RT-PCR Kit®, which is intended for the specific identification and quantification of low-risk (HPV 6, 11, 44) and high-risk (HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82) from female subjects in Turkey. RESULTS: From the total of 5975 samples, 2777 were positive for at least one HPV genotype, with an overall frequency of 46.4%. 1695 patients were positive for only one or more LR-HPV genotypes (61%) and 812 patients were positive for one or more HR-HPV genotypes (29%). The frequency of LR-HPV genotypes was 31.4%, while the frequency of HR-HPV genotypes was 118.8%. Our tecnology had a positive advantage to calculate the concentration of each genotypes. Although genotype 52 ranked fifth in frequency, it showed the highest mean concentration, with a value of 5.38 log (copies/sample). CONCLUSION: The presence and genotype of viruses before HPV vaccination have also gained importance. The data obtained would provide guidance for prevention strategies, mainly of vaccination. We decided to add a new estimate to the effectiveness of currently available HPV vaccines and the development of screening programs to prevent and decrease the incidence of CC in Turkey. Further studies would be planned to measure and define the high infection level that can lead to the development of cervical neoplasia. Using this tecnology could give us a clinical desicion to degree the cytological changes.

Should pancreas cyst fluids be divided into two for cytological diagnosis and biochemical tests?

BACKGROUND/AIMS: The aim of the present study was to investigate whether pancreas cyst fluids should be divided into two for cytological diagnosis and biochemical tests. MATERIALS AND METHODS: The present study was conducted with fluids aspirated from 12 pancreas cysts. The fluids were divided into two and sent to the cytopathology (fluid 1) and biochemistry (fluid 2) laboratories. Fluid 1 was centrifuged at the cytopathology laboratory. Cytology slides were prepared from the deposit, and the supernatant was sent to the biochemistry laboratory. Fluid 2 was centrifuged at the biochemistry laboratory, and amylase, carcinoembryonic antigen, and cancer antigen 19.9 levels were determined in the supernatant. These procedures were repeated for fluid 1 from the cytopathology laboratory. The remaining fluid 2 was sent to the cytopathology laboratory. Fluid 1-like slides were prepared from fluid 2 in the cytopathology laboratory. Cytological diagnoses of fluid 1 and fluid 2 were compared, and the Pearson correlation coefficient for biochemical test results was identified. RESULTS: 92% of fluid 1 and 50% of fluid 2 were diagnostic. Biochemical test results of fluid 1 and fluid 2 were similar, and the Pearson correlation coefficient was high. CONCLUSION: Our results showed that pancreatic cyst fluids did not need to be divided into two for cytological diagnosis and biochemical tests. Following centrifugation of the whole fluid at the cytopathology laboratory, the deposit and the supernatant can be used for cytological diagnosis and for biochemical tests, respectively. With this protocol, the sensitivity of cytological diagnoses and biochemical tests of pancreatic cyst fluids may increase.

Thromboelastography for the monitoring of the antithrombotic effect of low-molecular-weight heparin after major orthopedic surgery.

OBJECTIVE: Low-molecular-weight heparins (LMWHs) are commonly used to prevent and manage postoperative thromboembolism. In general, monitoring of anticoagulant activity by anti-Xa testing is not done properly. Thromboelastography (TEG) evaluates the viscoelastic properties of blood during coagulation. The clinical application of TEG variables in monitoring LMWH treatment is not yet well defined. METHODS: This prospective study was designed to systematically examine the correlation between anti-Xa and basic TEG parameters in monitoring LMWH treatment. We furthermore evaluated for the first time the usefulness of a composite TEG parameter, coagulation index (CI). Thirty patients undergoing unilateral or bilateral total knee replacement, admitted to the intensive care unit on a therapeutic dosage of subcutaneous enoxaparin (30-mg injections administered twice daily), were included into the study. TEG parameters and anti-Xa levels were measures at baseline and 4, 12, and 24 hours after the injection. RESULTS: This study demonstrates a significant correlation between CI and plasma anti-Xa activity in surgical patients treated with enoxaparin. Although the correlation was significant between r time and anti-Xa level only at Hour 4, CI was significant for each time interval (p<0.05). CI increased immediately after T0, peaking at Hour 4, and remained elevated (relative to baseline) at Hour 24 but still did not return to admission levels. CONCLUSION: The current study may be an important first step in order to use CI to measure LMWH activity. Meanwhile, the value and usefulness of TEG in predicting bleeding or thrombotic complications following major orthopedic surgery merit further investigation.

Gene polymorphisms of adducin GLY460TRP, ACE I/D, AND AGT M235T in pediatric hypertension patients.

BACKGROUND: Hypertension is a major global public health problem that affects both pediatric and adult populations. ACE I/D, AGT M235T, and ADD Gly460Trp polymorphisms are thought to be associated with primary hypertension. In the present study, we examined the frequency of these polymorphisms in a pediatric population with secondary hypertension. MATERIAL AND METHODS: Included in the study were 58 hypertensive and 58 normotensive pediatric patients. ACE I/D and AGT M235T polymorphisms are determined by conventional PCR; ADD Gly460Trp polymorphism was investigated using PCR amplification of genomic DNA. RESULTS: There were significant differences between the control group and pediatric hypertensive group in terms of ACE I/D (P<0.05) and AGT M235T (P<0.05) polymorphisms, but there were no differences in ADD Gly460Trp (P>0.05) polymorphism. CONCLUSIONS: We suggest that RAS gene polymorphisms (ACE-I/D, AGT M235T) are significantly associated with susceptibility to diseases that lead to secondary hypertension.

Optical and mechanical clot detection methodologies: a comparison study for routine coagulation testing.

BACKGROUND: Automated coagulation analyzers are preferred to meet increasing coagulation test volume. Two distinct technological families exist based on optical and mechanical clot detection methodologies. Which one is superior to the other is still a conflict and needs new studies. METHODS: We have compared prothrombin and activated partial thromboplastin results obtained with mechanical method with those obtained by photo-optical method used routinely in our specialized laboratory. RESULTS: The instrumental results showed good precision ranging between 0.7% and 1.8% coefficient of variation. Statistical analysis demonstrated an excellent correlation between the photo-optical and mechanical analyzers for PT (R² 0.97), and aPTT (R² 0.85). CONCLUSION: Correlation between the two clot-detection systems was maintained even when measuring turbid samples (R² ≥ 0.97 for two tests).

Discard first tube for coagulation testing.

Tissue thromboplastin may contaminate the first tube sample due to the trauma of the venipuncture, and therefore, affect the accuracy of coagulation testing. This practice was stopped by Clinical and Laboratory Standards Institute after several studies. However, most of the studies have verified these conclusions and refuted the need for a discard tube when drawing samples for coagulation tests in healthy groups. The purpose of our study was to evaluate the clinical importance of discarding a tube for prothrombin time (PT) determinations on large samples with international normalized ratio (INR) values between and over targeted therapeutic range. Patients receiving oral anticoagulation therapy (OAT) managed by our cardiology service were selected for this study. Tube 1 was always treated as the discard tube. Tube 2 was allocated to be analyzed along with the tube 1 for coagulation tests. Individual values were grouped into four cohorts according to the INR range. The ranges were as follows: less than 2.0, 2.1-3.0, 3.1-4.5 and more. Three hundred and seventy-six samples were drawn for PT/INR and activated partial thromboplastin time testing. We found statistically significant differences between tube 1 and tube 2 (P < 0.05), and satisfactory correlation coefficients were obtained by linear regression analysis (0.86 or greater in all cases). This study consisted of a high number of samples. Our data suggest that drawing a discard tube is still necessary for coagulation testing. Consideration should be given to revising the international guidelines related to the necessity of a discard tube for repeated evaluation of coagulation tests especially receiving long-term OAT.

Induction of HGF and VEGF in hepatic regeneration after hepatotoxin-induced cirrhosis in mice.

BACKGROUND/AIMS: Liver cirrhosis is the irreversible end-result of fibrous scarring and hepatocellular regeneration, characterized by diffuse disorganization of normal hepatic structure by regenerative nodules and fibrotic tissue. In this study, we elucidated the role of hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) in liver regeneration. METHODOLOGY: The study was conducted as an experimental laboratory investigation using a mouse model of lethal liver cirrhosis induced by carbon tetrachloride (CCl4), dimethylnitrosamine (DMN) and D-galactosamine (D-gal) administrations. RESULTS: Liver morphology showed fibrosis/cirrhosis in all groups, but to a different extent, as confirmed by the rise in serum transaminase levels. The immunolocalization of VEGF and HGF, and homogenate levels of HGF and serum levels of VEGF, were also analyzed. Liver fibrosis/cirrhosis was more severe in CCl4-treated mice. In cirrhotic livers, immunostaining for HGF was weak and the HGF content of liver tissue was lower. Strong immunoreactivity for VEGF was observed when hepatotoxins were administered, however as cirrhosis became apparent immunoreactivity was reduced. Blood VEGF levels increased gradually. CONCLUSIONS: Our results suggest possible involvement of VEGF in angiogenesis of cirrhotic liver. VEGF might be required for reconstruction of hepatic cells and sequentially participates in liver regeneration by facilitating hepatocyte proliferation. HGF production is supposed to be induced in the necrotic liver during regeneration and severe tissue damage followed by cirrhosis might account for low homogenate HGF levels.

Detection of immunoglobulin G antibodies to Toxoplasma gondii: Evaluation of two commercial immunoassay systems.

BACKGROUND: Toxoplasmosis is a disease, which can cause severe congenital infection and is normally diagnosed by the detection of Toxoplasma gondii (T gondii)-specific antibodies in the serum of infected patients. Several different tests allow to distinguish recent from past infections and to quantify anti-T gondii-specific IgG, and the results can be used as markers for a chronic or recently seroconverted toxoplasma. METHODS: In the present study, the recent Cobas 6000 Toxo IgG assay (Roche Diagnostics, Indianapolis, IN, USA) for the serological diagnosis of toxoplasmosis was compared with the Axsym Toxo IgG assay (Abbott Laboratories, Diagnostics Division, Abbott Park, IL, USA) employing a panel of negative, low- or high-reactive serum samples that were selected after routine screening in a laboratory of clinical analyses. RESULTS: The overall agreement between two methods was 99% (r=0.99, p<0.001). Of 91 analyzed samples, only one presented discrepant result, being positive in the Cobas 6000 Toxo IgG assay and negative in the Axsym Toxo IgG assay. By using an immunofluorescent assay as a confirmation test, this positive result was assayed to be negative. CONCLUSIONS: Both assays performed in each analyzer were proven to be fast and fully automated procedures for reproducible measurement of IgG antibodies to T gondii. The new method, used for the determination of anti-T gondii IgG antibodies, should be evaluated with a further analysis with increased number of serum samples to get a broad performance of this newer test.

Types and frequencies of pre-analytical mistakes in the first JCI:2002 accredited hospital in Turkey.

BACKGROUND: In light of increasing attention focused on patient safety and the need to reduce laboratory mistakes, it is crucial for clinical laboratories to collect statics on error rates over the whole testing cycle, particularly on the pre-analytical phase. METHODS: Memorial Hospital has a laboratory service providing stat and routine tests. From January 2005 to December 2007, a total of 766,095 specimens were received in our laboratory. The frequencies and types of pre-analytical errors and the potential targets for quality improvement strategies were evaluated. RESULTS: The relative frequencies of pre-analytical errors were ordered in a gradually decreasing trend through years; 0.50% in 2005; 0.25% in 2006 and 0.07% in 2007 (p < 0.0001; between 2005 and 2007). The most common mistake was incorrect/missing identification (42 %). CONCLUSIONS: We believe that the use of standardized routines and regular staff training, combined with an exchange of the existing paper-based referral management system with an electronic system, can increase the safety in the pre-analytical processes.