Prevalence of human papillomavirus (HPV) and HPV-16 genotyping by real-time PCR in patients with several cervical pathologies

PurposeThis study was planned to evaluate the prevalence of HPV (excepting type 16) and HPV 16 by real-time PCR in colposcopy patients and to interprete the results with age, age of first sexual intercourse (FSI), parity and Pap smear results.MethodsOne hundred and two colposcopy patients (50 and 52 of the patients were classified as colposcopy positive and negative, respectively) applying to Gynecology clinic were included. HPV (excepting type 16) and HPV 16 were detected by real-time PCR using the L1 region. Real-time nested amplifications of MY09/11 products were done by GP5+/GP6+ primers and Cyanine-5 labeled HPV and HPV 16 DNA specific probe after HPV DNA extraction by phenol chloroform isoamylalcohol.ResultsHPV (excepting type 16) and HPV 16 were positive in 12% and 18% of the colposcopy positive patients respectively. HPV (excepting type 16) and HPV 16 were positive in 5.7% and 3.8% of the colposcopy negative patients, respectively.ConclusionThere was a statistically significant difference between colposcopy positive and colposcopy negative patients comparing HPV 16 with total HPV positivity (p = 0.021 for type 16 and p = 0.010 for total HPV) but there was not a statistically significant difference between colposcopy positive and colposcopy negative patients when we compared HPV (excepting type 16) positivity (p = 0.314). In conclusion, HPV detection and typing may be helpful for cervical cancer screening and prevention.     

HBV-related events after allogeneic hematopoetic stem cell transplantatıon in a center from Turkey

To investigate the frequency of hepatitis B virus (HBV)-related events after allogeneic HCT in a moderate endemic area for HBV infection. The data of 197 patients who underwent allogeneic hematopoetic stem cell transplatation (HCT) from September 2003 through December 2010 were reviewed retrospectively with respect to HBV-related events. Resolved HBV infection was described as negative HBsAg, positive HBcAb, and positive HBsAb. Latent HBV infection was defined in patients with HBcAb positivity in the abscence of HBV DNA and HBsAb. Hepatitis B naive patients are defined as the patiens with no serological or molecular marker related to HBV. Seropositive patients were the patients with positive HBsAg and HBV-DNA. Median age was 28 (range, 15–64) years, with 128 male and 69 female patients. Median follow-up of the cohort was 8 (range, 0.5–78) months. We detected HBV-related events in 7 (3.6 %) recipients after allogeneic HCT. Five (71.4 %) of these events were HBV reactivation, while two cases (28.6 %) had acute hepatitis B infection. Four of the five reactivations were in the seropositive group (80 %), while one ocurred in a patient with resolved hepatitis. Two patients who developed acute hepatitis B were HBV naive and previously immunized patients, respectively. Hepatitis B virus reactivation remains a problem in seropositive patients and might require more effective treatment strategies.     

5-fluorouracil loaded chitosan microspheres for chemoembolization

In this study, chitosan microspheres were prepared by a suspension cross-linking technique. A petroleum ether/mineral oil mixture was used as the suspension medium which includes an emulsifier, e.g. Tween-80. Glutaraldehyde was used as the cross-linker. 5-Fluorouracil was incorporated in the matrix for the possible use of the microspheres in chemoembolization. The size and size distribution of the chitosan microspheres varied in the size range of 100-200 microns, by changing the emulsifier concentration, stirring rate, chitosan/solvent ratio and drug/chitosan solution ratio. In summary, the size and size distribution of the microspheres decreased when the emulsifier concentration and stirring rate were increased. Smaller microspheres with narrower size distributions were obtained when the chitosan/solvent ratio and drug/chitosan ratio were lower. It was possible to load the chitosan microspheres with 5-FU to a concentration of 10.4 mg 5-FU/g chitosan. Around 60% of the loaded drug was released within the first 24 h, then the release rate became much slower.     

Complete genome sequence and phylogenetic analysis of hepatitis B virus isolated from Turkish patients with chronic HBV infection

Hepatitis viruses are the leading causes of chronic liver disease resulting in chronic hepatitis, cirrhosis, and hepatocellular carcinoma in the world and also in Turkey. Although Turkey has an intermediate rate of hepatitis B virus (HBV) infection with a prevalence reported as 5%, a complete HBV genome sequence has not been published. In this study, the molecular characterization and phylogenetic analysis are described of 11 complete HBV genomes isolated from 11 naïve patients (5 male, 6 female; ages: 18–54 years old, median 35 years old) with chronic HBV infection. Of 11 patients, 7 and 4 were HBeAg positive/anti‐HBe negative and HBeAg negative/anti‐HBe positive, respectively. All patients had no co‐infection with HCV, HDV, or HIV. HBV DNA was extracted from the sera of the patients. The complete genome was amplified by PCR and cloned into a TA vector. The PCR products were sequenced directly and the complete HBV genome sequences were determined. Ten HBV genomes were 3182 base pairs in length. There was a 183 bp deletion (between nucleotides 2987–3169) in pre‐S region in one HBeAg positive patient. There were two pre‐core stop codons (G1896A) in two HBeAg negative and three core promoter dual mutations (T1762/A1764) in one HBeAg positive and two HBeAg negative patients' HBV genomes. Phylogenetic analysis of all complete genomes yielded that all Turkish sequences were clustered in genotype D branch (ten in subgenotype D1 and one in subgenotype D2). The analysis of S gene amino acid sequences revealed that surface gene subtypes of one and ten HBV strains were subtype ayw3 and ayw2, respectively. This study indicates that Turkish patients with chronic hepatitis B infection show very little genotypic heterogeneity. Genotype D of HBV DNA and subtype ayw2 of surface gene represent almost the whole Turkish patient population infected with HBV. J. Med. Virol. 76:476–481, 2005. © 2005 Wiley‐Liss, Inc.     

Molecular detection of cytomegalovirus,herpes simplex virus 2, human papillomavirus 16–18 in Turkish pregnants

ObjectiveHuman cytomegalovirus (CMV) is the most common cause of viral intrauterine infections in the world. Herpes simplex virus type 2 (HSV-2) and human papillomavirus (HPV) are the main agents of viral sexually transmitted diseases, which cause genital ulcers and genital warts, respectively. HPV infection has been linked to the majority of the anogenital malignancies. The aim of this study was to detect the existence of CMV, HSV-2 and HPV type 16–18 in Turkish pregnants by using sensitive molecular assays.MethodsOne hundred thirty-four women (18–41 years old; mean age ± SD: 27 ± 8) applied to outpatient clinic of Obstetrics and Gynecology, in between 18th - 22nd weeks of their pregnancy and a control group of 99 healthy women (15–39 years old; mean age ± SD: 24 ± 8) were included in the study. Cervical smear samples were used for DNA extraction. CMV, HSV-2 and HPV 16–18 detections were carried out by real time PCR and in house PCR method, respectively.ResultsThree patients (3/134; 2.2%) were found to be positive for each HPV and HSV-2. Dual infection with HPV and HSV was found in just one patient. HPV 18 was detected in all positive samples. CMV was found to be positive in two patients (2/134; 1.4%).ConclusionHPV, HSV and CMV must be screened due to high prevalence of these viruses in pregnants by using sensitive molecular methods.     

The correlation between EBV viral load in the palatine tonsils of patients with recurrent tonsillitis and concurrent serum titers of VCA–IgG

Epstein–Barr virus (EBV) infection usually occurs in early childhood and can persist in palatine tonsil lymphocytes to induce tonsillitis at a later date. We have examined the presence of EBV in palatine tonsils and relationship between EBV-DNA quantity in tonsil tissues and VCA–IgG quantity in autologous sera. Tonsils were obtained from 36 patients, male 20 (55.6%), female 16 (44.4%) (mean age 7.96 ± 6.97 years), who underwent tonsils removal because of recurrent tonsillitis. Tissues were processed for real-time PCR and patient’s sera were assayed to determine VCA–IgG by VCA–IgG ELISA. In 27 out of 36 cases (75%), positive EBV-DNA reaction was found. However, statistical analysis showed no correlation between EBV-DNA quantity and VCA–IgG quantity. We conclude that tonsils of children can be colonized by EBV and that virus may have a direct and indirect role in recurrent tonsillitis and nasopharyngeal carcinoma.     

Hepatitis B-related events in autologous hematopoietic stem cell transplantation recipients

AIM: To investigate the frequency of occult hepatitis B, the clinical course of hepatitis B virus (HBV) reactivation and reverse seroconversion and associated risk factors in autologous hematopoietic stem cell transplantation (HSCT) recipients. METHODS: This study was conducted in 90 patients undergoing autologous HSCT. Occult HBV infection was investigated by HBV-DNA analysis prior to transplantation, while HBV serology and liver function tests were screened prior to and serially after transplantation. HBV...     

Diversity of human rotavirus G9 among children in Turkey

Between September 2004 and December 2005 a prospective study was conducted to understand the epidemiology of rotavirus infection among children with diarrhea attending two hospitals in Ankara, Turkey. Rotavirus was detected in 39.7% of the 322 stool samples and affected mainly children in the age group of 6-23 months. More than 70% and 39% of these cases occurred in children <2 and <1 year of age, respectively. In the temperate climate of Ankara rotavirus infection was prevalent throughout the year. Serotype G1P[8] was dominant followed by G9P[8]. In 38 samples a total of 5 electropherotypes were detected. All G9P[8] were of long electropherotype except one of short electropherotype. A proportion of G1 and G9 strains were in combination with P[6], P[4] or P nontypable. Mixed serotypes were responsible for 2.4% of the infections. A phylogenetic tree constructed with the deduced amino acid sequences of the VP7 gene showed that 16 Turkish G9 strains clustered with rotaviruses of lineage III. One G9 strain formed a new lineage, lineage IV with the Sri Lankan G9 rotaviruses. In the phylogenetic tree of the VP8* gene, the Turkish G9P[6] rotaviruses clustered with human strains of lineage Ia. Increased diversity of the G/P type combination and the presence of infection throughout the year in Turkey was a situation similar to developing countries. The occurrence of rotavirus infection at later age and low level of mixed infections in Turkey represented the situation of developed countries. This study suggests that diverse G9 rotaviruses are emerging in Turkey.     

5-Fluorouracil loaded chitosan microspheres for chemoembolization

In this study, chitosan microspheres were prepared by a suspension crosslinking technique. A petroleum ether/mineral oil mixture was used as the suspension medium which includes an emulsifier, e.g. Tween-80. Glutaraldehyde was used as the cross-linker. 5-Fluorouracil was incorporated in the matrix for the possible use of the microspheres in chemoembolization. The size andsize distributionof thechitosanmicrospheres variedinthesizerangeof 100-200mum, by changing the emulsifier concentration, stirring rate, chitosan/ solvent ratio and drug/chitosan solution ratio. In summary, the size and size distribution of the microspheres decreased when the emulsifier concentration and stirring rate were increased. Smaller microspheres with narrower size distributions were obtained when the chitosan/solvent ratio and drug/chitosan ratiowerelower. Itwas possibletoload thechitosanmicrospheres with 5-FU to a concentration of 10.4mg 5-FU/g chitosan. Around 60%of the loaded drug was released within the first 24h, then the release rate became much slower.     

Does Epstein-Barr virus infection have an influence on the development of laryngeal carcinoma? Detection of EBV by Real-Time Polymerase Chain Reaction in tumour tissues of patients with laryngeal carcinoma

Epstein-Barr virus (EBV) is a well-known carcinogenic virus, and the association of EBV with some tumours suggests that there may also be an association between laryngeal carcinoma and EBV.ObjectiveThe aim of this study is to determine the role of EBV in the aetiology of laryngeal carcinoma.MethodProspective investigation the EBV with real time polymerase chain reaction in tumour tissues of 25 patients with laryngeal carcinoma and 17 patients with benign laryngeal lesions, and investigation of the relationship between the presence of viral DNA and patients’ smoking habits, alcohol consumption, localization and differentiation of the tumour.ResultsThere was no significant difference between the control group and patient group in terms of EBV polymerase chain reaction positivity (p > 0.05). Also we couldn't find a statistically significant relationship between EBV positivity and differentiation of the tumour, localization of the tumour, smoking and alcohol consumption habits (p > 0.05).ConclusionOur results suggest that, although EBV is present in some of the squamous cell laryngeal carcinomas, its presence has no effect on the pathogenesis of laryngeal carcinomas.