Progressive increase of human papillomavirus carriage rates in potentially malignant and malignant oral disorders with increasing malignant potential.

INTRODUCTION: We investigated the potential role of human papillomaviruses (HPVs) in potentially malignant oral disorders, oral leukoplakia (OL) and oral lichen planus (OLP), and in oral squamous cell cancer (OSCC) in an Eastern Hungarian population with a high incidence of OSCC. METHODS: Excised tumor samples (65 OSCC patients) and exfoliated cells from potentially malignant lesions (from 44 and 119 patients with OL and OLP, respectively) as well as from healthy controls (72 individuals) were analysed. OLPs were classified based on clinical appearance, 61 patients had erosive-atrophic lesions (associated with higher malignancy risk, EA-OLP) and 58 had non-erosive non-atrophic lesions (with lower risk of becoming malignant, non-EA-OLP), respectively. Exfoliated cells collected from apparently healthy mucosa accompanied each lesion sample. HPV was detected by MY/GP polymerase chain reaction (PCR) and genotyped by restriction analysis of amplimers. Copy numbers in lesions were determined using real-time PCR. Prevalence rates, copy number distributions, and association with risk factors and diseases were analysed using chi-square test, t-test, and logistic regression, respectively. RESULTS: We detected HPVs significantly more frequently in lesions than in controls (P < or = 0.001 in all comparisons). HPV prevalence increased gradually with increasing severity of lesions (32.8, 40.9, and 47.7% in OLP, OL, and OSCC, respectively). Copy number distribution patterns roughly corresponded to prevalence rates, but OLP and OL were comparable. HPV prevalence differed significantly between EA-OLP and non-EA-OLP groups (42.6 vs. 22.4%); EA-OLP group showed a prevalence similar to that found in OL. CONCLUSION: HPVs may be involved in the development or progression of not only OSCC but also of potentially malignant oral lesions.

Survivin promoter polymorphism and cervical carcinogenesis.

BACKGROUND: Survivin, a novel member of the inhibitor of apoptosis family, plays an important role in cell cycle regulation. A common polymorphism at the survivin gene promoter (G/C at position 31) was shown to be correlated with survivin gene expression in cancer cell lines. AIM: To investigate whether this polymorphism could be involved in the development of human papillomavirus (HPV)-associated cervical carcinoma. METHODS: Survivin promoter polymorphism was detected in patients with cervical cancer, in patients with equivocal cytological atypia and in a control population using polymerase chain reaction (PCR-restriction fragment length polymorphism (RFLP) and PCR-single strand conformation polymorphism analysis. HPV was typed in patients with cervical cancer and cytological atypia using PCR-RFLP. RESULTS: No statistically significant differences were found in the genotype distributions of the survivin promoter variants among our study groups. CONCLUSIONS: The survivin promoter polymorphism at position 31 may not represent an increased risk for the development of cervical cancer, at least in the population studied here.

High co-prevalence of genogroup 1 TT virus and human papillomavirus is associated with poor clinical outcome of laryngeal carcinoma.

BACKGROUND: The aetiology and factors leading to the progression of laryngeal cancer are still unclear. Although human papillomavirus (HPV) has been suggested to play a role, reports concerning the effect of HPV infection on tumour development are controversial. Recently, transfusion transmitted virus (TTV) was suggested to play a role in certain infections as a causative or coinfecting agent. AIMS: To investigate whether the development and progression of laryngeal squamous cell carcinoma is associated with coinfection with TTV and HPV. METHODS: The prevalence of TTV and HPV was investigated using the polymerase chain reaction in tissue samples from 40 healthy individuals, 10 patients with recurrent papillomatosis, five patients with papillomatosis with malignant transformation, and 25 patients with laryngeal carcinoma. The obtained prevalence data were compared and analysed statistically. RESULTS: In the 11 patients with carcinoma who had metastasis or relapse there was a high rate of coinfection with genogroup 1 TTV and HPV (eight of 11), whereas in the 14 without tumour progression no coinfection was found. Coinfection was associated with significantly lower tumour free survival in patients with carcinoma (p < 0.001). Furthermore, four of five patients who had papillomatosis with malignant transformation were coinfected with genogroup 1 TTV and HPV. CONCLUSIONS: Although the nature of cooperation between HPV and TTV needs to be investigated further, coinfection with genogroup 1 TTV and HPV appears to be associated with poor clinical outcome in laryngeal cancer.

The characteristics of human papillomavirus DNA in head and neck cancers and papillomas.

AIM: To determine the prevalence, type, physical state, and viral load of human papillomavirus (HPV) DNA in cases of head and neck cancer and recurrent respiratory papillomatosis (RRP). METHODS: The prevalence and type of HPV DNA was determined in 27 fresh frozen tissue specimens from patients with head and neck cancers and 16 specimens from 10 patients with RRP by MY09/MY11 and GP5+/GP6+ nested polymerase chain reaction (PCR) and subsequent restriction enzyme cleavage. The physical state of HPV DNA was analysed by E1, E2, and E1E2 specific PCRs and Southern blot hybridisation (SBH). RESULTS: HPV DNA was detected in 13 of 27 cancers and 10 of 10 papillomas. Both low risk HPV-6 and HPV-11 and high risk HPV-16 were present in cancers in low copy numbers, whereas papillomas exclusively harboured low risk HPV-6 and HPV-11. E1E2 PCRs failed to determine the physical state of HPV in cancers except one case where HPV-6 DNA was integrated. In contrast to cancers, all papillomas showed the episomal state of HPV DNA and a relatively higher viral load. CONCLUSIONS: Based on the prevalence, type, physical state, and copy number of HPV DNA, cancers and papillomas tend to show a different HPV DNA profile. The 100% positivity rate of low risk HPV types confirms the role of HPV-6 and HPV-11 in the aetiology of RRP.

Persisting TT virus (TTV) genogroup 1 variants in renal transplant recipients.

TT virus (TTV) genogroup 1 infection has an increased prevalence in solid organ transplant recipients. In this study, the presence of TTV in renal transplant recipients was examined by two PCR methods, one capable of detecting most TTV genotypes (UTR-PCR), the other specific to genogroup 1 (N22-PCR). The N22-PCR detected TTV in 57% (53/92) of the renal transplant patients and in 20% (13/66) of the healthy individuals, while the prevalence of TTV with the UTR-PCR was above 90% in both the control and the patient groups. The N22-PCR was used in longitudinal studies of 31 renal transplant recipients, these PCR products were sequenced and aligned. TTV status was not associated with the patients' age at transplantation, male to female ratio and the time lag between kidney transplantation and the TTV test. During the follow-up consistent TTV status was found in 26 patients, while two initially TTV positive patients converted to negative and three initially negative patients converted to positive. The TTV variants varied among the tested patients, but were the same in the consecutive samples of each patient, indicating that TTV infection was persistent in renal transplant recipients and novel infection occurred rarely in the post-transplant period.

Demonstration of Trichomonas vaginalis in tracheal aspirates in infants with early respiratory failure.

Trichomonas vaginalis was isolated from the tracheal aspirates of two premature newborns with early respiratory failure who were delivered vaginally by mothers with T. vaginalis infection. The babies were treated successfully with antiparasitic drugs.

Neonatal pneumonia caused by Trichomonas vaginalis.

The authors present two cases of newborn babies infected by Trichomonas vaginalis (hereafter referred to as T. vaginalis) and suffering from severe congenital breathing difficulties and needing artificial respiration. Microscopic examination of the tracheal discharge revealed characteristically moving, flagellated, pear-shaped unicellular organisms. Cultures on CPLM medium proved the presence of T. vaginalis. During pregnancy the mothers' clinical status was negative and both of them mentioned leukorrhoea of changing intensity. They were regularly involved in antenatal care. The infection caused by T. vaginalis could be detected in the two mothers later by culture procedures.

9-Anthroylnitrile binding to serine-181 in myosin subfragment 1 as revealed by FRET spectroscopy and molecular modeling.

It has been shown that one of the 12 serine residues within the 23 kDa segment of myosin subfragment 1 can be covalently modified with a fluorescent probe 9-anthroylnitrile (ANN) [Hiratsuka, T. (1989) J. Biol. Chem. 264 (30), 18188-18194]. To identify the exact binding site of the probe, the distances between the bound ANN as donor and acceptors in known positions (Lys-553 or Cys-707) of the myosin head were determined by using fluorescence resonance energy transfer. Comparison of the spectroscopic results with distances obtained from the atomic model of subfragment 1 revealed that ANN binds to Ser-181. The result was in good agreement with the assumptions of Andreev and co-workers [Andreev, O. A., et al. (1995) J. Muscle Res. Cell Motil. 16 (4), 353-367]. This conclusion was further supported by protein modeling calculations. The results presented herein might bring ANN into the focus when the molecular mechanism and effects of the binding of ATP and its subsequent hydrolysis are studied.

Transcriptional activity of human papillomavirus type 16 variants having deletions in the long control region.

Transcription of the E6 and E7 viral oncogenes of human papillomavirus (HPV) type 16 is regulated by the P97 major early promoter, and enhancer and silencer elements found in the long control region (LCR). In this study, we tested the transcriptional activity of natural HPV 16 variants having long deletions in the LCR. The HPV 16 LCR regions were amplified from invasive cervical cancer specimens, and cloned into the reporter vector pALuc. Transcriptional activity of the different clones was measured by luciferase test after transient transfection into HeLa cells. The deletions found in the LCR encompassed parts of the enhancer and either the YY1-specific silencer alone or together with the CDP-specific silencer. The transcriptional activity of these deletion variants were usually reduced compared with that of the corresponding full-length clones. However, a deletion variant lacking the whole enhancer and both silencer regions retained substantial enhancer activity on the P97 promoter. These results point to the existence of a novel context-dependent enhancer element in the 5' LCR of HPV 16.

Human cytomegalovirus load in the peripheral blood determined by quantitative competitive polymerase chain reaction.

Primary human cytomegalovirus infection and the viral reactivation from latency are major complications in organ transplant recipients. In the peripheral blood the replicating virus can be detected either by nucleic acid based tests or by demonstrating the HCMV structural proteins in antigenemia test. We developed a quantitative competitive PCR method to assess the HCMV load in the peripheral blood. The viral load in nine healthy blood donors and in four renal transplant recipients with negative antigenemia test was in the same range: 5-124 (median: 18) HCMV copies/10(6) beta-globin copies for healthy blood donors and 16-48 (median: 37) HCMV copies/10(6) beta-globin copies for the transplant recipients. Three antigenemia positive renal transplant recipients had a HCMV load of 2.2 x 10(5)/10(6) beta-globin, 1.5 x 10(4)/10(6) beta-globin and 6.5 x 10(3)/10(6) beta-globin, respectively. In conclusion, the quantitative measurement of HCMV load in the peripheral blood correlated well with the routine HCMV antigenemia test. The DNA-based test, however can detect earlier the reactivation of the HCMV infection.

[Neonatal respiratory insufficiency caused by maternal infection with trichomonas vaginalis]. / Anyai Trichomonas vaginalis okozta (újszülöttkori) veleszületett légzészavar.

Authors presented cases of two premature newborns with severe respiratory insufficiency due to congenital maternal Trichomonas vaginalis infection requiring mechanical ventilation. They focused on this special perinatal problem and also on diagnostical and therapeutical modalities of this newly recognised syndrome in the early neonatal period.

Integration status of virus DNA and p53 codon 72 polymorphism in human papillomavirus type 16 positive cervical cancers.

BACKGROUND: The aim of this study was to determine the physical state of HPV16 DNA and to reveal any association between the physical state of virus DNA and pathologic or prognostic factors in HPV16 positive cervical cancers. The other aim was to estimate the role of p53 codon 72 polymorphism in disease progression. MATERIALS AND METHODS: The presence and physical state of HPV16 DNA was analysed by Southern blot hybridisation and E1-E2 specific PCRs in the primary tumours and pelvic lymph nodes of 85 cervical carcinoma patients. Results Integrated HPV16 DNA was found in 32 out of 41 (78%) primary tumours and 2 out of 22 (95%) lymph nodes carrying HPV16 DNA. No significant association was found between integration of virus DNA and course of the disease. There was a trend towards an association between disease recurrence and the presence of the p53 codon 72 arginine homozygous genotype (OR = 3.41, p = 0.23). CONCLUSION: The physical state of HPV16 DNA does not seem to play a major role as a prognostic indicator in Hungarian cervical cancer patients, while the p53 codon 72 genotype may have an impact on the clinical outcome of the disease.

Additional human papillomavirus types detected by the hybrid capture tube test among samples from women with cytological and colposcopical atypia.

The type specificity of the human papillomavirus (HPV) Hybrid Capture Tube (HCT) test was evaluated by using typing with PCR (MY09-MY11)-restriction fragment length polymorphism (RFLP) and sequencing. All samples HCT test positive for only low-risk HPV (n = 15) or only high-risk HPV (n = 102) were confirmed, whereas 9 of 12 HCT test double-positive samples contained only high-risk HPV types as determined by PCR-RFLP. Several high-risk HPV types (HPV-53, -58, -62, -66, -CP8304, and -MM4) not included in the HCT test were indeed detected, indicating a broader detection range with retained distinction between low-risk and high-risk HPV types.

Frequency of p53 codon 72 genotypes in human papillomavirus associated squamous intraepithelial lesions and cervical cancer.

BACKGROUND: A recent study suggested that the p53Arg (at residue 72) homozygous genotype could be a potential genetic risk factor for cervical cancer among white women. To confirm this result we examined the proportion of p53 genotypes in a larger number of patients with cervical cancer and in patients with squamous intraepithelial lesions (SIL) compared to a control population. MATERIALS AND METHODS: We used allele-specific primers to amplify the p53Arg and p53Pro sequences and we examined the proportion of genotypes in the study populations using chi 2-test. RESULTS: The distributions of p53Arg homozygous, heterozygous and p53Pro homozygous genotypes were 63%, 27% and 10% in cervical cancer patients, 53%, 36% and 8% in individuals with SIL, and 60%, 36% and 4% in control population. Using chi-square test, no significant difference was found between genotype frequencies in the study groups. CONCLUSION: Thus, the p53Arg homozygous genotype does not seem to increase the risk of cervical cancer in Hungarian women.

Functional significance of sequence variation in the E2 gene and the long control region of human papillomavirus type 16.

The long control region (LCR) and the E2 protein of human papillomaviruses (HPV) are the most important viral factors regulating transcription of the viral oncogenes E6 and E7. Sequence variation within these genomic regions may have an impact on the oncogenic potential of the virus. Sequence variation in the LCR and in the E2 gene of human papillomavirus type 16 (HPV-16) isolates originating from cervical cancer patients from East Hungary was studied. In 30 samples, sequencing and/or single-strand conformation polymorphism analysis revealed variants belonging to the European variant lineage of HPV-16. These variants differed from the reference European clone only slightly in their E2 and LCR sequences. Three samples represented variants belonging to the Asian-American group. These differed from the published reference sequence at several positions in the LCR and E2 regions. Compared to the reference clone, the LCR clones of the European isolates showed very similar transcriptional activities, while that of an Asian-American isolate had approximately 1.7-fold increased activity. Most of the increased activity of the Asian-American isolate could be ascribed to nucleotide changes found at the 3' end of the LCR (nt 7660-7890). The transcriptional transactivation potentials of the HPV-16 E2 isolates differed only slightly from each other, and the differences seemed to be independent of the taxonomic position of the isolates.

Consistent polymerase chain reaction-single-strand conformation polymorphism pattern of human herpesvirus-8 in the course of classical Kaposi's sarcoma assumes its clonal origin.

There is emerging evidence that Kaposi's sarcoma-associated herpesvirus (KSHV or HHV-8) has a central role in the pathogenesis of Kaposi's sarcoma (KS). The occurrence of HHV-8 in classical KS biopsies is reported irrespective of its clinical stage (patch, plaque, nodular). HHV-8 was detected in 25 of 28 formalin-fixed paraffin-embedded classical KS samples by nested polymerase chain reaction. In addition, in six patients multiple tumors were available (n = 21). Single-strand conformation polymorphism (SSCP) analysis of the amplicons showed uniform SSCP pattern of samples belonging to the same patient regardless of whether the KS was multiplex or developed again years after the first excision. Most of the SSCP patterns were confirmed by further sequence analysis. The presence of the same sequence variant of HHV-8 in various samples of the same patient supports the clonal origin of classical Kaposi's sarcoma.