Association of asymptomatic bacterial vaginosis with persistence of female genital human papillomavirus infection.

More data are needed on the role of abnormal vaginal microbiota in the natural history of cervical human papillomavirus (HPV) infections. Our purpose was to study the prevalence of mixed flora (MF), bacterial vaginosis (BV) and yeast infection in women with known HPV outcomes during the 72-month follow-up (FU). Asymptomatic pregnant women (N = 329) were enrolled in the third trimester of their pregnancy. Pap smears and HPV genotyping samples were taken at baseline and at 12-, 24-, 36- and 72-month FU visits, with one additional sample at 2 months for HPV. HPV testing was done with nested PCR and Multimetrix assay to determine the point prevalence and persistence of HPV. Conventional Pap smears were scored for MF, BV and yeast infection. Covariates of the outcomes were analyzed using generalized estimating equation (GEE) and Poisson regression. Of the women, 76.6% (252/329) tested HPV-positive at least once during the FU. BV was detected in 12.2% (40/329), MF in 57.4% (189/329) and yeast infection in 22.9% (73/329) of the women. HPV-positive women had significantly more leucocytes in their Pap smear (p = 0.023) than the HPV-negative ones. MF (OR 2.75, 95% CI 1.77-4.27) and yeast infection (p = 0.007) were linked with HPV positivity. BV but not yeast infection was a significant covariate of HPV persistence (p = 0.024; OR 2.15, 95% CI 1.13-4.08). MF and yeast infection were associated with prevalent cervical HPV infection. In the longitudinal setting, BV predicted HPV persistence, implicating that treatment of asymptomatic BV in women with cervical HR-HPV infections might be justified.

Genotype-specific concordance of oral and genital human papillomavirus infections among marital couples is low.

Data on genotype-specific concordance of oral-oral and genital-oral HPV infections among marital couples are key to understand HPV transmission between spouses. Genotype-specific concordance of HPV infections (oral/genital) and their co-variates among 131 marital couples were determined during 6-year follow-up (FU). Seven oral scrapings were taken from both spouses, accompanied by six genital samplings from the women and one (at baseline) from the male partners. HPV-genotyping was performed by nested PCR and a Luminex®-based Multimetrix Assay. Demographic data were collected with questionnaires at baseline and study conclusion. Prevalence of oral HPV varied from 10.3 to 27.0 % and 15.8 to 31.3 % in women and men, respectively. At baseline, 37.6 % of the male genital samples were HPV-positive while in female genital samples, HPV prevalence varied from 13.3 to 59.4 %. Only 15 couples had HPV genotype-specific concordance (oral-oral n = 7; male oral-female genital n = 9; female oral-male genital n = 2). In the nested case-control setting, higher number of deliveries (OR 0.145, 95%CI 0.030-0.706, p = 0.017) and higher number of intercourse (OR 0.488, 95%CI 0.243-0.978, p = 0.043) decreased the likelihood of concordant HPV infections while practicing oral sex increased the risk (OR 0.299, 95%CI 0.120-0.748, p = 0.010). In multivariate analysis, the likelihood of concordance was decreased by higher number of pregnancies of the female partner (p = 0.020) and by higher frequency of intercourse reported by the male spouse (p = 0.027). To conclude, asymptomatic HPV infections were common in both spouses while genotype-specific concordance was low. This supports the view that HPV profile of the spouses has been established before the current marital relationship.

Identification of quantitative trait loci influencing inflammation-mediated alveolar bone loss: insights into polygenic inheritance of host-biofilm disequilibria in periodontitis.

BACKGROUND AND OBJECTIVE: The relative contribution of genetic and environmental factors to the onset and progression of periodontitis is inconclusive. Despite the high prevalence, phenotypic heterogeneity and significant local and systemic implications of this disease, early detection and individualized therapy are problematic. Using a murine model of periodontitis in a panel of 17 recombinant inbred mice, the current study addressed the heritability of, and oral dysbiosis associated with, inflammation-mediated alveolar bone loss (iABL), the hallmark of periodontitis. MATERIAL AND METHODS: Quantitative trait locus (QTL) genomics and quantitative PCR for over 99% of known murine oral microbiota were used. RESULTS: It was found that iABL is a polygenic trait with 32.7% heritability. One suggestive QTL, nicknamed inflammation-mediated alveolar bone loss locus (iABLL), was identified on chromosome 2. Eleven genes involved in innate immune responses and bone metabolism, particularly related to macrophage and osteoblast function, namely Etl4, Pdss1, Cobll1, 9330158F14Rik, Xirp2, Stk39, Mettl5, Metapl1, Itga6, Pdk1 and Sp3, were found in the iABLL using cis expression QTL and nonsynonymous single nucleotide polymorphism analyses. Specific oral microbiome shifts in saliva and tongue mucosa are associated with disease in this model. CONCLUSION: Our results indicate that complex host-biofilm interactions generate pathogenic states that extend beyond subgingival biofilms and periodontal tissues. Although no temporal relationship between the onset of iABL and microbiome changes were established, our findings suggest that host factors may be responsible for pathogenic shifts in subgingival biofilms when persistent and undisturbed.

Oral human papillomavirus infection in men might contribute to HPV serology.

The prospective Finnish Family HPV Study evaluated the dynamics of human papillomavirus (HPV) infection within families. Here, we focused on HPV serology in men. Seroprevalence at baseline, seroconversion and decay of low-risk (LR)-HPV6 and 11, and high risk (HR)-HPV16, 18 and 45 L1 antibodies in 122 men at 12, 24 and 36 months were determined using Luminex-based multiplex HPV serology, and correlated with demographic data. At baseline, seropositivity to HPV6, 11, 16, 18 and 45 was observed in 41.0, 11.5, 23.0, 13.9 and 5.7 % of the men, respectively. In univariate analysis, LR-HPV seropositivity was related to smoking status, history of genital warts and being seropositive to HR-HPV. Oral HR-HPV DNA and baseline LR-HPV seropositivity predicted HR-HPV seropositivity. Seroconversion to HPV6, 11, 16, 18 and 45 antigens during follow-up was found in 24.6, 11.5, 5.7, 5.7 and 0.8 %, respectively. Seroconversion to LR-HPV was negatively related to a higher number of children and oral sex, and positively associated with seroconversion to HR-HPV. In multivariate analysis, the same predictors remained significant except for the number of children. In univariate generalised estimating equations (GEE) for HR-HPV, being seroconverted to LR-HPV was the only predictor, but lost its significance in multivariate analyses. Decay of all HPV L1 antibodies was rare and observed in 0-2 %. The HPV antibody profile in men was dominated by response to HPV6, also showing the highest cumulative seroconversion. Oral HPV infection might affect HPV serology: (1) HPV DNA in oral mucosa is associated with baseline HR-HPV seropositivity and (2) practising oral sex significantly reduces longitudinal seroconversion to HPV6 and/or 11.

The clearance of oral high-risk human papillomavirus infection is impaired by long-term persistence of cervical human papillomavirus infection.

Persistence of high-risk (HR-) human papillomavirus (HPV) infection of the uterine cervix increases the risk of cervical cancer. Oral HPV infections are among potential covariates of long-term genotype-specific persistent cervical HR-HPV infections. It is not known whether this persistence reflects inability of the host to reject HPV infections in general. A case-control setting was designed to estimate the covariates of long-term persistent cervical HR-HPV infections using multivariate generalized estimating equation (GEE) models. HPV was detected with PCR using GP05+/GP06+-primers and genotyped for 24 HPVs with a Multimetrix-kit. The cases (n=43) included women who had genotype-specific persistent cervical HR-HPV infection for at least 24 months (24M+) and controls were women who tested repeatedly HPV-negative in their cervical samples (n=52). These women represent a sub-cohort of the Finnish Family HPV Study. The cases differed significantly from the HPV-negative controls in several aspects: they were younger, had a longer mean time to incident oral HPV infection (40.7 versus 23.6 months), longer duration of oral HPV persistence (38.4 versus 14.1 months), and longer time to clearance of their oral HPV infection (50.0 versus 28.2 months). In multivariate GEE analysis, the second pregnancy during the follow up was the only independent predictor with significant protective effect against 24M+ persistent cervical HR-HPV infections, OR of 0.15 (95% CI 0.07-0.34). To conclude, long-term persistent cervical HR-HPV infections are associated with a prolonged clearance of oral HR-HPV infections while new pregnancy protects against persistent cervical HR-HPV infections.

Stable marital relationship protects men from oral and genital HPV infections.

Human papillomavirus (HPV) infections are associated with sexual behavior. Changes in the sexual habits of couples and their impact on male genital and oral HPV infections were determined during 7 years of follow-up (FU). At baseline and 7 years FU, urethral, semen/penile, and oral samples were collected from 46 men and cervical and oral samples of their spouses for HPV DNA detection. Demographic data and risk factors of spouses were recorded by questionnaire at both time points and analyzed for concordance. HPV genotyping was done with the Multimetrix® kit. At baseline, 29.5 % of the male genital and 11 % of their oral samples tested positive. Incident genital HPV infection was found in 23 % and oral infection in 10.9 % of men. Genotype-specific persistence was detected in one man (HPV53) in genital samples. Moderate to almost perfect concordance of changes in sexual habits during FU among spouses were found. Changing partners [p = 0.028; odds ratio (OR) = 15; 95 % confidence interval (CI) 1.355-166.054] and marital status (p = 0.001; 95 % CI 0.000-0.002) increased the risk of incident genital HPV infections. The overall outcome of genital HPV disease in men was linked to the frequency of sexual intercourse (p = 0.023; 95 % CI 0.019-0.026) and changes in marital status (p = 0.022; 95 % CI 0.019-0.026), while oral HPV infections were associated with the number of sexual partners (p = 0.047; 95 % CI 0.041-0.052). Taken together, asymptomatic genital HPV infections among the men were common. The risk of incident genital HPV infections increased among men reporting a change of sexual partner during FU, implicating that a stable marital relationship protects against oral and genital HPV infection.

Smoking increases oral HPV persistence among men: 7-year follow-up study.

Persistent high-risk human papillomavirus (HR-HPV) infection is the key event in the progression of HPV lesions, and more data are urgently needed on asymptomatic oral HPV infections in men. Asymptomatic fathers-to-be (n = 131, mean age 28.9 years) were enrolled in the cohort, sampled by serial oral scrapings at baseline and at 2-month, 6-month, 12-month, 24-month, 36-month, and 7-year follow-up visits to accomplish persistent and cleared HPV infections. HPV genotyping was performed using nested PCR and Multimetrix® assay. Covariates of persistent and cleared oral HPV infections were analysed using generalised estimating equation (GEE) and Poisson regression. Altogether, 17 HPV genotypes were detected in male oral mucosa point prevalence, varying from 15.1 % to 31.1 %. Genotype-specific HPV persistence was detected in 18/129 men the mean persistence time ranging from 6.0 to 30.7 months. History of genital warts decreased (p = 0.0001; OR = 0.41, 95 % CI 0.33-0.51) and smoking increased (p = 0.033, OR = 1.92, 95 % CI 1.05-3.50) the risk of persistent species 7/9 HPV infections. Of the 74 HPV-positive men, 71.6 % cleared their infection actuarial and crude clearance times, varying between 1.4 and 79.6 months. No independent predictors were identified for species 7/9 clearance. At the last follow-up-visit, 50.1 % of the fathers had oral mucosal changes, correlating only with smoking (p = 0.046). To conclude, most of the persisting oral infections in males were caused by HPV16. Smoking increased while previous genital warts decreased oral HR-HPV persistence. No predictors of HR-HPV clearance were disclosed.

Effects of tacrolimus on an organotypic raft-culture model mimicking oral mucosa.

BACKGROUND: Tacrolimus ointment has shown efficacy in treating T-cell-mediated inflammatory oral mucosal diseases, including lichen planus. However, the safety of topical tacrolimus has been questioned, based on its possible association with malignant transformation. AIM: To evaluate the safety aspects of tacrolimus in a three-dimensional in vitro model of oral mucosa containing both multilayered epithelium and connective tissue (raft culture). METHODS: Raft cultures mimicking oral mucosa were topically exposed to tacrolimus, and the effects on cell proliferation and adhesion, epidermal growth factor receptors (EGFR, ERBB2, ERBB3, ERBB4), and apoptosis were evaluated with immunohistochemistry and terminal dUTP nick-end labelling, respectively. Results. The epithelium of the cultures was found to be slightly thinner, but no changes in cell proliferation or adhesion, apoptosis, or expression of epidermal growth factor receptors were detected. CONCLUSIONS: Our results suggest that short-term topical tacrolimus exposure of in vitro constructed oral mucosa does not induce changes in a number of factors known to be involved in malignant transformation.

ERBB receptors in developing, dysplastic and malignant oral epithelia.

Some oral squamous cell carcinomas (OSCCs) overexpress epidermal growth factor receptor (EGFR) but little is known about the receptor system overall during oral carcinogenesis. We studied all four ERBB receptors (EGFR, ERBB2-4) in developing (n=2), normal (n=7), dysplastic (n=23) and malignant (n=26) oral epithelia by means of immunohistochemistry. The investigations were supplemented by conducting reverse transcription-polymerase chain reactions in relation to 13 OSCC samples. All four ERBB receptors were detected in developing oral epithelium and, to a lesser degree, in mature oral epithelium. An increase in EGFR immunoreactivity was seen in 61% and 54% of dysplasias and OSCCs, respectively. The corresponding percentages for ERBB2 were 48 and 12, for ERBB3 48 and 43. ERBB4 nuclear staining was increased in 30% of dysplasias and 26% of OSCCs. Changes in ERBB receptor mRNA levels were not statistically significant. The results show that ERBB receptor profiles are specific to each tumour. Increased nuclear translocation of ERBB4 in some OSCCs may alter transcription of target genes and be associated with cancer progression. This information may be useful for clinicians as EGFR inhibitors are becoming treatment options in modern oncology.

Different mechanisms of syndecan-1 activation through a fibroblast-growth-factor-inducible response element (FiRE) in mucosal and cutaneous wounds.

Syndecan-1 expression is enhanced in cutaneous and mucosal wounds. We have previously demonstrated that wounding-induced syndecan-1 expression in the skin occurs transcriptionally, through a fibroblast-growth-factor-inducible element (FiRE). Here, we show that FiRE is also activated in mucosal wounds. However, both the expression patterns and the activation mechanisms of FiRE are different from those in the skin. In the mucosa in vivo, the activation starts and ends earlier than in cutaneous wounds. FiRE is first detected at around 12 hours in keratinocytes, and the activation declines by the third day after wounding occurs. The activation is seen on the migrating sheet of epithelial mucosa, as in the case of cutaneous wounding. In contrast to the situation in vivo, organ-cultured mucosal wounds exhibit no FiRE activity, while organ-cultured cutaneous wounds show robust activity. Activation in mucosal wounds is enhanced, however, by the application of epidermal growth factor. This suggests that exogenous growth factor activity is required for activation of syndecan-1 in mucosal wounds but not in cutaneous wounds.