Distribution of oximetry data in treatment and control groups.

Distribution of oximetry data in treatment and control groups.</p

SpO₂ measurement area in perineum region.

Dyspareunia is genital pain before, during or after penile-vaginal sexual intercourse. The prevalence of dyspareunia ranges from 8 to 22%. Sexual intercourse concomitant with a pelvic organic lesion is likely to cause pain in most cases. However, in these cases, the pain depends not only on sexual intercourse. In its basic definition, dyspareunia in women is considered an idiopathic affection without a typical organic constitution. It is only present with penile-vaginal penetration. Long-term hypoxia in perineal muscles can cause muscle and perimuscular changes, leading to chronic pain not sufficiently responding to standard therapy. During the entrance examination to our previous study on dyspareunia, we noted significantly lower pulse oximetry levels in the perineal area of affected women. We aimed to compare pulse oximetry oxygen saturation (SpO2) of dyspareunia-affected women to healthy, pain-free women. A retrospective study was performed. The study participants were women who had participated in our previously published study on dyspareunia. This retrospective study was approved by the Ethical Committee. The study included 62 women: 31 dyspareunia-affected women in the treatment group and 31 healthy women in the control group. Method: During their examinations, women in the dyspareunia and control groups were measured for SpO2. The procedure was performed in the vulvo-perineal rear region, involving the commissure and the bulbospongiosus muscle. Median and mean SpO2 were compared between the treatment and control groups. Testing for sample size accuracy was performed retroactively. Results: There were 31 participants in each group. The SpO2 data were skewed and did not follow a Gaussian distribution. The Mann-Whitney U test was run to determine differences in perineum oximetry between the treatment group and controls. The median SpO2 was 91 in the treatment group and 92 in the control group. This difference was statistically significant, p = 0.002. Sample size accuracy was assured by post hoc calculation. Conclusions: Idiopathic dyspareunia is inherent in cohabitation muscle pain that standard therapy could not explain nor treat. We detected clinically meaningfully decreased levels of SpO2 in affected patients. We compared pelvic oximetry between dyspareunia-affected women in the treatment and control groups. This comparison showed significant hypoxia in the perineal muscle area (p = 0.002). Our results may help us understand the source of this pain and guide treatment accordingly.</div

Data sets used in this study.

<p>Cervical cells collected three years before half of the patients developed dysplasia (BDy), normal versus dysplastic cervical cells (Dy), normal cervical tissue and invasive cervical cancer (Cvx CA), non-neoplastic breast tissue and breast cancer (BC), normal endometrium and endometrial cancer (EC) and metastatic endometrial cancer (EC (Meta)), ovarian cancer tissue (OvC; The Cancer Genome Atlas, TCGA), colon cancer tissue (ColC), lung cancer tissue (LC) and normal and cancer associated lung fibroblasts (LC Fibro), and white blood cell (WBC) samples from <i>BRCA1</i> mutation carriers and controls (<i>BRCA1</i>). The number of samples, their distribution in terms of normal and cancer, cell-type, age-range (years) and reference to data access is given. GSE numbers are GEO accessions. NA, not available.</p

Heatmap of the top 50 most frequently hypomethylated MESCs in cancers.

<p>The CpGs show stable fully methylated states across all normal samples. Methylation values: blue = high methylation (β-value>0.7), skyblue = hemi methylation (0.25<β-value<0.7), yellow = low methylation (β-value<0.25). Sample labels at the top of the heatmaps: normal (grey) and cancer (black).</p

Survival analysis of the MESCs demethylation instability index in various cancers.

<p>Kaplan-Meier survival curves between the upper (blue) and lower (green) tertiles of the demethylation instability index (DeMI) at MESCs in (A) Endometrial cancer, (B) Ovarian cancer, (C) Cervical cancer, and (D–E) two Breast Cancer cohorts. The hazard ratio (HR), 95% confidence interval (CI) and P-values are from the multivariate Cox regression model, with “n” denoting the number of samples in cohort. Clinical endpoint used is indicated on the y-axis (OS = overall survival, RFS = relapse free survival).</p

Anti-correlation between the TET mRNA expression level and methylation β-value.

<p>Heatmap of the 250 most (upper half) and least (lower half) anti-correlated hypomethylated CpGs in cervical cancer samples ranked according to their TET3 mRNA expression levels from the lowest (green) to the highest (red) with the indication of MESCs (purple) and nonMESCs (white). The odds ratio (OR) and P-value (P) are obtained from Fisher's exact test estimating the enrichment of MESCs among hypomethylated CpGs that are significantly anti-correlated with TET3 mRNA expression level.</p

Methylation changes between normal, primary, and metastatic endometrial cancer.

<p>(A) Boxplots comparing the frequency of PCGT (0→1/2) DNA methylation changes, and the frequency of combined MESC (1→0) and MESC (2→0/1) DNA methylation changes (“combined DeMI index”), between normal endometrium (N), primary endometrial cancer (C), and between normal endometrium and metastatic endometrial cancers (MET). One-tailed Wilcoxon rank sum test P-values for the instability indices between cancer and metastases are indicated. (B) Receiver operating characteristic (ROC) curves measuring the dissimilarity in the combined DeMI index between primary and metastatic endometrial cancers with corresponding Area Under Curve (AUC) and 95% CI.</p

Differential dynamics of hypermethylated and hypomethylated PCGTs and MESCs.

<p>Bar charts representing percentages of significantly hypermethylated (blue) and hypomethylated (orange) PCGT and MESC CpGs in (A) each stage of cervical carcinogenesis: Cervix ‘Before Dysplasia’, ‘Dysplasia’, and ‘Invasive Cancer’, all relative to normal cervical cells or tissue; and in (B) ‘Breast CA’, ‘Endo CA’, ‘Colon CA’, and ‘Lung CA’, all relative to their respective normal controls. The significance of the binomial test assessing skew of hypermethylated versus hypomethylated is indicated by ‘*’, ‘**’, and ‘***’ for P-value<0.05, 0.01, and 0.001 respectively. (C) and (D) are the scatterplots of the age-adjusted linear regression t-statistics against their corresponding −log10(P-values) testing the association with the normal and lung cancer fibroblasts on the colon-PMD PCGTs and colon-PMD MESCs respectively.</p

Definition of epigenetic instability indices and dynamics of PCGT and MESC methylation in cancer.

<p>(A) Definition of epigenetic instability indices. Shown are the six possible DNA methylation changes between normal and cancer tissue. Thresholds used to define unmethylated (yellow), hemimethylated (skyblue) and fully methylated (blue) CpGs are described in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002517#s4" target="_blank">Materials and Methods</a>. Stable MESC (or PCGT) CpGs are defined by MESC (or PCGT) CpGs, which have the same methylation state in all normal samples. The Epigenetic Instability Index (EpI) is then defined as the fraction of stable CpGs altered in cancer. We defined 4 separate indices to describe the transitions: 0→1/2, 1→2, 1→0, 2→1/0. The index describing alterations from a fully methylated to either a hemi or unmethylated state is called the Demethylation instability index (DeMI). (B) Dynamics of PCGT and MESC DNA methylation in cancer. Diagram illustrates the differential dynamics of PCGT and MESC CpG DNA methylation in cancer. Most PCGTs start out unmethylated (white lolly-pops) in normal cells but acquire DNA methylation (black lolly-pops) in normal cells 3 years before the emergence of dysplasia (BDy). PCGT hypermethylation increases further with Dysplasia (Dy) and cancer, but is not a strong determinant of invasion or poor outcome (metastasis). In contrast, most MESCs start out either fully or hemi methylated in normal cells, and gradually lose methylation during the progressive stages of cancer, with hypomethylation at MESCs a key determinant of metastases and poor outcome.</p

Methylation profile of PCGTs and MESCs in cervix data.

<p>Scatter plots of mean β-values in normal cervical tissue (x-axes) vs. cervical cancer tissue (y-axes) of (A) all CpGs with PCGTs highlighted in brown and (B) all CpGs with MESCs highlighted in purple. (C) is the bar chart indicating the enrichment odds ratios (OR) and P-values (Fisher-test), testing for enrichment of PCGTs among CpGs unmethylated in normal cervix (mean β-value<0.2) and with a higher mean β-value in (i) normal samples which develop dysplasia (BDy), (ii) non-invasive dysplastic samples (Dy), and (iii) invasive cervical cancer (CA); (D) the bar chart indicating the enrichment of MESCs among those CpGs methylated in the normal cervix (mean β-value>0.4) and with a lower mean β-value in tissue representing the three stages of cervical carcinogenesis (BDy, Dy, and CA).</p