Diffusion-Weighted MRI in the Evaluation of Early-Stage Breast Cancer Treated with a Short Preoperative Radiotherapy: Preliminary Results.

Objective: To assess tumor response with diffusion-weighted MRI (DW-MRI) after a short preoperative radiotherapy in early-stage breast cancer (BCa). Materials and Methods: This was a prospective, single-center pilot study. 3T-MRI were performed before and after radiotherapy. The longest diameter (LD) and the apparent diffusion coefficient (ADC) value of a region of interest (ROI) of the tumors were recorded. Histopathology and immunohistochemistry, including the Ki-67 index of the core biopsy and of the surgical specimen, were the reference standards. Results: Nineteen patients with 22 early-stage BCa were included. The mean ROI ADC value was 1.093 ± 0.278 × 10-3 mm2/s before radiotherapy and 1.490 ± 0.429 × 10-3 mm2/s (p-value < 0.001) after radiotherapy. The Ki-67 index was 9.2 ± 9.1% at the percutaneous biopsy before radiotherapy and 4.9 ± 7.5% (p-value = 0.005) after radiotherapy at the surgical specimen. After neoadjuvant radiotherapy, a 4.7% decrease in LD and a 36.3% increase in ROI-ADC of the tumors were measured at MRI and a 46.7% decrease in Ki-67 index was observed at histology of the surgical specimen in comparison with the percutaneous core biopsy. Conclusion: In early-stage BCa, a significant increase in ROI-ADC at DWI and a significant decrease in Ki-67 index were observed after a short preoperative radiotherapy, suggesting early tumor response.

Depolarization imaging for fast and non-invasive monitoring of cervical microstructure remodeling in vivo during pregnancy.

The cervix plays a crucial role in conception, maintenance of pregnancy, and childbirth. The mechanical properties of a pregnant woman's cervix change dramatically during gestation due to a remodeling of its microstructure, necessary for delivery. However, external factors can accelerate this process and lead to prematurity, the primary cause of perinatal mortality worldwide, due to the inefficiency of existing diagnostic methods. This study shows that polarized light is a powerful tool to probe the cervical microstructure during pregnancy. A wide-field multispectral polarimetric imaging system was fabricated to explore in vivo the cervix of full-term pregnant women. The polarimetric properties of the cervix change significantly with pregnancy progression. In particular, a set of several depolarization parameters (intrinsic and extrinsic) showed a strong linear correlation with gestational age in the red part of the visible spectral range. This trend can be attributed, among other things, to a decrease in collagen density and an increase in hydration of cervical connective tissue. Wide field depolarization imaging is a very promising tool for rapid and non-invasive analysis of cervical tissue in vivo to monitor the steady progression of pregnancy, providing the practitioner with useful information to improve the detection of preterm birth.

Pre-OPerative accelerated radiotherapy for early stage breast cancer patients (POPART): A feasibility study.

Pre-operative 5-fraction breast radiotherapy followed by immediate breast-sparing surgery and sentinel node procedure was feasible in 14 patients with 15 clinical early-stage breast cancers. However wound problems occurred frequently and was documented in 5 of the 14 patients: 2 patients with a mastitis needing antibiotics, 2 patients developed a fistula with exudate needing antibiotics and local disinfection and 1 patient developed a fistula needing surgical reintervention. Other acute and late iatrogenic events were rather limited. Two patients had a pathological lymph node involvement, which underlines the importance to perform the sentinel node procedure before pre-operative radiotherapy.

Broad-Range Papillomavirus Transcriptome as a Biomarker of Papillomavirus-Associated Cervical High-Grade Cytology.

Human papillomaviruses (HPVs) are responsible for >99% of cervical cancers. Molecular diagnostic tests based on the detection of viral DNA or RNA have low positive predictive values for the identification of cancer or precancerous lesions. Triage with the Papanicolaou test lacks sensitivity; and even when combined with molecular detection of high-risk HPV, this results in a significant number of unnecessary colposcopies. We have developed a broad-range detection test of HPV transcripts to take a snapshot of the transcriptome of 16 high-risk or putative high-risk HPVs in cervical lesions (HPVs 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 73, and 82). The purpose of this novel molecular assay, named HPV RNA-Seq, is to detect and type HPV-positive samples and to determine a combination of HPV reads at certain specific viral spliced junctions that can better correlate with high-grade cytology, reflecting the presence of precancerous cells. In a proof-of-concept study conducted on 55 patients, starting from cervical smears, we have shown that HPV RNA-Seq can detect papillomaviruses with performances comparable to a widely used HPV reference molecular diagnostic kit; and a combination of the number of sequencing reads at specific early versus late HPV transcripts can be used as a marker of high-grade cytology, with encouraging diagnostic performances as a triage test.

Mueller polarimetric imaging of biological tissues: classification in a decision-theoretic framework.

Mueller polarimetry is increasingly recognized as a powerful modality in biomedical imaging. Nevertheless, principled statistical analysis procedures are still lacking in this field. This paper presents a complete pipeline for polarimetric bioimages, with an application to ex vivo cervical precancer detection. In the preprocessing stage, we evaluate the replacement of pixels by superpixels. In the analysis stage, we resort to decision theory to select and tune a classifier. Performances of the retained classifier are evaluated. Decision theory provides a rigorous and versatile framework, allowing generalization to other pathologies, to other imaging procedures, and to classification problems involving more than two classes.

In vivo imaging of uterine cervix with a Mueller polarimetric colposcope.

Mueller polarimetric imaging enables the detection and quantification of modifications of the collagen fibers in the uterine cervix due to the development of a precancerous lesion. This information is not accessible through the use of the classic colposcope, a low magnification microscope used in current practice for cervical cancer screening. However, the in vivo application of Mueller polarimetric imaging poses an instrumental challenge: the device should be sufficiently compact, while still being able to perform fast and accurate acquisition of Mueller matrices in real-world conditions. In this study, the first wide field Mueller Polarimetric Colposcope (MPC) for the in vivo analysis of uterine cervix is presented. The MPC has been fabricated by grafting a miniaturized Mueller polarimetric imager on a classic colposcope. This new imaging tool performs the fast acquisition of Mueller polarimetric images, thus eliminating any blurring effects due to patient movements. It can be easily used by a practitioner with little change to their existing practice. Finally, the MPC was tested in vivo on a number of patients in the field.

Ex vivo Mueller polarimetric imaging of the uterine cervix: a first statistical evaluation.

Early detection through screening plays a major role in reducing the impact of cervical cancer on patients. When detected before the invasive stage, precancerous lesions can be eliminated with very limited surgery. Polarimetric imaging is a potential alternative to the standard screening methods currently used. In a previous proof-of-concept study, significant contrasts have been found in polarimetric images acquired for healthy and precancerous regions of excised cervical tissue. To quantify the ability of the technique to differentiate between healthy and precancerous tissue, polarimetric images of seventeen cervical conization specimens (cone-shaped or cylindrical wedges from the uterine cervix) are compared with results from histopathological diagnoses, which is considered to be the "gold standard." The sensitivity and specificity of the technique are calculated for images acquired at wavelengths of 450, 550, and 600 nm, aiming to differentiate between high-grade cervical intraepithelial neoplasia (CIN 2-3) and healthy squamous epithelium. To do so, a sliding threshold for the scalar retardance parameter was used for the sample zones, as labeled after histological diagnosis. An optimized value of ∼83% is achieved for both sensitivity and specificity for images acquired at 450 nm and for a threshold scalar retardance value of 10.6 deg. This study paves the way for an application of polarimetry in the clinic.

Optimization of Picrosirius red staining protocol to determine collagen fiber orientations in vaginal and uterine cervical tissues by Mueller polarized microscopy.

Polarized microscopy provides unique information on anisotropic samples. In its most complete implementation, namely Mueller microscopy, this technique is well suited for the visualization of fibrillar proteins orientations, with collagen in the first place. However, the intrinsic optical anisotropy of unstained tissues has to be enhanced by Picrosirius Red (PR) staining to enable Mueller measurements. In this work, we compared the orientation mapping provided by Mueller and second harmonic generation (SHG) microscopies on PR stained samples of vaginal and uterine cervix tissues. SHG is a multiphoton technique that is highly specific to fibrillar collagen, and was taken as the "gold standard" for its visualization. We showed that Mueller microscopy can be safely used to determine collagen orientation in PR stained cervical tissue. In contrast, in vaginal samples, Mueller microscopy revealed orientations not only of collagen but also of other anisotropic structures. Thus PR is not fully specific to collagen, which necessitates comparison to SHG microscopy in every type of tissue. In addition to this study of PR specificity, we determined the optimal values of the staining parameters. We found that staining times of 5 min, and sample thicknesses of 5 µm were sufficient in cervical and vaginal tissues.

Determination of collagen fiber orientation in histological slides using Mueller microscopy and validation by second harmonic generation imaging.

We studied the azimuthal orientations of collagen fibers in histological slides of uterine cervical tissue by two different microscopy techniques, namely Mueller polarimetry (MP) and Second Harmonic Generation (SHG). SHG provides direct visualization of the fibers with high specificity, which orientations is then obtained by suitable image processing. MP provides images of retardation (among other polarimetric parameters) due to the optical anisotropy of the fibers, which is enhanced by Picrosirius Red staining. The fiber orientations are then assumed to be those of the retardation slow axes. The two methods, though fully different from each other, provide quite similar maps of average fiber orientations. Overall, our results confirm that MP microscopy provides reliable images of dominant fiber orientations at a much lower cost that SHG, which remains the "gold standard" for specific imaging of collagen fibers using optical microscopy.

Polarimetric imaging of uterine cervix: a case study.

We present a preliminary investigation of macroscopic polarimetric imaging of uterine cervix. Orthogonal state contrast (OSC) images of healthy and anomalous cervices have been taken in vivo at 550 nm. Four ex vivo cervix samples have been studied in full Muller polarimetry, at 550 nm and 700 nm, and characterized in detail by standard pathology. One sample was totally healthy, another one carried CIN lesions at very early stage (CIN1) in its visible exocervical region, while for the other two samples more advanced (CIN3) lesions were present, together with visible glandular epithelium (ectropion). Significant birefringence has been observed in the healthy regions of all six samples, both in vivo and ex vivo. Standard treatments of the Mueller images of the ex vivo samples allowed to quantify both retardation and depolarization. Retardation reached 60° in healthy regions, and disappeared in the anomalous regions of the other three ex vivo samples. The depolarization power was largest in healthy regions, and lower in CINs and ectropion. Possible origins of the observed effects are briefly discussed.

Total adhesions treated by hysteroscopy: must we stop at two procedures?

OBJECTIVE: To study the anatomic and fertility results after treatment for Asherman syndrome involving more than two separate surgical procedures. DESIGN: Retrospective case series. SETTING: Tertiary center. PATIENT(S): Twenty-three women who had Asherman syndrome and required more than two hysteroscopic operative procedures. INTERVENTION(S): Third or higher-order operative hysteroscopy procedure. MEAN OUTCOME MEASURE(S): Fertility rate. RESULT(S): The women's mean age was 34 years (± 5.8 years) when treatment for adhesions began. All women initially had adhesions classified as severe with total amenorrhea. Twelve patients had three separate procedures to treat the adhesions, nine had four treatments, and two had five treatments. One woman was lost to follow-up. At the conclusion of treatment, more than 80% of the women had either no adhesions at all or only mild adhesions. The overall pregnancy rate was 40.9%; there were nine pregnancies and six term infants (27.2%). All but one of these pregnancies were spontaneous. The mean time to pregnancy was 10.5 months (± 4.7 months). CONCLUSION(S): The number of hysteroscopic procedures envisioned to treat Asherman syndrome should not be a limiting factor. It is appropriate to treat women, especially those younger than 35 years, until uterine anatomy permits the visualization of both ostia.