Soemmerring's Rings Developed around IOLs, in Human Donor Eyes, Can Present Internal Transparent Areas.

Soemmerring's rings consist of a ring of lens epithelial derived cells that grow along the periphery of an aphakic lens capsule, or around an intraocular lens. These rings when visualized frontally, appear opaque, however, in some cases the cells that compose these rings are organized in the same fashion as those in normal transparent adult lenses. Thus, our purpose was to test whether any part of the adult Soemmerring's ring could be transparent and how this related to morphological factors. To study this, 16 Soemmerring's rings were extracted from donor eye globes. After imaging, they were thickly sectioned sagittally in order to analyze the degrees of transparency of different areas. All samples were also histologically analyzed using alpha smooth muscle actin, Vimentin, wheat germ agglutinin and DAPI. Our results showed that many samples had some transparent areas, mostly towards the center of their cross-section. Of the factors that we analyzed, only lens fiber organization at the bow region and an increased area of mature lens fiber cells had a significant relation to the degree of transparency at the center. Thus, we can conclude that as Soemmerring's rings mature, they can develop organized and transparent areas of lens cells.

Detection of somatic mutations in peritoneal lavages and plasma of endometrial cancer patients: A proof-of-concept study.

Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries. Although most patients are diagnosed at early stages, 15-20% will relapse despite local treatment. Presently, there are no reliable markers to identify patients with worse outcomes who may benefit from adjuvant treatments, such as chemotherapy, and liquid biopsies may be of use in this setting. Peritoneal lavages are systematically performed during endometrial surgery but little data are available about their potential as liquid biopsies. We analyzed KRAS and PIK3CA mutations in paired surgical biopsies, blood and cytology-negative peritoneal lavages in a cohort of 50 EC patients. Surgical biopsies were submitted to next-generation sequencing (NGS) while circulating-free DNA (cfDNA) purified from plasma and peritoneal lavages was analyzed for KRAS and PIK3CA hotspot mutations using a sensitive quantitative polymerase chain reaction (PCR) assay. NGS of biopsies revealed KRAS, PIK3CA or concomitant KRAS + PIK3CA mutations in 33/50 (66%) EC patients. Of those, 19 cases carried hotspot mutations. Quantitative PCR revealed KRAS and/or PIK3CA mutations in the lavages of 9/19 (47.4%) hotspot EC patients. In contrast, only 2/19 (10.5%) blood samples from hotspot EC patients were positive. Mutations found in cfDNA consistently matched those in paired biopsies. One of the two patients positive in plasma and lavage died in less than 6 months. In conclusion, mutational analysis in peritoneal lavages and blood from early stage EC is feasible. Further studies are warranted to determine if it might help to identify patients with worse prognosis. Human genes discussed: KRAS, KRAS proto-oncogene, GTPase; PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha.

Breast density measured volumetrically in a clinical environment: cross-sectional study with photon counting technology.

OBJECTIVE: Mammographic breast density (BDen), the ratio of glandular volume (GVol) to breast volume (BVol), is the second most prevalent risk factor for breast cancer (BC). Newly developed photon counting technology allows precise and systematic measurements in clinical practice. Our objective is to see how these parameters change with age in women with and without cancer. MATERIALS AND METHODS: This retrospective study analyzed results of BDen, GVol, and BVol in 64,182 mammograms performed with photon counting technology on 32,448 consecutive women from April 2014 to December 2015. Only their first study was included. We excluded women with incomplete data or with breast implants. RESULTS: Mean age of women without BC diagnosed during the study period was 52.1 ± 9.9. BC and was found in 263 women (0.81%). Mean age was 53.0 ± 10.4. BDen, GVol, and BVol were 14%, 24%, and 2% greater in women with BC (P < 0.001 for BDen and GVol and P = 0.02 for BVol). BDen and GVol diminished following similar patterns across age in both groups, with soft slopes before and after a steep drop from 50 to 60, probably due to menopause. CONCLUSION: BDen diminishes with age in women with or without BC, but it is generally higher in women with BC. GVol could be a more robust indicator associated with BC risk than BDen. This technology can ease the way to studies of interventions to diminish BDen (or GVol) in the hope of diminishing BC incidence or predict if longitudinal changes are indicative of impending cancer.

Changes in breast density during hormone treatment with transdermal estrogens alone or in combination with progesterone.

A retrospective observational study to assess whether hormonal treatment (HT) with transdermal estrogens alone or in combination with micronized progesterone increases breast density and to compare these changes to those of a control group of 4120 patients were not given HT. We included 150 patients whose baseline breast density was assessed with photon-counting spectral mammography and 1 year after hormone treatment. The reduction in breast density was compared using an analysis of covariance. The difference in breast density between mammographies in the HT group was -0.40 ± 5.5 and -0.85 ± 4.2 in the control group. The changes in density according to the type of HT, we found that women on treatment with estrogen alone presented a difference of 0.44 ± 5.8, and -1.35 ± 5 (p = 0.13) in women on combined treatment. After adjusting changes in density for age and average number of days between mammographies, we observed a difference of -0.36 95% confidence intervals (CI) [-1.04 to -0.31] in the women on HT and -0.71 95% CI [-1.65 to -0.21] in the control group. No increased breast density was observed in women on HT treatment, nor did we observe an increase according to HT type. The difference in breast density loss was smaller in the HT group versus the control group.

Impaired Expression of Ectonucleotidases in Ectopic and Eutopic Endometrial Tissue Is in Favor of ATP Accumulation in the Tissue Microenvironment in Endometriosis.

Endometriosis is a prevalent disease defined by the presence of endometrial tissue outside the uterus. Adenosine triphosphate (ATP), as a proinflammatory molecule, promotes and helps maintain the inflammatory state of endometriosis. Moreover, ATP has a direct influence on the two main symptoms of endometriosis: infertility and pain. Purinergic signaling, the group of biological responses to extracellular nucleotides such as ATP and nucleosides such as adenosine, is involved in the biology of reproduction and is impaired in pathologies with an inflammatory component such as endometriosis. We have previously demonstrated that ectonucleotidases, the enzymes regulating extracellular ATP levels, are active in non-pathological endometria, with hormone-dependent changes in expression throughout the cycle. In the present study we have focused on the expression of ectonucleotidases by means of immunohistochemistry and in situ activity in eutopic and ectopic endometrial tissue of women with endometriosis, and we compared the results with endometria of women without the disease. We have demonstrated that the axis CD39-CD73 is altered in endometriosis, with loss of CD39 and CD73 expression in deep infiltrating endometriosis, the most severe, and most recurring, endometriosis subtype. Our results indicate that this altered expression of ectonucleotidases in endometriosis boosts ATP accumulation in the tissue microenvironment. An important finding is the identification of the nucleotide pyrophophatase/phosphodiesterase 3 (NPP3) as a new histopathological marker of the disease since we have demonstrated its expression in the stroma only in endometriosis, in both eutopic and ectopic tissue. Therefore, targeting the proteins directly involved in ATP breakdown could be an appropriate approach to consider in the treatment of endometriosis.

Histopathology Findings of Corneal Buttons in Congenital Aniridia Patients.

PURPOSE: To evaluate the corneal button of primary penetrating keratoplasty of patients diagnosed with congenital aniridia. The study took place at the Instituto Universitario Barraquer and the Centro de Oftalmología Barraquer. METHODS: A retrospective analysis of cases diagnosed with congenital aniridia was carried out. We analyzed 13 corneal buttons of 11 eyes with congenital aniridia. We only included those patients who underwent penetrating keratoplasty for the first time. The corneal buttons were analyzed for histological characteristics of the presence of vascularization, the presence or not of Bowman's layer, the thickness of the stroma and Descemet's membrane, and endothelium layer alterations. RESULTS: We found alterations in the epithelium and stroma in all patients, although this loss of architecture was not seen in Descemet's membrane and the endothelial population. CONCLUSION: Patients with advanced congenital aniridic keratopathy may be good candidates for deep or superficial anterior lamellar keratoplasty for the preservation of normal endothelium and Descemet's membrane, along with limbal stem cell transplantation, to address epithelial and stromal pathology.

Endoresection technique with/without brachytherapy for management of high posterior choroidal melanoma: extended follow-up results.

PURPOSE: To evaluate the long-term efficacy and safety of endoresection for high posterior choroidal melanoma. METHODS: Retrospective nonrandomized interventional case series. Forty-one patients had endoresection as primary treatment for posterior choroidal melanoma. Of these, 21 patients had adjuvant brachytherapy. The inclusion criteria were tumor thickness ≥8 mm, base diameter <15 mm, and posterior tumors not extending anterior to the equator. Main outcomes measures were enucleation rate, visual outcome, surgical complications, local recurrence, metastasis, and mortality. RESULTS: Mean follow-up time was 102.5 months. Mean preoperative best-corrected visual acuity was 20/100. Mean tumor thickness was 9.8 mm (range, 7.7-13.5 mm; standard deviation, 1.7 mm), mean base diameter was 9.9 mm (range, 5-15 mm; standard deviation, 2.8 mm). At the latest visit, 36 patients (87.8%) still retained the eye. Mean postoperative best-corrected visual acuity was 20/1,625. Retinal detachment was the main postoperative complication (28.9%). At completion of follow-up, 12% of patients had phthisis bulbi, and 3 developed chronic hypotony. Five patients (12.2%) had local tumor recurrence; none of them had received brachytherapy as initial treatment. At 5 years of follow-up, 3 patients (7.3%) had liver metastasis. On Kaplan-Meier analysis at 10 years, all-cause mortality was 7.3% and specific mortality because of melanoma was 2.4%. CONCLUSION: Endoresection of high posterior melanomas was not associated with a higher risk of metastasis, death, or local recurrence than other reported techniques used to treat similar melanomas.

Converging and evolving immuno-genomic routes toward immune escape in breast cancer.

The interactions between tumor and immune cells along the course of breast cancer progression remain largely unknown. Here, we extensively characterize multiple sequential and parallel multiregion tumor and blood specimens of an index patient and a cohort of metastatic triple-negative breast cancers. We demonstrate that a continuous increase in tumor genomic heterogeneity and distinct molecular clocks correlated with resistance to treatment, eventually allowing tumors to escape from immune control. TCR repertoire loses diversity over time, leading to convergent evolution as breast cancer progresses. Although mixed populations of effector memory and cytotoxic single T cells coexist in the peripheral blood, defects in the antigen presentation machinery coupled with subdued T cell recruitment into metastases are observed, indicating a potent immune avoidance microenvironment not compatible with an effective antitumor response in lethal metastatic disease. Our results demonstrate that the immune responses against cancer are not static, but rather follow dynamic processes that match cancer genomic progression, illustrating the complex nature of tumor and immune cell interactions.

Atrophic congenital melanocytic nevus mimicking rapidly involuting congenital hemangioma.

Congenital melanocytic nevi (CMN) are benign skin lesions present in some newborns. We describe a newborn boy with a giant CMN on the scalp. It appeared as an atrophic patch with a central nodule that was suggestive of a rapidly involuting congenital hemangioma or some other vascular tumor. Histopathology confirmed the diagnosis of CMN. Magnetic resonance imaging was normal, excluding leptomeningeal melanocytosis. We did not find any other cases of CMN with this presentation in the literature.

Reference standards for gene fusion molecular assays on cytological samples: an international validation study.

AIMS: Gene fusions assays are key for personalised treatments of advanced human cancers. Their implementation on cytological material requires a preliminary validation that may make use of cell line slides mimicking cytological samples. In this international multi-institutional study, gene fusion reference standards were developed and validated. METHODS: Cell lines harbouring EML4(13)-ALK(20) and SLC34A2(4)-ROS1(32) gene fusions were adopted to prepare reference standards. Eight laboratories (five adopting amplicon-based and three hybridisation-based platforms) received, at different dilution points two sets of slides (slide A 50.0%, slide B 25.0%, slide C 12.5% and slide D wild type) stained by Papanicolaou (Pap) and May Grunwald Giemsa (MGG). Analysis was carried out on a total of 64 slides. RESULTS: Four (50.0%) out of eight laboratories reported results on all slides and dilution points. While 12 (37.5%) out of 32 MGG slides were inadequate, 27 (84.4%) out of 32 Pap slides produced libraries adequate for variant calling. The laboratories using hybridisation-based platforms showed the highest rate of inadequate results (13/24 slides, 54.2%). Conversely, only 10.0% (4/40 slides) of inadequate results were reported by laboratories adopting amplicon-based platforms. CONCLUSIONS: Reference standards in cytological format yield better results when Pap staining and processed by amplicon-based assays. Further investigation is required to optimise these standards for MGG stained cells and for hybridisation-based approaches.