Specific mesothelial signature marks the heterogeneity of mesenchymal stem cells from high-grade serous ovarian cancer.

Mesenchymal stem/stromal cells (MSCs) are the precursors of various cell types that compose both normal and cancer tissue microenvironments. In order to support the widely diversified parenchymal cells and tissue organization, MSCs are characterized by a large degree of heterogeneity, although available analyses of molecular and transcriptional data do not provide clear evidence. We have isolated MSCs from high-grade serous ovarian cancers (HG-SOCs) and various normal tissues (N-MSCs), demonstrated their normal genotype and analyzed their transcriptional activity with respect to the large comprehensive FANTOM5 sample dataset. Our integrative analysis conducted against the extensive panel of primary cells and tissues of the FANTOM5 project allowed us to mark the HG-SOC-MSCs CAGE-seq transcriptional heterogeneity and to identify a cell-type-specific transcriptional activity showing a significant relationship with primary mesothelial cells. Our analysis shows that MSCs isolated from different tissues are highly heterogeneous. The mesothelial-related gene signature identified in this study supports the hypothesis that HG-SOC-MSCs are bona fide representatives of the ovarian district. This finding indicates that HG-SOC-MSCs could actually derive from the coelomic mesothelium, suggesting that they might be linked to the epithelial tumor through common embryological precursors.

Effects of age and heart failure on human cardiac stem cell function.

Currently, it is unknown whether defects in stem cell growth and differentiation contribute to myocardial aging and chronic heart failure (CHF), and whether a compartment of functional human cardiac stem cells (hCSCs) persists in the decompensated heart. To determine whether aging and CHF are critical determinants of the loss in growth reserve of the heart, the properties of hCSCs were evaluated in 18 control and 23 explanted hearts. Age and CHF showed a progressive decrease in functionally competent hCSCs. Chronological age was a major predictor of five biomarkers of hCSC senescence: telomeric shortening, attenuated telomerase activity, telomere dysfunction-induced foci, and p21(Cip1) and p16(INK4a) expression. CHF had similar consequences for hCSCs, suggesting that defects in the balance between cardiomyocyte mass and the pool of nonsenescent hCSCs may condition the evolution of the decompensated myopathy. A correlation was found previously between telomere length in circulating bone marrow cells and cardiovascular diseases, but that analysis was restricted to average telomere length in a cell population, neglecting the fact that telomere attrition does not occur uniformly in all cells. The present study provides the first demonstration that dysfunctional telomeres in hCSCs are biomarkers of aging and heart failure. The biomarkers of cellular senescence identified here can be used to define the birth date of hCSCs and to sort young cells with potential therapeutic efficacy.

Myocyte turnover in the aging human heart.

RATIONALE: The turnover of cardiomyocytes in the aging female and male heart is currently unknown, emphasizing the need to define human myocardial biology. OBJECTIVE: The effects of age and gender on the magnitude of myocyte regeneration and the origin of newly formed cardiomyocytes were determined. METHODS AND RESULTS: The interaction of myocyte replacement, cellular senescence, growth inhibition, and apoptosis was measured in normal female (n=32) and male (n=42) human hearts collected from patients 19 to 104 years of age who died from causes other than cardiovascular diseases. A progressive loss of telomeric DNA in human cardiac stem cells (hCSCs) occurs with aging and the newly formed cardiomyocytes inherit short telomeres and rapidly reach the senescent phenotype. Our data provide novel information on the superior ability of the female heart to sustain the multiple variables associated with the development of the senescent myopathy. At all ages, the female heart is equipped with a larger pool of functionally competent hCSCs and younger myocytes than the male myocardium. The replicative potential is higher and telomeres are longer in female hCSCs than in male hCSCs. In the female heart, myocyte turnover occurs at a rate of 10%, 14%, and 40% per year at 20, 60, and 100 years of age, respectively. Corresponding values in the male heart are 7%, 12%, and 32% per year, documenting that cardiomyogenesis involves a large and progressively increasing number of parenchymal cells with aging. From 20 to 100 years of age, the myocyte compartment is replaced 15 times in women and 11 times in men. CONCLUSIONS: The human heart is a highly dynamic organ regulated by a pool of resident hCSCs that modulate cardiac homeostasis and condition organ aging.

Are PSA density and PSA density of the transition zone more accurate than PSA in predicting the pathological stage of clinically localized prostate cancer?

PURPOSE: To assess whether PSA density (PSAD) and PSA density of the transition zone (PSADTZ) are more accurate than PSA alone in predicting the pathological stage of prostate cancer. MATERIALS AND METHODS: One hundred and nine consecutive patients with clinically localized prostate cancer and preoperative PSA values over the whole range, treated with radical retropubic prostatectomy and limited pelvic lymph node dissection were included in this prospective study. Total prostate and transition zone volumes were measured by transrectal ultrasound using the prolate ellipsoid method. PSA, PSAD, and PSADTZ were compared to percentage of positive biopsy cores (% PC), biopsy and surgical Gleason score, and pathological stage, using univariate and multivariate analysis. RESULTS: Pathological stage was pT2a, pT2b, pT3a, and pT3b in 25.6%, 37.7%, 25.6%, and 11.1% of patients, respectively. Lymph node metastases were found in 4.6% of patients. PSA, PSAD, and PSADTZ were significantly related to % PC, biopsy, and surgical Gleason score and pathological stage (P < 0.001), and were equally able to predict higher pathological stage, i.e., seminal vesicle invasion and lymph node metastases. Only by adding % PC in multivariate analysis was it possible to discriminate intra- from extracapsular tumors. CONCLUSIONS: The results of the present study demonstrate that PSAD and PSADTZ failed to outperform PSA in preoperative stage prediction of prostate cancer, possibly because the formula used to calculate them does not eliminate the contribution to total PSA of the nonmalignant portion of the gland.

Prevention of postoperative recurrence of Crohn's disease by infliximab.

The prevention of the recurrence of Crohn's disease after surgery remains difficult. The monoclonal antibody anti-TNF-alpha, infliximab, is very effective in inducing and maintaining the remission of uncomplicated, active Crohn's disease. We present here the case of a 23-year-old white woman who underwent resection for a sigmoid stricture caused by Crohn's disease. Surgery removed the involved colon, and pathology confirmed the stricture to be fibrotic. Two weeks after the operation she was given infliximab at the dose of 5 mg/kg body weight and followed in time. Since then, she has been disease free for approximately 4 years after surgery on clinical, radiological and endoscopic/histological grounds (Crohn's Disease Activity Index < or = 110 on all occasions). Up to now, she has had no increase in inflammatory indices, no anaemia and no abnormal blood tests. In contrast, all of five control patients operated in the same period with colonic or ileocolonic resection for symptomatic strictures and treated with mesalamine or no medication developed endoscopic or clinical recurrence (abdominal pain or diarrhoea) by year 3. This is the first case, to our knowledge, in which infliximab has been successfully used to prevent the postsurgical recurrence of Crohn's disease, an event so far considered to be inescapable. We believe that, with this aim in mind, clinical trials with this drug are warranted.

Neovascularization and mast cells with tryptase activity increase simultaneously with pathologic progression in human endometrial cancer.

OBJECTIVE: In vitro and in vivo studies have linked mast cell (MC) degranulation and activation with angiogenesis and neovascularization. This assumption is partially supported by the close anatomic association between MC and the vasculature and the recruitment of these cells during tumor growth. The aim of this study was to correlate the extent of angiogenesis with the number of MC expressing tryptase in human endometrial adenocarcinoma. STUDY DESIGN: Tissues from human endometrial hyperplasia and endometrial adenocarcinoma were investigated immunohistochemically, using 2 murine monoclonal antibodies against the endothelial cell marker CD31 and the MC marker tryptase. RESULTS: Angiogenesis, measured as microvessel counts, was highly correlated with MC tryptase-positive cell counts and that these parameters increase in agreement with tumor progression. CONCLUSION: These results suggest that angiogenesis in endometrial cancer increases with tumor progression and that angiogenic tryptase secreted by host MC cooperate in its induction.

A pixel-based autofocusing technique for digital histologic and cytologic slides.

The present paper describes a method for autofocusing specifically studied for the acquisition of digital slides, i.e. full histologic and cytologic slides, utilising low-cost equipment. At first, experimentations with some of the most used focus measures and algorithms have been made, in order to choose the most suitable for histologic and cytologic images. Then, a study of the specific features of digital slides has been preliminarily carried out in order to understand the constraints of the domain. These included the capability of autofocusing in an unattended way on thousands of microscope fields, while fast performance is not a strict requirement. Based on the findings, an algorithm based on a dynamic focus position space, with a variance-based focus measure has been adapted to the specific situation. A qualitative and quantitative evaluation of the proposed algorithm allowed us to show that the proposed algorithm is suitable for the acquisition of digital slides, and furthermore it can be implemented starting from a basic microscope with an inexpensive motorised stage. The algorithm is currently implemented into a complete digital slide acquisition system, which is in turn being used for a Quality Assurance Programme in cervicovaginal cytologic screening.

Adipose tissue derived stem cells: in vitro and in vivo analysis of a standard and three commercially available cell-assisted lipotransfer techniques.

INTRODUCTION: Autologous fat grafting is commonly used to correct soft-tissue contour deformities. However, results are impaired by a variable and unpredictable resorption rate. Autologous adipose-derived stromal cells in combination with lipoinjection (cell-assisted lipotransfer) seem to favor a long-term persistence of fat grafts, thus fostering the development of devices to be used in the operating room at the point of care, to isolate the stromal vascular fraction (SVF) and produce SVF-enhanced fat grafts with safe and standardized protocols. Focusing on patients undergoing breast reconstruction by lipostructure, we analyzed a standard technique, a modification of the Coleman's procedure, and three different commercially available devices (Lipokit, Cytori, Fastem), in terms of 1) ability to enrich fat grafts in stem cells and 2) clinical outcome at 6 and 12 months. METHODS: To evaluate the ability to enrich stem cells, we compared, for each patient (n=20), the standard lipoaspirate with the respective stem cell-enriched one, analyzing yield, immunophenotype and colony-forming capacity of the SVF cells as well as immunophenotype, clonogenicity and multipotency of the obtained adipose stem cells (ASCs). Regarding the clinical outcome, we compared, by ultrasonography imaging, changes at 6 and 12 months in the subcutaneous thickness of patients treated with stem-cell enriched (n=14) and standard lipoaspirates (n=16). RESULTS: Both methods relying on the enzymatic isolation of primitive cells led to significant increase in the frequency, in the fat grafts, of SVF cells as well as of clonogenic and multipotent ASCs, while the enrichment was less prominent for the device based on the mechanical isolation of the SVF. From a clinical point of view, patients treated with SVF-enhanced fat grafts demonstrated, at six months, a significant superior gain of thickness of both the central and superior-medial quadrants with respect to patients treated with standard lipotransfer. In the median-median quadrant the effect was still persistent at 12 months, confirming an advantage of lipotransfer technique in enriching improving long-term fat grafts. CONCLUSIONS: This comparative study, based on reproducible biological and clinical parameters and endpoints, showed an advantage of lipotransfer technique in enriching fat grafts in stem cells and in favoring, clinically, long-term fat grafts.

Introducing videoconferencing into educational oncopathology seminars: technical aspects, user satisfaction and open issues.

We used set-top videoconferencing equipment connected by ISDN at 384 kbit/s for six educational seminars held between the University of Udine (the local site) and the National Cancer Institute in Aviano (the remote site), 60 km away. User satisfaction was evaluated by questionnaire. The median length of seminars was 58 min (range 48-61 min), followed by a 20 min (15-26 min) discussion. Eighty-two users answered the questionnaire (a 43% response rate): 56 in Udine (a median of 11 per seminar) and 26 in Aviano (a median of 5 per seminar). Answers to the questions were similar at the two sites. Videoconferencing did not affect the users' experience of attending the seminars, as both interest and clarity were similar at the local and remote site. The results suggested that videoconferencing is a viable method for delivering seminars in oncopathology, where image quality is important.

Preliminary results from a crowdsourcing experiment in immunohistochemistry.

BACKGROUND: Crowdsourcing, i.e., the outsourcing of tasks typically performed by a few experts to a large crowd as an open call, has been shown to be reasonably effective in many cases, like Wikipedia, the Chess match of Kasparov against the world in 1999, and several others. The aim of the present paper is to describe the setup of an experimentation of crowdsourcing techniques applied to the quantification of immunohistochemistry. METHODS: Fourteen Images from MIB1-stained breast specimens were first manually counted by a pathologist, then submitted to a crowdsourcing platform through a specifically developed application. 10 positivity evaluations for each image have been collected and summarized using their median. The positivity values have been then compared to the gold standard provided by the pathologist by means of Spearman correlation. RESULTS: Contributors were in total 28, and evaluated 4.64 images each on average. Spearman correlation between gold and crowdsourced positivity percentages is 0.946 (p < 0.001). CONCLUSIONS: Aim of the experiment was to understand how to use crowdsourcing for an image analysis task that is currently time-consuming when done by human experts. Crowdsourced work can be used in various ways, in particular statistically agregating data to reduce identification errors. However, in this preliminary experimentation we just considered the most basic indicator, that is the median positivity percentage, which provided overall good results. This method might be more aimed to research than routine: when a large number of images are in need of ad-hoc evaluation, crowdsourcing may represent a quick answer to the need.