Andrea Galassi

Tissue Factors Influencing Growth and Maintenance of Endometriosisa

The unpredictable response of endometriosis to steroids and its recurrence after therapy, led us to hypothesize a possible further control of this pathology by factors other than steroids. The presence of estrogen, progesterone and epidermal growth factor receptors (ER, PR, EGFr) was evaluated using immunohistochemistry before and after therapy with Danazol or a gonadotropin-releasing hormone analogue (GnRHa), Buserelin. EGFr, ER and PR were present in 100% of endometrial specimens, and in 71%, 29% and 49% of endometriotic implants, respectively. Danazol and GnRHa reduced immunohistochemical staining for EGFr antisera in the endometrial and endometriotic specimens. About 21% of endometriosis were EGFr positive and ER negative, suggesting a potential role of epidermal growth factor in growth and maintenance of endometrial ectopia.

Basement membrane components in normal hyperplastic and neoplastic endometrium

The major basement membrane (BM) components, Iaminin and type IV collagen, were studied by immunochemistry in normal, hyperplastic, and neoplastic endometrium. By immunoperoxidase technique, proliferative and secretive endometrium showed capillary and epithelial cell basement membranes with linear staining with antibodies to both laminin and type IV collagen. Immunostaining of laminin and type IV collagen showed that capillaries were surrounded by a continuous perivascular sheath of these matrices in specimens of adenomatous hyperplasia and in nearly all specimens of endometrial adenocarcinoma. Laminin and type IV collagen were found to accumulate around glandular epithelial cells of adenomatous hyperplastic endometrium, but in several specimens these linear surrounding formations were defective and discontinuous. In several areas of well‐differentiated endometrial adenocarcinomas BM‐like structures were found around glandular epithelial cells as shadows without staining for laminin and type IV collagen. These basement membrane components accumulate around stromal cells to encircle each cell with a gradual, progressive, and cyclic process depending on the phase of the menstrual cycle. Laminin and type IV collagen were clearly detected around stromal cells at days 20 to 22 of the menstrual cycle and more thickly at days 26 to 28. The accumulation of these matrices around stromal cells is a progesterone/progestin‐related process. In the well‐differentiated adenocarcinoma a mid‐term treatment with progestin (Danatrol Maggioni‐Winthrop, SPA, Milan, Italy) was found to be effective on laminin and type IV collagen accumulation around stromal cells.

Basement Membrane in Human Endometrium: Possible Role of Proteolytic Enzymes in Developing Hyperplasia and Carcinoma

Basement membranes (BM) are elements of the extracellular matrix that are essential for growth and differentiation of tissues. Several collagenolytic enzymes of tumor cells are involved in degradation of the extracellular matrix; growth and inhibitor factors [e.g. Epidermal Growth Factor (EGF), Transforming Growth Factors alpha and beta (TGF-alpha, beta)] seem to be involved in the extracellular matrix formation and degradation. To establish a possible association between the presence of collagenase (C), urokinase-type plasminogen activator (uPA) and the neoplastic growth of the endometrium, 44 endometrial specimens (14 proliferative, 11 secretive, 7 adenomatous hyperplasia, 12 adenocarcinoma) were studied using immunohistochemistry with antisera for C, uPA, EGF receptors and TGF-alpha. Immunostaining for collagenase revealed a positive reaction in moderately differentiated adeno-carcinoma without staining the normal and hyperplastic endometrium. A progressive increase in uPA immunostaining was observed in proliferative and neoplastic endometrium. TGF-alpha and its receptor (EGFr) were stained in proliferative and more clearly in hyperplastic and carcinomatous endometrium. In conclusion, BM play an important role in proliferation and differentiation of human endometrium; their degradation influences estrogen transportation from blood to the stroma. Endometrial BM degradation is associated with the presence of collagenolytic enzymes and growth factors.

Factors Regulating Interaction between Trophoblast and Human Endometrium

Implantation is a crucial step in human reproduction. Disturbances of this process are responsible for pregnancy failure after both in vivo and in vitro fertilization. The endometrium provides the implanting embryo with a unique substratum where the embryo communicates with biochemical signals, attaches itself, penetrates and grows without blood circulation. The highly proliferative phase of the cytotrophoblast, during early human embryogenesis, may be due to endogenous production of growth factors that may establish autocrine/short range paracrine stimulator loops which explain the tumor-like properties of these tissues. Endometrial BM penetration and stroma invasion may be due to the proteolytic capability of the human embryo. It is suggested that collagenase and the urokinase-like plasminogen activator are responsible for this activity. To clarify the molecular mechanisms involved in human embryo implantation several models are suggested: culture of blastocysts, culture of endometrial cells, and endometrial explant co-culture. Human blastocysts cultured with whole perfused human uteri make it possible to recognize some aspects of the entire implantation process and give us the possibility of improving the benefits provided by new technologies in reproductive medicine and reducing embryonic loss at an early stage.

Filamentous Inclusions in Nonneoplastic and Neoplastic Pancreas: An Ultrastructural and Immunogold Labeling Study

Filamentous inclusions (FI) are unusual, irregularly shaped cytoplasmic inclusions, which are mostly found in acinar cell carcinomas of the pancreas and are consequently thought to be an abnormal zymogen granule type. This study describes identical inclusions in acinar, centroacinar, and small duct epithelial cells from nonneoplastic pancreas, as well as those found in tumor cells from a mixed acinar-endocrine pancreatic carcinoma. An ultrastructural and immunogold labeling demonstration indicates that these inclusions are aggregates of intermediate filaments immunoreacting with the anti-cytokeratin AE1/AE3 mixture and with V9 clone anti-vimentin monoclonal antibodies. Their pleomorphic appearance, variable immunoreactivity, and frequent association with lipid droplets and secondary lysosomes, mostly of the angulate type, led to the hypothesis that the FI undergo a degenerative remodeling pathway similar to that proposed for hepatic Mallory bodies. A survey of the literature on FI and human tumors suggests that they are a variably expressed ultrastructural feature of tumor cells originating from exocrine cell-containing tissues, namely the pancreas and gastrointestinal tract.

The Effect of Progestin on Factors Influencing Growth and Invasion of Endometrial Carcinoma

Progesterone (P) and progestins play an important role in the control of endometrial growth. We have investigated P and progestin effects on endometrial estrogen extraction, on basement membrane (BM) synthesis and on the presence of the epidermal growth factor receptor (EGFr) in normal and pathologic endometrium. E2 uptake, evaluated in human isolated perfused uteri is significantly decreased by P. BMs investigated using immunohistochemistry, with antisera to collagen IV and laminin, were found around stromal cells only in the luteal phase or during P or progestin administration. Glandular BM, discontinuous in hyperplastic and carcinomatous endometria, were restored to integrity only in typical hyperplasia after therapy with progestin. Endometrial EGFr is modified by P: revelation of this antigen is increased in proliferative phase and decreased in secretory phase. Similarly this molecule was present in hyperplastic and carcinomatous endometria. Only in benign hyperplasia did we observe no staining for the same antigen after progestinic therapy. These data suggest that P or progestins may also have an indirect influence through mechanisms such as estrogen uptake and tissue factor activity with important differences between normal and pathologic endometrium.

Model Agnostic Solution of CSPs via Deep Learning: A Preliminary Study

Deep Neural Networks (DNNs) have been shaking the AI scene, for their ability to excel at Machine Learning tasks without relying on complex, hand-crafted, features. Here, we probe whether a DNN can learn how to construct solutions of a CSP, without any explicit symbolic information about the problem constraints. We train a DNN to extend a feasible solution by making a single, globally consistent, variable assignment. The training is done over intermediate steps of the construction of feasible solutions. From a scientific standpoint, we are interested in whether a DNN can learn the structure of a combinatorial problem, even when trained on (arbitrarily chosen) construction sequences of feasible solutions. In practice, the network could also be used to guide a search process, e.g. to take into account (soft) constraints that are implicit in past solutions or hard to capture in a traditional declarative model. This research line is still at an early stage, and a number of complex issues remain open. Nevertheless, we already have intriguing results on the classical Partial Latin Square and N-Queen completion problems.

Dating the endometrial biopsy by flow cytometry

OBJECTIVE To determine endometrial changes throughout the menstrual cycle. DESIGN Flow cytometric analysis of endometrial cells versus chronological dating. SETTING Women volunteers with a normal menstrual cycle participated in this study that was done in an academic research environment. PATIENTS Two hundred thirty regular menstruating women with adequate luteal phase underwent endometrial biopsy at different days of their menstrual cycle; 138 biopsies were analyzed both histologically and by flow cytometry. MAIN OUTCOME MEASURE Percentage of biopsies correctly classified in terms of chronological dating. RESULTS Flow cytometry correctly classified the chronological day of biopsies in 59% of cases, the proliferative versus secretory days in 91% of cases, and the early, mid, and late secretory phases of the menstrual cycle were correctly predicted in 86% of cases. CONCLUSION Flow cytometry is a good method to determine growth and differentiation of the endometrium during the menstrual cycle. Detection of phases of the whole endometrial cell population offer information on the biological adequacy of endometrial response to the biochemical environment.

A Game-Based Competition as Instrument for Teaching Artificial Intelligence

This paper reports about teaching Artificial Intelligence (AI) by applying the experiential approach called “learning by doing”, where traditional, formal teaching is integrated with a practical activity (a game competition, in our case), that is relevant for AI discipline and allows for an active and playful participation of students.