Mismatch repair system in endometriotic tissue and eutopic endometrium of unaffected women

9Objective: To test the immunohistochemical staining pattern of some mismatch repair (MMR) system proteins in endometriotic tissue (ET) and eutopic endometrium. Methods: This was a retrospective study conducted at the Pathology and Obstetrics and Gynecology Departments of the Udine University Hospital. We analyzed 528 samples obtained from 246 patients affected by endometriosis and 71 samples from 71 patients with normal endometrium. A tissue microarray model was used to analyze the immunohistochemical expression of MMR system proteins. Results: Significant loss of MMR proteins was found in the stromal component of ETs. We found MSH2 to be expressed at a higher level than any other MMR system proteins in eutopic endometrium and ETs, to be significantly correlated to Ki-67 expression in both stromal and glandular components of ETs, and to be expressed at a significantly higher level in ETs than in eutopic endometrium. When considering the subgroup of endometriosis with high recurrence rate and glandular cytoplasmic staining for aurora A kinase, we found MMR proteins expressed at a significantly higher level in these ETs than in other ETs and eutopic endometrium of unaffected women. Conclusions: We found significant loss of MMR proteins (known to be associated with microsatellite instability) in the stromal component of ETs. The group of ETs with glandular cytoplasmic staining for aurora A kinase had higher MMR protein expression, suggesting an increased activity of this system. Our result suggests a novel role of increased MSH2 expression in cellular proliferation of endometriosis.openopenGrassi, T.; Calcagno, A.; Marzinotto, S.; Londero, A.P.; Orsaria, M.; Canciani, G.N.; Beltrami, C.A.; Marchesoni, D.; Mariuzzi, L.Grassi, T.; Calcagno, A.; Marzinotto, S.; Londero, Ambrogio P.; Orsaria, M.; Canciani, G. N.; Beltrami, Carlo Alberto; Marchesoni, Diego; Mariuzzi, Laur

Histomorphometrical evaluation of the effects of Aminogam® gel in oral healing process of post-surgical soft tissue

Wound healing is a dynamic process that involves a complex interaction of inflammatory cells, cytokines and mediators of extracellular matrix [1]. One of the processes that occur during tissue regeneration is angiogenesis and it is considered to have a pivotal role in wound repair. Previous studies have shown that a topical application of proteins and sodium hyaluronate to wounds can expedite the repair of damaged tissue [2]. The aim of this preliminary study is to evaluate the efficacy of Aminogam\uae gel (A\uae) (ErreKappa Euroterapici SpA, Milano), a topical medication which contains 4 amino acids (glycine, leucine, proline, lysine) and sodium hyaluronate, used to improve and accelerate gingival flap healing following molar extraction by analyzing collagen fibers amount, orientation and microvascular distribution (MVD). Ten patients (mean age 49ys) were included in the study. Two teeth (38 and 48) were extracted at an interval of 30 days. The \u201ctest\u201d site (AM) was treated with A\uae while the \u201ccontrol\u201d site (no AM) was not. Dental extraction was performed and the flaps were sutured with a consequent excess of tissue for histological processing (T0). A\uae had been applied only at the AM site for 10 days post-extraction. At suture removal, a gingivoplasty was performed and the exceeding tissue was histologically analysed (T1). Paraffin blocks were cut and slides were stained with haematoxylin-eosin and Sirius Red. No signs of inflammatory infiltrate or necrosis were observed. Sirius Red staining highlighted a lower degree of organized collagen fibers at T1 vs T0. At T0 the fibers were organized in closely packed and well-oriented bundles. At T1-no AM fibers were thin and formed a disorganized grid. At T1-AM fibers appeared thicker and the tissue appeared more mature compared to T1-no AM. Immunohistochemistry against CD31 was performed to mark endothelial cells and to calculate MVD by stereological method [3]. MVD resulted highest at T1-AM. The T1 data normalized on T0 presented a statistically significant difference (p=0.012) between AM and no AM group. In conclusion, A\uae gel seems to increase new blood vessels formation and to promote collagen deposition and organization. References [1] Gurtner et al. Nature. 2008;453(7193):314-21. [2] Zhu et al. BMC Oral Health. 2015;15:60. [3] Canullo et al. J Clin Periodontol. 2016;38:86-94

CONCEPTT: Continuous Glucose Monitoring in Women with Type 1 Diabetes in Pregnancy Trial: A multi-center, multi-national, randomized controlled trial - Study protocol.

BACKGROUND: Women with type 1 diabetes strive for optimal glycemic control before and during pregnancy to avoid adverse obstetric and perinatal outcomes. For most women, optimal glycemic control is challenging to achieve and maintain. The aim of this study is to determine whether the use of real-time continuous glucose monitoring (RT-CGM) will improve glycemic control in women with type 1 diabetes who are pregnant or planning pregnancy. METHODS/DESIGN: A multi-center, open label, randomized, controlled trial of women with type 1 diabetes who are either planning pregnancy with an HbA1c of 7.0 % to ≤10.0 % (53 to ≤ 86 mmol/mol) or are in early pregnancy (<13 weeks 6 days) with an HbA1c of 6.5 % to ≤10.0 % (48 to ≤ 86 mmol/mol). Participants will be randomized to either RT-CGM alongside conventional intermittent home glucose monitoring (HGM), or HGM alone. Eligible women will wear a CGM which does not display the glucose result for 6 days during the run-in phase. To be eligible for randomization, a minimum of 4 HGM measurements per day and a minimum of 96 hours total with 24 hours overnight (11 pm-7 am) of CGM glucose values are required. Those meeting these criteria are randomized to RT- CGM or HGM. A total of 324 women will be recruited (110 planning pregnancy, 214 pregnant). This takes into account 15 and 20 % attrition rates for the planning pregnancy and pregnant cohorts and will detect a clinically relevant 0.5 % difference between groups at 90 % power with 5 % significance. Randomization will stratify for type of insulin treatment (pump or multiple daily injections) and baseline HbA1c. Analyses will be performed according to intention to treat. The primary outcome is the change in glycemic control as measured by HbA1c from baseline to 24 weeks or conception in women planning pregnancy, and from baseline to 34 weeks gestation during pregnancy. Secondary outcomes include maternal hypoglycemia, CGM time in, above and below target (3.5-7.8 mmol/l), glucose variability measures, maternal and neonatal outcomes. DISCUSSION: This will be the first international multicenter randomized controlled trial to evaluate the impact of RT- CGM before and during pregnancy in women with type 1 diabetes. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01788527 Registration Date: December 19, 2012

Sprouty Proteins Inhibit Receptor-mediated Activation of Phosphatidylinositol-specific Phospholipase C

PLCγ03B3 binds Spry1 and Spry2. Overexpression of Spry decreased PLCγ03B3 activity and IP3 and DAG production, whereas Spry-deficient cells yielded more IP3. Spry overexpression inhibited T-cell receptor signaling and Spry1 null T-cells hyperproliferated with TCR ligation. Through action of PLCγ03B3, Spry may influence signaling through multiple receptors

Nephron endowment assessment in pre-transplantation kidneys using a digital histomorphometric approach

Introduction Kidney function largely depends on nephron number. However, the role of (donor\u2019s) nephron endowment (NE) has been poorly investigated in kidney transplantation (KTx) probably because technically difficult and no reliable surrogate markers exists. The aim of this study was developing an easy and reliable algorithm for computation of NE in kidneys addressed to KTx. Materials and methods Fourty-one kidneys removed from 26 brain-dead donors (12 female, mean age of 62.6 years) were analyzed. Computed tomography was performed on kidneys to stereologically compute the cortical volume. Tissue sections from 6 standardized areas of each kidney were harvested and processed for histomorphometry. On each section stereological method was employed to evaluate glomerular density (GD), size and volume. NE was computed for each kidney by multiplying the cortical volume for GD and dividing by the average glomerular volume. Number of functional (FG) and atrophic (AG) glomeruli was also assessed. Results Mean volume of the cortical portion was 48.2cm3\ub117.7. Glomeruli represented 9.53%\ub12.19 of cortex. Mean glomerular volume was 5.75E-6\ub12.04E-06 cm3. Glomerular number was: 858,550\ub1373,245 (NE), 650,606\ub1310,400 (FG) and 207,944\ub199,837 (AG). GD was higher at the upper pole compared to medium and inferior areas but no significantly. Conclusions Nephron count is feasible with relatively limited resources by measuring GD in a single spot and applying the algorithm we have developed. The integration of traditional pathology with cutting-edge digital technologies and bioinformatics is a reproducible promising tool for diagnostics also in the area of KTx towards a better understanding of the mechanisms influencing the outcom

Inducible t-cell costimulator ligand plays a dual role in melanoma metastasis upon binding to osteopontin or inducible t-cell costimulator

Recently, we demonstrated that inducible T-cell costimulator (ICOS) shares its unique ligand (ICOSL) with osteopontin (OPN), and OPN/ICOSL binding promotes tumor metastasis and angiogenesis in the 4T1 breast cancer model. Literature showed that OPN promotes melanoma metastasis by suppressing T-cell activation and recruiting myeloid suppressor cells (MDSC). On the opposite, ICOS/ICOSL interaction usually sustains an antitumor response. Here, we engineered murine B16F10 melanoma cells, by transfecting or silencing ICOSL. In vitro data showed that loss of ICOSL favors anchorage-independent growth and induces more metastases in vivo, compared to ICOSL expressing cells. To dissect individual roles of the three molecules, we compared data from C57BL/6 with those from OPN-KO, ICOS-KO, and ICOSL-KO mice, missing one partner at a time. We found that OPN produced by the tumor microenvironment (TME) favors the metastasis by interacting with stromal ICOSL. This activity is dominantly inhibited by ICOS expressed on TME by promoting Treg expansion. Importantly, we also show that OPN and ICOSL highly interact in human melanoma metastases compared to primary tumors. Interfering with this binding may be explored in immunotherapy either for nonresponding or patients resistant to conventional therapies