Sjogren's syndrome (SS) is characterized by the features of systemic autoimmunity and exocrine gland dysfunction and inflammation. Deregulated cytokine production is known to contribute to the etiology of SS but the underlying molecular mechanism is still remains to be unclear. TNF-alpha-induced protein 3 or TNFAIP3 is involved in the negative feedback regulation of nuclear factor-kappa B (NF-kappa B) signaling in response to specific pro-inflammatory stimuli in different cell types. To define the contribution of TNFAIP3 to SS, the levels of TNFAIP3 expression in human salivary gland epithelial cells (SGEC) derived from active primary SS patients were analyzed. Histological analysis was performed on paraffin-embedded human Sjogren's samples and healthy tissues. In separate experiments, immunofluorescence staining, western blot analysis and quantitative real-time PCR for TNFAIP3 was conducted in SGEC from SS and healthy subjects. Our findings clearly demonstrate changes in levels of the protein and gene expression between healthy controls and SS patients, depicting a very weak positivity for TNFAIP3 in SS samples. TNFAIP3 was found down-regulated in SGECs derived from SS patients in comparison with controls, and the cells with down-regulated TNFAIP3 expression exhibited enhanced NF-kappa B activities. In addition, to investigate the role of TNFAIP3 in the activation of NF-kappa B, we depleted TNFAIP3 expression by siRNA in healthy SGEC after treatment with or without TNF-alpha. Intriguingly, the silencing of TNFAIP3 by its siRNA in healthy SGEC increased NF-kappa B activation that could explain the deregulated cytokines production observed in SS.

The spatial distribution of mast cells inside the tumor stroma has been little investigated. In this study, we have evaluated tumor mast cells distribution through the analysis of the morphological features of the spatial patterns generated by these cells, including size, shape, and architecture of the cell pattern. We have compared diffuse large B cells lymphoma (DLBCL) and systemic mastocytosis in two different anatomical localizations (lymph nodes for DLBCL and, respectively, bone marrow for mastocytosis). Results have indicated that, despite the high difference in size exhibited by the mast cells patterns in the two conditions, the spatial relationship between the mast cells forming the aggregates resulted similar, characterized by a significant tendency of the mast cells to self-organize in clusters.

Purpose: To develop a Risk Score (RS) to predict distant recurrence among premenopausal women with node-negative endocrine responsive early breast cancer. Methods: The Cox model was used to develop the RS using clinical and histopathological features from 378 women participating in the IBCSG Trial VIII who received endocrine therapy alone or following chemotherapy. The performance of the resulting model was validated on a cohort of 1005 patients from a single institution who received endocrine therapy alone. Results: In a multivariable analysis, the risk of distant recurrence was associated with tumor size, ER, Ki- 67 and peritumoral vascular invasion. In the validation cohort, patients with high RS were at greater risk of distant recurrence compared to patients with low RS (HR, 17.41; 95% CI, 5.72e52.95). Conclusion: In premenopausal women with node-negative endocrine-responsive early breast cancer, the RS identifies patients at higher risk of distant recurrence.

Gastric cancer shows intratumoral heterogeneity for human epidermal growth factor receptor 2 expression. We evaluated whether the number of tissue blocks analyzed or the antibodies used may influence the immunohistochemical results in gastrectomy specimens. Clinicopathologic data from 148 patients receiving gastric surgery for cancer were collected. One tissue block for each of 88 primary tumors and 60 paired primary tumors and metastases was examined for human epidermal growth factor receptor 2 status by immunohistochemistry using 3 different antibodies (HercepTest, CB11, and 4B5) and by fluorescent in situ hybridization. Two additional tissue blocks of the primary tumor were tested by immunohistochemistry if the results were negative on the first tissue block. The concordance among the 3 antibodies was 94.5% (testing 1 tissue block). Two cases showed a clinically significant discrepancy between primary tumor (score 0) and lymph nodes metastases (score 3+). Additional block analysis increased both the sensitivity (from 63% to 83%) and the accuracy (from 91% to 94%) of immunohistochemistry as compared with fluorescent in situ hybridization. The multiblock approach could potentially identify a greater number of human epidermal growth factor receptor 2–positive gastric cancers, particularly those with higher levels of intratumor heterogeneity.

BACKGROUND: Calcium glucarate (CGT) is a promising chemopreventive agent. This study evaluated the in vivo efficacy of CGT in preventing 7,12-dimethylbenz(alpha) anthracene (DMBA)-induced oral carcinogenesis in the hamster. Matherials and Methods: Seventy-six Syrian hamsters were used, divided into four groups: group 1, untreated animals; 2, CGT controls; 3, DMBA-treated; 4, DMBA- and CGT-treated. Hamsters were painted three times weekly with 0.5% solution of DMBA and were fed a diet supplemented with CGT (64 mmol/kg, 2%). Animals were sacrificed at week 9 and 12 and pathology and histomorphometric analyses were performed. RESULTS: At week 9, four dysplastic lesions and six carcinomas were identified in group 3 while only three dysplasias and five carcinomas were detected in group 4. At week 12, five animals of group 3 displayed a dysplasia, which was only detected in one animal of group 4. Squamous carcinomas were identified in all animals of both group 3 and 4. However, in group 3 four of the animals displayed multifocal lesions and carcinomas displayed histological features indicative of increased aggressiveness. CONCLUSION: The results obtained suggest that CGT can exert an inhibitory effect on oral carcinogenesis in tha hamster and that further studies are warranted to evaluate its potential use as a chemopreventive agent in humans.

Background: Clusterin (CLU) is a ubiquitous multifunctional factor involved in neoplastic transformation. The CLU transcript variants and protein forms play a crucial role in balancing cells proliferation and death. Methods: We investigated the regulation of CLU transcript variants expression in an in vivo model system consisting of both neoplastic tissues and fine needle aspiration biopsy (FNAB) samples isolated from patients undergoing thyroidectomy. Results: The immunohistochemical analyses showed an overall CLU up-regulation in papillary carcinoma. A specific CLU2 transcript variant increase was registered using qPCR in papillary carcinomas while CLU1 decreased. In addition, the analysis of CLU transcripts expression level showed an increase of the CLU2 transcript in the TIR 3 patients with histologically confirmed thyroid cancer. Conclusions: Our results suggest the existence of a specific alteration of CLU2:CLU1 ratio towards CLU2, thus providing the first circumstantial evidence for the potential use of CLU transcript variants as effective biomarkers for a more accurate assessment of the so called “indeterminate” thyroid nodules.

Diffuse large B cell lymphoma (DLBCL) is recognized as the most common non-Hodgkin lymphoma subtype. Advanced high-resolution digital scans of pathology slides have enabled the development of computer-based image analysis algorithms that may assist pathologists in quantifying immunohistochemical stains. In this retrospective study, we reviewed data from 29 patients affected by DLBCL. In order to evaluate the number of tumor cells and microenvironment T cells, we performed an analysis of CD20, Ki67, and CD3 counts, assessed with the Positive Pixel Count algorithm embedded in the Aperio ImageScope software. A lower tumor cell count was observed in patients with a non-germinal center immunophenotype, high LDH, splenomegaly and an IPI ≥ 3. A lower number of CD3 was observed in patients with bulky disease, an IPI ≥ 3 and disease stage 3-4. Overall, these data confirm that quantitative analysis of the tumor cells and of the tumor microenvironment by means of computer-driven quantitative image analysis may add new information in DLBCL diagnosis.

Bortezomib and Lenalidomide have been shown to be effective in the control of multiple myeloma (MM) progression. We have investigated their role in the in vitro expression of Osterix by primary osteoblast cultures from MM patients and found that Osterix RNA was constitutively down-regulated in these cells. Treatment of osteoblasts with Bortezomib resulted in an increase of Osterix RNA and in enhanced activity of both BMP-2 and Runx2. Instead, Lenalidomide was unable to modify Osterix transcription. These findings provide additional evidence suggesting that, at least in vitro, Bortezomib promotes the osteoblast maturation whereas Lenalidomide is ineffective. (C) 2009 Elsevier Ltd. All rights reserved.

Objective: Cri du Chat Syndrome (CCS) is a rare chromosomal disorder resulting from deletion of the short arm of chromosome 5, characterized by distinctive catlike cry since born due to typical larynx (small and narrow) and epiglottis anomalies. We report on still unreported clinic-pathological features CCS: diffuse Gingival Fibromatosis (GF) with Confocal Laser Scanning Microscopic (CLSM) examination.Case Presentation: A 21-year-old female, affected by CCS showed diffuse GF on the palatal gingiva in the molar region (linked to dental dislocation and malocclusion), which was surgically removed by Diode Laser, formalin-fixed, stained with hematoxylin-eosin and picrosirius red and analyzed at CLSM Nikon E-600 with double Laser inducing fluorescence (green and red).Results: Microscopically, the lesion consisted in large parallel collagenous fibers, abundant blood vessels with plump endothelial cells and chronic inflammatory reaction, and large polygonal cells in the vascular interstitial spaces, with large nuclei, resembling undifferentiated totipotent mesenchymal cells (stem-like cells). At CLSM large and variably oriented collagenous fibers displayed intense fluorescence due to cross-linking between such fibers, which generally characterize fibromatosis, and the vascular structures. The latter showed lower fluorescence intensity, and were surrounded by loose collagenous fibers and trapezoidal large mesenchymal cells.Conclusions: GF seems a characteristic feature of CCS, with typical fluorescent pattern at CLSM, in which purported totipotent mesenchymal cells may play a pathogenetic role.

Most sporadic colorectal cancers (CRCs) develop through the adenoma-carcinoma sequence pathway and are initiated by adenomatous polyposis coli (APC) gene mutations. Estrogen receptor beta (ERbeta) is recognized to progressively reduce its expression in adenomatous and carcinomatous tissues in humans. Moreover, ERbeta deficiency enhances small intestinal tumorigenesis in rodents. In the Apc(Min/+) mouse model, we evaluated intestinal polyp development and ERbeta expression plus other biological parameters influencing tumor growth (epithelial cell proliferation, apoptosis and migration) following the addition of a combination of the ERbeta-selective agonist silymarin (SIL) and/or lignin (LIG) to a high-fat/low-fiber diet. Forty-five Apc(Min/+) mice were divided in four groups: animals fed on the tumorigenic high-fat/low-fiber diet, the tumorigenic diet supplemented with SIL (0.02%) or purified LIG (6.24%) or SIL (0.005%) + LIG (6.24%). In these animals, we assessed polyp number and volume and their degree of dysplasia together with ERbeta messenger RNA (mRNA) and protein levels and epithelial cell proliferation, migration and apoptosis. The latter group of parameters was evaluated in normal and adenomatous mucosa and the results compared with those found in wild-type (WT) mice fed on the control diet. The addition of SIL or LIG to the diet and even more the specific combination of the two significantly counteracted intestinal tumorigenesis and increased ERbeta mRNA and protein levels. Cell proliferation and apoptosis were rebalanced and cell migration accelerated, restoring values similar to those observed in WT animals. Our results further support a protective effect of ERbeta in CRC suggesting the use of the combination of SIL-LIG as a potential approach against CRC development.

Hereditary haemorrhagic telangiectasia (HHT) is rare, and characterised by vascular dysplasia that leads to various symptoms including visceral arteriovenous malformations and mucocutaneous telangiectatic lesions. Our aim was to describe the clinical features and options for the treatment of multiple oral lesions, and to illustrate the efficacy of the diode laser in the treatment of early (<2mm) and advanced lesions (2mm or more). We report 24 patients with 1200 oral telangiectatic lesions, which were often associated with regular bleeding (from monthly to daily), superinfection, pain, and swelling, and treated with multiple sessions of laser according to the number and size of the lesions. Early lesions were treated with a single laser impulse in ultrapulsed mode, and advanced lesions with repeated laser impulses in pulsed mode (t-on 200ms/t-off 500ms), at a power of 8W. Early lesions healed completely after laser photocoagulation with no operative or postoperative complications, while advanced lesions improved with a remarkable reduction in size but more discomfort. Protective occlusal plates were sometimes used to reduce the incidence of new lesions caused by dental trauma. The treatment of oral telangiectatic lesions is still being debated, and it is important to improve quality of life for patients. Diode laser surgery could be an effective treatment for oral lesions in those with hereditary haemorrhagic telangiectasia.

Background: Breast cancer (BC) cells secrete soluble factors that accelerate osteoclast (OC) differentiation, leading to the formation of osteolytic bone metastases. In the BOLERO-2 trial, BC patients with bone involvement who received Everolimus had a delayed tumor progression in the skeleton as a result of direct OC suppression through the inhibition of mTOR, in addition to the general suppressor effect on the cancer cells. Here, we explored the effect of Everolimus, as mTOR inhibitor, on the pro-OC paracrine activity of BC cells. Methods: Both MDA-MB-231 and MCF-7 BC cell lines were incubated with sub-lethal amounts of Everolimus, and their conditioned supernatants were assessed for their capacity to differentiate OCs from PBMC from healthy donors, as well as to interfere with their bone resorbing activity shown on calcium phosphate slices. We also measured the mRNA levels of major pro-OC factors in Everolimus-treated BC cells and their secreted levels by ELISA, and evaluated by immunoblotting the phosphorylation of transcription factors enrolled by pathways cooperating with the mTOR inhibition. Finally, the in vivo pro-OC activity of these cells was assessed in SCID mice after intra-tibial injections. Results: We found that Everolimus significantly inhibited the differentiation of OCs and their in vitro bone-resorbing activity, and also found decreases of both mRNA and secreted pro-OC factors such as M-CSF, IL-6, and IL-1β, whose lower ELISA levels paralleled the defective phosphorylation of NFkB pathway effectors. Moreover, when intra-tibially injected in SCID mice, Everolimus-treated BC cells produced smaller bone metastases than the untreated cells. Conclusions: mTOR inhibition in BC cells leads to a suppression of their paracrine pro-OC activity by interfering with the NFkB pathway; this effect may also account for the delayed progression of bone metastatic disease observed in the BOLERO-2 trial. Keywords: BOLERO-2 trial, Breast cancer cells, mTOR, Osteoclastogenesis, Everolimus

Objective: We report on the clinic-pathological features of Familial Tumoral Calcinosis (FTC), a rare disease of early childhood and adulthood, caused by mutations in fibroblast growth factor 23 and GalNAc transferase 3. It is a bone metabolism disorder with abnormal phosphate and calcium (calcinosis) deposits around the joints, in visceral and soft tissues. Case Presentation: A 17 year-old girl complaining for long-standing night leg pain, resistant to FANS therapy, had been diagnosed with osteogenesis imperfecta and was therefore undergoing bisphosphonates therapy. She was referred to our Dental Clinic for diffuse dental anomalies, maxillary hypoplasia and tooth roots inclusions and underwent combined surgical and orthodontic treatment. The surgical samples were used for conventional and Confocal Laser Scanning Microscopic (Nikon E-600) (CLSM) examination. Results: Microscopically several metaplastic micro and macro-calcificationin soft and periodontal tissue location were detected, along with a typical islands of homogenous, non tubular, dentino-osteoid calcified structures in dentinal tissues. Also, dentinal dysplasia with osteoid-like material, without incremental lines but with strong basophilia, intermingled with remnants of mature mucous connective tissue, was demonstrated. The diagnosis of FTC was confirmed by genetic analysis. Conclusions: CLSM helps to demonstrate distinct odontoblast and osteoblast anomalies in FTC that lead to the accumulation of atypical calcified tissues, responsible for the several clinical signs detected in the patient and formerly attributed to osteogenesis imperfecta.

Glucose-6-phosphate isomerase (GPI), also known as phosphoglucose isomerase, was initially identified as the second glycolytic enzyme that catalyzes the interconversion of glucose-6-phosphate to fructose-6-phosphate. Later studies demonstrated that GPI was the same as the autocrine motility factor (AMF), and that it mediates its biological effects through the interaction with its surface receptor (AMFR/gp78). In this study, we assessed the role of GPI/AMF as a prognostic factor for clear cell renal cell carcinoma (ccRCC) cancer-specific (CSS) and progression-free survival (PFS). In addition, we evaluated the expression and localization of GPI/AMF and AMFR, using tissue microarray-based immunohistochemistry (TMA-IHC), indirect immunofluorescence (IF), and confocal microscopy analysis.Primary renal tumor and nonneoplastic tissues were collected from 180 patients who underwent nephrectomy for ccRCC. TMA-IHC and IF staining showed an increased signal for both GPI and AMFR in cancer cells, and their colocalization on plasma membrane. Kaplan-Meier curves showed significant differences in CSS and PFS among groups of patients with high versus low GPI expression. In particular, patients with high tissue levels of GPI had a 5-year survival rate of 58.8%, as compared to 92.1% for subjects with low levels (P < 0.0001). Similar findings were observed for PFS (56.8% vs 93.3% at 5 years). At multivariate analysis, GPI was an independent adverse prognostic factor for CSS (HR = 1.26; P = 0.001), and PFS (HR = 1.16; P = 0.01).In conclusion, our data suggest that GPI could serve as a marker of ccRCC aggressiveness and a prognostic factor for CSS and PFS.

An altered metabolism is involved in the development of clear cell - renal cell carcinoma (ccRCC), and in this tumor many altered genes play a fundamental role in controlling cell metabolic activities. We delineated a large-scale metabolomic profile of human ccRCC, and integrated it with transcriptomic data to connect the variations in cancer metabolism with gene expression changes. Moreover, to better analyze the specific contribution of metabolic gene alterations potentially associated with tumorigenesis and tumor progression, we evaluated the transcription profile of primary renal tumor cells. Untargeted metabolomic analysis revealed a signature of an increased glucose uptake and utilization in ccRCC. In addition, metabolites related to pentose phosphate pathway were also altered in the tumor samples in association with changes in Krebs cycle intermediates and related metabolites. We identified NADH dehydrogenase (ubiquinone) 1 alpha subcomplex 4-like 2 (NDUFA4L2) as the most highly expressed gene in renal cancer cells and evaluated its role in sustaining angiogenesis, chemoresistance, and mitochondrial dysfunction. Finally, we showed that silencing of NDUFA4L2 affects cell viability, increases mitochondrial mass, and induces ROS generation in hypoxia.

Invasive lobular carcinoma (ILC) is the most common “special type” of breast cancer. Although conflicting literature data are available on the outcome of ILC, recently reported data indicate that ILC carries a poorer prognosis if compared to invasive ductal carcinomas. We evaluated clinical and biological features of 981 consecutive patients with pT1-3, pN1-3 M0 ILC. Median follow-up was 7.4 years for survival. A total of 541 patients were classified as classic (55.8%), 146 alveolar (14.9%), 145 mixed non-classic (14.8%), 104 solid (10.6%), and 38 trabecular (3.9%). A statistically significant difference in the outcome was observed at multivariate analysis for patients with solid (HR 2.44, 95% CI 1.39–4.29 for OS; HR 1.92, 95% CI 1.29–2.88 for DFS) and mixed non-classic (HR 1.99, 95% CI 1.12–3.53 for OS) versus patients with classical ILC. A statistically significant difference in the risk of distant metastases was observed at multivariate analysis for patients with Luminal B (HR 2.56, 95% CI 1.38–4.76), HER2 positive (HR 7.80, 95% CI 1.55–39.3), and triple negative (HR 7.61, 95% CI 2.63–22.1) subtypes versus patients with Luminal A ILC. Age ≥70 years, tumor size and degree of nodal involvement were additional independent predictors of reduced overall survival. The outcome of ILC significantly correlated with histological and immunohistochemically defined molecular subtypes. New tailored strategies should be explored in these subgroups of patients with poor outcome.

Prostate cancer (PCa) is the second leading cause of cancer-related death in men; however, the molecular mechanisms leading to its development and progression are not yet fully elucidated. Of note, it has been recently shown that conditional stk11 knockout mice develop atypical hyperplasia and prostate intraepithelial neoplasia (PIN). We recently reported an inverse correlation between the activity of the STK11/AMPK pathway and the MAPK/p38 cascade in HIF1A-dependent malignancies. Furthermore, MAPK/p38 overactivation was detected in benign prostate hyperplasia, PIN and PCa in mice and humans. Here we report that STK11 expression is significantly decreased in PCa compared to normal tissues. Moreover, STK11 protein levels decreased throughout prostate carcinogenesis. To get insight into the role of STK11-MAPK/p38 activity balance in PCa, we treated PCa cell lines and primary biopsies with a well-established MAPK14-MAPK11 inhibitor (SB202190), which has been extensively used in vitro and in vivo. Our results indicate that inhibition of MAPK/p38 significantly affects PCa cell survival in a STK11-dependent manner. Indeed, we found that pharmacologic inactivation of MAPK/p38 does not affect viability of STK11-proficient PCa cells due to the triggering of the AMPK-dependent autophagic pathway, while it induces apoptosis in STK11-deficient cells irrespective of androgen receptor (AR) status. Of note, AMPK inactivation or autophagy inhibition in STK11-proficient cells sensitize SB202190-treated PCa cells to apoptosis. On the other end, reconstitution of functional STK11 in STK11-deficient PCa cells abrogates apoptosis. Collectively, our data show that STK11 is a key factor involved in the early phases of prostate carcinogenesis, and suggest that it might be used as a predictive marker of therapeutic response to MAPK/p38 inhibitors in PCa patients.

BACKGROUND: Many authors have considered dental implants to be unrelated to increased risk of medication-related osteonecrosis of the jaw (MRONJ). Nevertheless, more recently, more cases of peri-implant MRONJ (PI-MRONJ) have been described, thus becoming a challenging health problem. Also, metastatic cancer deposits are not infrequently found at peri-implant sites and this may represent an additional complication for such treatments. We present the case of a breast cancer patient with PI-MRONJ, presenting a clinically and radiologically undetected metastasis within the necrotic bone, and highlight the necessity of an accurate histopathological analysis. CASE REPORT: A 66-year-old female patient, who had received intravenous bisphosphonates for bone breast cancer metastases, came to our attention for a non-implant surgery-triggered PI-MRONJ. After surgical resection of the necrotic bone, conventional and immunohistochemical examinations were performed, which showed breast cancer deposits within the necrotic bone. CONCLUSIONS: Cancer patients with metastatic disease, who are undergoing bisphosphonate treatment, may develop unusual complications, including MRONJ, which is a site at risk for hosting additional metastatic deposits that may be clinically and radiologically overlooked. Such risk is increased by previous or concomitant implant procedures. Consequently, clinicians should be prudent when performing implant surgery in cancer patients with advanced-stage disease and consider the possible occurrence of peri-implant metastases while planning adequate treatments in such patients.

Diffuse Large B-cell Lymphoma (DLBCL) is the most common form of Non-Hodgkin lymphoma characterized by clinical and biological heterogeneity attributable both to the tumor cells and the complex tumor-microenvironment surrounding them. Tumor-associated macrophages (TAMs) and mast cells are two major components of the tumor inflammatory infiltrate with a definite role in enhancing tumor angiogenesis. In this study, we have investigated CD68 and tryptase expression and their relationship with microvascular density (MVD) in chemo-resistant and chemosensitive patients affected by DLBCL. CD68 and tryptase expression as well as MVD were increased in chemo-resistant patients when compared with chemosensitive patients. Tryptase expression showed a positive correlation with MVD, supporting a role for mast cell in DLBCL tumor angiogenesis, while CD68 correlation with MVD was not significant, indicating a different role for TAMs than angiogenesis in DLBCL.

Lymphatic Malformation (LM) according to ISSVA Classification, is a rare benign disorder with unknown aetiology. LM may grow slowly over years or develop rapidly over the course of days becoming a bulky lump, infected or bleeding. We propose our three steps Diode Laser protocol for LM management, based on its persistent vascular blood component. 1. Histological and cytological examination, to evaluate the vascular blood component (10-40%), shows mature lymphocytes with red blood cells and endothelial cells. 2. Diode Laser Photocoagulation (DLP) in pulsed mode (on 100ms / off 400ms) at 10W and 800nm with a 300μm fibre kept 2-3mm from the tissues, to reduce the lesion. 3. Diode Laser surgical excision in pulsed mode (on 50ms / off 200ms) at 8W and 800nm with a 300 μm fibre in close contact with tissues, and histological intraoperative margins control on frozen sections. Even if it has inconstant results (lesions decreasing rate is 10% to 40% proportionally to vascular blood component), DLP simplifies the last and the most important step. Use of Diode Laser also in surgical excision reduces intra and postoperatory complications.

Introduction and objectives. Lymphangioma, or Lymphatic Malformation (LM) according to ISSVA Classification, is a rare benign disorder with unknown etiology. LM may grow slowly over years or develop rapidly over the course of days becoming a bulky lump, infected or bleeding. Surgical excision is the gold standard treatment for adult LM with 10-15% recurrence rate and 15-30%complication rate. Other treatment modalities have also been proposed: simple aspiration with high risk for recurrence; sclerotherapy (alcohol, steroids, bleomycin or interferon) without results but only complications like fever, pain or lesion enlargement. The aim of this study is to show our three steps Laser protocol for LM management, based on its persistent vascular blood component. Methods. We select 11 patients aged between 12 and 60 years with clinical diagnosis of LM of tongue, lower lip and cheek divided in circumscriptum (&lt;3x3cm) and major lesions (&gt;3x3cm). LM are fluctuant and mobile suggesting a differential diagnosis from hemangioma, metastatic lesions and lymphoma. LM laser protocol includes three steps: 1. Histological and cytological examination, to evaluate the vascular blood component (10-40%), shows mature lymphocytes with red blood cells and endothelial cells. 2. Diode Laser Photocoagulation (DLP) in pulsed mode (on 190-250ms / off 250-450ms) at 14-20W and 800nm, to reduce the lesion. 3. Diode Laser surgical excision with histological intraoperative margins control on frozen sections. Results and conclusions. Histological analysis, highlighting the vascular blood component in all LM, validates photocoagulation and surgical combined approach with Diode Laser. Even if it has inconstant results (lesions decreasing rate is 10% to 40% proportionally to vascular blood component), DLP simplifies the last and the most important step. Use of Diode Laser also in surgical excision resets bleeding, infection and lymphorrhea to zero, accelerates healing time and improves aesthetic results.

INTRODUCTION: The acronym PHACES describes the association of posterior fossa malformations, facial hemangiomas, arterial anomalies (cardiovascular or cerebrovascular), coarctation of the aorta and cardiac defects, eye abnormalities, and sternal or ventral defects. In this study we report on 6 patients affected by the PHACES syndrome and showing 34 intraoral hemangiomas (IH), treated by diode laser photocoagulation (DLP). CASE PRESENTATION: IH appeared as red-bluish soft masses, smooth or lobulated, from a few millimetre to several centimetres in size, covered by intact mucosa and blanching on pressure. IHs were treated by DLP with 320μm fibres at a wavelength of 800±10nm. The diode laser techniques applied were: Transmucosal DLP (DLTP), a no-contact technique in which laser energy is delivered by a flexible optic quartz fiber, which is kept 2-3mm apart from the lesion, and Intralesional DLP (DLIP), in which the fibre is introduced into the lesion through a transmucosal access. DLTP was used for 20 flat, superficial IHs and, after a variable number of laser sessions (average=3) depending on the size of the lesion, 65% completely regressed, while in the remaining 35% shrinkage of the lesion was achieved with minor and few complications. The remaining 14 deep/multi-lobulated IHs were treated by DLIP, resulting in complete regression of 79% of them. CONCLUSIONS: DLP techniques are an effective and minimally invasive procedure for IH in patients with PHACES, in consideration of the multiple lesions to treat, of the necessity of multiple interventions and the higher compliance of the patients.

Objective: This study was designed to evaluate whether a sterile gel formulation of sodium hyaluronate and amino acids Gly-Pro-Leu-Lys (AMINOGAM®) is effective in accelerating post-surgical bone defects regeneration, implant rehabilitation and complication management. Materials and Methods: We selected 56 patients aged between 16 and 64 years and we evaluated different healing in 80 bone defects divided in 2 groups: - Test group: 40 defects treated with intracavitary intraoperative filling of gel and application 4 times/die upon the stitches till the complete mucosal healing (sandwich technique). - Control group: 40 untreated defects. Each group was divided in two subgroups: minor (&lt;1cm2) and major defects (&gt;1cm2). Outcomes were evaluated by clinical and radiographic follow-up with OPT rx and CT scan through densitometric analysis. Defects treatment after bone healing in both groups was completed with histological pre-implantar examination and insertion of 52 implants in test group and 49 implants in control group. Clinical and radiographic examinations of implants were performed at 12 months after functional loading. Gel preparation enhances angiogenesis, fibroblast and osteoblast proliferation, collagen biosynthesis, production of growth factors as evidenced by MTT test and alkaline phosphatase histochemical staining. In vivo and in vitro studies suggested that hyaluronic acid plays important roles in bone wound healing by enhancement of osteoblast differentiation through the down-regulation of BMP-2 antagonists. Lysine and proline are important metabolic factors regulating collagen matrix synthesis during osteogenesis. Results: Soft tissues outcomes in test group show similar results in both subgroups with immediate haemostatic effect, pain and swelling decrease and infective complication dejection (0%) compared to control group (6%). Gel preparation reduces the removing stitches and wound complete healing time at 6 days in minors defects and 14 days in major ones of test group compared to 14 and 23 days of control group. Hard tissues outcomes show faster healing time: 20% difference between ossification level in test and control group at 2 months. The difference decrease in following months until 3% at 12 months. Pre-implantar bone specimen get with 2,5mm trephine drill shows a more dense and mature lamellar bone with twisted fiber and different calcification level also evaluated through densitometic analysis, allowing implant insertion at 30-45 days in minor defects and at 60 days in major ones of test group compared to 60 and 90 days of control group. Osseointegration rate in regenerated defects is 100%. Conclusions: Sterile gel based on sodium hyaluronate and amino acids is a new cheap and useful medical device able in resetting post-surgical morbidity to zero. It allows a quickly bone defects healing time with an earlier implant insertion and a faster osseointegration thanks to more quality bone evaluated by histological analysis and grey scale densitometry. Therefore the whole rehabilitation treatment is considerably shortened and free from complication.

INTRODUCTION: Pyogenic granuloma (PG) is a relatively common benign mucocutaneous lesion. The term is a misnomer as the lesion does not contain pus nor it is granulomatous. Etiology of the lesions is unknown, but predisposing factors that have been reported include pregnancy, trauma, vascular malformation and chronic inflammation. PG are usually solitary lesions. The most common intraoral site is marginal gingiva, but lesions have been reported on palate, buccal mucosa, tongue, and lips. Extraoral sites commonly involve the skin of face, neck, upper and lower extremities, and mucous membrane of nose and eyelids. In this report, we seek to highlight the therapeutic advantages achieved with diode laser in intraoral PG treatment compared with surgical excision. MATERIAL AND METHODS: We report the cases of 85 patients presenting intraoral dull red, sessile, or pedunculated smooth surfaced nodule that may easily bleed, crust, or ulcerate. 62 were treated with surgical excision and 23 with diode laser treatment. The laser session consisted in diode laser photocoagulation ensued by diode laser excision of the lesion, preceded by treated areas cooling to avoid the tissue demage. According to the literature were used the following specification: wavelength 808nm, 10W power. Histological evaluation showed hyperplastic stratified squamous epithelium with stroma consisted of a large number of budding and dilateted capillaries and a dens chronic inflammatory cell infiltrate. Diagnosis of PG was confirmed. RESULTS AND CONCLUSION: Rapid healing can be observed within a few days of treatment, and as blood vessels are sealed, there is an improvement of haemostasis and coagulation compared with surgical excision. Post-operative pain discomfort, edema and bleeding are notably reduced. In conclusion, the use of diode laser offers a new tool that can change the way in which existing treatments are performed.

Clusterin (CLU) is a nearly ubiquitous multifunctional protein synthesized in different functionally divergent isoforms, sCLU and nCLU, playing a crucial role by keeping a balance between cell proliferation and death. Studying in vivo CLU expression we found a higher mRNA expression both in neoplastic and hyperplastic tissues in comparison to normal endometria; in particular, by RT-qPCR we demonstrated an increase of the specific sCLU isoform in the neoplastic and hyperplastic endometrial diseases. On the contrary, no CLU increase was detected at the protein level. The CLU gene transcriptional activity was upregulated in the hyperplastic and neoplastic tissues, indicating the existence of a fine post-trans-criptional regulation of CLU expression possibly responsible for the protein decrease in the malignant disease. A specific CLU immunoreactivity was present in all the endometrial glandular cells in comparison to the other cellular compartments where CLU immunoreactivity was lower or absent. In conclusion, our results suggest the existence of a complex regulatory mechanism of CLU gene expression during the progression from normal to malignant cells, possibly contributing to endometrial carcinogenesis. Moreover, the specific alteration of the sCLU:nCLU ratio associated with the pathological stage, suggests a possible usage of CLU as molecular biomarker for the diagnosis/prognosis of endometrial proliferative diseases.

Clusterin (CLU) is a nearly ubiquitous multifunctional protein synthesized in different functionally divergent isoforms, sCLU and nCLU, playing a crucial role by keeping a balance between cell proliferation and death. Studying in vivo CLU expression we found a higher mRNA expression both in neoplastic and hyperplastic tissues in comparison to normal endometria; in particular, by RT-qPCR we demonstrated an increase of the specific sCLU isoform in the neoplastic and hyperplastic endometrial diseases. On the contrary, no CLU increase was detected at the protein level. The CLU gene transcriptional activity was upregulated in the hyperplastic and neoplastic tissues, indicating the existence of a fine post-trans-criptional regulation of CLU expression possibly responsible for the protein decrease in the malignant disease. A specific CLU immunoreactivity was present in all the endometrial glandular cells in comparison to the other cellular compartments where CLU immunoreactivity was lower or absent. In conclusion, our results suggest the existence of a complex regulatory mechanism of CLU gene expression during the progression from normal to malignant cells, possibly contributing to endometrial carcinogenesis. Moreover, the specific alteration of the sCLU:nCLU ratio associated with the pathological stage, suggests a possible usage of CLU as molecular biomarker for the diagnosis/prognosis of endometrial proliferative diseases.

Aim: Sarcosine has been identified as a differential metabolite that is greatly increased during progression from normal tissue to prostate cancer and metastatic disease. In this study we assessed the role of serum sarcosine in metastatic castration-resistant prostate cancer (mCRPC) patients. Patients & methods: Data from 52 mCRPC patients treated with docetaxel-based chemotherapy were retrospectively analyzed. Receiver operating characteristic curves, and Kaplan-Meier and Cox multivariate analyses were performed. Results: Median sarcosine values were significantly higher in mCRPC versus non-mCRPC patients (0.81 vs 0.52 nmol/µl; p < 0.0001). A significant correlation resulted between serum sarcosine levels and the duration of hormone sensitivity (Spearman's correlation coefficient: -0.51; p = 0.001). At multivariate analysis sarcosine was an independent prognostic factor of outcome in terms of overall and progression-free survival. Conclusion: Serum sarcosine values were significantly increased in patients with metastatic disease. Moreover, this biomarker is a risk factor for progression and survival in chemotherapy-treated mCRPC patients.

Introduction & Objectives: High-throughput analysis of low-molecular-weight metabolites, represents a new tool for the global assessment of a cellular state, taking into account genetic regulation, altered kinetic activity of enzymes, and changes in metabolic reactions and regulation. Recently, sarcosine, an N-methyl derivative of glycine, was identified as a differential metabolite highly increased during prostate cancer (PCa) progression. In the present study we assessed the utility of sarcosine detected in serum as a biomarker for PCa and reported the association between the sarcosine levels and clinical-pathological parameters. Materials & Methods: In prospective fashion, sarcosine was measured in serum samples from subjects 289 PCa patients and 312 patients with no evidence of malignancy (NEM), confirmed by 8–12 core prostate biopsies. Sarcosine was measured using the Sarcosine Assay Kit (Biovision, Mountain View, CA, USA) following the manufacturer’s instructions. Nonparametric statistical tests and receiver operating characteristics (ROC) analyses were performed to assess the diagnostic performance. Results: According to A. Sreekumar et al. study, we initially restricted the analysis to patients with PSA values in the grey zone of 2-10 ng/ml, but we didn’t find a better diagnostic performance of sarcosine (AUC=0.57; 95% CI: 0.52, 0,62) vs total PSA (AUC=0.56; 95% CI: 0.51, 0,61) (p=0.7) and vs % fPSA. (AUC=0.68; 95% CI: 0.63, 0,73) (p=0.004). After we investigated the performance of sarcosine in other range of PSA (PSA<4 ng/ml; 410 ng/ml). ROC analysis on subjects with PSA<4 mg/l showed a higher predictive value of sarcosine vs total PSA (sarcosine AUC= 0.668; 95% CI: 0.58 to 0,74 vs total PSA AUC=0,53; 95% CI: 0.45 to 0,62) (p=0.039). ROC analyses for the other two ranges of PSA, showed the predictive superiority of %PSA vs sarcosine. Sarcosine values were not associated with tumour stage (pT2 vs pT3) or grade (Gleason score <7 vs ≥7). Conclusions: We reported the first independent study to validate the role of serum sarcosine in the diagnosis of prostate cancer. We provided evidence that serum sarcosine had a higher predictive value than tPSA and % fPSA in patients with PSA<4 ng/ml. Moreover serum sarcosine was not associated with tumor stage and grade, weakening its possible role in predicting cancer aggressiveness, as initially postulated

Objectives: The aim of this study was to carry out clinical and radiographic outcomes of bone healing using a new medical device, a sterile gel formulation of sodium hyaluronate and amino acids Gly-Leu-Pro-Lys (AMINOGAM®) in treatment of third stage bisphosphonates-related osteonecrosis of the jaws (BRONJ). Materials and methods: We selected 32 third stage BRONJ patients divided in two groups according to systemic pathology: - Neoplatic diseases group that includes 21 patients; - Non-neoplastic diseases group of 11 patients. According to AAOMS guidelines, all patients suspended biphosphonate therapy three-six months before the surgery and were subjected to antibiotic therapy: three courses of 1g ceftriaxone intramuscular injection/die and 250mg metronidazolo oral tablet two times/die for 8 days with 10 days rest between each course. Surgical treatment provides local anesthesia without vasoconstrictor, segmental resection, Piezosurgery osteoplasty, intracavitary intraoperative use of gel to fill up residual bone defect and a first application upon the stitches (sandwich technique). Our procedure includes using of gel 4 times/die till to complete mucosal healing. Finally a clinical and radiographic follow-up by orthopantomograph and CT examinations at 3, 6, 12 and 24 months was carried out. Results: Clinical outcomes showed complete hard and soft tissue healing in all post-surgical sites, with a difference between two groups: neoplastic deseases group needed a longer soft wound healing time of 5 days compared to non-neoplastic diseases group. Radiographic outcomes show radiolucent areas decreasing due to gel direct osteoinductive effect with a faster osteoregeneration time in non-neoplastic deseases group: 15% difference between ossification level at 3 and 6 months. Gel preparation of sodium hyaluronate and amino acids enhances angiogenesis, fibroblast and osteoblast proliferation, collagen biosynthesis and production of growth factors as evidenced by MTT test and alkaline phosphatase histochemical staining. In vivo and in vitro studies have suggested that hyaluronic acid plays important roles in bone wound healing by enhancement of osteoblast differentiation through the down-regulation of BMP-2 antagonists. Lysine and proline regulate collagen matrix synthesis during osteogenesis. Conclusions: Sodium hyaluronate and amino acids gel formulation decreases postoperative pain, swelling and infective complications after surgery by surgical wound mechanical protectection. This new medical device is biocompatible, extremely cheap, safe and useful in all surgical procedure in order to obtain a faster healing of oral hard and soft tissues, specially in BRONJ that are often prone to difficult, slow and complicate recovery.

Diffuse large B cell lymphoma (DLBCL) is recognized as the most common form of non-Hodgkin lymphoma (NHL), accounting for about 40 % of all cases of NHL. Among the cellular components of the tumor inflammatory infiltrate, T cells and mast cells have been demonstrated to be correlated with tumor angiogenesis. In this report, we have investigated CD3 and tryptase expression and their relationship with microvascular density (MVD) in DLBCL patients. Moreover, we determined the significance of CD3 expression in bulky and non-bulky disease. CD3 expression was significantly lower in bulky disease patients when compared to non-bulky ones. CD3 showed a positive correlation with tryptase and MVD, while multiple regression analysis efficaciously predicted MVD depending on CD3 and tryptase as predictors, supporting a complex interplay between these cells in sustaining tumor angiogenesis in DLBCL patients.

Cervico-facial actinomycosis is an infectious, suppurative, and granulomatous disease due to Actinomyces species. Usually, the diagnosis is confirmed by microbiological cultures; however, the need for careful anaerobic handling of specimens often makes it difficult to obtain an effective microbial growth. Therefore, we conducted a retrospective study on biopsy samples from patients with a clinical suspicion of cervico-facial actinomycosis, in order to determine whether accurate histopathological examination could reliably confirm the diagnosis. A retrospective revision of formalin-fixed, paraffin-embedded archival material from 68 cases of cervico-facial lesions, with negative culture for anaerobic/microaerophilic microorganisms, was performed. Twelve serial sections for each case were cut from the paraffin blocks, individually collected on positively charged slides to obtain good section-to-slide adhesion, and stained with hematoxylin and eosin (H&amp;E) and periodic acid-Schiff (PAS). Histopathological examination of the serial sections allowed the identification of bacterial colonies consistent with actinomycetes in 22 cases (32 %). The proposed histopathological examination allowed the retrospective diagnosis of cervical actinomycosis in one-third of clinical specimens that remained misdiagnosed following traditional H&amp;E examination.

Objective: We report on a case of Tuberous Sclerosis (TS) with gingival angiofibromatosis (GA), diagnosed by histopathological analysis with Confocal Laser Scanning Microscopy (CLSM) and treated with High-Power Diode Laser gingivectomy. Case presentation: The patient underwent gingivectomy and gingivoplasty with High-Power Diode Laser in pulsed modality and the surgical sample was formalin-fixed, paraffin-embedded and stained with hematoxylin-eosin and Pricrosirius red. Results: Microscopically, thickened acanthotic epithelium with elongated rete ridges, densely packed, whorly collagen fibers, fibroblasts, variably sized vascular structures, and a few chronic inflammatory cells were detected. At CLSM examination, (Nikon Eclipse E-600 with green/red Laser inducing fluorescence) the collagen fibers, showing intense fluorescence, also manifested variable spatial orientation, due to cross-links among the bundles, ad typical of fibromatosis. Also, variably sized blood vessels and large and polygonal interstitial cells displayed fluorescence of lower intensity. The vascular component consisted of small groups of venous-like structures, frequently showing dilated lumina, thin walls and plump endothelial lining. Conclusions: The histopathological analysis with CLSM of GA occurring in TS highlightes distinctive features, such as low fluorescence areas and a typical vascular component which may represent distinctive features of such lesion.