Factors influencing adherence to clinical guidelines in the management of cancer-associated thrombosis.

International academic and regulatory guidelines consistently recommend the long-term use of low-molecular-weight heparins (LMWHs) as the standard for the treatment of cancer-associated thrombosis (CAT). However, both physicians and patients are reluctant to follow established guidelines. Insufficient compliance with treatment recommendations among care physicians represents a loss of opportunity for patients at very high risk of recurrence of venous thromboembolism (VTE) and death. Few data are available regarding adherence to CAT clinical practice guidelines. Based on published data, we aimed to review the gap between guidelines and practice to draw a more precise picture of current practice in order to precisely identify the extent to which patient management is currently lacking with respect to treatment guidelines. Published observational studies, registries and surveys on cancer-associated VTE treatment were reviewed. In spite of evidence from randomized controlled trials (RCTs) showing the usefulness of long-term LMWH, only approximately 50% of patients are managed according to established guideline recommendations. Patient profiles and co-morbidities influence compliance with standard guidelines. A better knowledge of physician and patient-related factors that influence therapeutic decisions may improve the implementation of clinical practice guidelines. Efficient awareness programs including a multidisciplinary approach are necessary to implement guidelines aimed at optimizing the therapeutic management of cancer-associated VTE.

PO-23 - Expression of heparanase in cancer as biomarker of malignancies: overexpression in an aggressive, poor survival gastric cancer "gastric signet ring cell carcinoma" compared with that of other gastric cancers.

INTRODUCTION: Gastric signet ring cell carcinoma (GSRC) is a distinct entity among of other gastric cancer. With unknown etiopathology, their incidence is increasing and it presents a low sensitivity to chemotherapy. AIM: Here, we studied the expression of the heparanase (HPA) in cancer tissues from GSRC patients and several cancer cell lines. HPA is an endo-ß-D-glucuronidase, capable of cleaving the lateral chains of heparan sulfate on cell surfaces and the extracellular matrix at acidic pH. Apart from its well-characterized enzyme activity, HPA also has independent enzymatic functions, such as up-regulation of vascular endothelial growth factor (VEGF) -A and VEGF-C and activation of intracellular signaling pathways involved in, survival, migration and proliferation of tumor cells. It can also induce an hypercoagulability by a non-enzymatic manner. MATERIALS AND METHODS: HPA was tested in several cancer cell lines from ovaries (OVCAR-3, SKOV-3), breast (MDAMB231, MCF7) colon (LS-174), lung (A549), uterus (HELA) and gastric (adenocarcinoma (AGS) and GSRC (KATO-III) using several techniques such as RT-PCR, Q-PCR, immunocytochemistry, flow cytometry and degradation of Fondaparinux at pH 5, evaluated by its anti Xa activity evaluated by Factor Xa amidolytic activity. The amount of HPA mRNA in the biopsy simples of gastric adenocarcinoma (n=10) and GSRC (n=11) in tumors and their environment were analyzed. RESULTS: HPA gene is expressed in all cancer cell lines, but its level varies depending on the tumor cell line. In biopsies of gastric cancer, the HPA gene is more expressed in the tumor regions (p=0.0002) and tumor environment (p=0.015) in GSRC than in gastric adenocarcinoma. B) The activity of HPA, evaluated by degradation of Fondaparinux at pH 5, 1) in the supernatants of 10(6) cancer cells: the residual activity of Fondaparinux after 2 hours incubation at 37°C with OVCAR-3 supernatant was of 70% of control value, and of 80% with KATO-III cell supernatants. 2) in patient's plasmas, this technique cannot be used because the site of degradation of fondaparinux by heparanase is masked by AT present in plasma. CONCLUSIONS: Heparanase was found in in many cancer cell lines and its level depends on origin of tumor cells and on its aggressivity. Taking into account the pro-metastatic functions, proangiogenic and procoagulant activity of HPA and its overexpression in gastric signet ring cell carcinoma of poor prognosis and its cell line, HPA can be considered as a biomarker of malignancy and as a therapeutic target in GSRC patients.

PO-47 - Microparticles derived from ovarian cancer cell line contained genomic and biologically active proteins, including tissue factor involved in coagulation.

INTRODUCTION: The microparticles (MPs) are sized vesicles of less than 1 µm, from different cell types upon activation or subsequent to apoptosis. They are involved in the thrombotic process, particularly in cancer. The role of MPs in ovarian cancer and their involvement in thrombosis being poorly understood. AIM: The aim of this study was to identify in vitro the generation of MPs by an human ovarian adenocarcinoma cell line (OVCAR-3). MATERIALS AND METHODS: OVCAR-3 cells were cultured in three conditions [without stimulation, with protein C (PC), and with activated protein C (APC)]. Then, the MPs present in the supernatant, were isolated by ultracentrifugation and were analyzed for their shape and properties by flow cytometry, electron microscopy, cryofracture analysis, DNA and RNA, and proteomic analysis. The level of tissue factor (TF) on MP was evaluated by TF-induced shortening of Ca(2+) plasma coagulation time. RESULTS: Our results demonstrated that 1) 92% of MPs derived from OVCAR-3 were less than 100 nm. 2) As tested by flow cytometry, the MPs contained b2 microglobulin, annexin, DNA fragments and TF that induces a shortening of Ca(2+) -induced plasma coagulation time. When OVCAR-3 were cultured for 18H with PCA, MPs were generated in greater amount than those generated by OVCAR-3 in its absence and their level of TF was increased of 20%. Curiously, in contrast with intact OVCAR-3 cells, the endothelial protein C receptor (EPCR) was not detected in MPs 3) Proteomic analysis show that the MPs contain proteins involved in cancer progression such as mucins (5A and 5B), keratin type-1, actin, annexin (A1, A2, A4), CD44, glypican, heat shock (70kDa and HS90a) proteins, Agrin associated with heparan sulfate proteoglycan abundant in the tumor-specific basement membrane, Ephrin type A receptor, coronin-1C, catenin α, integrin ß-1 and also p-selectin responsible of platelet activation. They also express several DNA associated proteins includingtranscription factors, various polymerases, nucleases, and histones involved in chromosome packaging and transcription in the cell nucleus. CONCLUSIONS: MPs derived from OVCAR-3 have an apoptotic character. They expressed several biologically active proteins, DNA and their associated proteins. Despite the absence of EPCR expression on MPs that was expressed on intact OVCAR-3 cells, they expressed procoagulant TF activity already found on intact ovarian cancer cells. This activity is greater extent in the presence of APC.

Adherence to treatment guidelines for cancer-associated thrombosis: a French hospital-based cohort study.

PURPOSE: French 2008 treatment guidelines recommend low-molecular-weight heparins (LMWH) for the treatment of cancer-associated thrombosis (CAT) with treatment duration of at least 3 months and up to 6 months and beyond if cancer remains active. Our aim was to assess adherence to guidelines in hospital clinical practice. METHODS: The French hospital database (PMSI) was used to identify patients with CAT admitted to three hospitals of the Paris region to be included in a retrospective cohort study. Adherence to guidelines was assessed in patients included from different treatment periods following the venous thromboembolism (VTE) episode i.e. first 10 days (T1), day 10 to 3 months (T2), months 3 to 6 (T3) and beyond 6 months (T4) when applicable. RESULTS: A total of 240 patients with CAT were included from January 2012 to December 2012 of whom 204 were analyzable. Treatment was adherent to guidelines in 55, 31 and 34 % of patients in T1, T2 and T3 treatment periods, respectively, while overall treatment adherence was found in 52 % of patients. Adherence rates were the highest among patients with pulmonary embolism (PE, 60.5 %), catheter-related thrombosis (62.5 %), class III/IV extended cancer (58.0 %) and metastatic malignancy (60.3 %) while only 40 % with deep vein thrombosis (DVT) received a treatment consistent with guidelines. CONCLUSION: Adherence to guidelines appears insufficient since only half of patients received an appropriate treatment. Adherence dropped significantly across treatment periods T2 and T3. VTE diagnosis and cancer characteristics influenced the anticoagulant prescription. Management of patients with CAT requires further education and information of health care professionals.

Modified technique for the exposure of the inferior alveolar nerve in rats.

The inferior alveolar nerve (IAN), a branch of the mandibular division of the trigeminal nerve, is a major component of the neurovascular bundle along with the inferior alveolar artery and vein. In rats, when exposed using an external oral approach while remaining intact, it can serve as an important tool to study the different effects of neuromediators and assess the role of different groups of nerve fibers. This paper describes a new technique to expose this nerve giving some experimental results to support its usefulness.

Comparison of CT scanograms and cephalometric radiographs in craniofacial imaging.

OBJECTIVE: To compare measurements from human skulls and their images from cephalometric radiographs (CR) and computed tomography (CT) scanograms, in order to gauge the potential clinical use of the latter. DESIGN: Based on specific inclusion criteria, including stable centric occlusion, 13 adult skulls were selected from a larger collection. The mandible was taped to the maxilla after securing the occlusion of teeth and condylar seating in the glenoid fossa. Lateral and posteroanterior cephalographs and CT 'scout views' were taken of each skull by standardized methods. Landmarks were identified on skulls and images. OUTCOME MEASURES: Linear measurements were made on all three records; angular measurements only on CR and CT images. Intraclass correlation coefficients (r) were calculated to assess similarity among records. Paired t-tests were used to compare differences between mean measurements. RESULTS: No statistically significant differences were noted between mean angular values on CR and CT views (P > 0.05). The highest correlations were observed for several vertical midline distances between CT and direct skull measures: 0.82 < r < 0.995-greatest for nasion-menton. For sagittal distances, the highest correlation was between the direct measure of condylion-pogonion and its CR image (r= 0.73). Correlations between CR and skull transverse measures were higher (0.46 < r < 0.80) than the corresponding skull vs. CT measures (0.06 < r < 0.38). CT and CR images are 2D slices and projections, respectively, of 3D structures. Vertical CT and skull measures correspond because the CT projection reflects a 1:1 ratio in the midsagittal plane; CT projected lateral images are smaller than the skull measures. The CR image reflects a distortion (approximately 8%) that brings Co-Pg closer to its anatomic distance, inadvertently contributing to better clinical planning, particularly in orthognathic surgery. The pattern of distortion of PA images was in opposite directions for CR and CT views. CONCLUSIONS: Cephalograms and CT scanograms are close in depicting angular relations of structures, but they differ in the accuracy of imaging linear measurements, because the location and size of an object within the imaged 3D structure varies with both records. Logistic and economic considerations favor the use of cephalographs.

Gagging prevention using nitrous oxide or table salt: a comparative pilot study.

PURPOSE: The objective of this study was to compare two methods for reducing gagging induced by stimulation of the soft palate: table salt and nitrous oxide inhalation sedation. MATERIALS AND METHODS: Fifteen healthy volunteers, eight men and seven women with a mean age of 20.6 years, were subjected to a gagging event three times using a large tablespoon to stimulate the soft palate: event 1 = spoon alone, event 2 = spoon and table salt on the tip of the tongue with a 30-minute break between events 1 and 2, and event 3 = spoon and nitrous oxide sedation on another day. Time in seconds was measured from the moment the spoon touched the soft palate until gagging was felt using a chronometer held by the subject. RESULTS: The mean time for eliciting the gagging reaction was 7.7 seconds for the spoon alone, 8.9 seconds for the spoon and table salt, and 24.0 seconds for the nitrous oxide sedation. Nitrous oxide inhalation sedation significantly (P < .001) reduced the gagging/retching reaction, whereas there was no significant time difference in gagging reaction between stimulation with the spoon alone or when table salt was added. CONCLUSION: Within the limits of this study, table salt did not seem to reduce the time to triggering the gag reflex, whereas nitrous oxide had a substantial effect.

A perfusion technique for the determination of pro-inflammatory mediators induced by intradental application of irritants.

INTRODUCTION: Several morphological and functional features contribute to the consideration of the tooth as a separate compartment having special type of innervation and special immune mechanisms. This study describes a new method allowing the intradental perfusion of rat incisors for the in vivo assessment of pulpal reaction to inflammatory agents. METHODS: Under deep anesthesia, the distal 2-3 mm of each of the rat lower incisors was cut and wrapped in a polyethylene tubing connected to a perfusion chamber made of tigone tubing (ID 1/8 in., volume 100-150 microl). Several groups of rats (n=5 each) were used for intradental application of either saline, capsaicin (100 microg in 100 microl), or endotoxin (ET, 20 microg in 100 microl) for a period of 40 min followed by filling the tooth chamber with saline and collecting the perfusate every 40 min for a period of 8 h. The collected perfusates were stored at -70 degrees C for subsequent determination of the concentration of prostaglandin E(2) (PGE(2)) and nerve growth factor (NGF) by enzyme-linked immunosorbent assay (ELISA). RESULTS: Dentinal injury produced a moderate increase in the levels of NGF and PGE(2) in incisors perfused with saline. Application of ET or capsaicin, however, produced a highly significant increase in the levels of both mediators. These effects peaked at 1.5-3 h for PGE(2) and at 5 h for NGF. Capsaicin showed the most significant effects. DISCUSSION: The reported results cannot be attributed to any factor other than the inflammation of the incisor's pulp, because the described chamber does not allow any spread or leak of the applied irritants. Further studies using other reagents can allow the determination of the variation of the levels of the various pro-inflammatory mediators and their modulation by treatment with anti-inflammatory drugs.