N Funel

Histopathologic evaluation of liver metastases from colorectal cancer in patients treated with FOLFOXIRI plus bevacizumab.

Background:The FOLFOXIRI regimen produces a high rate of radiological and histopathological responses. Bevacizumab added to chemotherapy showed an improvement in pathological response and necrosis of colorectal liver metastases (CLMs). FOLFOXIRI plus bevacizumab produced promising early clinical results and is under investigation in several randomised trials, although no data are currently available on its effects on response of CLMs and on liver toxicities.Methods:Starting from 499 patients enrolled in first-line phase II/III trials, we selected on the basis of tissue sample availability 18 patients treated with FOLFOXIRI/XELOXIRI and 24 patients treated with FOLFOXIRI plus bevacizumab who underwent secondary resection of CLMs. The 28 untreated patients who underwent primary resection of CLMs were included as control group. Responses of CLMs and chemotherapy-induced toxicities were assessed.Results:Among the patients, 63% of those treated with FOLFOXIRI plus bevacizumab, as compared with 28% of those treated with only FOLFOXIRI/XELOXIRI, showed a histopathological response (P=0.033). In the two groups, 52% and 12.5%, respectively, showed necrosis ⩾50% (P=0.017). The incidence of liver toxicities was not significantly increased in patients treated with FOLFOXIRI plus bevacizumab.Conclusion:The addition of bevacizumab to FOLFOXIRI produces high rates of pathologic responses and necrosis of CLM without increasing liver toxicity.

Adjuvant chemotherapy seems beneficial for invasive intraductal papillary mucinous neoplasms

AIMS The incidence of intraductal papillary mucinous neoplasm (IPMN) is rising and these neoplasms now represent up to 25% of resected pancreatic neoplasms. The optimal postoperative management of resected invasive IPMN is still debated in the absence of large prospective clinical trials and of validated prognostic factors in this setting. The objective of our study was to identify potential prognostic factors and to investigate the role of adjuvant therapies for patients radically resected for invasive IPMN. METHODS We retrospectively reviewed clinical and pathological data regarding a large series of patients with invasive IPMN who underwent surgical resection in the last six years at University Hospital of Pisa. RESULTS Sixty-four patients were considered for the analysis, thirty-three of whom received adjuvant chemotherapy with gemcitabine. In our series node involvement and high tumoral grade emerged as the major pathologic prognostic factors. Patients treated with adjuvant chemotherapy with gemcitabine experienced a longer disease-free survival than those who received surgery alone. CONCLUSIONS Gemcitabine-based chemotherapy seems beneficial as adjuvant treatment for patients with resected invasive IPMN.

Ectopic expression of FSH receptor isoforms in neoplastic but not in endothelial cells from pancreatic neuroendocrine tumors

FSH receptor (FSHR) expression is restricted to gonads, where it drives FSH-dependent cell differentiation; in addition, FSHR plays an important role in the regulation of ovarian angiogenesis. Recently, FHSR expression has been shown in blood vessels of various tumors. However, pancreatic neuroendocrine tumors (p-NET), which have high-degree blood supply, were not included in that study. The aim of this study was to evaluate FSHR expression in p-NET. FSHR expression was evaluated in tumor samples from 30 patients with p-NET by immunohistochemistry and Western blot; fluorescence microscopy was used to localize FSHR in specific cells from tissue samples. von Willebrand factor (vWF) and chromograninA (chrA) was used as blood vessel and NET cells marker, respectively, to co-localize FSHR. FSHR expression was detected in all p-NET by immunohistochemistry. Western blot confirmed FSHR expression on p-NET although different FSHR isoforms, ranging from 240 kD to 55 kD were found in the samples studied. Surprisingly, FSHR co-localized with chrA but not with vWF, suggesting that neoplastic cells of neuroendocrine origin rather than blood vessels expressed FSHR. No relationship was found between degree of FSHR expression and histology of p-NET. FSHR may be aberrantly expressed in neoplastic cells from p-NET and not in tumor blood vessels; however, its biological significance as well as its clinical relevance remains to be elucidated.

Sub-micrometric liposomes as drug delivery systems in the treatment of periodontitis.

Periodontitis is a complex disease and bacterial infection is one of the most common factors involved in this disease. Current strategies for the local delivery of antibiotics do not allow a complete clearance of bacteria filling dentinal tubules and this limits their therapeutic efficacy. Therefore, there is a strong need for the development of new delivery strategies aimed at improving the efficacy of antibiotic therapy for periodontitis with special reference to their ability to penetrate into the tubules. The aim of the present study is to develop liposome-based delivery systems of sub-micron dimension, able to diffuse into the dentinal tubules. A further aim of the research is to develop a protocol for enhanced diffusion based on the use of magnetic liposomes and magnetic fields. Liposomes were produced by hydration of a pre-liposomal formulation. The vesicles were stabilised with PEG and their re-sizing was achieved by extrusion. Magnetite nanoparticles were synthesized inside the vesicles, i.e., the chemical reaction involving FeCl2, FeCl3 and NH3 occurred within the core of the newly formed liposomes. Dynamic Light Scattering analysis was performed for size characterization. A mathematical model was implemented to predict the diffusion of the liposomes in dentinal tubules. Ex-vivo validation was performed on extracted human teeth. We produced PEG-ylated liposomes (average size 204.3 nm) and PEG-ylated magnetic liposomes (average size 286 nm) and an iron content of 4.2μg/ml. Through mathematical modelling, we deduced that sub-micrometer vesicles are able to penetrate into dentinal tubules. This penetration is considerably more effective when the vesicles are magnetized and subjected to an external magnetic field which accelerates their movement within the tubules. The liposome-based delivery systems developed by the present study are able to penetrate deeply into the tubules, sometimes reaching their terminal ends.

Correlation of basal EGFR expression with pancreatic cancer grading but not with clinical outcome after gemcitabine-based treatment

Philip et al. [1] recently published the results of a phase III study in 745 patients with locally advanced/metastatic pancreatic ductal adenocarcinoma (PDAC) treated either with gemcitabine and the anti-epidermal growth factor receptor (EGFR) cetuximab or with gemcitabine alone. This trial was based on evidence from earlier studies that suggested the combination had sufficient activity to warrant further testing but failed to meet its primary end point of improving overall survival (OS). However, the combination of gemcitabine with the epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) erlotinib translated into a significant improvement of OS [2]. Although increase in tumor control rates and association between rash and outcome seen in the gemcitabine + erlotinib study suggested that therapeutic benefit is confined to a subset of patients, in both trials analysis of EGFR expression failed to reveal a correlation with outcome. No correlation with OS was also observed in patients treated with single-agent gemcitabine [1]. However, previous studies showed a correlation of EGFR expression with grading and prognosis in PDAC patients who underwent only surgical treatment [3]. Therefore, we carried out an immunohistochemical analysis of EGFR in 100 Caucasian patients who underwent PDAC resection and were treated with gemcitabine in order to evaluate the correlation with (i) histological grading and (ii) outcome. Furthermore, we evaluated by uni-/multivariate analysis the role of stage (I–II/ III–IV), lymph node/neural infiltration (yes/no) and resection margin (R0/R1). EGFR staining was detectable at membrane and cytoplasmatic level (Figure 1A) in 84% and 82% of the patients, respectively. The values of EGFR expression, as evaluated by a total score from the analysis of both the number of positive cells and the staining intensity [4], ranged between 0 and 10 (median = 7) (Figure 1B and C). EGFR expression was significantly higher in grade 3 PDAC (P = 0.036, ‘Mann–Whitney test’). The histological differentiation resulted as a prognostic factor, with median OS of 24.8 [95% confidence interval (CI) 15.8–33.7] versus 11.1 months (95% CI 0.2–22.1) in patients with grade 1/2 and grade 3 tumors, respectively (P = 0.018, ‘log-rank test’) (Figure 1D). Similarly, patients with grade 1/2 and grade 3 PDAC had median Progression Free Survival (PFS) of 16.6 (95% CI 11.4–21.8) and 7.3 (95% CI 5.5–9.2) months, respectively (P = 0.007). Among other possible prognostic factors, only lymph node-positive status was correlated with significantly shorter OS and PFS (P = 0.016 and 0.006, respectively). In the Cox proportional hazards model, both grade 3 and lymph node-positive status resulted independent predictive parameters of death (Hazard Ratio (HR) = 2.0, 95% CI 1.1–3.8, P = 0.036 and HR = 3.1, 95% CI 1.1–8.6, P = 0.033, respectively) and progression risk (HR = 2.2, 95% CI 1.2–4.0, P = 0.012 and HR = 3.3, 95% CI 1.3–8.3, P = 0.012, respectively). In contrast, no correlation was observed between EGFR expression and both PFS and OS (Figure 1E), as well as with stage, lymph node/neural infiltration and resection margin. The lack of correlation between EGFR expression and outcome in these patients might be caused by several factors, including modulation of EGFR expression/phosphorylation or related pathways in the relapse/metastatic tumors, which might then differ from resected specimens. Previous studies showed that resistance to cetuximab–gemcitabine in PDAC xenografts was caused by persistent mitogenactivated protein kinase activation and impaired EGFR internalization, associated with constitutive ErBb3 signaling [5]. In conclusion, EGFR expression is correlated with PDAC grading, but it lacks predictive value with respect to outcome in gemcitabine-treated patients both in the palliative and in the adjuvant setting ([1], present study). Future trials should provide the platforms for additional necessary translational research to identify other biomarkers/targets that can improve gemcitabine activity in PDAC.

NSC‑631570 alkaloid mixture modulates the human equlibrative transporter nucleoside 1 and deoxycytidine kinase gene expressions in pancreatic ductal adenocarcinoma cell cultures

Pancreatic ductal adenocarcinoma (PDAC) is treated by palliative chemotherapy for advanced disease. Gemcitabine is the standard drug in both adjuvant and palliative treatment, but yields a marginal impact on disease outcome. Several attempts to improve the efficacy of gemcitabine by addition of a second cytotoxic or targeted agent have not shown a significant survival advantage. NSC-631570, showed greater median survival in combination with gemcitabine compared to gemcitabine alone in the palliative treatment of unresectable PDAC. However, the authors did not study the interactions between alkaloids and molecular determinant expressions involved in the metabolism of gemcitabine. Therefore, the aim of present study was to evaluate the modulation of the expression of two pivotal genes (hENT1 and dCK) involved in gemcitabine activity. In vitro studies on cell lines and primary cell Cultures from PDAC patients were treated with NSC-631570 at IC50 concentration for 48h. the quantitation of gene expression was performed using the standard curve method and the ΔΔCT calculation. Alkaloids mixture positively modulates the expression of hENT1 mRNA in all PDAC cell cultures. The 2 (-ΔΔCt) analyses revealed a mean increase of 2.8 fold with respect to untreated control cells. In two cell lines cells alkaloids positively affects mRNA expression of dCK gene as well. Based on the previous clinical data the Alkaloids-gemcitabine combination appears a promising regimen and the results of the present study provide the experimental basis for the further clinical testing of the NSC-631570-gemcitabine schedule in PDAC patients.

Beta-cell hyperplasia and direct acinar-beta-cell trans-differentiation in insulin-resistant partially eNOS-deficient mice.

and expressed as steady-state plasma glucose (SSPG); regional fat deposition was quantifi ed by CT abdomen and MRS liver. At baseline, similarly-obese IR vs. IS subjects (BMI 30) demonstrated signifi cantly increased waist circumference, diastolic blood pressure, fasting triglyceride concentrations, visceral adipose tissue (VAT), and intrahepatic fat. After mean weight gain of 3.25 kg over four weeks, followed by one week of weight maintenance, tests were repeated. The group as a whole, experienced signifi cant increases in SSPG (38%), total cholesterol (15 mgdL), triglycerides (45%), waist circumference (3.2 cm). IS but not IR individuals exhibited signifi cant increases in waist circumference, VAT, %VAT, VAT/SAT ratio, and intrahepatic fat. Furthermore, multiple linear regression demonstrated signifi cant group (IR/IS) interactions in the relationship between change in regional fat mass and worsening of SSPG: specifi cally, in IS but not IR individulas, increase in intrahepatic fat correlated with increase in SSPG (r=0.77, p=0.027, and as did increase in VAT (r=0.63, p=0.06). These results show that short-term weight gain induces insulin resistance in both IR and IS individuals, but that IS are more prone to experience signifi cant expansion of intraabdominal and intrahepatic fat, which in turn correlate with development of insulin resistance, pointing to a causal relationship. Supported By: ADA (1-11-CT-35)

Molecular mechanisms underlying the synergistic interaction of the novel anticancer drug celandine with gemcitabine in preclinical models of pancreatic cancer.

233 Background: Celandine (Ukrain) increased median survival in combination with gemcitabine compared to gemcitabine alone (10.4 vs. 5.2 months; p<0.001) in unresectable PDAC (Gansauge et al.; Langenbecks Arch Surg 2002). There is compelling evidence that gene transcripts of determinants of gemcitabine activity could be used to tailor PDAC chemotherapy (Giovannetti et al., Cancer Res 2006). METHODS In vitro studies were performed in two ATCC cell lines (PL45 and Mia PaCa-2) and two primary cell cultures obtained from PDAC patients who underwent surgical resections (PPTCC78 and PPTCC109). Cells were treated with celandine at IC50concentration levelsfor 48 h. The total RNA extraction was performed with TRIzol protocol. All the amplifications were carried out with normalization of gene expression against the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) housekeeping control gene, and the quantitation of gene expression was performed using the direct ratio, the standard curve method and the ΔΔCT calculation, in which the amount of target, normalized to the endogenous control and relative to the calibrator (untreated control cells) was calculated as 2(-ΔΔCt). RESULTS Celandine positively modulates the expression of hENT1 mRNA in all PDAC cell cultures treated with IC50 (p<0.001). The 2(-ΔΔCt) analysis revealed a mean increase of 2.8-fold (p=0.001) with respect to untreated control cells. In PL45 and Mia PaCa-2 cells celandine positively affects mRNA expression of dCK gene as well. CONCLUSIONS To date a few options based on gemcitabine are available for treatment of PDAC. Most gemcitabine-based chemotherapy regimens resulted in a very limited disease control, and studies attempting to widen the therapeutic armamentarium against this disease are warranted. Based on the previous clinical data the celandine-gemcitabine combination appears a promising regimen and the results of the present study provide the experimental basis for the further clinical testing of the celandine-gemcitabine schedule in PDAC patients. No significant financial relationships to disclose.

Evaluation of prognostic factors and of the role of adjuvant chemotherapy in patients with invasive carcinoma derived from intraductal papillary mucinous neoplasms (IPMNs) of the pancreas.

e14605 Background: The natural history of invasive carcinomas derived from IPMNs is not well defined; few data on prognostic factors have been reported and the role for adjuvant treatment is not st...