Background: Neoadjuvant chemoradiotherapy followed by surgery is the mainstay treatment for local... more Background: Neoadjuvant chemoradiotherapy followed by surgery is the mainstay treatment for locally advanced rectal cancer, leading to significant decrease in tumor size (downsizing) and a shift towards earlier disease stage (downstaging). Extensive histopathological work-up of the tumor specimen after surgery including tumor regression grading and lymph node status helped to visualize individual tumor sensitivity to chemoradiotherapy, retrospectively. As the response to neoadjuvant chemoradiotherapy is heterogeneous, however, valid biomarkers are needed to monitor tumor response. A relevant number of studies aimed to identify molecular markers retrieved from tumor tissue while the relevance of blood-based biomarkers is less stringent assessed. MicroRNAs are currently under investigation to serve as blood-based biomarkers. To date, no screening approach to identify relevant miRNAs as biomarkers in blood of patients with rectal cancer was undertaken. The aim of the study is to investigate the role of circulating miRNAs as biomarkers in those patients included in the TiMiSNAR Trial (NCT 03465982). This is a biomolecular substudy of TiMiSNAR Trial (NCT03962088). Methods: All included patients in the TiMiSNAR Trial are supposed to undergo blood collection at the time of diagnosis, after neoadjuvant treatment, after 1 month from surgery, and after adjuvant chemotherapy whenever indicated.
In literature, most of the comparative studies of robotic (RRC) versus laparoscopic (LRC) right c... more In literature, most of the comparative studies of robotic (RRC) versus laparoscopic (LRC) right colectomy are biased by the type of the anastomotic technique adopted. With this study, we aim to understand whether there is a role for robotics in performing right colectomies, comparing RRC versus LRC, both performed with intracorporeal anastomosis. In this retrospective cohort study, all consecutive patients who underwent minimally invasive right colectomy (robotic or laparoscopic) with intracorporeal anastomosis in three Italian high-volume centers between February 1, 2007 and December 31, 2017 were included. Patients were grouped according to the method of surgery: RRC or LRC. A total of 389 patients were included in the study (305 RRC vs. 84 LRC). Patients' baseline characteristics were comparable between the groups. Operative time was significantly longer in RRC (250 min, IQR 209-305) group than LRC group (160 min, IQR 130-200) (p < 0.001). The median number of lymph nodes ...
Robot-assisted surgery has been reported to be a safe and effective alternative to conventional l... more Robot-assisted surgery has been reported to be a safe and effective alternative to conventional laparoscopy to treat patients with rectal cancer in a minimally invasive manner. Never the less, substantial data concerning functional outcomes and long term oncological adequacy are still lacking. We aimed to assess the current role of robotics in rectal surgery focusing on functional and oncological outcomes. A comprehensive review of the English-language literature was conducted for articles dealing with robotic-assisted rectal resections. All relevant papers were evaluated on functional implications such as postoperative sexual and urinary dysfunction and oncological outcomes. Robotics showed a general trend toward lower rates of sexual and urinary postoperative dysfunction and earlier recovery compared to laparoscopy. The rates of 3-year local recurrence, disease free survival and overall survival compare favourably with those of laparoscopy. At present, the literature fails to prov...
Robotic surgical system provides many unique advantages which might compensate the limitations of... more Robotic surgical system provides many unique advantages which might compensate the limitations of usual laparoscopic surgery. By using robotic surgical system, we performed robot-assisted laparoscopic pancreaticoduodenectomy (PD). A Sixty-two year old female patient with an ampullary mass underwent robot assisted PD due to imcomplete treatment of endoscopic ampullectomy. The removal of specimen and reconstruction were performed through small upper midline skin incision. Robot working time was about 8 hours, and blood loss was about 800 ml without blood transfusion. She returned to an oral diet on postoperative day 3. Grade B pancreatic leak was noted during the postoperative period, but was successfully managed by conservative management alone. We successfully performed da Vinci-assisted laparoscopic PD, and robot surgical system provided three-dimensional stable visualization and wrist-like motion of instrument facilitated complex operative procedures. More experiences are necessar...
To provide some insight into molecular mechanisms of 5 fluorouracil (5-FU) clinical resistance in... more To provide some insight into molecular mechanisms of 5 fluorouracil (5-FU) clinical resistance in colorectal cancer, we hypothesized that different in vitro exposure schedules of human colorectal cancer cell lines mimicking clinical infusion or bolus regimens could lead to differential gene expression. Resistant HCT-8 colon cancer cell lines (HCT-8/FUI/15R and HCT-8/FUB/2R) were selected from parental sensitive HCT-8 cells by long-term and short-term exposure schedules, respectively. Expression levels of the 437 genes evaluated by the Atlas Select cDNA Expression Human Tumor Array were not substantially different between HCT-8/FUB/2R and HCT-8 cell lines except for three genes downregulated in the resistant subline. Several genes were differentially expressed in HCT-8/FUI/15R cells compared to the parental cell line: 43 genes, including three chemoresistance-related genes, were upregulated, and three genes were downregulated. HCT-8/FUB/2R cells were substantially more resistant to 5...
The rapid diffusion of new technologies in surgery, together with high expectations of both patie... more The rapid diffusion of new technologies in surgery, together with high expectations of both patients and the mass media, has led to many gastrointestinal procedures being approached using robots. Robotic technology seems to resolve many of the drawbacks of laparoscopic advanced procedures, such as anastomotic reconstructions, accurate lymphadenectomy, and vascular sutures. In addition, a deeper tridimensional steady vision with excellent high definition, the EndoWrist technology offering seven degrees of freedom, tremor filtration, scaled motion, optimal working ergonomics, and avoidance of the 'fulcrum effect', are the main strengths of the da Vinci ® system. The use of near-infrared technology and the possibility of tutoring through a double-console will most likely add many more advantages of this technology over laparoscopy alone. However, none of the gastrointestinal robotic interventions has reached a level of evidence-based efficacy that enables it to be routinely applied. The main limitations of robotic gastrointestinal procedures are represented by the learning curve, the higher costs of robotic surgery compared to traditional and laparoscopic surgery, and the longer operation times, including setup and organizational troubles. Moreover, while the limits of robotics for benign diseases are mainly represented by technical issues, oncologic outcomes remain the foundation of any procedures to cure malignancies, and long-term follow-up is still lacking. On the other hand, a word of caution should be presented on the adoption of robotics in too many surgical units without the correct and formal technical background and third-party control to guarantee the best outcomes for patients at minimum risk. Therefore, the robotic treatment of gastrointestinal diseases requires a thorough analysis of the published evidence, in order to determine the correct indications and patient selection. This review aims to examine the evidence for the use of robotic surgery in both malignancies and benign disease arising from the gastrointestinal area. Future developments in robotics and ongoing areas of research are also analyzed.
Helicobacter pylori up-regulates cyclo-oxygenase-2 (COX-2) expression, which in turn is involved ... more Helicobacter pylori up-regulates cyclo-oxygenase-2 (COX-2) expression, which in turn is involved in tumourigenesis. Recently, a causal link between COX-2 and multidrug resistance 1 (MDR-1) gene expression, implicated in cancer chemoresistance, has been demonstrated. Thus, the expression of COX-2 and the downstream enzyme involved in PGE2 biosynthesis, microsomal PGE-synthase1 (mPGES1), was correlated with P-gp, the product of MDR-1, and the anti-apoptotic protein, Bcl-xL, in gastric biopsies from patients with H pylori infection and in patients with gastric cancer. In a retrospective analysis of endoscopic and pathology files, 40 H pylori-negative patients (Hp-), 50 H pylori-positive patients who responded to eradication therapy (Hp+R), 84 H pylori-positive patients who did not respond to eradication therapy (Hp+NR), and 30 patients with gastric cancer (18 intestinal and 12 diffuse types) were selected. COX-2, mPGES1, P-gp, and Bcl-xL were detected by immunohistochemistry. COX-2, mPGES1, P-gp, and Bcl-xL expression was undetectable in gastric mucosa from Hp- patients. By contrast, COX-2 and mPGES1 expression was detected in 42% and 44% of Hp+R patients, respectively, and in up to 66% (range 63-66%) of Hp+NR patients (p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The expression of COX-2 and mPGES1 correlated significantly (p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001) with that of P-gp and Bcl-xL. High levels of COX-2, mPGES1, P-gp, and Bcl-xL expression were found in intestinal-type gastric cancer samples. In conclusion, H pylori-dependent induction of COX-2 and mPGES1 is associated with enhanced production of P-gp and Bcl-xL that may contribute to gastric tumourigenesis and resistance to therapy.
Purpose: Up-regulation of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2... more Purpose: Up-regulation of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) enzymes has been reported in colorectal cancer. We aimed at evaluating the possible interaction between the nitric oxide and COX-2 pathways, and its effect on promoting tumor angiogenesis. Experimental Design: Expression of iNOS, COX-2, vascular endothelial growth factor (VEGF), and CD31 was analyzed in tumor samples and corresponding normal mucosa obtained from 46 surgical specimens. We also evaluated iNOS activity, prostaglandin E 2 (PGE 2), cyclic GMP and cyclic AMP production in the same specimens. Nitrite/nitrate levels, and PGE 2 and VEGF production were assessed in HCT116 and HT29 colon cancer cell lines after induction and selective inhibition of the two enzyme pathways. Results: A significant correlation was found between iNOS and COX-2 immunohistochemical expression. PGE 2 production significantly correlated with iNOS activity and cGMP levels. A significant correlation was also found among PGE 2 production, microvessel density, and VEGF expression. Coinduction of both iNOS and COX-2 activities occurred after lipopolysaccharide (LPS) and epidermal growth factor (EGF) treatment in HCT116 and HT29 cells. Inhibition of iNOS by 1400W significantly reduced both LPS-and EGF-induced PGE 2 production. Treatment with LPS, EGF, and arachidonic acid significantly increased VEGF production in the iNOS-negative/COX-2-positive HT29 cells. This effect was completely reversed by treatment with the selective COX-2 inhibitor celecoxib. Conclusions: Our data showed a prominent role of nitric oxide in stimulating COX-2 activity in colorectal cancer. This interaction is likely to produce a cooperative effect in promoting angiogenesis through PGE 2-mediated increase in VEGF production.
COX-2 is an immediate-to-early response gene undetectable in most normal tissues but promptly ind... more COX-2 is an immediate-to-early response gene undetectable in most normal tissues but promptly induced by proinflammatory and mitogenic stimuli in inflamed and neoplastic tissues (7). An increase in COX-2 mRNA levels and protein ...
To investigate the potential involvement of the nitric oxide (NO) pathway in colorectal carcinoge... more To investigate the potential involvement of the nitric oxide (NO) pathway in colorectal carcinogenesis, we correlated the expression and the activity of inducible nitric oxide synthase (iNOS) with the degree of tumor angiogenesis in human colorectal cancer. Tumor samples and adjacent normal mucosa were obtained from 46 surgical specimens. Immunohistochemical expression of iNOS, vascular endothelial growth factor (VEGF), and CD31 was analyzed on paraffin-embedded tissue sections. iNOS activity and cyclic GMP levels were assessed by specific biochemical assays. iNOS protein expression was determined by Western blot analysis. iNOS and VEGF mRNA levels were evaluated using Northern blot analysis. Both iNOS and VEGF expressions correlated significantly with intratumor microvessel density (r(s) = 0.31, P = 0.02 and r(s) = 0.67, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001, respectively). A significant correlation was also found between iNOS and VEGF expression (P = 0.001). iNOS activity and cyclic GMP production were significantly higher in the cancer specimens than in the normal mucosa (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001 and P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001, respectively), as well as in metastatic tumors than in nonmetastatic ones (P = 0.002 and P = 0.04, respectively). Western and Northern blot analyses confirmed the up-regulation of the iNOS protein and gene in the tumor specimens as compared with normal mucosa. NO seems to play a role in colorectal cancer growth by promoting tumor angiogenesis.
Antimicrobial resistance has decreased eradication rates for Helicobacter pylori infection worldw... more Antimicrobial resistance has decreased eradication rates for Helicobacter pylori infection worldwide. To determine whether sequential treatment eradicates H. pylori infection better than standard triple-drug therapy for adults with dyspepsia or peptic ulcers. Randomized, double-blind, placebo-controlled trial. Two Italian hospitals between September 2003 and April 2006. 300 patients with dyspepsia or peptic ulcers. (13)C-urea breath test, upper endoscopy, histologic evaluation, rapid urease test, bacterial culture, and assessment of antibiotic resistance. A 10-day sequential regimen (40 mg of pantoprazole, 1 g of amoxicillin, and placebo, each administered twice daily for the first 5 days, followed by 40 mg of pantoprazole, 500 mg of clarithromycin, and 500 mg of tinidazole, each administered twice daily for the remaining 5 days) or standard 10-day therapy (40 mg of pantoprazole, 500 mg of clarithromycin, and 1 g of amoxicillin, each administered twice daily). The eradication rate a...
Rapid diagnostic tools for Helicobacter pylori are important in endoscopy. To assess the accuracy... more Rapid diagnostic tools for Helicobacter pylori are important in endoscopy. To assess the accuracy of a new 5-min rapid urease test (UFT300, ABS Cernusco, sul Naviglio, Italy) compared with the 1-h Pyloritek (Serim Laboratories, Elkhart, IN) and the 24-h CLO test (Kimberly-Clark Ballard Medical Products, Roswell, GA), consecutive dyspeptic patients referred to our unit for endoscopy were prospectively studied. All patients underwent a 13C-urea-breath test, histology and the UFT300 (ABS; Cernusco, sul Naviglio, Italy). Two additional rapid urease tests were performed. Patients were deemed infected when both 13C-UBT and histology were positive. Rapid urease tests were read at 1, 5, and 60 min, respectively. Of the 375 enrolled patients, 45.3% were infected with H. pylori. The sensitivity of the new 5-min rapid urease test 300 was 90.3, 94.5, and 96.2% at 1, 5, and 60 min, respectively (specificity 100%). The Pyloritek and the new 5-min rapid urease test were comparable, but the CLO test was not reliable at 5 and 60 min. In conclusion, the new 5-min rapid urease test is comparable to the Pyloritek test, but the CLO test is significantly less sensitive at early time points. Reading the test results at 1 min may increase false-negative results with decreasing sensitivity.
A correlation between d over baseline (DOB) values of the [ 13 C]urea breath test (UBT) and Helic... more A correlation between d over baseline (DOB) values of the [ 13 C]urea breath test (UBT) and Helicobacter pylori clarithromycin resistance has been reported, suggesting a possible predictive role of UBT in therapeutic outcome. However, available data are limited and conflicting. This study aimed to clarify this issue, assessing the possible relationship between H. pylori resistance towards different antibiotics (clarithromycin, metronidazole and levofloxacin) and UBT values. The data showed similar DOB values between susceptible and resistant strains for clarithromycin (46.9±32.3 vs 45.7±30.6; P50.8), metronidazole (46.4±29.6 vs 47.4±37.9; P50.8), and levofloxacin (45.0±30.2 vs 54.2±38.4; P50.08). Likewise, comparable DOB values were observed between susceptible and multidrug-resistant strains (45.4±29.6 vs 54.8±44.8; P50.1). In conclusion, our data failed to find a significant correlation between UBT values and H. pylori antibiotic resistance.
Background: Neoadjuvant chemoradiotherapy followed by surgery is the mainstay treatment for local... more Background: Neoadjuvant chemoradiotherapy followed by surgery is the mainstay treatment for locally advanced rectal cancer, leading to significant decrease in tumor size (downsizing) and a shift towards earlier disease stage (downstaging). Extensive histopathological work-up of the tumor specimen after surgery including tumor regression grading and lymph node status helped to visualize individual tumor sensitivity to chemoradiotherapy, retrospectively. As the response to neoadjuvant chemoradiotherapy is heterogeneous, however, valid biomarkers are needed to monitor tumor response. A relevant number of studies aimed to identify molecular markers retrieved from tumor tissue while the relevance of blood-based biomarkers is less stringent assessed. MicroRNAs are currently under investigation to serve as blood-based biomarkers. To date, no screening approach to identify relevant miRNAs as biomarkers in blood of patients with rectal cancer was undertaken. The aim of the study is to investigate the role of circulating miRNAs as biomarkers in those patients included in the TiMiSNAR Trial (NCT 03465982). This is a biomolecular substudy of TiMiSNAR Trial (NCT03962088). Methods: All included patients in the TiMiSNAR Trial are supposed to undergo blood collection at the time of diagnosis, after neoadjuvant treatment, after 1 month from surgery, and after adjuvant chemotherapy whenever indicated.
In literature, most of the comparative studies of robotic (RRC) versus laparoscopic (LRC) right c... more In literature, most of the comparative studies of robotic (RRC) versus laparoscopic (LRC) right colectomy are biased by the type of the anastomotic technique adopted. With this study, we aim to understand whether there is a role for robotics in performing right colectomies, comparing RRC versus LRC, both performed with intracorporeal anastomosis. In this retrospective cohort study, all consecutive patients who underwent minimally invasive right colectomy (robotic or laparoscopic) with intracorporeal anastomosis in three Italian high-volume centers between February 1, 2007 and December 31, 2017 were included. Patients were grouped according to the method of surgery: RRC or LRC. A total of 389 patients were included in the study (305 RRC vs. 84 LRC). Patients' baseline characteristics were comparable between the groups. Operative time was significantly longer in RRC (250 min, IQR 209-305) group than LRC group (160 min, IQR 130-200) (p < 0.001). The median number of lymph nodes ...
Robot-assisted surgery has been reported to be a safe and effective alternative to conventional l... more Robot-assisted surgery has been reported to be a safe and effective alternative to conventional laparoscopy to treat patients with rectal cancer in a minimally invasive manner. Never the less, substantial data concerning functional outcomes and long term oncological adequacy are still lacking. We aimed to assess the current role of robotics in rectal surgery focusing on functional and oncological outcomes. A comprehensive review of the English-language literature was conducted for articles dealing with robotic-assisted rectal resections. All relevant papers were evaluated on functional implications such as postoperative sexual and urinary dysfunction and oncological outcomes. Robotics showed a general trend toward lower rates of sexual and urinary postoperative dysfunction and earlier recovery compared to laparoscopy. The rates of 3-year local recurrence, disease free survival and overall survival compare favourably with those of laparoscopy. At present, the literature fails to prov...
Robotic surgical system provides many unique advantages which might compensate the limitations of... more Robotic surgical system provides many unique advantages which might compensate the limitations of usual laparoscopic surgery. By using robotic surgical system, we performed robot-assisted laparoscopic pancreaticoduodenectomy (PD). A Sixty-two year old female patient with an ampullary mass underwent robot assisted PD due to imcomplete treatment of endoscopic ampullectomy. The removal of specimen and reconstruction were performed through small upper midline skin incision. Robot working time was about 8 hours, and blood loss was about 800 ml without blood transfusion. She returned to an oral diet on postoperative day 3. Grade B pancreatic leak was noted during the postoperative period, but was successfully managed by conservative management alone. We successfully performed da Vinci-assisted laparoscopic PD, and robot surgical system provided three-dimensional stable visualization and wrist-like motion of instrument facilitated complex operative procedures. More experiences are necessar...
To provide some insight into molecular mechanisms of 5 fluorouracil (5-FU) clinical resistance in... more To provide some insight into molecular mechanisms of 5 fluorouracil (5-FU) clinical resistance in colorectal cancer, we hypothesized that different in vitro exposure schedules of human colorectal cancer cell lines mimicking clinical infusion or bolus regimens could lead to differential gene expression. Resistant HCT-8 colon cancer cell lines (HCT-8/FUI/15R and HCT-8/FUB/2R) were selected from parental sensitive HCT-8 cells by long-term and short-term exposure schedules, respectively. Expression levels of the 437 genes evaluated by the Atlas Select cDNA Expression Human Tumor Array were not substantially different between HCT-8/FUB/2R and HCT-8 cell lines except for three genes downregulated in the resistant subline. Several genes were differentially expressed in HCT-8/FUI/15R cells compared to the parental cell line: 43 genes, including three chemoresistance-related genes, were upregulated, and three genes were downregulated. HCT-8/FUB/2R cells were substantially more resistant to 5...
The rapid diffusion of new technologies in surgery, together with high expectations of both patie... more The rapid diffusion of new technologies in surgery, together with high expectations of both patients and the mass media, has led to many gastrointestinal procedures being approached using robots. Robotic technology seems to resolve many of the drawbacks of laparoscopic advanced procedures, such as anastomotic reconstructions, accurate lymphadenectomy, and vascular sutures. In addition, a deeper tridimensional steady vision with excellent high definition, the EndoWrist technology offering seven degrees of freedom, tremor filtration, scaled motion, optimal working ergonomics, and avoidance of the 'fulcrum effect', are the main strengths of the da Vinci ® system. The use of near-infrared technology and the possibility of tutoring through a double-console will most likely add many more advantages of this technology over laparoscopy alone. However, none of the gastrointestinal robotic interventions has reached a level of evidence-based efficacy that enables it to be routinely applied. The main limitations of robotic gastrointestinal procedures are represented by the learning curve, the higher costs of robotic surgery compared to traditional and laparoscopic surgery, and the longer operation times, including setup and organizational troubles. Moreover, while the limits of robotics for benign diseases are mainly represented by technical issues, oncologic outcomes remain the foundation of any procedures to cure malignancies, and long-term follow-up is still lacking. On the other hand, a word of caution should be presented on the adoption of robotics in too many surgical units without the correct and formal technical background and third-party control to guarantee the best outcomes for patients at minimum risk. Therefore, the robotic treatment of gastrointestinal diseases requires a thorough analysis of the published evidence, in order to determine the correct indications and patient selection. This review aims to examine the evidence for the use of robotic surgery in both malignancies and benign disease arising from the gastrointestinal area. Future developments in robotics and ongoing areas of research are also analyzed.
Helicobacter pylori up-regulates cyclo-oxygenase-2 (COX-2) expression, which in turn is involved ... more Helicobacter pylori up-regulates cyclo-oxygenase-2 (COX-2) expression, which in turn is involved in tumourigenesis. Recently, a causal link between COX-2 and multidrug resistance 1 (MDR-1) gene expression, implicated in cancer chemoresistance, has been demonstrated. Thus, the expression of COX-2 and the downstream enzyme involved in PGE2 biosynthesis, microsomal PGE-synthase1 (mPGES1), was correlated with P-gp, the product of MDR-1, and the anti-apoptotic protein, Bcl-xL, in gastric biopsies from patients with H pylori infection and in patients with gastric cancer. In a retrospective analysis of endoscopic and pathology files, 40 H pylori-negative patients (Hp-), 50 H pylori-positive patients who responded to eradication therapy (Hp+R), 84 H pylori-positive patients who did not respond to eradication therapy (Hp+NR), and 30 patients with gastric cancer (18 intestinal and 12 diffuse types) were selected. COX-2, mPGES1, P-gp, and Bcl-xL were detected by immunohistochemistry. COX-2, mPGES1, P-gp, and Bcl-xL expression was undetectable in gastric mucosa from Hp- patients. By contrast, COX-2 and mPGES1 expression was detected in 42% and 44% of Hp+R patients, respectively, and in up to 66% (range 63-66%) of Hp+NR patients (p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The expression of COX-2 and mPGES1 correlated significantly (p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001) with that of P-gp and Bcl-xL. High levels of COX-2, mPGES1, P-gp, and Bcl-xL expression were found in intestinal-type gastric cancer samples. In conclusion, H pylori-dependent induction of COX-2 and mPGES1 is associated with enhanced production of P-gp and Bcl-xL that may contribute to gastric tumourigenesis and resistance to therapy.
Purpose: Up-regulation of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2... more Purpose: Up-regulation of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) enzymes has been reported in colorectal cancer. We aimed at evaluating the possible interaction between the nitric oxide and COX-2 pathways, and its effect on promoting tumor angiogenesis. Experimental Design: Expression of iNOS, COX-2, vascular endothelial growth factor (VEGF), and CD31 was analyzed in tumor samples and corresponding normal mucosa obtained from 46 surgical specimens. We also evaluated iNOS activity, prostaglandin E 2 (PGE 2), cyclic GMP and cyclic AMP production in the same specimens. Nitrite/nitrate levels, and PGE 2 and VEGF production were assessed in HCT116 and HT29 colon cancer cell lines after induction and selective inhibition of the two enzyme pathways. Results: A significant correlation was found between iNOS and COX-2 immunohistochemical expression. PGE 2 production significantly correlated with iNOS activity and cGMP levels. A significant correlation was also found among PGE 2 production, microvessel density, and VEGF expression. Coinduction of both iNOS and COX-2 activities occurred after lipopolysaccharide (LPS) and epidermal growth factor (EGF) treatment in HCT116 and HT29 cells. Inhibition of iNOS by 1400W significantly reduced both LPS-and EGF-induced PGE 2 production. Treatment with LPS, EGF, and arachidonic acid significantly increased VEGF production in the iNOS-negative/COX-2-positive HT29 cells. This effect was completely reversed by treatment with the selective COX-2 inhibitor celecoxib. Conclusions: Our data showed a prominent role of nitric oxide in stimulating COX-2 activity in colorectal cancer. This interaction is likely to produce a cooperative effect in promoting angiogenesis through PGE 2-mediated increase in VEGF production.
COX-2 is an immediate-to-early response gene undetectable in most normal tissues but promptly ind... more COX-2 is an immediate-to-early response gene undetectable in most normal tissues but promptly induced by proinflammatory and mitogenic stimuli in inflamed and neoplastic tissues (7). An increase in COX-2 mRNA levels and protein ...
To investigate the potential involvement of the nitric oxide (NO) pathway in colorectal carcinoge... more To investigate the potential involvement of the nitric oxide (NO) pathway in colorectal carcinogenesis, we correlated the expression and the activity of inducible nitric oxide synthase (iNOS) with the degree of tumor angiogenesis in human colorectal cancer. Tumor samples and adjacent normal mucosa were obtained from 46 surgical specimens. Immunohistochemical expression of iNOS, vascular endothelial growth factor (VEGF), and CD31 was analyzed on paraffin-embedded tissue sections. iNOS activity and cyclic GMP levels were assessed by specific biochemical assays. iNOS protein expression was determined by Western blot analysis. iNOS and VEGF mRNA levels were evaluated using Northern blot analysis. Both iNOS and VEGF expressions correlated significantly with intratumor microvessel density (r(s) = 0.31, P = 0.02 and r(s) = 0.67, P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001, respectively). A significant correlation was also found between iNOS and VEGF expression (P = 0.001). iNOS activity and cyclic GMP production were significantly higher in the cancer specimens than in the normal mucosa (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001 and P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.0001, respectively), as well as in metastatic tumors than in nonmetastatic ones (P = 0.002 and P = 0.04, respectively). Western and Northern blot analyses confirmed the up-regulation of the iNOS protein and gene in the tumor specimens as compared with normal mucosa. NO seems to play a role in colorectal cancer growth by promoting tumor angiogenesis.
Antimicrobial resistance has decreased eradication rates for Helicobacter pylori infection worldw... more Antimicrobial resistance has decreased eradication rates for Helicobacter pylori infection worldwide. To determine whether sequential treatment eradicates H. pylori infection better than standard triple-drug therapy for adults with dyspepsia or peptic ulcers. Randomized, double-blind, placebo-controlled trial. Two Italian hospitals between September 2003 and April 2006. 300 patients with dyspepsia or peptic ulcers. (13)C-urea breath test, upper endoscopy, histologic evaluation, rapid urease test, bacterial culture, and assessment of antibiotic resistance. A 10-day sequential regimen (40 mg of pantoprazole, 1 g of amoxicillin, and placebo, each administered twice daily for the first 5 days, followed by 40 mg of pantoprazole, 500 mg of clarithromycin, and 500 mg of tinidazole, each administered twice daily for the remaining 5 days) or standard 10-day therapy (40 mg of pantoprazole, 500 mg of clarithromycin, and 1 g of amoxicillin, each administered twice daily). The eradication rate a...
Rapid diagnostic tools for Helicobacter pylori are important in endoscopy. To assess the accuracy... more Rapid diagnostic tools for Helicobacter pylori are important in endoscopy. To assess the accuracy of a new 5-min rapid urease test (UFT300, ABS Cernusco, sul Naviglio, Italy) compared with the 1-h Pyloritek (Serim Laboratories, Elkhart, IN) and the 24-h CLO test (Kimberly-Clark Ballard Medical Products, Roswell, GA), consecutive dyspeptic patients referred to our unit for endoscopy were prospectively studied. All patients underwent a 13C-urea-breath test, histology and the UFT300 (ABS; Cernusco, sul Naviglio, Italy). Two additional rapid urease tests were performed. Patients were deemed infected when both 13C-UBT and histology were positive. Rapid urease tests were read at 1, 5, and 60 min, respectively. Of the 375 enrolled patients, 45.3% were infected with H. pylori. The sensitivity of the new 5-min rapid urease test 300 was 90.3, 94.5, and 96.2% at 1, 5, and 60 min, respectively (specificity 100%). The Pyloritek and the new 5-min rapid urease test were comparable, but the CLO test was not reliable at 5 and 60 min. In conclusion, the new 5-min rapid urease test is comparable to the Pyloritek test, but the CLO test is significantly less sensitive at early time points. Reading the test results at 1 min may increase false-negative results with decreasing sensitivity.
A correlation between d over baseline (DOB) values of the [ 13 C]urea breath test (UBT) and Helic... more A correlation between d over baseline (DOB) values of the [ 13 C]urea breath test (UBT) and Helicobacter pylori clarithromycin resistance has been reported, suggesting a possible predictive role of UBT in therapeutic outcome. However, available data are limited and conflicting. This study aimed to clarify this issue, assessing the possible relationship between H. pylori resistance towards different antibiotics (clarithromycin, metronidazole and levofloxacin) and UBT values. The data showed similar DOB values between susceptible and resistant strains for clarithromycin (46.9±32.3 vs 45.7±30.6; P50.8), metronidazole (46.4±29.6 vs 47.4±37.9; P50.8), and levofloxacin (45.0±30.2 vs 54.2±38.4; P50.08). Likewise, comparable DOB values were observed between susceptible and multidrug-resistant strains (45.4±29.6 vs 54.8±44.8; P50.1). In conclusion, our data failed to find a significant correlation between UBT values and H. pylori antibiotic resistance.
Uploads
Papers by Federico Perna