Potential Benefits of Sequential Inhibitor-Mutagen Treatments of RNA Virus Infections

Lethal mutagenesis is an antiviral strategy consisting of virus extinction associated with enhanced mutagenesis. The use of non-mutagenic antiviral inhibitors has faced the problem of selection of inhibitor-resistant virus mutants. Quasispecies dynamics predicts, and clinical results have confirmed, that combination therapy has an advantage over monotherapy to delay or prevent selection of inhibitor-escape mutants. Using ribavirin-mediated mutagenesis of foot-and-mouth disease virus (FMDV), here we show that, contrary to expectations, sequential administration of the antiviral inhibitor guanidine (GU) first, followed by ribavirin, is more effective than combination therapy with the two drugs, or than either drug used individually. Coelectroporation experiments suggest that limited inhibition of replication of interfering mutants by GU may contribute to the benefits of the sequential treatment. In lethal mutagenesis, a sequential inhibitor-mutagen treatment can be more effective than the corresponding combination treatment to drive a virus towards extinction. Such an advantage is also supported by a theoretical model for the evolution of a viral population under the action of increased mutagenesis in the presence of an inhibitor of viral replication. The model suggests that benefits of the sequential treatment are due to the involvement of a mutagenic agent, and to competition for susceptible cells exerted by the mutant spectrum. The results may impact lethal mutagenesis-based protocols, as well as current antiviral therapies involving ribavirin

Sensitisation of TRPV4 by PAR2 is independent of intracellular calcium signalling and can be mediated by the biased agonist neutrophil elastase

Proteolytic activation of protease-activated receptor 2 (PAR2) may represent a major mechanism of regulating the transient receptor potential vanilloid 4 (TRPV4) non-selective cation channel in pathophysiological conditions associated with protease activation (e.g. during inflammation). To provide electrophysiological evidence for PAR2-mediated TRPV4 regulation, we characterised the properties of human TRPV4 heterologously expressed in Xenopus laevis oocytes in the presence and absence of co-expressed human PAR2. In outside-out patches from TRPV4 expressing oocytes, we detected single-channel activity typical for TRPV4 with a single-channel conductance of about 100 pS for outward and 55 pS for inward currents. The synthetic TRPV4 activator GSK1016790A stimulated TRPV4 mainly by converting previously silent channels into active channels with an open probability of nearly one. In oocytes co-expressing TRPV4 and PAR2, PAR2 activation by trypsin or by specific PAR2 agonist SLIGRL-NH2 potentiated the GSK1016790A-stimulated TRPV4 whole-cell currents several fold, indicative of channel sensitisation. Pre-incubation of oocytes with the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA)-AM did not reduce the stimulatory effect of PAR2 activation on TRPV4, which indicates that the effect is independent of intracellular calcium signalling. Neutrophil elastase, a biased agonist of PAR2 that does not induce intracellular calcium signalling, also caused a PAR2-dependent sensitisation of TRPV4. The Rho-kinase inhibitor Y27362 abolished elastase-stimulated sensitisation of TRPV4, which indicates that Rho-kinase signalling plays a critical role in PAR2-mediated TRPV4 sensitisation by the biased agonist neutrophil elastase. During acute inflammation, neutrophil elastase may sensitise TRPV4 by a mechanism involving biased agonism of PAR2 and activation of Rho-kinase

Prognostic factors and clinical outcome of patients with recurrent early-stage epithelial ovarian cancer: an Italian multicenter retrospective study.

OBJECTIVE: The objective of this study was to assess the clinical outcome of patients with recurrent early-stage ovarian cancer. METHODS: The hospital records of 87 patients were reviewed. The median follow-up of survivors from recurrence was 87.6 months. RESULTS: The 25%, 50%, and 75% quantiles of time to recurrence were 15, 25, and 44 months, respectively. The pelvis was the most common site of failure (39.1%), followed by abdomen (18.3%) and retroperitoneal nodes (18.3%). Treatment at recurrence consisted of chemotherapy in 46 patients, surgery plus chemotherapy in 29, surgery in 3, surgery plus radiotherapy in 2, and other therapies in 7. A macroscopically complete cytoreduction was obtained in 29 (85.2%) of the 34 patients who underwent secondary surgery. Five- and 7-year survival rates after recurrence were 34.3% and 29.6%. By log-rank test, survival after recurrence was related to patient age (≤60 vs. >60 years; P = 0.001), time to recurrence (>15 vs. ≤15 months; P = 0.049), site of recurrence (retroperitoneum vs pelvis vs other; P = 0.004), and surgery at recurrence (yes vs. not; P = 0.001), but not to substage, histotype, grade, prior adjuvant chemotherapy, examination that detected recurrence, and chemotherapy at recurrence. On multivariate analysis, patient age (hazard ratio, 1.836; 95% confidence interval, 1.060-3.180) and surgical treatment at recurrence (hazard ratio, 1.972; 95% confidence interval, 1.084-3.587) were independent prognostic variables for survival after recurrence. CONCLUSIONS: Patient age and surgery at recurrence were independent prognostic variables for patients with recurrent early-stage ovarian cancer. When feasible, salvage surgery appears to give a survival advantage in this clinical setting

The clinical outcome of epithelial ovarian cancer patients with apparently isolated lymph node recurrence: a multicenter retrospective Italian study

The clinical outcome of epithelial ovarian cancer patients with apparently isolated lymph node recurrence: A multicenter retrospective Italian study Angiolo Gadducci a,⁎, Stefania Cosio a, Paolo Zola b, Benedetta Sostegni c, Anna Maria Ferrero b, Giancarlo Teti a, Renza Cristofani d, Enrico Sartori c a Department of Procreative Medicine, Division of Gynecology and Obstetrics, University of Pisa, Via Roma 56, Pisa, 56127, Italy b Department of Gynecology and Obstetrics, University of Turin, Mauriziano Hospital, Turin, Italy c Department of Gynecology and Obstetrics, University of Brescia, Brescia, Italy d Department of Experimental Pathology, University of Pisa, Pisa, Italy a r t i c l e i n f o a b s t r a c t Article history: Received 3 September 2009 Available online 1 December 2009 Keywords: Epithelial Ovarian cancer Prognostic variable Lymph node recurrence Chemotherapy Secondary surgical cytoreduction Objectives. To assess the clinical outcome of epithelial ovarian cancer patients who developed an apparently isolated lymph node recurrence after primary therapy. Methods. The authors retrospectively assessed 69 patients with epithelial ovarian cancer who were clinically or pathologically free of disease after primary therapy and who subsequently developed an apparently isolated lymph node recurrence. The median follow-up of survivors was 74.5 months. Results. Median age was 58 years, FIGO stage was III–IV in 52 (75%) patients, residual disease after primary surgery was N1 cm in 36 (52%), first-line chemotherapy consisted of paclitaxel-/platinum-based chemotherapy in 44 (64%), time to recurrence was N12 months in 43 (62%), recurrence was pelvic and/or para-aortic in 41 (59%), and treatment at recurrence consisted of chemotherapy alone in 44 (64%), surgery plus chemotherapy in 22 (32%), surgery alone in one patient, surgery plus irradiation in one, and irradiation alone in one patient. Survival after recurrence was significantly related to the type of treatment (chemotherapy alone versus surgery plus chemotherapy, median: 20.8 months versus not reached, p=0.0002), and patient age (N58 versus b58 years, median: 26.8 versus 44.0 months, p=0.02). Overall survival was significantly related to the type of treatment (chemotherapy alone versus surgery plus chemotherapy, median: 45.4 months versus not reached, p=0.0001), patient age (N58 versus b58 years, median: 45.4 versus 62.9 months, p=0.03) and time to recurrence (b12 months versus N12 months, median: 45.4 versus 66.9 months, p=0.01). Cox model showed that treatment at recurrence was the strongest independent prognostic variable for both survival after recurrence (hazard ratio [HR]=0.277, p=0.0003) and overall survival (HR=0.249, p=0.0002). Conclusion. Patients who underwent surgery plus chemotherapy had a 72% reduction in the risk of death after recurrence and a 75% reduction in the risk of death after initial diagnosis when compared with those treated with chemotherapy alone. Secondary cytoreductive surgery appears to be able to prolong survival in epithelial ovarian cancer patients with apparently isolated lymph node recurrence. © 2009 Elsevier Inc. All rights reserved

Prognostic factors in early-stage ovarian cancer.

The purpose of this study was to identify the main prognostic factors in patients with early-stage epithelial ovarian cancer. Data were extracted from 222 patients with initial stage (I–IIA) invasive epithelial ovarian cancer treated with primary surgery followed or not followed by adjuvant therapy, from 1 January 1980 to 31 December 2008, at the Division of Obstetrics and Gynecology, Spedali Civili, Brescia, Italy; the median follow-up was 79 months (SD ± 35,945, range 20–250 months). The negative prognostic factors that were statistically significant (p<0.050) in univariate analysis were grade 2, 3, and X (clear cell in our study); stage IB, IC, IIA; positive peritoneal cytology, age equal to/greater than 54; dense adhesions; capsule rupture (pre-operative or intra-operative) and endometrioid histotype (only for disease-free survival (DFS)). Positive cytology was strongly associated with peritoneal relapses, while adhesions were associated with pelvic relapses. A positive prognosis was associated with the mucinous histotype. Conservative treatment had been carried out in 52% of patients under 40 years of age, and we detected only two relapses and three completions of surgery after a few weeks among 31 women in total. Our study indicated a possible execution in patients with patients with cancer stage IA G1–G2 (p=0.030) or IC G1 (p=0.050), provided well staged. Adjuvant chemotherapy improved the survival of cancers that were not IA G1. The positive prognostic role of taxanes must be emphasised, when used in combination with platino

Adjuvant treatment and analysis of failures in patients with high-risk FIGO Stage Ib-II endometrial cancer: An Italian multicenter retrospective study (CTF study)

Objectives The purpose of this retrospective study was to assess the clinical outcome of patients with high-risk, early-stage endometrioid endometrial cancer (stage Ib or II with myometrial invasion > 50%, grade 2-3). Methods We assessed 192 patients who underwent hysterectomy, bilateral salpingo-oophorectomy and pelvic lymphadenectomy, had histologically negative pelvic nodes, and had negative CT findings for aortic node involvement. Results Tumor relapsed in 36 patients after a median time of 21.2 months. The recurrence was vaginal in 7 (19.4%), distant in 16 (44.4%), aortic in 8 (22.2%), and involved multiple sites in 5 (13.9%). There was a trend to a lower vaginal recurrence rate in the 143 patients who received adjuvant radiotherapy (+ chemotherapy) compared with the 46 who did not (2.1% versus 8.7%). Distant or aortic recurrences were lower in the 37 patients who received adjuvant chemotherapy (+ radiotherapy) than in the 152 who did not (2.7% versus 18.4%, p = 0.02). Of the 29 patients who received sequential adjuvant chemotherapy and radiotherapy, none developed local recurrence and only one had distant recurrence. There was a trend for a better 5-year progression-free survival and overall survival for the patients who received chemotherapy (+ radiotherapy) compared with those who did not (86.0% versus 71.3%, and 92.3% versus 75.6%, respectively). Conclusions Our data appear to suggest that adjuvant chemotherapy reduces the risk of distant or aortic recurrences and that sequential adjuvant chemotherapy and radiotherapy achieve an excellent local and distant control of disease in these clinical settings

Adjuvant treatment and analysis of failures in patients with high-risk FIGO Stage Ib-II endometrial cancer: an Italian multicenter retrospective study (CTF study).

Abstract OBJECTIVES: The purpose of this retrospective study was to assess the clinical outcome of patients with high-risk, early-stage endometrioid endometrial cancer (stage Ib or II with myometrial invasion >50%, grade 2-3). METHODS: We assessed 192 patients who underwent hysterectomy, bilateral salpingo-oophorectomy and pelvic lymphadenectomy, had histologically negative pelvic nodes, and had negative CT findings for aortic node involvement. RESULTS: Tumor relapsed in 36 patients after a median time of 21.2 months. The recurrence was vaginal in 7 (19.4%), distant in 16 (44.4%), aortic in 8 (22.2%), and involved multiple sites in 5 (13.9%). There was a trend to a lower vaginal recurrence rate in the 143 patients who received adjuvant radiotherapy (+chemotherapy) compared with the 46 who did not (2.1% versus 8.7%). Distant or aortic recurrences were lower in the 37 patients who received adjuvant chemotherapy (+radiotherapy) than in the 152 who did not (2.7% versus 18.4%, p=0.02). Of the 29 patients who received sequential adjuvant chemotherapy and radiotherapy, none developed local recurrence and only one had distant recurrence. There was a trend for a better 5-year progression-free survival and overall survival for the patients who received chemotherapy (+radiotherapy) compared with those who did not (86.0% versus 71.3%, and 92.3% versus 75.6%, respectively). CONCLUSIONS: Our data appear to suggest that adjuvant chemotherapy reduces the risk of distant or aortic recurrences and that sequential adjuvant chemotherapy and radiotherapy achieve an excellent local and distant control of disease in these clinical settings