Treatment of diffuse malignant peritoneal mesothelioma (DMPM) by cytoreductive surgery and HIPEC.

AIM: Diffuse malignant peritoneal mesothelioma (DMPM) is a rare and locally aggressive tumor with poor prognosis, related in most cases to asbestos exposure. It is increasing in frequency, but currently no standard therapy is available. The biology of this disease is still poorly understood. Several highly specialized centers have recently reported improved survival by means of an innovative local-regional approach. The purpose of this article is to evaluate the survival benefit and the morbidity rate of patients affected by DMPM treated at our institution by cytoreductive surgery (CRS) associated with hyperthermic intraperitoneal perioperative chemotherapy (HIPEC). METHODS: This study includes 42 patients affected by DMPM treated by an uniform approach consisting of cytoreductive surgery associated with HIPEC using cisplatin and doxorubicin. The primary end point was overall survival and morbidity rate. The secondary end point was evaluation of prognostic variables for overall survival. RESULTS: The median follow-up period was 72 months (range 1-235 months). Thirty-five patients (83.3%) presented epithelial tumors and 7 were affected by multicystic mesothelioma. The mean peritoneal cancer index (PCI) was 13. Thirty-eight patients (90.4%) had complete cytoreduction (CC-0/1). The overall morbidity rate was 35.7% associated to a perioperative mortality of 7.1%. Median overall survival rate was 65 months with a 1- and 5-year survival rates of 63% and 44%, respectively. CONCLUSION: The treatment of DMPM by CRS+HIPEC in selected patients is a feasible technique that allows to achieve encouraging results in terms of overall survival rate, with an acceptable morbidity rate. Further investigations are needed to clarify the role and the timing of this promising technique.

Prevalence and risk factors for Chlamydia trachomatis infection in young women in north-west of Italy.

AIM: The aim of the study was to estimate the prevalence of Chlamydia trachomatis infection, risk factors and best predictors of infection in young sexually active women in north-west of Italy. METHODS: One thousand one hundred and eighty 18-24 years old women of family planning clinics and three STI Clinics over Turin city area underwent vaginal swabs to detect infection and completed a questionnaire. Logistic regression and multivariate analysis identified risk factors and a receiver operating characteristic (ROC) curve was used to assess the model accuracy. RESULTS: Overall prevalence of infection was 10.4% 2.71 times higher (P<0.001) among women afferent to STI clinics than family planning clinics. Higher among women of Eastern European, Asian and South American ethnic origin (P=0.012) compared to Western European or African ethnic origin. Age at first intercourse (P=0.006), absence of a stable partner (P<0.001) partner with urogenital complaints (P<0.001), number of lifetime partners (P<0.001) number of partners in the last 6 months (P<0.001) history of occasional intercourse (P<0.001) and of IST (P<0.007) resulted associated with chlamydial infection. Multivariate analysis showed setting, partner with urogenital complaints and number of lifetime partners as best predictors of infection. ROC curve on variables from multivariate analysis showed an AUC of 0.732. CONCLUSION: The study showed high rates of Chlamydial infection among sexually active women between 18-24 years in north-west area of Italy. Predictors of infection are related to sexual activity and to population sub-groups. Selective screening protocols should be supported by wider and more representative studies in order to increase knowledge and involve public opinion.

Reduction of opioid dependence by the CB(1) antagonist SR141716A in mice: evaluation of the interest in pharmacotherapy of opioid addiction.

Several compounds, mainly opioid agonists such as methadone, are currently used for long term medication of heroin addicts. Nevertheless, these maintenance treatments have the disadvantage to induce a dependence to another opiate. As interactions between opioid and cannabinoid systems have been demonstrated, the ability of the CB(1) antagonist, SR141716A to reduce morphine-induced addiction was investigated. The effects of SR141716A on the rewarding responses of morphine were evaluated in the place conditioning paradigm. No significant conditioned preference or aversion were observed after repeated treatment with the CB(1) antagonist alone. However, SR141716A was able to antagonize the acquisition of morphine-induced conditioned place preference. SR141716A was co-administered with morphine for 5 days, and the withdrawal syndrome was precipitated by naloxone administration. A reduction in the incidence of two main signs of abstinence: wet dog shakes and jumping was observed while the other were not significantly modified. In contrast, an acute injection of the CB(1) antagonist just before naloxone administration was unable to modify the incidence of the behavioural manifestations of the withdrawal, suggesting that only chronic blockade of CB(1) receptors is able to reduce morphine-induced physical dependence. Several biochemical mechanisms could explain the reduction of opioid dependence by CB(1) antagonists. Whatever the hypotheses, this study supports the reported interaction between the endogenous cannabinoid and opioid systems, and suggests that SR 141716A warrants further investigations for a possible use in opioid addiction.

Generation of monoclonal antibodies specifically directed against the proximal zinc finger of HIV type 1 NCp7.

The HIV-1 NCp7 contains two spatially close zinc fingers, required for the production of infectious particles. To investigate in more detail the function of the zinc finger domain, monoclonal antibodies were generated with a cyclic analog of the NCp7 proximal zinc finger. This analog was shown to bind zinc ions and to preserve the highly folded structure of the native peptide (Dong C-Z et al.: J Am Chem Soc 1995;117:2726-2731). We report here two monoclonal antibodies (2B10 and 4D3), which are the first monoclonal antibodies directed against CCHC NCp7 zinc fingers. Dot-blot experiments revealed that a few nanograms of synthetic NCp7 can be detected on a nitrocellulose membrane. Whereas 2B10 appears specific for an epitope located in sequence 19-27 of NCp7, 4D3 appears to be structurally specific. Immunocomplex affinities were evaluated, using BIAcore technology, to be up to 1 and 10 nM, respectively, for 2B10 and 4D3 in 100 mM NaCl. These antibodies were able to recognize NCp7 in the Gag polyprotein precursor and were shown to immunoprecipitate NCp7 from a cell supernatant. Moreover, NCp7-Vpr interaction mediated by the zinc fingers is inhibited by 2B10, emphasizing the role of these domains in the protein-protein complex. These results indicate that 2B10 and 4D3 behave as useful tools for studying both NC protein functions during the course of virion morphogenesis and the role played by its zinc finger domain at various steps in the retroviral life cycle.

Sexually transmitted diseases and pelvic inflammatory disease.

AIM: The aim of this study was to determine the prevalence in the Turin area of the pathogens most implicated in pelvic inflammatory disease (PID), with particular regard to which risk factors the population taken into consideration is exposed to. METHODS: From January 1st 1997 to December 31(st) 2001, 13809 women, aged between 14-54, all subjects being fertile and sexually active, were examined for the first time at St. Anna Hospital in Turin for the diagnosis of sexually transmitted diseases (STDs). A total of 5559 unselected patients were divided into 2 groups according to the presence (1721) or absence (3838) of subjective symptoms related to PID. Both groups underwent a cervico-vaginal bacteriological test for common pathogens, Candida spp., T. vaginalis, bacterial vaginosis, C. trachomatis, Mycoplasma spp., N. gonorrhoeae. The prevalence of each micro-organism was coupled with the anamnestic data collected from a pre-determined questionnaire submitted to all patients. The questionnaire collected personal data: age at the time of first sexual intercourse; the number of partners in the last 6 months; the type of contraceptives used. Statistical analysis was performed using a chi squared test. RESULTS: In our analysis 2 factors proved to be decisive for a correct PID diagnosis: a subjective symptomatology and an anamnesis mainly focused on risk factor evaluation. This result is in accordance with what has been emphasized many times in the literature, i.e. many of these infections have only a few or no symptoms at all. CONCLUSION: Greater attention to the anamnestic data collection would therefore be the key to focusing the clinical investigations on those who are at a major risk to contracting STDs.

Contamination risk for operators performing semi-closed HIPEC procedure using cisplatin.

AIMS: Aim of this study was to assess operators' safety while performing a semi-closed HIPEC procedure for peritoneal carcinomatosis using cisplatin drugs. METHODS: Environmental air, theater personnel urine, operators' gloves and hand skin contamination were assessed during two non-consecutive working days. Six operating surgeons, two anesthesiologists and two theater nurses were included in the study. Glove samples were collected from the inner surface of the external glove and from the external surface of the inner glove from operating surgeons wearing a double pair of gloves. Personnel urine samples were collected before, after and 24 h from the procedure. RESULTS: Air and urine samples permanently resulted below detectable levels for cisplatin presence on all the tested sources and sessions. Cisplatin contamination was detected on the inner surface of the external gloves and on the outer surface of the inner gloves, but in a lower concentration for the latter. Skin wipe samples were below detectable levels for platinum presence. CONCLUSION: The results suggest that two pairs of gloves are adequate to protect the skin from antiblastic drugs. No sign of direct contact or systemic absorption of drugs was ever detected from the inspected samples. Semi-closed HIPEC technique appears to be a safe procedure for operators.

Evidence of interactions between the nucleocapsid protein NCp7 and the reverse transcriptase of HIV-1.

The human immunodeficiency virus (HIV-1) nucleocapsid protein NCp7 containing two CX2CX4HX4C-type zinc fingers was proposed to be involved in reverse transcriptase (RT)-catalyzed proviral DNA synthesis through promotion of tRNA3Lys annealing to the RNA primer binding site, improvement of DNA strand transfers, and enhancement of RT processivity. The NCp7 structural characteristics are crucial because mutations altering the finger domain conformation led to noninfectious viruses characterized by defects in provirus integration. These findings prompted us to study a putative RT/NCp7 protein-protein interaction. Binding assays using far Western analysis or RT immobilized on beads clearly showed the formation of a complex between NCp7 and RT. The affinity of NCp7 for p66/p51RT was 0.60 microM with a 1:1 stoechiometry. This interaction was confirmed by chemical cross-linking and co-immunoprecipitation of the two proteins in a viral environment. Competition experiments using different NCp7 mutants showed that alteration of the finger structure disrupted RT recognition, giving insights into the loss of infectivity of corresponding HIV-1 mutants. Together with structural data on RT, these results suggest that the role of NCp7 could be to enhance RT processivity through stabilization of a p51-induced active form of the p66 subunit and open the way for designing new antiviral agents.

Interactions of the C-terminus of viral protein R with nucleic acids are modulated by its N-terminus.

The basic viral protein R (Vpr) performs several functions during the human immunodeficiency virus HIV-1 retroviral cycle, including G2 mitosis arrest and nuclear import of the preintegration complex allowing lentivirus to replicate in nondividing cells. Accordingly, this protein was found in the nucleus of infected cells. In the virus, Vpr is incorporated through interaction with both nucleocapsid protein 7 (NCp7) and p6, two small proteins encoded by the C-terminal part of the Gag precursor. NCp7 is also involved in genomic RNA encapsidation during the budding process suggesting a possible interaction of Vpr with nucleic acids, either directly or via the NCp7 intermediate. Gel shift experiments were carried out with RNA and DNA using synthetic Vpr and peptide derivatives. The results show that Vpr binds to nucleic-acid inducing aggregates. This process, which requires the C-terminal basic domain of the protein (in particular the helical 70-80 domain), is regulated by the N-terminal region of Vpr. Moreover, NCp7 was shown to enhance RNA recognition by Vpr, a feature that could be required for Vpr encapsidation and during nuclear import of the preintegration complex.