Impact of exclusive e-cigarettes and heated tobacco products use on muco-ciliary clearance.

BACKGROUND: Tobacco smoking impairs mucociliary clearance (MCC) efficiency as shown by prolonged saccharin test transit time (STTT). Avoiding exposure to tobacco smoke from combustible cigarettes may restore MCC function and former smokers have been shown to exhibit similar STTT as never smokers. The impact on STTT of switching from smoking to combustion-free tobacco products such as e-cigarettes (ECs) and heated tobacco products (HTPs) is not known. METHODS: We report STTT of exclusive EC and HTP users. Test results were compared with those obtained in current, former, and never smokers. RESULTS: STTT were obtained from 39 current, 40 former, 40 never smokers, and from 20 EC and 20 HTP users. Comparison of STTT values showed significant difference among the five study groups (p < 0.00001) with current smokers having a median [interquartile range (IQR)] STTT of 13.15 min, which was significantly longer compared with that of all other study groups. In particular, compared with former (7.26 min) and never smokers (7.24 min), exclusive EC users and exclusive HTP users had similar STTT at 7.00 and 8.00 min, respectively. CONCLUSION: Former smokers who have switched to exclusive regular use of combustion-free nicotine delivery systems (i.e., ECs and HTPs) exhibit similar saccharin transit time as never and former smokers. This suggests that combustion-free nicotine delivery technologies are unlikely to have detrimental effects on MCC function.

Dietary inflammatory index and mortality: a cohort longitudinal study in a Mediterranean area.

BACKGROUND: Higher Dietary Inflammatory Index (DII®) scores are associated with increased morbidity and mortality. However, little is known about the effects of DII on mortality in Mediterranean countries. Therefore, in the present study, we aimed to investigate the potential association between DII scores and overall, cancer and cardiovascular disease (CVD) mortality in people living in a Mediterranean area. METHODS: DII scores were calculated using a validated food-frequency questionnaire. DII scores were then categorised into tertiles. Mortality was ascertained via death certificates. The association between DII scores with overall and cause-specific mortality was assessed via a multivariable Cox's regression analysis and reported as hazard ratios (HRs) with their 95% confidence intervals (CIs). RESULTS: The study included 1565 participants (mean age 65.5 years; females 44.7%). After a median follow-up of 12 years (2005-2017), 366 (23.4%) participants died. After adjusting for 17 potential confounders, people with higher DII scores had an increased risk of death compared to those in the lowest (most anti-inflammatory) tertile (HR = 1.38; 95% CI = 1.04-1.82 for the second tertile; HR = 1.38; 95% CI = 1.03-1.86 for the third tertile). Each 1 SD increase in DII score increased the risk of death by 13%. No association was found between DII scores and cancer or CVD death when considered separately. CONCLUSIONS: Higher DII scores were associated with a significantly higher mortality risk, whereas the association with cause-specific mortality was less clear. These findings highlight the potential importance of diet in modulating inflammation and preventing death.

One stage percutaneous transhepatic biliary stenting for malignant jaundice: a safe, quick and economical option of treatment.

OBJECTIVE: Patients with proximal malignant jaundices are often diagnosed in an advanced stage and need biliary decompression treatments, such as percutaneous transhepatic biliary drainage (PTBD) and bare metal stenting (BMS), to improve the hepatic function. Whether it is better to perform those two procedures together or in a separate time, it is not well understood. The aim of this study was to investigate the effectiveness and cost-benefit of a combined "one-stage" PTBD/BMS procedure in patients with malignant jaundices. PATIENTS AND METHODS: Forty-five patients with malignant jaundice treated with "one-stage" PTBD/BMS were retrospectively enrolled to evaluate technical success, complications, survival, and length of hospitalization. RESULTS: A full technical success of the procedures was reported for all patients, with only one major complication among 45 treated patients. A better performance in terms of hospitalization rate was achieved by the one-stage procedure compared to the two-stage, also resulting in global saving of costs. A high survival rate was observed at the 3rd and 6th month (97.7% and 86.6%, respectively), with a median overall survival time of 271,58 days. CONCLUSIONS: Our study shows that performing PTBD/BMS as a "one-stage" procedure is useful, safe, and cost-effective with a high percentage of technical success and a similar occurrence of complications compared to the two-stage procedure.

Early-adolescent male C57BL/6J and DBA/2J mice display reduced sensitivity to acute nicotine administration.

BACKGROUND: The initial response to nicotine is an important predictor of subsequent use. Multiple factors may alter this response including genetics and age of first use. Here we investigated the influence of age, genetics, and their interaction on nicotine sensitivity. We then examined whether these factors influence the relationship between initial behavioral responses and voluntary nicotine consumption in adulthood. METHODS: We measured initial nicotine responses, including nicotine-induced locomotor depression and hypothermia following an acute intraperitoneal injection (0, 0.5, or 1 mg/kg), during early-adolescence, middle-adolescence, late-adolescence, or adulthood. Thirty-five days after the initial testing, mice were assessed for voluntary oral nicotine consumption. RESULTS: Early-adolescent mice were more resistant to nicotine-induced hypothermia and locomotor depression than later ages, further hypothermia was influenced by genetics. In the DBA/2J strain, early-adolescent mice were insensitive to nicotine-induced hypothermia, but this response developed at later ages. In contrast, C57BL/6J mice were sensitive at all ages, but sensitivity increased across developmental age. There was little evidence of a relationship between initial behavioral response and choice nicotine consumption. CONCLUSION: By understanding how age of exposure and genetics influence initial nicotine behavioral responses, we have a greater understanding of factors that make adolescents differentially sensitive to the effects of this drug.

Translating the combination of gene therapy and tissue engineering for treating recessive dystrophic epidermolysis bullosa.

The combination of gene therapy and tissue engineering is one of the most promising strategies for the treatment of recessive dystrophic epidermolysis bullosa (RDEB). RDEB is a rare genetic disease characterised by mutations in the COL7A1 gene, encoding type VII collagen (COLVII), which forms anchoring fibrils at the dermal-epidermal junction of the skin. This disease causes severe blistering and only palliative treatments are offered. In this study, the base of a strategy combining gene therapy and a tissue-engineered skin substitute (TES), which would be suitable for the permanent closure of skin wounds, was set-up. As a high transduction efficiency into fibroblasts and/or keratinocytes seems to be a prerequisite for a robust and sustained correction of RDEB, different envelope pseudotyped retroviral vectors and the transduction enhancer EF-C were tested. When green fluorescent protein (GFP) was used as a reporter gene to evaluate the retroviral-mediated gene transfer, the fibroblast infection efficiency was 30 % higher with the Ampho pseudotyped vector as compared with the other pseudotypes. At least a 3.1-fold and a 1.3-fold increased transduction were obtained in fibroblasts and keratinocytes, respectively, with EF-C as compared with polybrene. A continuous and intense deposit of haemagglutinin (HA)-COLVII was observed at the dermal-epidermal junction of self-assembled TESs made of cells transduced with a HA-tagged COL7A1 vector. Furthermore, HA-tagged basal epidermal cells expressing keratin 19 were observed in TESs, suggesting stem cell transduction. This approach could be a valuable therapeutic option to further develop, in order to improve the long-term life quality of RDEB patients.

Adolescent chronic variable social stress influences exploratory behavior and nicotine responses in male, but not female, BALB/cJ mice.

Anxiety disorders and nicotine use are significant contributors to global morbidity and mortality as independent and comorbid diseases. Early-life stress, potentially via stress-induced hypothalamic-pituitary-adrenal axis (HPA) dysregulation, can exacerbate both. However, little is known about the factors that predispose individuals to the development of both anxiety disorders and nicotine use. Here, we examined the relationship between anxiety-like behaviors and nicotine responses following adolescent stress. Adolescent male and female BALB/cJ mice were exposed to either chronic variable social stress (CVSS) or control conditions. CVSS consisted of repeated cycles of social isolation and social reorganization. In adulthood, anxiety-like behavior and social avoidance were measured using the elevated plus-maze (EPM) and social approach-avoidance test, respectively. Nicotine responses were assessed with acute effects on body temperature, corticosterone production, locomotor activity, and voluntary oral nicotine consumption. Adolescent stress had sex-dependent effects on nicotine responses and exploratory behavior, but did not affect anxiety-like behavior or social avoidance in males or females. Adult CVSS males exhibited less exploratory behavior, as indicated by reduced exploratory locomotion in the EPM and social approach-avoidance test, compared to controls. Adolescent stress did not affect nicotine-induced hypothermia in either sex, but CVSS males exhibited augmented nicotine-induced locomotion during late adolescence and voluntarily consumed less nicotine during adulthood. Stress effects on male nicotine-induced locomotion were associated with individual differences in exploratory locomotion in the EPM and social approach-avoidance test. Relative to controls, adult CVSS males and females also exhibited reduced corticosterone levels at baseline and adult male CVSS mice exhibited increased corticosterone levels following an acute nicotine injection. Results suggest that the altered nicotine responses observed in CVSS males may be associated with HPA dysregulation. Taken together, adolescent social stress influences later-life nicotine responses and exploratory behavior. However, there is little evidence of an association between nicotine responses and prototypical anxiety-like behavior or social avoidance in BALB/cJ mice.

TNFα deficiency results in increased IL-1ß in an early onset of spontaneous murine colitis.

Inflammatory bowel disease (Crohn's disease (CD) and ulcerative colitis (UC)) is a multifactorial disease resulting from immune dysregulation in the gut. The underlying colitis is characterized by high levels of inflammatory cytokines, including TNFα. Biological intervention for IBD patients using anti-TNFα antibodies is often an effective therapeutic solution. However, TNFα neutralization fails to induce remission in a subgroup of IBD patients, primarily in UC patients. There is a dearth of suitable animal models representing TNFα non-responders. Here we have combined one of the best UC models currently available, namely Winnie and the TNFαKO mouse to generate a TNFα-deficient Winnie to study early onset colitis. The induced TNFα deficiency with underlying colitis does not influence general health (viability and body weight) or clinical parameters (colon weight, colon length and histological colitis) when compared with the Winnie genotype alone. The molecular characterization resulted in identification of Il1ß as the major elevated cytokine during early phases of colitis. Further, in vitro functional assay using bone marrow-derived dendritic cells confirmed IL-1ß as the major cytokine released in the absence of TNFα. This study has generated a successful model of colitis that remains TNFα non-responsive and has demonstrated that IL-1ß expression is a major pathway for the progression of colitis in this system. These data also suggest that IL-1ß can be a potential target for clinical intervention of UC patients who fail to respond to TNFα neutralization.

Cytokine modulation in patients with idiopathic pulmonary fibrosis undergoing treatment with steroids, immunosuppressants, and IFN-γ 1b.

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease of unknown etiology and pathogenic mechanisms. From an etiopathogenic point of view, alveolar macrophages play a key role in accumulation of fibroblasts and deposition of collagen and extracellular matrix by releasing specific cytokines and inflammatory mediators. IPF seems to be also associated with circulating fibrocytes, which might be involved with an abnormal pulmonary vascular repair and remodeling. Based on its hypothesized pathologic mechanisms, anti-inflammatory, anti-fibrotic and immunosuppressive therapies are often used. For these reasons, Interferon-g (IFN-g) has been used to exploit its activity on macrophages and fibroblasts. The aim of this study was to investigate the response to corticosteroids and/or IFN-g 1b treatments based on pulmonary function tests and on inflammatory cytokine patterns of expression on bronchoalveolar lavage (BAL), at baseline and during and after the therapies. Unlike previous studies, we analyzed a period of therapy longer than 1 year. Our results demonstrated the effectiveness of IFN-γ in a group of IPF patients in whom the treatment was prolonged for over a year. These data suggest a positive role of IFN-γ; treatment in patients in the initial stage of the disease.

A combined approach for predicting the cytotoxic effect of drug-nanoaggregates.

We present a combined spectroscopic and computational approach aimed to elucidate the mechanism of formation and activity of etoposide nanoaggregates upon release from dextran-etoposide conjugates. Etoposide is an anticancer drug that inhibits cell growth by blocking Topoisomerase II, the key enzyme involved in re-ligation of the DNA chains during the replication process. In silico and spectroscopic analysis indicate that released etoposide nanoaggregates have a different structure, stability, and bioactivity, which depend on the pH experienced during the release. Molecular dynamics simulation and in silico docking of etoposide dimers suggest that the aggregation phenomena inhibit etoposide bioactivity, yet without drastically preventing Topoisomerase II binding. We correlated the diminished cytotoxic activity exerted by dextran-etoposide conjugates on the A549 lung cancer cells, compared to the free drug, to the formation and stability of drug nanoaggregates.

Effect of low energy light irradiation by light emitting diode on U937 cells.

Photobiomodulation (PBM) can induce a set of different biological modulators either in vitro or in vivo. Experimental evidence has highlighted the role of light effects on the mechanisms related to inflammation, apoptosis and autophagy. The goal of this project was the evaluation of PBM on U937, an established cell line of histiocytic lymphoma origin. Several aspects of modulation of proinflammatory pathways were analyzed and autophagic and proapoptotic mechanisms related to low laser light exposure of cells were studied. As a source of low energy light emission, we used an NIR-LED device, characterized by an 880 nm-wavelength as light source. Flow cytometry analysis was performed on supernatants of controls and treated U937 cells to detect inflammatory cytokine levels. In order to evaluate NF-kB and caspase3 expressions, Western blot analysis was performed according to standard procedures. In this report, we show the effect of PBM on a monocyte/macrophage established tumor cell line (U-937). We demonstrate that LED exposure, in the presence or absence of lipopolysaccharide (LPS), activates cell degranulation, increased expression of Interleukin-8 (IL-8) and modulation of beta galactosidase activity. Evidence shows that the well-known pro-inflammatory nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) and the apoptotic marker (caspase3/cleaved-caspase3 ratio) are up-regulated in response to a proinflammatory biochemical pathway.

A numerical analysis of the aortic blood flow pattern during pulsed cardiopulmonary bypass.

In the modern era, stroke remains a main cause of morbidity after cardiac surgery despite continuing improvements in the cardiopulmonary bypass (CPB) techniques. The aim of the current work was to numerically investigate the blood flow in aorta and epiaortic vessels during standard and pulsed CPB, obtained with the intra-aortic balloon pump (IABP). A multi-scale model, realized coupling a 3D computational fluid dynamics study with a 0D model, was developed and validated with in vivo data. The presence of IABP improved the flow pattern directed towards the epiaortic vessels with a mean flow increase of 6.3% and reduced flow vorticity.

In vitro combined treatment with cetuximab and trastuzumab inhibits growth of colon cancer cells.

OBJECTIVES: Overexpression or constitutive activation of epidermal growth factor receptors (EGFR) is involved in growth of human cancers. We investigated effects of EGFR and HER-2 blockade in colon cancer cell lines using cetuximab and trastuzumab, with the aim of developing novel approaches to cancer therapy. MATERIALS AND METHODS: We studied effects of treatment on cell growth, cell cycle distribution, induction of apoptosis, changes in EGFR and HER-2 mRNA-protein expression and EGFR and HER-2 gene copy number in Caco-2, HT-29 and HCT-116 cells. RESULTS: Treatment of cells resulted in no effect in one of the three cell lines and in inhibition of cell proliferation in a time- and dose-dependent manner in the other two, with modulation of EGFR and HER-2 mRNA and protein levels. Differences in sensitivity to cetuximab and trastuzumab were observed. Treatment induced specific changes in cell cycle distribution in both cell lines affected, while apoptosis was not increased. Fluorescence in situ hybridization analysis revealed abnormal copy number of two genes resulting from aneuploidy; this was not responsible for different sensitivity to combination between the two cell lines. CONCLUSIONS: Targeting EGFR and HER-2 simultaneously could have useful applications in colorectal cancer treatment. To improve pharmacological efficacy of cetuximab and trastuzumab combination, molecular mechanisms involved in their activity need to be elucidated.

Tumor-induced alterations in lipid metabolism.

Alterations of lipid metabolism have been increasingly recognized as a hallmark of cancer cells. Cancer cells esterify fatty acids predominantly to phospholipids, an essential component of cell membranes. The main pathway along which proliferating cells gain lipids for membrane synthesis is the endogenous mevalonate pathway. Increased synthesis of mevalonate and mevalonate-derived isoprenoids supports increased cell proliferation through activating growth-regulatory proteins and oncoproteins and promoting DNA synthesis. The importance of a better knowledge of metabolic changes in lipogenic enzymes pathways, as well as of the role of each biochemical pathway in carcinogenesis, provides the rationale for in-depth study of the oncogenic signaling important for the initiation and progression of tumors. The dependence of tumor cells on a dysregulated lipid metabolism suggests that the proteins involved in this process may be excellent chemotherapeutic targets for cancer treatment. Here, we confirm the vital link between lipogenesis and cell proliferation, and our recent findings suggest that nutritional intervention is an effective and safe way to reduce cell proliferation in experimental models of carcinogenesis. The olive oil diet significantly reduces the protein activities of lipogenic enzymes associated with cell growth. The use of natural dietary components could potentially assist in the management of subjects with metabolic disorders-related tumors.

Access to prenatal cytogenetic diagnosis in Catania: a retrospective survey.

BACKGROUND: To determine the incidence of chromosome abnormalities (CAs) in prenatal cytogenetic diagnosis, to describe and compare indications in Italian and migrant women, and to assess the level of compliance with published national guidelines. METHODS: A retrospective analysis of 7806 amniotic fluid samples (AFS) and 228 chorionic villi samples (CVS) was conducted. RESULTS: Advanced maternal age was the most common indication. CAs incidence was 3.1 per 100 AFS, and 12.6 per 100 CVS. Only parental chromosome rearrangement and ultrasound abnormalities were significantly associated with CA occurrence (RR= 20.15 95%CI: 11.96-33.96; RR= 4.33; 95%CI: 2.95-6.36, respectively). Both in amniocentesis and in chorionic villi sampling CA incidence was significantly higher when performed according to the national guidelines, than for other reasons. Incidence data for trisomy 21, trisomy 18 and inversions were significantly higher than those reported in a previous Italian report. CONCLUSIONS: Increased maternal age may explain, at least in part, the increase by time of CAs, although an excess was shown in our population independently from it. Our results show that advanced maternal age may not be sufficient as a single criterion for prenatal diagnosis, and suggesting a future revision of national clinical indications is suggested.

Myocardial infarction in young adults: risk factors, clinical characteristics and prognosis according to our experience.

OBJECTIVES: Myocardial infarction is a relatively unusual phenomenon in young subjects. The aim if this work is to characterize the risk profile and factors influencing outcomes of these patients since it makes possible to manage prevention interventions. PATIENTS AND METHODS: We examined cardiovascular risk factors, clinical presentation, angiographic picture and outcome of a group of young patients hospitalized for a myocardial infarction. We enrolled 121 young patients consecutively admitted to our hospital for a myocardial infarction and examined them not only at the initial stage, but also after 3 months and one year; finally a long-term telephonic follow up was performed, when possible. RESULTS: We found some peculiarity making these patients quite different from the older ones who develop a myocardial infarction: cigarettes smoking, family history of ischemic heart disease and hyperlipidemia were the most frequent cardiovascular risk factors, while diabetes and hypertension were less represented; moreover coronary angiography showed more frequently a less extensive coronary atherosclerosis. Patients who developed a cardiovascular event at follow-up presented a significantly higher prevalence of hypertension and obesity and a significantly lower frequency of healthy coronary arteries and of previous revascularization. CONCLUSIONS: Myocardial infarction in young adults presents several peculiarities, represented not only by the risk profile, but also by the angiographic picture and the prognosis. Considering the long life expectancy of the involved population, the essential role of preventive interventions should be strongly underlined.

Are biopsy specimens predictive of HER2 status in gastric cancer patients?

BACKGROUND: Trastuzumab has been recently proposed as a treatment for patients with HER2-positive advanced/metastatic gastric cancer (GC). Since most patients have inoperable disease at diagnosis, accurate assessment of HER2 status on biopsy specimens is essential to select the patients who may benefit from therapy. AIM: The aim of this study is to establish whether HER2 status assessed on biopsy material could be reliable for treatment decisions using anti-HER2 agents. METHODS: The HER2 status was evaluated in 61 consecutive pairs of biopsy and surgical GCs samples by immunohistochemistry and chromogenic in situ hybridization. RESULTS: The overall concordance of HER2 status between biopsy and surgical specimens was 91.8 % with a predictive positive value of 71.4 % and a negative predictive value of 94.4 %. Of five discordant cases, there were three negative and two positive false biopsy results. All the false negative cases showed heterogeneous expression of HER2 protein in surgical samples. Two cases displayed overexpression of the receptors without corresponding gene amplification. CONCLUSIONS: HER2 status as evaluated on biopsy samples is a fairly good predictor of HER2 status of surgically-excised GCs. The most important influence for discordant results is tumor heterogeneity. However, HER2 overexpression, especially without coexisting gene amplification, may only be a temporary change in a tumor population. This may explain those cases with positive HER2 evaluation on biopsy material and a negative result on corresponding surgical specimen.

Identification of predictors of Salmonella shedding in adult horses presented for acute colic.

BACKGROUND: Colic has been associated with shedding of Salmonella. Horses with salmonellosis typically develop diarrhea, fever, and leukopenia. Overlooking additional predictors may result in failure to detect shedding horses and increase environmental contamination. OBJECTIVES: Evaluate associations between signalment and clinicopathologic data during early hospitalization and Salmonella shedding in horses treated for acute colic. ANIMALS: Horses with acute colic admitted to a referral hospital. A total of 59 horses shedding Salmonella compared to 108 Salmonella-negative horses. METHODS: Retrospective case-control study evaluating patient and Salmonella culture data. Associations between variables and Salmonella shedding were identified using logistic regression. Two multivariable models were developed pertaining to (1) information available within 24 hours of admission and (2) clinical findings that developed later during hospitalization. RESULTS: Variables retained for multivariable model 1 indicated that Warmbloods and Arabians had increased odds for shedding Salmonella, as did horses requiring surgery (OR, 2.52; 95% CI, 1.10-5.75) or having more severe gastrointestinal disease (OR, 2.59; 95% CI, 1.08-6.20). Retained variables for model 2 demonstrated that horses that were treated surgically (OR, 1.60; 95% CI, 0.70-3.62), developed fever >103°F (OR, 2.70; 95% CI, 0.92-7.87), had abnormal leukocyte count (OR, 1.38; 95% CI, 0.61-3.09), or became inappetent and lethargic (OR, 16.69; 95% CI, 4.08-68.24) had increased odds for shedding Salmonella. CONCLUSIONS AND CLINICAL IMPORTANCE: In horses with acute colic that present without signs of diarrhea, fever, or leukopenia, additional predictors associated with shedding Salmonella could be used to more promptly identify horses likely to shed organisms.

Endothelial activation and injury by cigarette smoke exposure.

Endothelial activation/injury following exposure to cigarette smoke may explain incidence of atherosclerosis and cardiovascular disease in smokers. We investigated cigarette smoke extract (CSE) effects relative to activation, injury, and survival of human umbilical vein endothelial cells (HUVEC) and compared circulating levels of specific endothelial activation markers between smokers and healthy non-smokers before and after smoking cessation. Viability and toxicity of HUVEC were tested by MTT and LDH assay. Release (by endothelial cells) and circulating levels (in smokers) of von Willebrand Factor (vWF), thrombomodulin (TM), was evaluated by ELISA. Incubation with increasing concentrations of CSE reduced the percentage of viable cells, being 33.9%, 23.9% after CSE 4%, 6% respectively. Dose- and time-dependent release of LDH was observed after incubation with CSE. vWF, TM release were assayed after CSE 2% HUVEC stimulation. Significant 42%, 61%, 76% increase in vWF concentration was detected respectively at 30', 60', 120'. Reduction in circulating levels of vWF, from a median value of 144.0% to 123.7%, was observed in the quitters group after smoking cessation. Exposure to cigarette smoke is cytotoxic and induces activation/injury of endothelium in vitro and in vivo. These findings may provide pathogenetic basis by which smoking can predispose to development of atherothrombosis and cardiovascular disease.

Therapeutic mammaplasties: full local control of breast cancer in one surgical stage with frozen section.

AIM: To evaluate the effectiveness of therapeutic mammoplasty with frozen section in achieving negative surgical margins in a single-stage surgery for breast cancer. METHODS: Fifty patients affected by early stage breast cancer treated by therapeutic mammaplasties were retrospectively reviewed in this study. Fifty-two therapeutic mammaplasties were accomplished. After resection the specimen was sent to pathologist for examination with frozen section. Tumour positive margins were defined as presence of cancerous cells at ≤ 2 mm from the edge of the specimen. In case of positive margins a second large re-excision was accomplished intra-operatively. All patients were followed every 4 months for the first 2 years and twice a year subsequently. RESULTS: Fifty-two procedures were evaluated (median follow-up of 72.6 months). The overall survival rate was 98% we had a single case of local recurrent disease (1.9%) that progressed to metastatic disease and patient's death. Frozen section as a diagnostic tool for identification of positive margins has been tested. In conclusion we report a sensitivity of 0.83 and a specificity of 0.93; the predictive positive value was 0.62 and the negative predictive value was 0.97, for a final accuracy of 0.94. CONCLUSION: Frozen section coupled to oncoplastic resections allows a proper control of local disease and can minimize any second surgical look for margins revision.

Gap junctions in human glioblastomas: implications for suicide gene therapy.

Glioblastoma is a very aggressive astrocytic tumor and most patients have 1-year survival time after diagnosis. A promising therapeutic strategy is the local delivery of the herpes simplex virus thymidine kinase gene in the tumor bed followed by ganciclovir treatment. The presence of functional gap junctions is highly relevant for the success of suicide gene therapy. Connexins are expressed in practically all tissues and form gap junctions that allow intercellular communication. Connexin 43 (Cx43) is the major connexin member being expressed in astrocytes but its status in glioblastoma is not well defined. We have investigated by immunofluorescence the presence of Cx43 in 74 human glioblastoma samples; its expression was detected in 77% of the samples analyzed. We report here that glioblastoma is a heterogenous disease as regards Cx43 expression with presentations, in which Cx43 expression is unaltered, reduced or totally lost. A predominant Cx43 cytoplasmic localization was observed in four out of eight primary glioblastoma cultures that we have established. This aberrant localization reduced gap junctionnal intercellular communication by 50 to 75% as compared with primary cell cultures displaying gap junctional plaques. However, the bystander effect evaluated after lentiviral delivery of the herpes simplex virus thymidine kinase gene and ganciclovir treatment was detected in all Cx43-positive primary cell cultures, and it was independant of the Cx43 localization. These findings may have important clinical implications for the design of anticancer cytotoxic therapies that rely on the gap junction-mediated bystander effect for their success.