Acute toxicity study of antibacterial organophilic bentonite incorporated with geranyl acetate in mice and geranyl acetate liberation in simulated gastric fluid.

In the present study, the antibacterial property of the organophilic bentonite (Bent-ODA) and organophilic bentonite incorporated with geranyl acetate ester (Bent-ODA-GA) was evaluated against bacteria Staphylococcus aureus, Escherichia coli and Salmonella typhimurium. Oral acute toxicity of Bent-ODA-GA was evaluated in mice, by a single oral dose of 300 and 2000 mg kg-1. Animals were observed for any toxicity clinical signs or mortality for 15 days according to OECD 423 guidelines. The release assay of GA presents in Bent-ODA in simulated gastric fluid, pH 3.5 and pH 6.5 was also performed. Bent-ODA-GA composite presented antibacterial activity against S. aureus and S. typhimurium bacteria with 10.7 ± 0.6 mm and 2.2 ± 0.1 mm inhibition halo, respectively, which make it possible to associate the composite antimicrobial feature due to the ester presence. The composite did not reveal any toxicity signs or mortality in any animal for acute toxicity treatment during the 15 days observation period. The LD50 of Bent-ODA-DA was estimated to be greater than 2000 mg kg-1. It was also observed that geranyl acetate is released from Bent-ODA in concentrations lower than 0.03 mg kg-1 for pH 3.5 and 0.004 mg kg-1 for pH 6.5, which are lower than those that could cause some toxic effects in animals.

Prehabilitation and heart failure: main outcomes in the COVID-19 era.

OBJECTIVE: The advent of the SARS-CoV-2 pandemic has resulted in an increase in sedentary behavior, with consequences on cardiopulmonary capacity, especially in the elderly population. Prehabilitation is a strategy usually used before a surgical procedure to improve functional capacity; however, it can be used for non-surgical patients and not in the acute phase of disease. The purpose of this study is to evaluate the effectiveness of a prehabilitation program, using telerehabilitation, in frail elderly patients with chronic heart failure. PATIENTS AND METHODS: This is a randomized, controlled, single-blind study. Fifteen patients with chronic heart failure were randomized into three groups: two active groups (telerehabilitation and in-person) and the control group. Patients in the active groups underwent a rehabilitation program divided into two 4-week periods, for 45-60 minutes per day, 2 days per week. RESULTS: In the Study Group, the quality of life significantly improved (EQoL-5D), and between the two groups a statistically significant difference in the motor dimension of SF-36 was identified. CONCLUSIONS: The telerehabilitation prehabilitation program for patients with chronic heart failure was confirmed to be effective and not inferior to a prehabilitation program performed in-person, avoiding the worsening of some domains of quality of life and motor performance, and leading to the improvement of others.

Prehabilitation and heart failure: perspective in primary outcomes, a randomized controlled trial.

OBJECTIVE: Prehabilitation, intended as a multidisciplinary approach where physical training is combined with educational and counselling training, in cardiology could optimizing care, and has been shown to be able to reduce morbidity and mortality in several diseases. The present study aims to assess the effectiveness of a prehabilitation program in elderly patients (over 65) with chronic heart failure and to evaluate functional and quality indices of life. PATIENTS AND METHODS: This is randomized, single blind controlled trial. Fourteen older adult patients diagnosed with chronic heart failure were enrolled. Patients were randomly assigned into the study or the control group. Patients in the study group underwent physical training organized into 10 twice-weekly meetings, nutritional and lifestyle counseling. RESULTS: In the Study Group, the quality of life improved significantly (EQoL-5D), and between the two groups there is a statistically significant difference in the motor dimension of SF-36. CONCLUSIONS: Because of our preliminary results, prehabilitation program should be included among the management strategies of in elderly patients with chronic heart failure to better manage their disease and to improve their Quality of Life.

Microencapsulation of eugenol molecules by ß-cyclodextrine as a thermal protection method of antibacterial action.

Eugenol is natural oil that has excellent antibacterial properties but cannot be used to fabricate many products that require thermal processing. One possible alternative to the use of the eugenol molecules in high-temperature processes is the encapsulation of these molecules in a structure that is not toxic and is resistant to thermal treatment. This work investigated the encapsulation process of eugenol molecules in ß-cyclodextrine and the antibacterial properties of eugenol-ß-cyclodextrine (the eugenol-ßCD complex) against Escherichia coli and Staphylococcus aureus. The FTIR, DSC, MEV and TGA results show that the encapsulation method is an excellent alternative to increase the thermal stability of eugenol molecules. A value of 241.32L.mol-1 was determined for the formation constant (Kc) of the eugenol-ßCD complex, which confirmed the success of the encapsulation process. The MEV analysis shows the formation of approximately 12µm microcapsules. After the thermal treatment of the eugenol-ßCD complex at a temperature of 80°C for 2h, the complex retained significant antibacterial action, which confirms the thermal protection of the eugenol molecules. The minimum inhibitory concentration (MIC) and agar diffusion results show that the microcapsules containing 17.08mmol.L-1 of eugenol exhibited excellent antibacterial action against Escherichia coli and Staphylococcus aureus after thermal treatment.

Quality and fatty acid profile of the milk of indigenous goats subjected to different local diets in Tunisian arid lands.

The study tested the hypothesis that certain pastoral forages and olive by-products, available in arid areas, may positively influence fatty acid composition and physicochemical properties of goat's milk. Thirty indigenous goats (body weight = 25.2 kg; age = 4.1 years) were allocated to three groups. During 60 days, the goats received ad libitum either dried olive leaves + Stipa tenacissima (group OL), khortane grass hay (group Ko) or oat hay (control diet, group OH). Milk samples were collected and analysed for total solids, fat, protein, lactose and ash content and fatty acid profile. Average milk yield did not statistically differ among groups. Milk total solids from OL group were higher in comparison with Ko and C groups (15.3, 14.7 and 14.5%, respectively; p < 0.05). Fat content was also higher for the OL group as compared to the other groups (5.44 vs. 5.01 and 4.66%, respectively, for Ko and OH). No significant differences were observed for the milk content of lactose, protein and ash. The percentage of saturated fatty acids of total milk fat was higher in OL and Ko groups compared to the C group (p < 0.001); the milk whereof was characterized by the highest percentage of monounsaturated (p < 0.01) and total unsaturated fatty acids. Milk fat of Ko and C groups showed significantly higher proportions of rumenic (CLA cis-9 trans-11) and vaccenic acids (C18:1 trans-11) compared to OL milk. The feeding system based on Stipa tenacissima and dried olive leaves resulted in the milk lowest proportion of trans-fatty acids and the highest proportion of polyunsaturated ω3-fatty acids (p < 0.05).

Application of glass particles doped by Zn(+2) as an antimicrobial and atoxic compound in LLDPE and HDPE.

This study demonstrates the potential application of glass particles doped with Zn(+2) (GZn) as an atoxic, antimicrobial additive when used in conjunction with high density polyethylene (HDPE) and linear low density polyethylene (LLDPE) polymers. Toxicity tests demonstrated that these modified glass particles were nontoxic to human cells, and atomic absorption analyses demonstrated the migration of ionic species in quantities less than 2.0ppm for both the HDPE/GZn and LLDPE/GZn compounds. Microbiological tests demonstrated the antimicrobial effect of the pure GZn compound as well as the polymeric HDPE/GZn and LLDPE/GZn compounds. In addition, at percentages of GZn higher than 2.00wt.% and at a time of 4h, the bactericidal performance is excellent and equal for both polymeric compounds.

Effect of whey concentration on protein recovery in fresh ovine ricotta cheese.

Ricotta cheese, particularly the ovine type, is a typical Italian dairy product obtained by heat-coagulation of the proteins in whey. The aim of this work was to investigate the influence of whey protein concentration, obtained by ultrafiltration, on yield of fresh ovine ricotta cheese. Ricotta cheeses were obtained by thermocoagulation of mixtures with protein content of 1.56, 3.10, 4.16, and 7.09g/100g from the mixing of skim whey and ultrafiltered skim whey. A fat-to-protein ratio of 1.1 (wt/wt) was obtained for all mixtures by adding fresh cream. The initial mixtures, as well as the final ricotta cheeses, were analyzed for their composition and by SDS-PAGE. Protein bands were quantified by QuantityOne software (Bio-Rad, Hercules, CA) and identified by liquid chromatography-tandem mass spectrometry. Significant differences in the composition of the ricotta cheese were observed depending on protein concentration. Particularly, ricotta cheese resulting from the mixture containing 7.09g/100g of protein presented higher moisture (72.88±1.50g/100g) and protein (10.18±0.45g/100g) contents than that prepared from the mixture with 1.56g/100g of protein (69.52±1.75 and 6.70±0.85g/100g, respectively), and fat content was lower in this sample (12.20±1.60g/100g) compared with the other treatments, with mean values between 15.72 and 20.50g/100g. Each protein fraction presented a different behavior during thermocoagulation. In particular, the recovery of ß-lactoglobulin and α-lactalbumin in the cheese increased as their content increased in the mixtures. It was concluded that concentrating ovine rennet whey improved the extent of heat-induced protein aggregation during the thermal coagulation process. This resulted in a better recovery of each protein fraction in the product, and in a consequent increase of ricotta cheese yield.

Antitumor effect of miR-197 targeting in p53 wild-type lung cancer.

Lung cancer is the leading cause of tumor-related death. The lack of effective treatments urges the development of new therapeutic approaches able to selectively kill cancer cells. The connection between aberrant microRNA (miRNA - miR) expression and tumor progression suggests a new strategy to fight cancer by interfering with miRNA function. In this regard, LNAs (locked nucleic acids) have proven to be very promising candidates for miRNA neutralization. Here, we employed an LNA-based anti-miR library in a functional screening to identify putative oncogenic miRNAs in non-small-cell lung cancer (NSCLC). By screening NIH-H460 and A549 cells, miR-197 was identified as a new functional oncomiR, whose downregulation induces p53-dependent lung cancer cell apoptosis and impairs the capacity to establish tumor xenografts in immunodeficient mice. We further identified the two BH3-only proteins NOXA and BMF as new miR-197 targets responsible for induction of apoptosis in p53 wild-type cells, delineating miR-197 as a key survival factor in NSCLC. Thus, we propose the inhibition of miR-197 as a novel therapeutic approach against lung cancer.

Evaluation of the toxic and genotoxic potential of acid mine drainage using physicochemical parameters and bioassays.

Carboniferous activity generates acid mine drainage (AMD) which is capable of unleashing toxic effects on the exposed biota. The aim of this study was to evaluate the toxic and genotoxic potential of untreated-AMD and AMD treated with calcinated sediment, using physicochemical parameters and bioassays. Results revealed that untreated-AMD presented low pH values and elevated concentrations of the metals Fe, Al, Mn, Zn and Cu. High acute toxicity was observed in Artemia sp. and Daphnia magna, and sub-chronic toxicity and genotoxicity in Allium cepa L. as well as scission of plasmid DNA exposed to untreated-AMD. Treatment of AMD with calcinated sediment promoted the reduction of acidity and the removal of metals, as well as a reduction in toxic and genotoxic effects. In conclusion, the calcinated sediment can be used as an alternative AMD treatment.

Chronic obstructive pulmonary disease: an emerging comorbidity in HIV-infected patients in the HAART era?

PURPOSE: The objective of our study was to evaluate the presence of respiratory symptoms and chronic obstructive pulmonary disease (COPD) in a human immunodeficiency virus (HIV)-infected outpatient population and to further investigate the role of highly active antiretroviral therapy (HAART) and other possibly associated risk factors. METHODS: We consecutively enrolled in a cross-sectional study HIV-infected patients and HIV-negative age, sex and smoking status matched controls. All participants completed a questionnaire for pulmonary symptoms and underwent a complete spirometry. RESULTS: We enrolled 111 HIV-infected patients and 65 HIV-negative age- and sex-matched controls. HIV-infected patients had a significantly higher prevalence of any respiratory symptom (p = 0.002), cough (p = 0.006) and dyspnoea (p = 0.02). HIV-infected patients also had a significantly higher prevalence of COPD in respect of HIV-negative controls (p = 0.008). Furthermore, HIV-infected individuals had significantly (p = 0.002) lower forced expiratory volume at one second (FEV1) and FEV1/forced vital capacity (FVC) ratio (Tiffeneau index) (p = 0.028), whereas the total lung capacity (TLC) was significantly higher (p = 0.018). In the multivariate analysis, significant predictors of respiratory symptoms were current smoking [adjusted odds ratio (AOR) 11.18; 95 % confidence interval (CI) 3.89-32.12] and previous bacterial pneumonia (AOR 4.41; 95 % CI 1.13-17.13), whereas the only significant predictor of COPD was current smoking (AOR 5.94; 95 % CI 1.77-19.96). HAART receipt was not associated with respiratory symptoms nor with COPD. CONCLUSIONS: We evidenced a high prevalence of respiratory symptoms and COPD among HIV-infected patients. HIV infection, current cigarette smoking and previous bacterial pneumonia seem to play a significant role in the development of respiratory symptoms and COPD. Thus, our results suggest that the most at-risk HIV-infected patients should be screened for COPD to early identify those who may need specific treatment.

The influence of particle size and AgNO3 concentration in the ionic exchange process on the fungicidal action of antimicrobial glass.

Antimicrobial materials have long been used as an effective means of reducing the risks posed to humans by fungi, bacteria and other microorganisms. These materials are essential in environments where cleanliness, comfort and hygiene are the predominate concerns. This work presents preliminary results for the development of a fungicidal vitreous material that is produced by the incorporation of a silver ionic specimen through ionic exchange reactions. Silver ions were incorporated into powdered glass via ionic exchange in an ionic medium containing silver species with different concentrations of AgNO3. The fungicidal efficiency of the samples was studied as a function of the AgNO3 concentration and the particle size of the glass using the agar diffusion test for the microbiological analysis of the fungus species Candida albicans. The samples were examined by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). The experimental results showed that the fungicidal effect was dependent on the AgNO3 concentration in the ionic exchange medium but was not dependent on the particle size of the glass.

Activity of endovesical gemcitabine in BCG-refractory bladder cancer patients: a translational study.

Intravesical gemcitabine (Gem) has shown promising activity against transitional cell carcinomas (TCC) of the bladder, with moderate urinary toxicity and low systemic absorption. The present phase II study evaluated the activity of biweekly intravesical treatment with Gem using a scheme directly derived from in vitro preclinical studies. Patients with Bacille Calmette-Guérin (BCG) -refractory Ta G3, T1 G1-3 TCC underwent transurethral bladder resection and then intravesical instillation with 2000 mg Gem diluted in 50 ml saline solution on days 1 and 3 for 6 consecutive weeks. Thirty-eight (95%) of the 40 patients showed persistent negative post-treatment cystoscopy and cytology 6 months after Gem treatment, while the remaining 2 patients relapsed at 5 and 6 months. At a median follow-up of 28 months, recurrences had occurred in 14 patients. Among these, four had downstaged (T) disease, three had a lower grade (G) lesion and three had a reduction in both T and G. Urinary and systemic toxicity was very low, with no alterations in biochemical profiles. In conclusion, biweekly instillation of Gem proved active in BCG-refractory Ta G3, T1 G1-3 TCC. Our results highlight the importance of preclinical studies using in vitro systems that adequately reproduce the conditions of intravesical clinical treatment to define the best therapeutic schedule.

ELISA and LC-MS/MS methods for determining cyanobacterial toxins in blue-green algae food supplements.

The use of natural products as a diet supplement is increasing worldwide but sometimes is not followed by adequate sanitary controls and analyses. Twenty samples of pills and capsules of lyophilised cyanobacteria (blue-green algae), commercialised in Italy as dietary supplements, were found positive at the Vibrio fischeri bioassay. Further analyses with ELISA and LC-MS/MS methods revealed the presence of four microcystin (MC) analogues, MC-LR, -YR, -LA, -RR and two demethylated forms of MC-RR. The highest total microcystin content was 4.5 and 1.4 microg g-1 in pills and capsules, respectively. The ELISA measurements, compared to the LC-MS/MS analyses, showed significantly lower concentrations of microcystins in pills, this confirming a possible ELISA underestimate of mixed microcystins, due to different sensitivities for some toxic analogues.

Identification and quantification method of spiramycin and tylosin in feedingstuffs with HPLC-UV/DAD at 1 ppm level.

The use of the two macrolides antibiotics Spiramycin (S) and Tylosin (T) as growth promoters in animal feeding has been recently withdrawn in the European Union due to a concern about the outbreaks of farmacoresistance fenomena as a possible hazard for humans. For feed additives monitoring purposes, an analytical method has been developed for their extraction, purification and identification in different animal feedingstuffs (pelleted beef, pig, poultry feeds and calves milk replacer) at a minimum performance required limit (MRPL) of 1 microg g(-1) (ppm). Such limit has been established according to the lowest dosage of additives still able to elicit an appreciable growth promoting effect. Blank feeds were spiked at two concentration levels, 1.0 and 2.5 ppm in six replicates. After methanolic extraction, samples were cleaned up on SPE CN columns and extracts analysed in HPLC-UV/DAD, using a gradient elution. Detection limits, calculated as the tree time mean noise of 20 blank feeds, were 176 and 118 ng g(-1) for S and T, respectively. Results show good repeatability (CV% not exceeding the value of 15) and mean recovery in the range of 99-74% and 81-53% for S and T, respectively, at 1 ppm. When the standards were injected up to 250 ng the chromatographic method can resolve the components of analytes (Spiramycin I, II and III; Tylosin A and B) but can not resolve the components on real feed samples at the spiked levels considered. For this reason the identification and quantification of analytes on matrix were carried out considering the main compound of the drugs (Spiramycin I and Tylosin A). As a verification, the overlapping of UV spectra in the range 220-350 nm between analytical standards and the compounds in the matrix were considered.

Tension-free vaginal tape and female stress incontinence: further evidence of effectiveness.

OBJECTIVES: Female stress incontinence is a frequent and distressing event with subsequent impact on quality of life. The tension-free vaginal tape (TVT) technique is considered to fulfill the criteria for an ideal surgical procedure and we therefore reviewed our cases to further analyze its effectiveness. MATERIALS AND METHODS: 57 patients suffering from stress incontinence with grade 1-2 cystocele underwent the TVT operation. During follow-up (mean 22.2 months), the short- and long-term results were investigated. RESULTS: 91.3% of patients achieved complete resolution of previous stress incontinence in the early postoperative period. Early complications occurred in 8.7% of patients. Follow-up (mean 22.2 months) highlighted the stabilization of positive outcomes in 87.7% of cases. No postoperative modification of sexual habits and a significant improvement in quality of life were reported by all patients. CONCLUSION: The results of our study confirm that the TVT technique for stress incontinence is an easy to perform procedure which is safe, effective and long-lasting.

Traceability of inbred and crossbred Cinta Senese pigs by evaluating the oxidative stress.

Previous studies on the oxidative stress in swine indicated a strong link between the values of reactive oxygen metabolites (ROMs), the subsequent antioxidant adaptive response (OXY) and the genetic selection. Such findings, mainly related to a cardiovascular inadequacy in lean, large muscle blocks and fast growing breeds, is associated with specific metabolic diseases such as porcine stress syndrome and mulberry heart disease. In this study, we investigated the oxidative stress parameters to trace the genetics of Cinta Senese (CS) pigs, a historical breed free-range reared in Siena countryside. Sera from CS (n = 24) and Large White x CS (LW x CS) (n = 24) groups around 120 kg body weight fed the same diet were sampled at slaughter. Sera from wild boars (WB) (n = 24) hunted in the same district were also considered. ROMs and OXY were evaluated in the three groups of swine. Significant differences by one-way anova (P < 0.05) between groups were found for both procedures. ROM levels were lower in WB (9.79 +/- 5.76 mm H2O2) and CS (18.02 +/- 7.42 mm H2O2), and highest in LW x CS (42.78 +/- 8.61 mm H2O2). OXY values ranged from 271.37 +/- 50.90 microm neutralized HOCl (WB) to 343.21 +/- 57.45 microm neutralized HOCl (LW x CS). Results indicated that the evaluation of the oxidative stress can effectively trace the CS pigs, thus aiding in preserving the overall specific traits of such a historical animal.

Extraction, clean-up and gas chromatography-mass spectrometry characterization of zilpaterol as feed additive in fattening cattle.

Zilpaterol is an adrenergic drug currently licensed in Mexico and South Africa as a feed additive for cattle close to consignment. In this study an analytical method to detect zilpaterol in commercial feeds was set up. The influence of extraction solvent and matrix was evaluated. The drug as a trimethylsilyl derivative was characterized by GC-MS, on a quadrupole detector, in the electron impact mode. Acidic extraction, solid-phase extraction C(18) non-endcapped clean-up and mass characterization on ions m/z 308, 291, 405, 390 provided zilpaterol recoveries >75.3% and repeatability <3.3% in feeds spiked in the range 30.0-120.0 ng/g. The limits of detection and quantification were 7.5 and 25.0 ng/g, respectively. Such limits are well below the dose of 5.0-20.0 microgram/g proposed as effective.

Evaluation of the oxidative stress in growing pigs by microplate assays.

The ability of pigs to neutralize the reactive oxygen species (ROS) plays a key role in their welfare. Two micromethods were developed to evaluate the oxidative stress and the anti-oxidative response in sera. We tested these methods for linearity, repeatability and reproducibility. A hydroxyl radicals (HR) test, based on the Fenton reaction, showed a linearity between 0.26 and 4.5 mM H2O2 (r = 0.997), with a coefficient of repeatability (CVr) of 1.9 and a coefficient of reproducibility (CVR) of 2.9. An anti-oxidant power (AOP) test, based on the capability of sera to neutralize a titred hypochlorous acid solution (HClO), showed a linearity between 55 and 880 microM HClO neutralized (r = 0.984), with CVr = 3.1 and CVR = 4.9. We assessed the applicability of such tests in field conditions on three different farms. Farm A recorded a positive anamnesis of Mulberry heart disease. Dietary Vitamin E supplementation was given at 50 p.p.m. in farms A and B and at 150 p.p.m. in farm C, respectively. The group from farm A showed HR sera levels higher than those of farm B and farm C (farm A, mean = 31.61 +/- 2.87 mM H2O2, n = 10; farm B, 28.53 +/- 2.30, n = 10; and farm C, 25.63 +/- 1.03, n = 10; P < 0.01). The AOP test was not able to discriminate between farms. The AOP levels found (mean = 472.50 microM HClO neutralized: n = 30) probably represent the maximum response of the organism to compensate the oxidative stress. HR and AOP are stable in sera stored at +4 degrees C. We discuss the application of such methods for the assessment of farmed animals' welfare.

Use of molecularly imprinted polymers in the solid-phase extraction of clenbuterol from animal feeds and biological matrices.

Clenbuterol molecularly imprinted polymers (MIPs) as chromatographic stationary phase for the solid-phase extraction (SPE) of the drug from biological samples have been prepared. Propylene columns filled with 500 mg of clenbuterol MIPs have been tested with respect to their loading capacity, memory effects, selectivity toward related drugs (mabuterol, clenproperol, clenisopenterol, ritodrine) and specificity toward interferences arising from heterogeneous matrices such as animal feeds, bovine urine and liver. Analytes were concentrated on Extrelut 20 columns and the residues resuspended in 70% acetonitrile. Application, washing and elution fractions were collected and analyzed by HPLC-diode array detection. Results indicate this MIP approach in SPE is extremely selective for clenbuterol, mabuterol, clenproperol and clenisopenterol (>95% found in the eluate), with a loading capacity of about 20 microg/100 mg of stationary phase. Ritodrine showed a recovery rate of 51%. The molecular recognition mechanism is so specific to allow clenbuterol detection and identification by conventional detectors at level of interest (ppb) also from complex matrices such as feeds, urine and liver.

Percutaneous US-guided radiofrequency ablation of hepatocellular carcinomas: results in 15 patients.

BACKGROUND: The majority of patients with hepatocellular carcinoma (HCC) cannot undergo surgery because of multifocality, location or advanced cirrhosis. Our experience with percutaneous radiofrequency ablation for treatment of patients suffering from unresectable hepatocellular carcinoma is described here. PATIENTS AND METHODS: Fifteen patients (ten men and five women) with eighteen primary hepatocellular tumors underwent percutaneous radiofrequency ablation. The mean diameter of the HCCs was 32 mm (ranging from 15 mm to 62 mm). The patients were treated under ultrasound guidance using either a 18-gauge internally cooled electrode or a 14-gauge electrode with four expandable hooks. RESULTS: Complete necrosis was achieved in 15 lesions after one session of RF ablation. The persistence of a small portion of viable tissue was seen in two lesions. One lesion was not evaluable. After a mean follow-up period of 9.2 months (range 3-24 months), eleven patients (76%) showed no sign of local or distant recurrence, one patient developed a new lesion and one of two patients with persistence of viable tissue obtained a complete necrosis after the injection of percutaneous ethanol. Moreover, a major complication (intraperitoneal bleeding requiring surgical treatment) and three minor complications (1 pleuric effusion and 2 perihepatic fluid collections that resolved spontaneously) were observed. CONCLUSION: RF ablation is a simple, well-tolerated and effective procedure for the treatment of unresectable hepatocellular carcinomas.