Microwave ablation of hepatic tumors with a third generation system: loco­regional efficacy in a prospective cohort study with intermediate term follow-up.

PURPOSE: To evaluate the safety and intermediate-term efficacy of percutaneous microwave ablation (MWA) in primary and secondary liver tumors using a third generation MWA device, under ultrasound guidance. PATIENTS AND METHODS: Sixty-two patients (median age 74 years, 73 % males) with 69 liver tumors were enrolled in this prospective observational study. Forty-seven patients (76 %) had hepatocellular carcinoma (HCC) and 15 (24 %) metastases. Median follow-up was 3.6 years. RESULTS: Median tumor diameter at contrast enhanced computed tomography was 23 mm (I-III quartiles, 18 - 31 mm). All procedures were performed percutaneously using a 2.45 GHz generator. Median ablation time was 10 minutes (I-III quartiles, 10 - 14 minutes). A single percutaneous antenna insertion was performed for 56/69 (81 %) of the tumors. Technical success was obtained in all tumors. Primary efficacy at 24 hours was achieved in 68/69 (99 %) tumors. The overall one-year cumulative local tumor progression rate was 15.1 % (95 % CI, 7.7 - 24.8 %) with no significant difference between HCC and metastases (p = 0.26). There was one procedure-related mortality (1.6 %) and one major bleeding (1.6 %). CONCLUSION: Microwave ablation is a valid option for thermal ablation of HCC and liver metastases with comparable complication rate to other local ablative procedures.

Effects of quercetin on exercise performance, physical activity and blood supply in a novel model of sustained hind-limb ischaemia.

BACKGROUND: There are currently few effective drugs to treat the leg symptoms of peripheral arterial disease (PAD). Previous studies have suggested that the nutraceutical, quercetin, can improve exercise performance and reduce pain sensitivity in healthy mice and improve blood supply in a rodent model of acute hind-limb ischaemia. These models may not be relevant to people with PAD. The aim of this study was to examine the effect of quercetin on exercise performance, physical activity and blood supply in a novel mouse model of sustained hind-limb ischaemia. METHODS: Hind-limb ischaemia was induced in 6-month-old male apolipoprotein E-deficient mice using a novel two-stage surgical procedure. Five days after induction of ischaemia, mice were allocated to commence dietary quercetin or a control diet for 4 weeks. The primary outcome was exercise performance evaluated using a treadmill test. Other outcomes included physical activity, estimated by an open field test, and hind-limb blood supply, assessed by laser Doppler monitoring. RESULTS: A sustained reduction in relative limb blood supply (P < 0.001) was achieved consistently in all 48 mice before allocation to a control (n = 24) or quercetin (n = 24) diet. Quercetin did not improve exercise performance (P = 0.785), physical activity (P = 0.151) or relative limb blood supply (P = 0.954) over the 4-week assessment period. CONCLUSION: These data suggest that quercetin does not improve exercise performance, physical activity or limb blood supply in mice with sustained hind-limb ischaemia, and therefore is unlikely be an effective treatment for PAD.

WLIP and tolaasin I, lipodepsipeptides from Pseudomonas reactans and Pseudomonas tolaasii, permeabilise model membranes.

The activity of the White Line Inducing Principle (WLIP) and tolaasin I, produced by virulent strains of Pseudomonas reactans and Pseudomonas tolaasii, respectively, was comparatively evaluated on lipid membranes. Both lipodepsipeptides were able to induce the release of calcein from large unilamellar vesicles. Their activity was dependent on the toxin concentration and liposome composition and in particular it increased with the sphingomyelin content of the membrane. Studies of dynamic light scattering suggested a detergent-like activity for WLIP at high concentration (> 27 microM). This effect was not detected for tolaasin I at the concentrations tested (< 28 microM). Differences were also observed in lipodepsipeptides secondary structure. In particular, the conformation of the smaller WLIP changed slightly when it passed from the buffer solution to the lipid environment. On the contrary, we observed a valuable increment in the helical content of tolaasin I which was inserted in the membrane core and oriented parallel to the lipid acyl chains.

Antioxidant drugs combined with alpha-interferon in chronic hepatitis C not responsive to alpha-interferon alone: a randomized, multicentre study.

OBJECTIVE: After non-response to the initial course of therapy, retreatment with alpha-interferon is not effective. The aim of this study was to ascertain whether the administration of N-acetyl cysteine and vitamin E could increase the response rate to retreatment with alpha-interferon. DESIGN: Prospective, multicentre clinical trial. SETTING: Twelve hospitals in Lombardy, Italy. PARTICIPANTS: 120 consecutive patients affected by biopsy-proven chronic hepatitis C who had been non-responders to a previous course of alpha-interferon, administered at the dosage of 3-6 million units (MU) three times a week (tiw) for 6 months. INTERVENTIONS: The patients were randomly assigned to one of two groups of treatment: group A, natural interferon-alphaN3, 6 or 9 MU tiw, when the body weight was < 60 kg or > or = 60 kg, respectively; group B, the same dosage of natural interferon-alphaN3 in association with oral administration of N-acetyl cysteine 1200 mg/day and vitamin E 600 mg/day. The period of treatment was 6 months in both groups. RESULTS: Neither end-therapy biochemical response nor sustained biochemical response rates were improved by the combination treatment, and in no case was clearance of the virus from serum observed. CONCLUSIONS: In this randomized study carried out on 120 patients with chronic hepatitis C not responsive to alpha-interferon, oral supplementation with N-acetyl cysteine and vitamin E did not improve the poor efficacy of retreatment with alpha-interferon alone.

Contrast enhanced ultrasound: Roles in immediate post-procedural and 24-h evaluation of the effectiveness of thermal ablation of liver tumors.

PURPOSE: To retrospectively assess the diagnostic accuracy of immediate post-procedural CEUS, 24-h CEUS, and 24-h CT in verifying the effectiveness of thermal ablation of liver tumors ablation, using the combined results of 3-month post-procedure CEUS and MDCT as the reference standard. MATERIALS AND METHODS: From our database, we selected patients who had immediate post-procedural CEUS and 24-h CEUS and MDCT examinations after undergoing thermal ablation of a liver tumor between January 2009 and March 2010. The study population consisted of 53 subjects and 55 tumors (44 HCC and 11 metastasis) were evaluated. Thirty-seven tumors were treated with radiofrequency and 18 with microwave ablation. Post-procedural CEUS, 24-h CEUS and MDCT, and 3-month follow-up CEUS and MDCT images were blindly reviewed by two radiologists, who measured the size of the ablation area on the post-procedural and 24-h studies. They also evaluated the ability of each of these three index tests to predict the outcome (residual tumor vs. no residual tumor) using imaging studies done at the 3-month follow-up as the reference standard. RESULTS: Mean tumor diameter on preablation CEUS (the day before treatment) was 20 ± 9 mm. Mean diameter of the necrotic area was 29 ± 9 mm on post-procedural CEUS, 34 ± 11 mm on 24-h CEUS, and 36 ± 11 mm on 24-h MDCT. Diameters of the necrotic area (mean and maximum) on post-procedural CEUS were significantly smaller than those measured on 24-h CEUS or 24-h MDCT, which were not significantly different. For predicting the presence of residual tumor at the 3-month follow-up, post-procedural CEUS, 24-h CEUS, and 24-h MDCT displayed sensitivity of 33%, 33%, and 42%; specificity of 92%, 97%, and 97%; negative predictive value of 84%, 85%, and 83%. The accuracy parameters of these three imaging modalities were not significantly different from one another. CONCLUSIONS: In patients undergoing thermal ablation for liver tumors, the immediate post-procedural CEUS seems comparable to 24-h CEUS and MDCT in terms of detecting residual disease.

Formulation and characterization of polyphenol-loaded lipid nanocapsules.

The purpose of this study was to design and characterize two flavonoid-loaded lipid nanocapsules (LNC) by applying the phase inversion process, and to enhance their apparent solubility and/or the stability. The flavonoid-loaded LNC were characterized by particle size, encapsulation efficiency, drug leakage rates, stability and spectroscopic studies. It was observed that quercetin-loaded LNC30 (3%) and LNC60 (2%) carried a particle size of 30.3 and 55.1 nm, respectively and significant higher entrapment efficiency. Encapsulation of quercetin (QC) in LNC enabled us to increase its apparent aqueous solubility by a factor of 100. And in view of calculations and results, it seems most probable that QC is arranged at this LNC interface between the oil phase and the hydrophilic polyethylene glycol moieties of the surfactant. In addition, colloidal suspensions proved to be stable in term of encapsulation for at least 10 weeks and QC was not oxidised. With simple chemical modification of (-)-epigallocatechin-3-gallate or (-)-EGCG, it was possible to reach very high encapsulation rates (95%). Thus we obtained stable colloidal suspensions of (-)-EGCG in water over 4 weeks while free (-)-EGCG solubilised in water exhibited 100% degradation within 4h. The initial problems (solubility and stability) of these flavonoids were resolved thanks to drug-loaded LNC.

Risk of tumour seeding after percutaneous radiofrequency ablation for hepatocellular carcinoma.

BACKGROUND: A recent small study reported a high rate of neoplastic seeding after cooled-tip radiofrequency ablation (RFA) for hepatocellular carcinoma (HCC) in patients who had undergone previous needle biopsy. Tumour seeding was associated with subcapsular tumour location, poorly differentiated tumours and a high alpha-fetoprotein (AFP) level. The aim of the present study was to determine the rate of neoplastic seeding after RFA in a large series of unselected patients with HCC who had a long follow-up. METHODS: A total of 1314 patients with 2542 nodules were treated in three centres. Median follow-up was 37 months. Needle biopsy had been performed before RFA in 241 patients (18.3 per cent). The influence of subcapsular location, high AFP level and previous biopsy on risk of tumour seeding was assessed. RESULTS: Neoplastic seeding was identified in 12 patients (0.9 per cent); the rate was comparable at the three centres (0.9, 0.7 and 1.4 per cent). Only previous biopsy was significantly associated with tumour seeding (P = 0.004). CONCLUSION: RFA with a cooled-tip needle was associated with a low risk of neoplastic seeding, even in unselected patients. The use of biopsy before RFA is to be discouraged, particularly when liver transplantation is a possibility at a later date.

The effect of transportation on the immune status of Bos indicus steers.

This study investigated the effect of 72 h of road transport on the immune status of Bos indicus steers (n = 10; age = 15 to 18 mo). Total and differential leukocyte numbers and lymphocyte function were determined at 2 d before transport (-48 h), immediately after 72 h of transport (72 h), and 6 d after transport (216 h). Phytohemagglutinin (PHA)-stimulated lymphocyte proliferation, interferon-gamma production, and tetanus-toxoid specific antibody levels were determined. Total leukocyte and eosinophil numbers showed a transient decrease at 72 h (immediately after transport; P < 0.05) and returned to baseline values by 6 d after transport. Lymphocyte numbers and antibody titers were unaffected by transportation. The PHA-stimulated lymphocyte proliferation decreased (P < 0.05) at 72 h and returned to baseline levels 6 d after transport. This study demonstrated that transportation of mature Bos indicus steers caused transient decreases in leukocyte numbers and lymphocyte function, although all measures recovered by 6 d after transport. Therefore, Bos indicus cattle may be vulnerable to infection during this period.

Liver metastases: results of percutaneous ethanol injection in 14 patients.

Percutaneous ethanol injection (PEI) was performed under ultrasound (US) guidance in the outpatient department in 14 patients with 21 metastases 1.0-3.8 cm in diameter. The primary cancer was colorectal adenocarcinoma (seven patients), adenocarcinoma of the stomach (four patients), abdominal leiomyosarcoma (one patient), gastrinoma of unknown origin (one patient), and bronchial carcinoid (one patient). The indicators of therapeutic efficacy were US, computed tomography, fine-needle biopsy, and serologic markers. No complications occurred after a total of 175 treatment sessions. Complete response was obtained in 11 lesions, nine of which were less than 2 cm in diameter, and in all endocrine metastases (four lesions in two patients) with a maximum, recurrence-free follow-up of 38 months. Carcinoembryonic antigen values decreased in all patients but one for a 2-6-month period. The natural course of metastatic disease strictly limits the applicability of PEI, a local treatment. Single, metachronous, nonoperable metastasis of adenocarcinoma and endocrine metastases seem to be the only indications for PEI.

Percutaneous ethanol injection: single session treatment.

OBJECTIVE: Percutaneous ethanol injection (PEI) has become one of the most widely used procedures for treating hepatocellular carcinoma (HCC) in patients with cirrhosis. PEI spread rapidly thanks to its ease of execution, safety, low resection cost, repeatability, therapeutic efficacy, and survival rates comparable to those of surgical resection. A new version of the procedure known as "single session" PEI, designed for treatment of large HCC under general anesthesia, was proposed in 1993. MATERIALS AND METHODS: In our study of 108 patients with HCC>5 cm in diameter, 24 patients (Group A), were treated for a single and encapsulated lesions measuring from 5-8.5 cm, 63 patients (Group B), had a single and infiltrating lesions measuring from 5-10 cm or multiple HCC; and 21 patients (Group C), had advanced disease. RESULTS: In Group A, complete necrosis was achieved in 58.3% of lesions, in Group B complete necrosis was achieved only in encapsulated lesions. In infiltrating lesions of Group B and C, complete necrosis was never obtained. Survival of patients at 3 years with encapsulated HCC was 57%, in infiltrating or multiple HCC 42%, and in advanced HCC 0%. One death due to bleeding of esophageal varices occurred in a Child's C patient. Major complications included intraperitoneal hemorrhage, an important hepatic decompensation, a transitory renal insufficiency, and two infarction of adiacent segment. CONCLUSION: Single session PEI is as effective in inducing liver tumor necrosis as traditional PEI. Its advantage are shorter treatment time and the capability of treating larger and multiple HCC. The One-shot technique proved a more aggressive therapeutic modality than traditional PEI.

Radiofrequency thermal ablation of hepatocellular carcinoma.

Radiofrequency (RF) ablation resulted in a higher complete necrosis than percutaneous ethanol injection (PEI), above all in infiltrating lesions, and requires fewer treatment sessions in the treatment of small size tumors. We achieved 90% of complete necrosis in hepatocellular carcinoma (HCC)<3 cm, 71% in medium (3-5 cm) and 45% in large (5.1-9 cm) HCC. However, near complete necrosis was obtained in the majority of the remaining tumors. PEI is preferable in lesions at risk with RF, i.e. adjacent to main biliary ducts or to intestinal loops. Our current 3-yr survival in child A patient with single HCC<5 cm is 85%.

Percutaneous ethanol injection of hepatic tumors: single-session therapy with general anesthesia.

OBJECTIVE: We studied the feasibility and the effectiveness of percutaneous ethanol injection, performed with general anesthesia in a single session, for treating malignant hepatic lesions. SUBJECTS AND METHODS: We treated 30 patients with sonographically guided percutaneous injection of ethanol. Twenty had hepatocellular carcinoma and cirrhosis, and 10 had hepatic metastases, principally from colon cancer. The mean volume of ethanol injected was 57 ml (range, 6-165 ml). RESULTS: CT showed complete necrosis (up to 8.2 cm) in seven of 10 patients with encapsulated hepatocellular carcinoma and about 90% necrosis in the remaining three patients. In four of these patients, the alpha-fetoprotein level fell from more than 200 ng/ml to less than 20 ng/ml during treatment. In 10 patients with infiltrating hepatocellular carcinoma, about 70-90% necrosis was achieved; in six of these patients, the alpha-fetoprotein level, which had been more than 200 ng/ml, decreased during treatment. In the 10 patients with metastases, more than 50% necrosis was always achieved. Levels of carcinoembryonic antigen decreased after treatment in all patients. In three patients who had cirrhosis with superficial hepatocellular carcinoma, peritoneal hemorrhage occurred but did not require transfusion. CONCLUSION: Our results show that percutaneous injection of ethanol in a single session with general anesthesia is feasible and effective and has several advantages over multisession therapy. These include shorter treatment time and the ability to treat larger and more numerous lesions.

Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B.

Sourdough lactic acid bacteria were selected for antifungal activity by a conidial germination assay. The 10-fold-concentrated culture filtrate of Lactobacillus plantarum 21B grown in wheat flour hydrolysate almost completely inhibited Eurotium repens IBT18000, Eurotium rubrum FTDC3228, Penicillium corylophilum IBT6978, Penicillium roqueforti IBT18687, Penicillium expansum IDM/FS2, Endomyces fibuliger IBT605 and IDM3812, Aspergillus niger FTDC3227 and IDM1, Aspergillus flavus FTDC3226, Monilia sitophila IDM/FS5, and Fusarium graminearum IDM623. The nonconcentrated culture filtrate of L. plantarum 21B grown in whole wheat flour hydrolysate had similar inhibitory activity. The activity was fungicidal. Calcium propionate at 3 mg ml(-1) was not effective under the same assay conditions, while sodium benzoate caused inhibition similar to L. plantarum 21B. After extraction with ethyl acetate, preparative silica gel thin-layer chromatography, and chromatographic and spectroscopic analyses, novel antifungal compounds such as phenyllactic and 4-hydroxy-phenyllactic acids were identified in the culture filtrate of L. plantarum 21B. Phenyllactic acid was contained at the highest concentration in the bacterial culture filtrate and had the highest activity. It inhibited all the fungi tested at a concentration of 50 mg ml(-1) except for P. roqueforti IBT18687 and P. corylophilum IBT6978 (inhibitory concentration, 166 mg ml(-1)). L. plantarum 20B, which showed high antimold activity, was also selected. Preliminary studies showed that phenyllactic and 4-hydroxy-phenyllactic acids were also contained in the bacterial culture filtrate of strain 20B. Growth of A. niger FTDC3227 occurred after 2 days in breads started with Saccharomyces cerevisiae 141 alone or with S. cerevisiae and Lactobacillus brevis 1D, an unselected but acidifying lactic acid bacterium, while the onset of fungal growth was delayed for 7 days in bread started with S. cerevisiae and selected L. plantarum 21B.

Intralesional ethanol in the treatment of unresectable liver cancer.

Percutaneous ethanol injection (PEI) under ultrasonography guidance has been widely tried in not advanced hepatocellular carcinoma (HCC). Ten years after the introduction of PEI, some conclusions of its indications can be drawn. In our series, 210 cirrhotic patients were treated; 141 with multisession PEI in an outpatient clinic, 57 with "single session" PEI under general anesthesia, and 12 with both. The 1-, 3-, and 5-year survival rates (by Kaplan-Meier method) were 953%, 65%, and 41% for Child class A patients with single HCC < or = 5 cm, and 88%, 47%, and 33% for patients with multiple HCC up to five lesions < or = 5 cm. In these patients the local recurrence rate was 15% and the new lesion rate at the 5-year follow-up was 74%. One death due to hemorrhage from esophageal varices in a Child class C patient treated by single session PEI occurred. The large number of cirrhotic patients demand for effective, safe, repeatable, low-cost treatment that can be offered at many centers. PEI meets all these requirements. PEI is proposed as the treatment of choice for the mentioned patients, excluding candidates for liver transplantation and surgical resection according to the predictive adverse factors currently in use. "Single session" technique widened the indications of traditional PEI to larger lesions.

Use of a rapid method for genotyping human platelet antigen systems in neonatal alloimmune thrombocytopenia.

We applied a polymerase chain reaction-sequence specific primer (PCR-SSP) method developed by other researchers to study 4 families of newborns with neonatal alloimmune thrombocytopenia (NAITP) in which serology had provided inconclusive human platelet antigen (HPA) typing data. This method allowed for the identification of the newborn HPAs which were incompatible with their respective mothers. They were HPA-2b, -1b, -3a, and -5b. This PCR-SSP is a useful tool for improving the ability to identify the incompatible HPA in NAITP.

Small hepatocellular carcinoma: treatment with radio-frequency ablation versus ethanol injection.

PURPOSE: To compare the effectiveness of radio-frequency (RF) ablation with that of percutaneous ethanol injection in the treatment of small hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Eighty-six patients with 112 small (< or = 3-cm-diameter) HCCs underwent RF ablation (42 patients with 52 tumors) or percutaneous ethanol injection (44 patients with 60 tumors). Therapeutic efficacy was evaluated with dual-phase spiral computed tomography performed at least 4 months after treatment. RESULTS: Complete necrosis was achieved in 47 of 52 tumors with RF ablation (90%) and in 48 of 60 tumors with percutaneous ethanol injection (80%). These results were obtained with an average of 1.2 sessions per tumor with RF ablation and 4.8 sessions per tumor with percutaneous ethanol injection. One major complication (hemothorax that required drainage) and four minor complications (intraperitoneal bleeding, hemobilia, pleural effusion, cholecystitis) occurred in patients treated with RF ablation; no complications occurred in patients treated with percutaneous ethanol injection. CONCLUSION: RF ablation results in a higher rate of complete necrosis and requires fewer treatment sessions than percutaneous ethanol injection. However, the complication rate is higher with RF ablation than with percutaneous ethanol injection. RF ablation is the treatment of choice for most patients with HCC.

Long term results of single session percutaneous ethanol injection in patients with large hepatocellular carcinoma.

BACKGROUND: The objective of this study was to evaluate the indications for percutaneous ethanol injection (PEI) performed in a single session under general anesthesia for treating patients with cirrhosis and large (tumors > 5 cm) hepatocellular carcinoma (HCC), and relevant survival curves. METHODS: Between November 1991 and November 1996, 108 patients were treated (a total of 128 procedures). They fell into 3 groups: 24 patients with single, encapsulated HCC measuring from 5-8.5 cm (Group A); 63 patients with single, infiltrating HCC measuring from 5-10 cm or multiple HCC (Group B); and 21 patients with advanced disease, either hepatic (Child's Class C) or neoplastic (symptomatic HCC or with portal thrombosis) type (Group C). The mean amount of ethanol injected was 62 mL. The average hospital stay was 3.8 days. The mean follow-up time was 40 months. RESULTS: The 1-, 2-, 3-, and 4-year survival rates were: 72%, 65%, 57%, and 44%, respectively, for Group A; 73%, 60%, 42%, and 18%, respectively, for Group B; and 46%, 25%, and 0%, respectively, for Group C. Mortality was 0.7% (bleeding from esophageal varices in a Child's Class C patient). The rate of major complications was 4.6% (1 case of peritoneal hemorrhage, 1 case of severe liver failure, 1 case of transient renal insufficiency, 1 case of peritoneal seeding, and 2 cases of infarctions of a segment adjacent to the tumor). CONCLUSIONS: Single session PEI has been proven to be a valid alternative in patients otherwise treated surgically or with transcatheter arterial chemoembolization who present with adverse prognostic factors or risks for these therapies, and may be an option for selected patients with advanced disease previously excluded from any therapy. Risk conditions are marked portal or pulmonary hypertension or esophageal varices at risk of bleeding, superficial tumors with severe coagulation disorders, hyperfibrinolysis, chronic renal insufficiency, and obstructive jaundice.

Percutaneous ethanol injection in the treatment of hepatocellular carcinoma in cirrhosis. A study on 207 patients.

In 207 cirrhotic patient carriers of hepatocellular carcinoma (HCC), percutaneous ethanol injection (PEI) was administered with ultrasound guidance. The patients were classified as Child's Class A, 136; B, 54; and C, 17. Their mean age was 63.5 years, and the male-female ratio was 3.5:1. There was a single HCC less than 5 cm in diameter in 162 patients; 45 had more than one HCC. The follow-up ranged from 5 to 71 months (mean, 25 months). No noteworthy complications occurred during or after 2485 treatments. The 1-year, 2-year, and 3-year survival percentages (by the Kaplan-Meier method) for the patients with one HCC were 90%, 80%, and 63%, respectively. The corresponding percentages by Child's class were 97%, 92%, and 76% for Class A; 88%, 68%, and 42% for B; and 40%, 0%, and 0% for C. The 1-year, 2-year and 3-year survival rates for patients with more than one HCC were 90%, 67%, and 31% respectively. These results were similar to those found by others and showed that PEI was a safe, reproducible, easy-to-do, and low-cost therapeutic technique. In terms of survival, these PEI results were better than the published results of no treatment and equivalent to those of surgery. In uncontrolled series, bias can play an important role. Therefore, additional trials would be useful.