Measurement of the

^{140}Ce(n,γ) is a key reaction for slow neutron-capture (s-process) nucleosynthesis due to being a bottleneck in the reaction flow. For this reason, it was measured with high accuracy (uncertainty ≈5%) at the n_TOF facility, with an unprecedented combination of a high purity sample and low neutron-sensitivity detectors. The measured Maxwellian averaged cross section is up to 40% higher than previously accepted values. Stellar model calculations indicate a reduction around 20% of the s-process contribution to the Galactic cerium abundance and smaller sizeable differences for most of the heavier elements. No variations are found in the nucleosynthesis from massive stars.

Image-guided microwave ablation of hepatic tumours: preliminary experience.

PURPOSE: Microwave thermal ablation (MWA) opens up a new scenario in the field of image-guided tumour ablation thanks to its potential advantages over validated radiofrequency ablation (RFA). In this pilot study, we assessed the technical success, safety and efficacy of MWA in treating hepatic malignancies. MATERIALS AND METHODS: After obtaining informed consent, we enrolled 15 inoperable patients, for a total of 19 lesions (ten metastases, nine hepatocellular carcinoma) with a mean diameter of 47 mm (range 14-78 mm). Mean follow-up was 8 (range 1-14) months. RESULTS: Technical success reached 100%. Complications (one major and one minor) occurred in two cases. Complete ablation, obtained in 68.4% of cases, showed no significant correlation with either cancer histological type or with lesion diameter. At follow-up, treatment failures occurred in 60% of cases; lesion diameter was the only prognostic factor for maintaining complete ablation. CONCLUSIONS: Our preliminary results should encourage further trials of this technique. MWA proved to be feasible and safe in treating advanced-stage liver tumours and represented an additional therapeutic attempt to be validated in further and larger efficacy studies.

Long-term outcome of radiofrequency thermal ablation (RFA) of liver metastases from colorectal cancer (CRC): size as the leading prognostic factor for survival.

PURPOSE: The aim of this study was to review some prognostic factors for survival after radiofrequency ablation (RFA) of metastases from colorectal cancer (CRC). MATERIALS AND METHODS: From 1996 to 2009, 262 patients with metastases from CRC were treated with RFA. Fourteen were lost to follow-up. The following predictors were analysed in the remaining 248: synchronous/metachronous metastases, single/multiple metastases, diameter of largest metastasis and absence/presence of extrahepatic metastases. Survival was measured from the date of metastasis diagnosis and from the date of RFA. RESULTS: Survival at 1, 2, 3 and 5 years was 93%, 78%, 62% and 35% from metastasis diagnosis, and 84%, 59%, 43% and 23% from the date of RFA. Median survival was 41 months in patients with largest metastasis ≤3 cm and 21.7 months for those with metastases >3 cm (p=0.0001); survival increased to 45.2 months in patients with largest metastasis ≤2.5 cm and fell to 18.5 months in those with metastasis >3.5 cm. Median survival of patients with extrahepatic metastases was significantly lower than that of patients without extrahepatic disease (23.3 vs. 32.6 months, p=0.018). CONCLUSIONS: In light of our long-term results obtained with commonly used equipment, small lesion size (diameter of largest lesion ≤3 or 2.5 cm) proved to be the most favourable prognostic factor for survival in patients with CRC metastases to the liver treated with RFA. This conclusion is probably related to the possibility of obtaining radical ablation and points to the usefulness of devices allowing ablation of larger volumes. In the presence of extrahepatic metastases, RFA has less impact on survival, even though it is potentially useful in patients at a higher risk of death due to hepatic rather than extrahepatic metastases.

Effect-directed profiling of 32 vanilla products, characterization of multi-potent compounds and quantification of vanillin and ethylvanillin.

Food testing is of great importance to the food industry and organizations to verify the authenticity claims, to prove the quality of raw materials and products, and to ensure food safety. The market prices of vanilla differed by a factor of about 20 in the last three decades. Therefore the risk of adulteration and counterfeiting of vanilla products is high. Instead of commonly used target analyses and sum parameter assays, a complementary non-target multi-imaging effect-directed screening was developed, which provided a new perspective on the wide range of vanilla product qualities on the market. Planar chromatography was combined with effect-directed assays, and the obtained biological and biochemical profiles of 32 vanilla products from nine different categories revealed a variety of active ingredients. Depending on the region, typical vanilla product profiles and activity patterns were obtained for pods, tinctures, paste (inner part), oleoresin and powders. However, some vanilla products showed additional active compounds and a different intensity pattern. The vanilla product profiles substantially differed from those of vanilla aroma or products containing synthetic vanillin or vanilla-flavored food products. Bioactive compounds of interest were online eluted and further characterized via HPTLC-HRMS, which allowed their tentative assignment. After purchase of the standards, these were successfully confirmed by co-chromatography. Quantification of vanillin across nine different product categories revealed levels ranging from 1 µg/g to 36 mg/g with a mean repeatability of 1.9%. The synthetic ethylvanillin was not detected in the investigated samples in significant concentrations. The assessment of differences in the activity patterns pointed to highly active compounds, which were not detected at UV/Vis/FLD but first via the biological and enzymatic assays. This effect-directed profiling bridges the gap from analytical food chemistry to food toxicology, and thus, makes an important contribution to consumer safety. In the same way, it would accelerate investigations for Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) according to Regulation (EC) No. 1907/2006.

The pulmonary nodule: clinical and radiological characteristics affecting a diagnosis of malignancy.

The role of computed tomography (CT) in the diagnosis of the solitary pulmonary nodule (SPN) is constantly expanding. CT helps to detect a growing number of increasingly small lesions, but, as with chest radiography, the primary goal in the evaluation of small pulmonary nodules is to exclude malignancy. Despite the availability of numerous, variously invasive, diagnostic tests, diagnostic accuracy tends to decline as the size of the nodule decreases. The role of the radiologist is therefore to help the clinician determine the most appropriate management strategy by using all available modalities [CT, magnetic resonance (MR) imaging, positron emission tomography (PET)] and evaluating the patient's clinical history and the imaging features leading to a diagnosis of benignity or malignancy. Imaging features include nodule size, margins, calcifications and fatty component, internal features (cavitations, pseudocavitations, air bronchogram, halo sign), as well as advanced techniques for characterisation (growth rate, contrast enhancement) and management (computer-aided diagnosis, Bayesian analysis, neural networks). The aim of this paper is to summarise the approach to pulmonary nodules from the point of view of the radiologist, oncologist and thoracic surgeon.

Molecular and Histological Changes in Post-Treatment Biopsies of Non-Squamous Non-Small Cell Lung Cancer: A Retrospective Study.

BACKGROUND: Recently, in advanced non-small cell lung cancer (NSCLC), standard chemotherapy was flanked by biological agents directed against genomic abnormalities, including EGFR and ALK alterations, that significantly improved patient outcome. Despite these achievements, tumour progression almost always occurs and a reassessment of the tumour genetic profile may contribute to modulating the therapeutic regimen. Resampling may provide tissue for additional tests to detect acquired resistance and/or new genetic alterations, but the currently available information is limited. PATIENTS AND METHODS: Histological and genetic reassessments of biopsy or surgical tissue samples from 50 non-squamous NSCLC patients before and after at least one systemic treatment were performed. EGFR, KRAS, BRAF, PIK3CA and HER2 mutations were sequenced, p.T790M was identified with real-time PCR, and ALK and MET genomic alterations by fluorescence in situ hybridization. RESULTS: Overall in baseline biopsies, 37/50 (74 %) tumours had genetic alterations, either single (52 %) or multiple (22 %). Among them, 16 were EGFR mutations and 6 ALK rearrangements. In the second tissue sampling, 54 % of cases had additional genomic changes, including newly acquired alterations (81 %) or losses (18 %). The commonest changes were MET amplification and p.T790M mutation. One case had a histological shift from adenocarcinoma to small cell carcinoma. CONCLUSIONS: The remarkable number of molecular changes following systemic therapy and the genetic complexity of some cases underline the value of histological and molecular re-evaluation of lung cancer to tailor the most appropriate therapy during disease progression.

HIV-induced syncytium formation requires the formation of conjugates between virus-infected and uninfected T-cells in vitro.

The transmission of HIV requires the interaction of the cell-surface CD4 receptor and the viral envelope glycoprotein. Experiments were performed to determine the role of other cell-surface molecules in the development of HIV-induced syncytia. Although CEM and MT-2 cells had similar cell-surface CD4 receptor densities, less than 1% of CEM cells and greater than 95% of MT-2 cells formed syncytia with H9 cells chronically infected with HIV-1 (H9-IIIB). When compared with CEM cells, MT-2 cells exhibited a 10-fold and threefold greater capacity to form homotypic and heterotypic conjugates with H9 cells, respectively. Increasing the conjugate formation capacity of CEM cells with the lectin wheat germ agglutinin led to a greater than 30-fold increase in the formation of syncytia with H9/IIIB cells. The formation of syncytia between MT-2 and H9/IIIB cells was magnesium-, energy-, temperature-, and actin-cytoskeleton-dependent, and could be inhibited (65%) by an anti-LFA-1 monoclonal antibody. The combination of anti-leukocyte function-associated antigen-1 (LFA-1) and anti-CD2 monoclonal antibodies resulted in a synergistic inhibition (89%) of syncytium formation. These results indicate that integrins and other cell-surface adhesion molecules regulate HIV-induced syncytium formation.

Effects of long-term zidovudine treatment on cell-mediated immune response and lymphokine production.

The effects of long-term zidovudine treatment on functional parameters of cell-mediated immunity were investigated in 15 symptomatic HIV-antibody-positive patients with clinical evidence of opportunistic infections. Mononuclear leukocytes were obtained before administering the drug, and after 3 and 6 months of treatment. The cells were stimulated with lectins in order to assess variations of mitogen-induced lymphocyte proliferation and production of gamma-interferon (IFN) and interleukin-2 (IL-2), and with Newcastle disease virus (NDV) to assess variations of alpha-IFN production. Mean proliferative responses to PHA and Con-A did not show any significant change between baseline, 3 months, and 6 months values (25,158 +/- 11,763, 24,662 +/- 8,955, and 34,924 +/- 16,283 D-cpm, respectively, for PHA, p greater than 0.05; and 5,470 +/- 1,890, 4,953 +/- 2,518, and 4,539 +/- 3,286 D-cpm, respectively, for Con-A, p less than 0.05). Mean response to PWM (9,707 +/- 4,429 D-cpm at entry) increased significantly after 3 months, but returned to baseline values at 6 months (17,039 +/- 5,123 and 10,314 +/- 3,855 D-cpm, respectively, p = 0.016). IL-2 production (5.51 +/- 4.0 I.U. at entry) rose particularly at 3 months and persisted at the same levels at 6 months (9.6 +/- 4.8 and 9.5 +/- 6 I.U., respectively, p less than 0.05). By contrast, a moderate but significant decrease in gamma- and alpha-IFN production was observed (65 +/- 2.2, 35.1 +/- 1.6, and 22 +/- 2 I.U., respectively, for gamma-IFN, p less than 0.05; 60 +/- 3, 64 +/- 2.6, and 12 +/- 1.5 I.U., respectively for alpha-IFN, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Discovery, synthesis, and bioactivity of bis(heteroaryl)piperazines. 1. A novel class of non-nucleoside HIV-1 reverse transcriptase inhibitors.

A variety of analogues of 1-[4-methoxy-3,5-dimethylbenzyl]-4-[3-(ethylamino)-2-pyridyl]piperazine hydrochloride (U-80493E) were synthesized and evaluated for their inhibition of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT). Replacement of the substituted aryl moiety with various substituted indoles provided bis(heteroaryl)piperazines (BHAPs) that were 10-100-fold more potent than U-80493E. The pyridyl portion of the lead molecule was found to be very sensitive to modifications. Extensive preclinical evaluations of several of these compounds led to the selection of 1-[(5-methoxyindol-2-yl)carbonyl]-4-[3-(ethylamino)-2- pyridyl]piperazine methanesulfonate (U-87201E, atevirdine mesylate) for clinical evaluation.

Nucleotide dimers suppress HIV expression in vitro.

A series of nucleotide homo- and heterodimers [3'-azido-3'-deoxythymidilyl-(5',5')-2',3'-di-deoxy-5' adenylic acid (AZT-P-ddA), 3'-azido-3'-deoxythymidilyl-(5',5')-2', 3'-dideoxy;-5'-adenylic acid, 2-cyanoethyl ester [AZT-P(CyE)-ddA], 3'-azido-3'-deoxythymidilyl-(5',5')-2',3'-dideoxy-5'-inosinic acid (AZT-P-ddI), and 3'-azido-3'-deoxythymidilyl-(5',5')-3'-azido-3'-deoxy-5'-thymid ilic acid (AZT-P-AZT)] were synthesized and compared with respect to their anti-HIV and cytotoxic properties to their component monomers in vitro. MT-2 cells were infected with HIV (TM) followed by the addition of drug. The dimers and their respective monomers inhibited HIV-induced syncytia formation, reverse transcriptase production, and the expression of HIV p24 antigen. However, on an equimolar basis, greater anti-HIV potency and enhanced cytotherapeutic indices were observed with the heterodimers when compared with their monomers. Nucleotide dimers, such as AZT-P-ddA, should be actively considered for further evaluation as anti-HIV agents.

Cellular pharmacology and anti-HIV activity of 2',3'-dideoxyguanosine.

The antiviral activity, uptake, and metabolism of 2',3'-dideoxyguanosine was investigated in human immunodeficiency virus- (HIV) infected and noninfected human cells. 2',3'-Dideoxyguanosine had anti-HIV activity (effective dose 50%: 0.1-1.0 microM) in H-9 and MT-2 cells. The addition of excess (greater than or equal to 30 microM) guanosine, deoxyguanosine, or 8-aminoguanosine had no effect on the anti-HIV activity of 2',3'-dideoxyguanosine. In [8-3H]2',3'-dideoxyguanosine-exposed cells, the intracellular radioactivity was twofold higher than the extracellular. When guanosine, deoxyguanosine, or 8-aminoguanosine was preincubated or added simultaneously to 2',3'-dideoxyguanosine, uptake of 2',3'-dideoxyguanosine was reduced by 28 to 34%, whereas addition of p-nitrobenzylmercaptopurine riboside (20 microM) had no effect. In metabolism studies using H-9 cells, dideoxyguanosine triphosphate could not be detected despite a 24-h incubation of 2',3'-dideoxyguanosine at effective anti-HIV concentrations. The addition of excess (greater than or equal to 30 microM) guanosine, deoxyguanosine, and 8-aminoguanosine, while inhibiting the catabolism of 2',3'-dideoxyguanosine, did not enhance the anabolic conversion of 2',3'-dideoxyguanosine to dideoxyguanosine triphosphate. Our failure to detect the formation of dideoxyguanosine triphosphate and the lack of reversal of antiviral effects by natural purine nucleosides raises questions on the role of this metabolite in the anti-HIV activity of 2',3'-dideoxyguanosine.

Uptake of 2',3'-dideoxyadenosine in human immunodeficiency virus-infected and noninfected human cells.

The uptake of 2',3'-dideoxyadenosine was examined in a human immunodeficiency virus (HIV) infected and uninfected T cell line (H9 cells), a B cell line (Namalwa), and in normal peripheral blood mononuclear cells. After a 10-minute incubation at ambient temperature, the intracellular 2',3'-dideoxyadenosine-derived radioactivity was 8- to 16-fold higher than the extracellular radioactivity. In metabolically inactive cells (0 degrees C), the intracellular and extracellular 2',3'-dideoxyadenosine-derived radioactivities were nearly equal. In infected and noninfected H9 cells, a large excess of p-nitrobenzylmercaptopurine riboside or pyrimidine nucleosides weakly inhibited the uptake of 2',3'-dideoxyadenosine (7-30%), whereas deoxycoformycin was a stronger inhibitor (50-80%). Purine nucleosides minimally enhanced the uptake (10-20%). The cellular uptake was not associated with the accumulation of dideoxyadenosine triphosphate. In normal peripheral blood mononuclear cells, the uptake of 2',3'-dideoxyadenosine was inhibited by all agents except 2'-deoxyadenosine (15% enhancement). In contrast to H9 cells, the formation and accumulation of dideoxyadenosine triphosphate paralleled the uptake of dideoxyadenosine. The results of these studies suggest that the major route of transport of 2',3'-dideoxyadenosine into cells is by simple diffusion and that different metabolic patterns exist among cell lines and normal peripheral blood mononuclear cells. An understanding of these cellular differences could aid in the development of therapeutic strategies directed against HIV.

The benzylthio-pyrimidine U-31,355, a potent inhibitor of HIV-1 reverse transcriptase.

U-31,355, or 4-amino-2-(benzylthio)-6-chloropyrimidine is an inhibitor of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and possesses anti-HIV activity in HIV-1-infected lymphocytes grown in tissue culture. The compound acts as a specific inhibitor of the RNA-directed DNA polymerase function of HIV-1RT and does not impair the functions of the DNA-catalyzed DNA polymerase or the Rnase H of the enzyme. Kinetic studies were carried out to elucidate the mechanism of RT inhibition by U-31,355. The data were analyzed using Briggs-Haldane kinetics, assuming that the reaction is ordered in that the template:primer binds to the enzyme first, followed by the addition of dNTP, and that the polymerase is a processive enzyme. Based on these assumptions, a velocity equation was derived that allows the calculation of all the essential forward and backward rate constants for the reactions occurring between the enzyme, its substrates, and the inhibitor. The results obtained indicate that U-31,355 acts as a mixed inhibitor with respect to the template:primer and dNTP binding sites associated with the RNA-directed DNA polymerase domain of the enzyme. The inhibitor possessed a significantly higher binding affinity for the enzyme-substrate complexes, than for the free enzyme and consequently did not directly affect the functions of the substrate binding sites. Therefore, U-31,355 appears to impair an event occurring after the formation of the enzyme-substrate complexes, which involves either inhibition of the phosphoester bond formation or translocation of the enzyme relative to its template:primer following the formation of the ester bond. Moreover, the potency of U-31,355 depends on the base composition of the template:primer in that the inhibitor showed a much higher binding affinity for the enzyme-poly (rC):(dG)10 complexes than for the poly (rA):(dT)10 complexes.

Development of an assay that detects transcriptionally competent human immunodeficiency virus type one particles.

To study the functional properties of HIV-1 reverse transcriptase (RT) from intact viral particles without the requirement for tissue culture expansion, a method that couples HIV-1 reverse transcription utilizing its endogenous RT (ERT) with polymerase chain reaction amplification (PCR) was developed. Detection of endogenous reverse transcripts from HIV particles by ERT-PCR was compared to HIV RNA PCR detection using avian myeloblastosis virus (AMV) RT from plasma samples from 45 HIV-1 infected patients. The HIV ERT-PCR method was capable of detecting plasma viremia with the same efficiency (29/29 patients) as the AMV RT HIV RNA PCR in patients with CD4 cell counts of less than 500/mm3. The determination of HIV-RT drug sensitivities using four well-characterized HIV-1 lab strains was assessed. The ERT-PCR method detected reduced sensitivity to TIBO R82150 (10 microM) in a TIBO resistant strain but not in the TIBO sensitive HTLV-IIIB viral mixture or an HTLV-IIIB clone. In summary, the HIV ERT-PCR method provides a useful approach for the detection of HIV and the characterization of RT sensitivities among HIV-1 strains.

Vitiligo associated with alpha-interferon in a patient with chronic active hepatitis C.

We report a case of vitiligo that occurred during the second month of interferon alpha 2a therapy for chronic active hepatitis C. The patient received the interferon for four months.

Age-related histochemical and immunohistochemical changes in human labial salivary glands.

We here in determined the histochemical and immunological changes that salivary glands experience as a result of the ageing process. Samples from the inferior lip of children, youngsters and young adults were analyzed by histochemical techniques, Periodic-acid-Schiff, (PAS) Alcian blue (AB) (pHs 1.0 and 2.5) and Toluidine blue for mucosubstances and immunohistochemical staining of S 100 protein and cytokeratin 20, avidin-biotin system (DAB). In children, the techniques used evidenced various reaction degrees in the acinar cells, even within a single acinous. They displayed a slight metachromasia and were alcianophilic at pH 2.5. In youngsters, and especially in adults, glands showed a notable PAS positivity and alcianophilia at both pH levels, and an intense metachromasia. PS 100 was positive in the basal area of the acini and serous demilunes of all groups, the reactivity being higher in adults. Cytokeratin 20 was better observed in ductal cells from children glands. These findings suggest modifications at cytological level and in the chemical composition of the secretory granules, indicating possible functional variations in lip salivary glands related to the ageing process.

Ultrasound diagnosis of pneumothorax.

PURPOSE: The purpose of this study was to show the effectiveness of ultrasound (US) in the evaluation of pneumothorax by comparison with X-rays and computed tomography (CT). MATERIALS AND METHODS: A series of 184 patients (130 men and 54 women), aged 26 to 82 years, underwent chest US after percutaneous needle biopsy. US findings were compared with CT postbiopsy selective slices and to X-rays. RESULTS: Pneumothorax was identified in 46 patients (25%) by CT, in 44 by US, with no false positives, and in 19 by X-rays. US sensitivity was 95.65%, specificity 100% and diagnostic effectiveness 98.91%. CONCLUSIONS: Chest US was found to be a valuable diagnostic tool in pneumothorax diagnosis, with diagnostic effectiveness well beyond X-rays and similar to CT.