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Background Currently, most laboratories identify yeasts routinely on the basis of morphology and biochemical reactivity. This approach has quite often limited discriminatory power and may require long incubation periods. Due to the increase of fungal infections and due to specific antifungal resistence patterns for different species, accurate and rapid identification has become more important. Several molecular techniques have been described for fast and reliable identification of yeast isolates, but interlaboratory exchangeability of identification schemes of molecular techniques has hardly been studied. Here, we compared amplified ITS2 fragment length determination by an ABI Prism 310 (Applied Biosystems, Foster City, Ca.) capillary electrophoresis system with that obtained by a CEQ8000 (Beckman Coulter, Fullerton, Ca.) capillary electrophoresis system. Results Although ITS2 size estimations on both systems differed and separate libraries had to be constructed for each system, both approaches had the same discriminatory power with regard to the 44 reference strains, identical identifications were obtained for 39/ 40 clinical isolates in both laboratories and strains from 51 samples were correctly identified using CEQ8000, when compared to phenotypic identification. Conclusion Identification of yeasts with ITS2-PCR followed by fragment analysis can be carried out on different capillary electrophoresis systems with comparable discriminatory power.

Computed tomography (CT) affords the best visualization of all organs in the body, simultaneously depicting air, soft tissue, and bones on the same scan. This ability makes CT an ideal imaging modality for tissue ablation, as it enables the physician to target any type of organ accurately and to avoid inadvertently puncturing others along the needle path. However, in the past, as the duration of image acquisition and reconstruction was long with CT, this time-consuming aspect appeared inconvenient for tissue ablation. Recent improvements in computerized data management have transformed CT into almost a real-time imaging technique, and even more recently, the advent of multislice CT has made imaging of a volume in a single acquisition feasible, thereby improving the scope of CT guidance for tissue ablation. CT can now be used throughout tissue ablation for guidance, monitoring, and follow-up.

Background Mycoplasmas are present worldwide in a large number of animal hosts. Due to their small genome and parasitic lifestyle, Mycoplasma spp. require complex isolation media. Nevertheless, already over 100 different species have been identified and characterized and their number increases as more hosts are sampled. We studied the applicability of amplified rDNA restriction analysis (ARDRA) for the identification of all 116 acknowledged Mycoplasma species and subspecies. Methods Based upon available 16S rDNA sequences, we calculated and compared theoretical ARDRA profiles. To check the validity of these theoretically calculated profiles, we performed ARDRA on 60 strains of 27 different species and subspecies of the genus Mycoplasma. Results In silico digestion with the restriction endonuclease AluI (AG^CT) was found to be most discriminative and generated from 3 to 13 fragments depending on the Mycoplasma species. Although 73 Mycoplasma species could be differentiated using AluI, other species gave undistinguishable patterns. For these, an additional restriction digestion, typically with BfaI (C^TAG) or HpyF10VI (GCNNNNN^NNGC), was needed for a final identification. All in vitro obtained restriction profiles were in accordance with the calculated fragments based on only one 16S rDNA sequence, except for two isolates of M. columbinum and two isolates of the M. mycoides cluster, for which correct ARDRA profiles were only obtained if the sequences of both rrn operons were taken into account. Conclusion Theoretically, restriction digestion of the amplified rDNA was found to enable differentiation of all described Mycoplasma species and this could be confirmed by application of ARDRA on a total of 27 species and subspecies.

Radiofrequency current can be locally delivered through electrode-needle inserted in the tumors and allows local destruction of tissues, up to a volume of 4 to 5 cm in diameter with one application. Such destruction guided with medical imaging proved a high local efficacy over 90% for tumors less than 25 mm in the liver, lung, and kidney. For larger tumors, efficiency is lower but new technical development might improve these results. Additionally RF has been efficiently used to palliate pain due to osteoid osteomas and malignant bone metastases with a high success rate and low complication rate. Radiofrequency, although very efficient in local tumor control, has neither proven to prolong survival or to be equivalent to surgery in randomized trial, even if some retrospective studies have done so. Further studies are needed to evaluate the exact benefit of this promising technique.

Hepatic resection offers the best chance of survival for patients with liver metastases (LM) of colorectal origin. However, some patients are not eligible for surgery because of a too small future liver remnant (FLR) which carries a high risk of severe postoperative liver failure. The operability status of these patients can be favorably changed by selective right portal vein embolization (PVE) which induces compensatory growth of the left liver. However, during liver regeneration following right PVE, the left LM growth rate is faster than that of the non-embolized normal liver parenchyma. This study aimed at examining an approach for those patients in which there is bilateral LM potentially resectable following portal vein embolization, but in which there is a risk of rapid liver metastasis growth in the non-embolized liver.Between October 1998 and January 2001, 5 patients underwent simultaneous right PVE and radiofrequency ablation (RFA) of a left LM, prior to a major right-sided hepatectomy for initially unresectable bilateral LM. All these patients had one LM in the left liver in addition with multiple LM in the right liver. Simultaneous right PVE and left RFA was performed percutaneously under intravenous sedation and analgesia. One month later, hepatectomy was undertaken. To allow histologic assessment of the RFA effectiveness, the previously treated left-sided tumor was also resected and analyzed.Simultaneous PVE-RFA was successful in all patients. No tumor growth on the RFA site was observed during the interval between PVE-RFA and surgery. Histologic examination showed complete tumor sterilization of the RFA necrotic zone. In the postoperative course, 1 patient died of acute liver failure. For the 4 remaining patients, morbidity was minimal (transient bile leak in one patient).Simultaneous percutaneous right PVE and left RFA is feasible. This procedure allowed good left-sided tumor control during liver growth following PVE in all five patients. It is the most logical procedure for patients with bilateral colorectal LM needing right PVE before resection, if the left concomitant LM is small and accessible to percutaneous RFA. This procedure should be preferred because it eliminates the risk of left LM growth during the 1-month interval between PVE and surgery.

A total of 61 clinical yeast isolates of Candida , Cryptococcus , Blastoschizomyces , and Saccharomyces spp. were used to compare two identification techniques, VITEK 2 and ITS2-fragment length polymorphism analysis (ITS2-FLP), with ID32C as the reference method. ID32C identified 58 isolates correctly. ITS2-FLP with Instagene DNA extraction identified 59 isolates. ITS2-FLP combined with boiling-freezing DNA extraction identified 55 isolates. VITEK 2 identified 41 isolates correctly.

After a rather long period of stagnation, intra-arterial therapeutic approaches for treating liver tumors are currently progressing rapidly. These new modalities will increase the resectability of initially unresectable liver tumors after dramatic responses. At the same time, resectability rates are increasing with the assistance of local ablative physical treatments such as radiofrequency, resulting in an improvement of patients' median survival rates and quality of life.

Selective embolization of portal branches of some liver segments will induce hypertrophy of non embolized segments of the liver. This induced hypertrophy allows us to perform hepatectomy in patients with initially insufficient volume of future remnant liver (FRL). This technique aims at patients with initial FRL volume below 25% of total liver volume, and patient with initial FRL volume below 40% of total liver volume when a diffuse liver disease is present. The mean hypertrophy obtained 1 month after pre-operative portal vein embolization (POPE) was 82 +/- 56% in our experience. Post-operative complications and long-term survival for patient operating after POPE is not different from patient operated with a classical hepatectomy. Radiofrequency is able to destroy hepatic tumors after insertion of a needle electrode inside the tumor under imaging guidance. Thermocoagulation of the targeted tissue is obtained after emission of a 400 to 500 KHz alternative able to induce frictional heating in the neighboring of the electrode. Spreading of this technique is at least partially explained by a relative high local efficacy (90% of small tumor targeted can be destroyed) and a low invasiveness. However, today no benefit in survival has been demonstrated for patients treated with radiofrequency ablation of liver tumors. Consequently, this technique should not be proposed for tumors that can be surgically resected.

New therapies are needed to improve the prognosis of patients with advanced hepatocellular carcinoma (HCC). Various gemcitabine-oxaliplatin combinations have been tested recently in patients with ovarian and pancreatic carcinoma, yielding interesting results with little toxicity. Therefore, the authors evaluated the activity and toxicity of two such combinations in patients with HCC.Twenty-one patients were enrolled prospectively in the study. Eleven patients received gemcitabine 1000 mg/m2 on Day 1 and oxaliplatin 100 mg/m2 on Day 2 (GEMOX-1), and 10 patients received gemcitabine 1500 mg/m2 on Day 1 followed by oxaliplatin 85 mg/m2 on Day 1 (GEMOX-2). Treatment was repeated every 2 weeks until disease progression developed or until unacceptable adverse effects occurred.All patients were assessable for response and toxicity. Four patients (19%) achieved objective responses (95% confidence interval, 13-26%), including 3 patients in the GEMOX-1 group and 1 patient in the GEMOX-2 group. Ten patients (48%) had stable disease, and 7 patients (33%) experienced disease progression. The median progression-free survival was 5 months, and the median overall survival was 12 months. Fifty-four percent of patients in the GEMOX-1 group and 50% of patients in the GEMOX-2 group had received previous systemic chemotherapy or cisplatin-based chemoembolization. Grade 3-4 hematologic toxicity, according to National Cancer Institute Common Toxicity Criteria, consisted of thrombocytopenia (GEMOX-1 vs. GEMOX-2, 18% vs. 40%) and neutropenia (0% vs. 30%). No Grade 3-4 nonhematologic toxicity was observed, except for 1 episode of Grade 3 diarrhea. Grade 1 neurotoxicity and Grade 2 neurotoxicity (specific scale), respectively, were observed in 4 patients and 7 patients receiving GEMOX-1 and in 7 patients and 1 patient receiving GEMOX-2.Gemcitabine-oxaliplatin combination therapy is feasible in patients with advanced HCC. The GEMOX-1 regimen was tolerated better than the GEMOX-2 regimen. Currently, the GEMOX-1 regimen is being evaluated in a Phase II study in previously untreated patients with HCC.

Small-sized tumors sited closer than 15 mm to the central bile ducts cannot be destroyed by radiofrequency (RF) because the resulting heating causes destruction and stenosis of the bile ducts.Thirteen patients underwent intraoperative RF to treat tumors closer than 6 mm to a central bile duct. Stenosis prevention was achieved by cooling the main bile duct(s) (right, left, or both) with a 4 degrees C saline solution quickly infused by a catheter introduced inside the bile duct through a choledochotomy. Median followup after cooling was 19.7 months (range 6 to 42 months).There was no mortality and no RF-related morbidity. Twelve patients (92.3%) did not present any biliary stenosis. In one patient (7.7%) biliary stenosis was detected 6 months after RF. The patient was cirrhotic but did not require any treatment. One of the 13 patients had a local recurrence.The intraoperative cooling of central bile ducts prevents biliary stenosis efficiently when using RF to destroy tumors closer than 5 mm to the bile duct. This new technique decreases the contraindications for the use of RF and allows more patients to be treated with RF with curative intent.

The taxonomic status of seven glucose-non-acidifying, non-proteolytic Acinetobacter strains characterized by forming small colonies on agar media was studied. With one exception, all strains were from human specimens. They could be distinguished from all described Acinetobacter (genomic) species by their ability to grow on ethanol and acetate as sole sources of carbon but not on 22 other substrates tested including DL-lactate or DL-4-aminobutyrate. DNA-DNA hybridization studies, 16S rRNA gene sequence analysis, amplified rDNA restriction analysis and DNA polymorphism analysis by AFLP showed that these strains represent a hitherto unknown species of the genus Acinetobacter, for which the name Acinetobacter parvus (type strain LMG 21765(T)=LUH 4616(T)=NIPH 384(T)=CCM 7030(T)) is proposed.

We describe the rates and potential risk factors of complications of radiofrequency ablation of hepatic tumors. SUBJECTS AND METHODS. Over a 5-year period, 312 patients underwent 350 sessions of radiofrequency ablation (124 intraoperative and 226 percutaneous) for treatment of 582 liver tumors including 115 hepatocellular carcinomas and 467 metastatic tumors. The chi-square test was used for a group-to-group comparison of the occurrence of adverse events.Thirty-seven (10.6%) adverse events and five (1.4%) deaths were related to radiofrequency treatment. The deaths were caused by liver insufficiency (n = 1), colon perforation (n = 1), and portal vein thrombosis (n = 3). Portal vein thrombosis was significantly (p0.00001) more frequent in cirrhotic livers (2/5) than in noncirrhotic livers (0/54) after intraoperative radiofrequency ablation performed during a Pringle maneuver. Liver abscess (n = 7) was the most common complication. Abscess occurred significantly (p0.00001) more frequently in patients bearing a bilioenteric anastomosis (3/3) than in other patients (4/223). We encountered five pleural effusions, five skin burns, four hypoxemias, three pneumothoraces, two small subcapsular hematomas, one acute renal insufficiency, one hemoperitoneum, and one needle-tract seeding. The 6.3% of minor complications did not require specific treatment or a prolonged hospital stay. Among the 5.7% major complications, 3.7% required less than 5 days of hospitalization for treatment or surveillance and 2% required more than 5 days for treatment.Radiofrequency ablation of liver tumors is a well-tolerated technique, but caution should be exercised when treating patients with a bilioenteric anastomosis, and radiofrequency ablation during vascular occlusion in cirrhotic livers should be avoided.

Three PCR techniques (amplification of the psaA , ply , and lytA genes) and a commercial kit (AccuProbe [GenProbe, San Diego, Calif.], based on hybridization with the 16S rRNA gene), all four of which claimed to be specific for Streptococcus pneumoniae , were used to identify 49 alpha-hemolytic streptococcal isolates suspected of being pneumococci. The definite phenotypic identification of these organisms as S. pneumoniae was difficult when optochin susceptibility and the presence of a capsule were taken as markers. Furthermore, Rsa I digestion of the amplified 16S rRNA gene was applied. All 49 strains were optochin resistant. Eleven of these were encapsulated and were identified as pneumococci by all tests. Twenty of the 38 unencapsulated strains were unambiguously identified as nonpneumococci by all tests. The identities of another 18 unencapsulated strains remained inconclusive due to highly variable reactions for all phenotypic and genotypic techniques applied. The AccuProbe test was positive for seven strains for which the results of the other tests were inconclusive. Rsa I restriction of the amplified 16S rRNA gene confirmed the AccuProbe result for all strains, while the result of the psaA -specific PCR was in concordance with encapsulation for all strains. The results presented here indicate that identification problems continue to exist for some strains, despite the application of genotypic and phenotypic tests in combination. We found the psaA -specific PCR to be the genotypic technique best suited for the identification of genuine pneumococci and optochin-resistant pneumococci.

437 Background: Local treatment of BM in mRCC pts comprises different approaches including thermal ablation techniques as well as radiofrequency ablation (RFA) and cryotherapy. We report our experience for the treatment of BM of mRCC pts using thermal ablation. Methods: The medical records of all pts with mRCC submitted to thermal ablation of BM from Feb 2008 to Oct 2013 were retrospectively analyzed. Intention of treatment was recorded in 3 categories: complete treatment of all BM in pts with oligometastatic disease or prevention of skeletal related events (SRE) risk and/or pain relief in pts with multimetastatic disease. The local control rate and prognostic factors for local failure (LF) were analyzed including patient characteristics, extension of disease, BM characteristics and type of treatment. Results: 22 pts with BM of mRCC were treated by thermal ablation techniques, with 32 procedures (3 pts had >=3 BM treated). Histology of primary tumor was clear cell in 20 and chromophobe carcinoma in 2 pts. At time of BM treatment, Heng prognostic score was good, intermediate and poor in 36, 59 and 5% of pts. Median age was 67 yrs (48-85). A majority of men were treated (16/22=73%). In 34%, pts had BM synchronous with the primary. BM presented cortical bone erosion in 69% and neurological structures in the vicinity in 50%. Intention of treatment was complete control of BM in 43% and prevention of risk of SRE and/or pain relief in 57% of pts. Cryotherapy was performed in 72% and RF in 28% of cases. Concomitant cimentoplasty and arterial embolization was performed in 70% and 30% respectively. Pain relief was achieved in 81% of pts. LF occurred in 19% of pts (n=6) (95% CI: 4-33%). The median follow-up was 13.4 months (95% CI: 11-21). Median OS was not reached. 86% of pts are still alive at 12 months. The unfavorable prognostic factors for LF were BM size >50 mm (p=0.035) and neurological structures in the vicinity (p=0.0083). Conclusions: Thermal ablation techniques are treatment options that should be considered in the curative and palliative treatment of BM in pts with mRCC. When indications are carefully discussed, these techniques have the potential to achieve good local control and acceptable survival.

316 Background: Although several studies have reported genomic characterisation of primary urothelial carcinoma (UC), little is known about the genomic alterations of the metastatic cisplatin-resistant UC. We sought to define the prevalence and co-occurrence of actionable genomic alterations in patients with metastatic cisplatin-resistant lethal UC. Methods: Patients with metastatic UC, who progressed after at least one line of standard therapy, were enrolled in a prospective molecular screening trial (MOSCATO 01) at Gustave Roussy. CT-Scan or ultrasound-guided biopsies were performed in metastases to carry out a comprehensive molecular characterization. DNA was extracted from fresh tumor samples and analyzed by comparative genomic hybridization (CGH) (≥ 30% tumor cells) and by Next Generation Sequencing (NGS) for up to 74 target genes (≥ 10% tumor cells). Whole-exome (WES) and RNA seq sequencing were performed retrospectively in selected cases. Results: From 12/2011 to 12/2013, 30 heavily pretreated patients with metastatic UC were included. Median age was 61 (37-73); all patients had been treated with platin-based chemotherapy with a median number of lines of 2.5 (1-5). A tumor biopsy could be performed in 26 patients (87%). CGH and targeted exome sequencing profiles were assessed in 19 (73%) and 23 (88%) of them, respectively. A total of 19 patients (73%) were profiled for both sequencing and CGH. WES and RNA seq was performed in 16 (62%) pairs of metastatic tissue and normal DNA (Integragen Inc, Hiseq platform). Our analyses identified potential therapeutic targets in 61 % of the tumours, including 31% with targets in the PI3K/AKT/mTOR pathway, 35% with targets in the FGF/FGFR pathway and 15% with targets in the RTK/MAPK pathway (including ERBB2). Others frequent aberrations were found in the chromatin regulatory genes (MLL gene family) and cell-cycle regulatory genes (E2F3, CDKN2A/B genes). FGFR gene fusions were found in 1 out of 16 screened patients (6%) Conclusions: Lethal platin-resistant UC exhibit high-level of genomic heterogeneity. The majority of these tumors harbour actionable genomic alterations that can be targeted with selective agents currently in clinical development (phase I trials) or registered.

The present study was carried out to estimate the prevalence of caseous lymphadenitis (CL) in sheep and goats slaughtered at the local abattoir in Elazig province located in the east of Turkey, between September and December 2000. A total of 2046 sheep and 2262 goat carcasses were examined during the study period and 118 abscessed lymph nodes, 89 from sheep and 29 from goats, were collected. Corynebacterium spp. strains were isolated from 81.4% of the abscesses, giving an overall prevalence of 2.2%. The prevalence was 3.5 and 1.1% in sheep and goats, respectively. PCR on DNA extracted from 96 suspicious isolates, using a pair of Corynebacterium pseudotuberculosis-specific primers, was positive for 93. Although cross-reaction with C. ulcerans, a human/bovine species, was observed, the PCR assay used in this study may successfully be applied for the diagnosis of CL in goats and sheep as an alternative to conventional methods, owing to its advantages of specificity and speed.

A gram-negative alkaline phosphatase- and pyrrolidone peptidase-positive rod-shaped bacterium (CCUG 45702) was isolated from two aerobic blood cultures from a female cancer patient. No identification could be reached using phenotypic techniques. Amplification of the tRNA intergenic spacers revealed fragments with lengths of 116, 133, and 270 bp, but no such pattern was present in our reference library. Sequencing of the 16S rRNA gene revealed its identity as Moraxella atlantae , a species isolated only rarely and published only once as causing infection. In retrospect, the phenotypic characteristics fit the identification as M. atlantae (formerly known as CDC group M-3). Comparative 16S rRNA sequence analysis indicates that M. atlantae , M. lincolnii , and M. osloensis might constitute three separate genera within the Moraxellaceae . After treatment with amoxicillin-clavulanic acid for 2 days, fever subsided and the patient was dismissed.

The aim of our study was to confirm the value of high-frequency and color Doppler sonography in evaluating the metastatic potential of melanomas.Sixty-nine histologically proven melanomas in 67 patients were prospectively studied before surgical resection. A morphologic study was performed with a 20-MHz probe. The maximal thickness of tumors was measured and compared with the Breslow index. A color Doppler sonographic analysis of neovascularization was performed with a 13-MHz probe to depict intratumoral vessels.Of the 69 melanomas, 65 were depicted on sonography. Tumor thickness ranged from 0.26 to 8.0 mm as measured by sonography and from 0.15 to 8.0 mm according to the Breslow index. Sonographic measurements correlated strongly with the Breslow index (r0.96). Surgical reexcision was necessary in 33 tumors with a Breslow index exceeding 1 mm, but reexcision could have been avoided in 31 of these cases had the initial surgical planning been based on high-frequency sonographic measurements. Among the 62 patients studied with color Doppler sonography, intratumoral vessels were depicted on color Doppler sonography in 21 melanomas, all of which were thicker than 2 mm. The median follow-up was 40 months. Eleven patients developed metastases, and all except one had a vascularized melanoma. In the univariate analyses, neovascularization visualized with sonography and the Breslow index were both significantly linked to the occurrence of metastases (p0.0001). In the multivariate Cox proportional hazards regression analysis, neovascularization was the only significant parameter.This study confirms that prognostic value of angiogenesis evaluated with color Doppler sonography could be used to identify melanomas with a high metastatic potential.

To describe the appearance of hepatic tumors treated with radio-frequency (RF) ablation on computed tomographic (CT) and magnetic resonance (MR) images and the pattern of residual tumor at the site of RF ablation and to assess prospectively the sensitivity, specificity, and positive and negative predictive CT and MR imaging values in the evaluation of RF treatment.Thirty-one patients with 50 tumors (nine hepatocellular carcinomas and 41 metastases) treated with RF ablation underwent CT and MR imaging on the same day at 2, 4, and 6 months; CT was performed every 3 months thereafter. CT and MR findings were interpreted separately and prospectively by two reviewers with consensus. For both imaging techniques, appearance of the treated area, treatment efficacy, and complications were assessed at each time. Sensitivity and specificity were determined by using the McNemar test.After a mean follow-up of 19 months, nine tumors showed local regrowth. At 2 months, MR imaging depicted more local regrowths (eight of nine; sensitivity, 89%) than did CT (four of nine; sensitivity, 44%) but without significant differences (P =.12). In two cases, only T2-weighted imaging depicted local regrowth. All nine lesions became conspicuous at 4-month follow-up with both techniques. At 2 months, thin peripheral rim enhancement and arterioportal shunting were found in 24% and 12%, respectively, of the treated tumors. These findings disappeared thereafter and are not linked to tumor regrowth.Despite the small number of patients, CT and MR imaging may depicted all local regrowth at 4 months or sooner. MR imaging may have an edge over CT in the early detection of local regrowth.

Background: The widespread use of high-throughput molecular techniques has allowed the identification of recurrent and actionable molecular traits across various tumor types. Translating these approaches to bedside may guide the decision-making for cancer patients candidates to early clinical trials. Methods: Patients with advanced solid tumors, refered to our early drug development department (DITEP), were prospectively enrolled in a prospective molecular screening program at Gustave Roussy (France). CT-Scan or ultrasound-guided biopsies were performed in metastatic or primary tumor sites to carry out a comprehensive molecular characterization. DNA was extracted from fresh tumor samples and analyzed by comparative genomic hybridization (CGH) (≥ 30% tumor cells required) and by Next Generation Sequencing (NGS) for up to 74 target genes (≥ 10% tumor cells required). A weekly molecular tumor board reviewed all the profiles to identify actionable traits for which the most relevant targeted therapy may be available through early clinical trials or marketed drugs. Treatment efficacy was evaluated by RECIST 1.1 criteria. Results: From November 2011 to December 2013, 420 heavily pretreated patients were included in the MOSCATO 01 trial. Out of them, a tumor biopsy could be performed in 368 patients (87%). CGH and NGS profiles were assessed in 284 (77%) and 315 (85%) of biopsied patients, respectively, with 283 patients (76% of biopsied patients) being profiled for both CGH and NGS. The median time for delivering results of 20 days. Actionable molecular aberrations were found in 161 patients (44%). Among them, 81 patients (50%) were matched to a targeted therapy and enrolled in ongoing phase I clinical trials. The most relevant genomic aberrations of interest were FGFR or FGF ligand amplification (n=24), PTEN/PI3K/AKT pathway activation (n=22), HER2 amplification (n=11), KRAS/NRAS/HRAS mutation (n=5), BRAF mutation (n=4), cyclin dependent kinase amplification or deletion (n=8), EGFR amplification or mutation (n=7), MET amplification (n=3), androgen receptor amplification (n=2), ALK translocation (n=2), and KIT amplification, MDM2 amplification, IGF1R amplification, and ROS1 rearrangement (all n=1). Out of the 81 patients profiled and treated according to the genomic profiles, we observed at the first tumor evaluation: 1 (1%) complete response (CR), 9 (11%) partial responses (PR), 52 (64%) stable disease (SD) and 14 (17%) progressive disease (PD). Whole exome analyses on selected patients are ongoing (currently n=24) and will be presented at the meeting. Conclusions: High throughput genomic analysis is feasible in daily practice and allows biological-orientation of patients in early clinical trials. The enrichment of phase I trials with patients harboring specific molecular traits leads to promising antitumor activity and is likely to exert a major influence on early drug development. Citation Format: Charles Ferté, Christophe Massard, Ecaterina Ileana, Antoine Hollebecque, Ludovic Lacroix, Samy Ammari, Maud Ngo-Camus, Rastislav Bahleda, Anas Gazzah, Andrea Varga, Sophie Postel-Vinay, Yohann Loriot, Nathalie Auger, Valerie Koubi-Pick, Bastien Job, Thierry De Baere, Frederic Deschamps, Philippe Vielh, Vladimir Lazar, Marie-Cécile Le Deley, Catherine Richon, vincent ribrag, eric deutsch, eric angevin, gilles vassal, Alexander Eggermont, Fabrice André, Jean-Charles Soria. Molecular screening for cancer treatment optimization (MOSCATO 01): a prospective molecular triage trial; Interim analysis of 420 patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr CT240. doi:10.1158/1538-7445.AM2014-CT240

11057 Background: Although several studies have reported genomic characterisation of primary urothelial carcinoma (UC), little is known about the genomic alterations of the metastatic cisplatin-res...

10563 Background: Functionalized hafnium oxide nanoparticles (NBTXR3) have been developed as selective radioenhancers, which may represent a breakthrough approach for the local treatment of solid t...

10050 Background: This feasibility study intends to characterize genomic alterations in recurrent tumors of an individual pediatric patient in order to select a targeted therapy approach. Methods: ...

A PCR assay for the detection of Actinobacillus pleuropneumoniae was developed based on the amplification of a dsbE-like gene. All of 157 field isolates of A. pleuropneumoniae reacted in the PCR by the amplification of a 342 bp product. No reaction was observed with related bacterial species or other bacterial species isolated from pigs, except for A. lignieresii. The lower detection limit of the PCR was 102 CFU per PCR test tube and was not affected by the addition of 106 CFU Escherichia coli. The PCR was evaluated on mixed bacterial cultures from nasal and tonsillar swabs as well as suspensions of nasal conchae and tonsils obtained from specific pathogen-free (SPF) pigs, experimentally infected pigs, and pigs from farrow-to-finish herds. The results of the new PCR were compared with a PCR based on the detection of the omlA gene coding for an outer membrane protein, with a commercially available PCR (Adiavet™APP, Adiagene, Saint-Brieuc, France), and with conventional culturing. No positive reactions were observed with any of the PCR methods in samples of SPF animals. In samples of the other animals, no or low significant differences between nasal swabs and suspensions as well as tonsillar swabs and suspensions were observed in any method. In general, more positive results were obtained from tonsillar samples in comparison to nasal samples. Interassay sensitivity and specificity values were assessed for each test by pair wise comparisons between assays. The agreement between tests was evaluated by calculating Cohen’s κ coefficient. From these analyses the three PCR assays showed a good agreement. The dsbE-based PCR proved to be highly sensitive (95 and 93%) and specific (82 and 74%) in comparison to the omlA-based PCR and the commercially available PCR, respectively. It was concluded that the dsbE-like gene-based PCR is a reliable diagnostic assay for demonstration of A. pleuropneumoniae. Furthermore, it was demonstrated that tonsillar swabs can be used for the detection of the pathogen in healthy carrier animals.

Leclercia adecarboxylata was isolated from a patient with a chronically inflamed gallbladder, together with Enterococcus sp. The organism was considered clinically significant and was susceptible to all antibiotics tested. Another strain of L. adecarboxylata was cultured from blood, together with Escherichia hermannii and E. faecalis , from a patient with sepsis.

Bacterial strains isolated from the genital tracts of humans (predominantly males), semen of boars, and uterine and vaginal secretions of sows were identified as Corynebacterium glucuronolyticum and were compared with the type strains of the recently proposed species Corynebacterium glucuronolyticum and Corynebacterium seminale . The two type strains as well as the clinical strains were shown by DNA-DNA hybridization and sequencing of the 16S rRNA gene to be related at the species level. All strains were classified as C. glucuronolyticum , because this name has nomenclatural priority over C. seminale .

The objective of our study was to evaluate the efficacy and safety of radiofrequency ablation of hepatic metastases performed either percutaneously for treatment of hepatic metastases in patients deemed ineligible for surgery or intraoperatively during partial hepatectomy to destroy unresectable metastases.Sixty-eight patients with 121 hepatic metastases (5 metastases per patient) that were mainly colorectal in origin underwent 76 sessions of radiofrequency ablation with cooled-needle electrodes under sonographic guidance. Twenty-one patients with 33 metastases of 5-20 mm in diameter (mean +/- SD,13 +/- 7 mm) underwent intraoperative radiofrequency ablation. Forty-seven patients with 88 metastases of 10 to 42 mm in diameter (mean +/- SD, 26 +/- 9 mm) were treated with percutaneous radiofrequency ablation. Procedure efficacy was evaluated with dynamic enhanced CT and MR imaging performed 2, 4, and 6 months after treatment and then every 3 months.Radiofrequency ablation allowed eradication of 91% of the 100 treated metastases that were followed up for 4-23 months (mean, 13.7 months). Tumor control was equivalent for percutaneous radiofrequency ablation (90%) and for intraoperative radiofrequency ablation (94%). Failure to achieve tumor control occurred mostly with the largest tumor nodules. One bilioperitoneum and two abscesses were the major complications encountered after treatment of 121 metastases with a follow-up of more than 2 months.Radiofrequency ablation appears to be a promising therapeutic modality capable of extending the possibilities of partial hepatectomy and of efficiently treating small metastases percutaneously.

Moraxella canis was isolated in large numbers from an ulcerated supraclavicular lymph node of a terminal patient, who died a few days later. Although the patient presented with septic symptoms and with a heavy growth of gram-negative diplococci in the lymph node, blood cultures remained negative. M. canis is an upper-airway commensal from dogs and cats and is considered nonpathogenic for humans, although this is the third reported human isolate of this species.

Purpose A strong embolic effect of iodized oil/drug mixtures injected in the hepatic artery appeared to be an efficient way of prolonging the contact time between drugs and tumor tissue. Therefore, the authors evaluated arterial and portal embolic effects after hepatic intra-arterial injection of iodized oils and various emulsions of iodized oil. Materials and Methods Twenty-five pigs were monitored for the Doppler resistance index (DRI) in the hepatic artery and wedge hepatic vein pressure (WHVP) during 1 hour after injection of pure iodized oil, ultra-fluid or fluid, and four different emulsions of iodized oil ultra-fluid, into the hepatic artery. Results Mean area under the curve (AUC) values of DRI increases varied from 20.3 to 24.2 after injection of pure iodized oils or water-in-oil emulsions, and were 13.2 for large-droplet oil-in-water emulsion and 8.2 for small-droplet oil-in-water emulsion. Mean AUC values of WHVP increases varied from 151.6 to 195.6 after injection of pure iodized oils or water-in-oil emulsions, and were 105.5 for large-droplet oil-in-water emulsion and 8.5 for small-droplet oil-in-water emulsion. There was a significant difference in DRI and WHVP modifications between small-droplet oil-in-water emulsions and all other products ( P = .001), between the two oil-in-water emulsions and the two water-in-oil emulsions ( P = .004), and between the two oil-in-water emulsions and pure iodized oils ( P = .002). Conclusion After hepatic intra-arterial injection, water-in-oil emulsions and pure iodized oils provided a stronger embolic effect than oil-in-water emulsion, both in the hepatic artery and in the portal vein.

3599 Background: Optiliv allowed complete macroscopic resection (R0-R1) of previously unresectable LM-CRC in 28% of the pts, despite failure of 1-3 prior chemotherapy protocols (Lévi et al. Proc ASCO GI 2013). Purpose: To assess tolerability and efficacy of Optiliv according to previous chemotherapy exposure. Methods: Pts received iv Cet (500 mg/m²) and chronomodulated or conventional HAI of Irinotecan (180 mg/m²), 5-Fluorouracil (2800 mg/m²), and Oxaliplatin (85 mg/m²) q2 wks. Liver surgery was performed according to q6wks multidisciplinary reviews. Pts were categorized according to Optiliv as 2nd (N=29 pts) vs 3rd-4th chemotherapy line (N=35 pts). Results: Pt characteristics were similar in both groups. Overall, there were 22F and 42 M, aged 33-76 years, with good PS (0/1/2: 40/22/2) and predominantly liver lesions. LM-CRC were bilateral in 51 pts (79.7%), with a median of 10 metastases (1-50), 6 segments involved (1-8), and largest diameter of 52 mm (15-172). 61 pts (2nd line, 27; 3rd-4th line, 34) received a median of 6 courses (1-15). Grade 3-4 toxicities per pt were similar in both groups except for abdominal pain (2nd line, 15% vs 3rd-4th line, 35%, p=0.07), diarrhea (7% vs 24%, p=0.09), thrombocytopenia and febrile neutropenia (0 vs 9%, p=0.25). Four CR were achieved in 2nd line (15%) vs none in 3rd-4th line. Respective objective response rates were 63% and 38% (p=0.05). R0-R1 resections were performed in 11/27 pts (41%) on 2nd line vs 6/34 pts (18%) on 3rd-4th line (p=0.06), resulting in respective median progression-free survival (PFS) of 14.2 months [7.8 - 20.7] vs 7.3 [5.5 - 9.1] (p=0.002). Median overall survival (OS) was not reached at 3 years in the pts on Optiliv as 2nd line vs 15.2 months in the more heavily pretreated pts (pnd lineas compared to 3rd-4th line. Optiliv now deserves upfront testing. Clinical trial information: NCT00852228.

2512 Background: Characterization of the genomic alterations (GA) that could drive tumor growth of an individual patient (pt) is now critical to better select targeted therapies in phase I trials. Methods: Pts with advanced solid tumors, who failed at least one line of standard therapy, were offered an on-purpose tumor biopsy for molecular characterization. Biopsies were mainly obtained using 18G needles under CT or ultra-sound control, from metastatic or primary tumor sites. DNA extracted from fresh tumor biopsies was analyzed by CGH (Agilent platform) (if ≥ 50% tumor cells in the sample) and by sequencing for 30 target genes (if ≥ 30% tumor cells in the sample). An expert panel of scientists and clinicians reviewed results to determine the biological signification of the GA and match such pts to the most relevant targeted therapy available (mainly in early clinical trials). PFS using therapy based on GA was compared to the PFS for the most recent therapy on which the pt had just experienced progression (PFS ratio). Results: From December 2011 to august 2012, 129 heavily pretreated pts (median of 3 previous lines) were consented. Among them, 111 (86%) had a dedicated tumor biopsy. An actionable target was identified in 52 patients (40%). Among them, 25 pts (23% of biopsied pts) have been treated with a targeted therapy. The median time between biopsy and molecular results was 21 days [17 – 28]. GA of interest encompassed FGF ligand or receptor amplification (n=9), cyclin amplification or deletion (n=4), KRAS/BRAF/NRAS mutation (n=3), PI3K amplification or PTEN deletion (n=3), EGFR amplification or mutation (n=3) (outside lung cancer), HER2 amplification (n=2) (outside breast cancer), EML4/ALK translocation (n=1). Among the 25 pts treated according to their GA, we observed 5 PR (20%), 14 SD (56%) and 3 PD (12%). Three pts (12%) were not evaluable because of early discontinuation of the therapy. PFS ratio was >1.3 among 9 out of 19 evaluable pt (47%). Conclusions: High throughput molecular analysis is feasible in daily practice. It allows enrichment of phase I trials with specific GA, and leads to promising anti-tumor activity (20% PR as compared to the classical 7-10% PR obtained in all comers phase I trials). Clinical trial information: NCT01566019.

246 Background: Chemoembolization is used to treat liver malignancies. However recurrence occurs frequently, possibly because of neoangiogenesis triggered by ischemia caused by the embolic agent. In this context, the combination of an embolic agent with an anti-angiogenic drug seems appealing. This study characterizes the in vitro loading and release profile of sunitinib eluting beads of different sizes and their pharmacokinetic profile in a rabbit model. Methods: 70-150 μm and 100-300 μm drug eluting beads (DC Bead, Biocompatibles UK) were loaded by incubation in a sunitinib hydrochloride solution. Drug was quantified by spectrophotometry at 430 nm. Drug release was measured over one-week periods and normalized using an internal standard in 30% ethanol in NaCl 0.9%. New-Zealand white rabbits were used. Eight animals received 0.2 ml of 100-300 μm DC Bead loaded with 6 mg of sunitinib in the hepatic artery (group 1) and 4 animals received 6 mg of sunitinib p.o. (group 2). Half of the animals were sacrificed after 6 hours and half after24 hours. Liver enzymes were measured at 0, 6 and 24 hours in both groups. Plasmatic sunitinib concentration was determined by tandem mass spectroscopy (LC MS/MS) at 0, 1, 2, 3, 4, 5, 6 and 24 hours. At sacrifice, the livers were harvested and sunitinib concentration in liver tissue was assessed by LC MS/MS. Results: High drug loading was obtained for both microsphere bead sizes. Particle shrinking was observed with adsorption of sunitinib. Almost complete release of sunitinib was detected under physiological conditions, with very similar release for 70-150 μm and 100-300 μm (t50%=1.2 h) DC Bead. Conclusions: Sunitinib eluting beads are well tolerated by rabbits when administered in the hepatic artery. No unexpected toxicity was observed. Very high drug concentration can be obtained at the site of embolization with minimal systemic passage.

466 Background: Multiple, large and bilateral LM-CRC can be downsized with chemotherapy (chemo) and resected with curative intent in ~15% pts (Adam et al. 2004). The complete resection rate (R0+R1) may be increased with Cet addition, triplet chemotherapy, chronomodulated delivery (Chrono) and hepatic artery infusion (HAI) (Bouchahda et al. 2009; Kemeny et al. 2009; Garufi et al. 2010; Goere et al. 2010). Purpose: To increase the R0+R1 rate from 15% to 30% in pts with unresectable KRAS wt LM-CRC despite failure of 1-3 prior chemo protocols. Methods: 64 pts received iv Cet (500 mg/m²) and Chrono (18 pts) or conventional (Conv, 46 pts) HAI of Irinotecan (180 mg/m²), 5-Fluorouracil (2800 mg/m²), and Oxaliplatin (85 mg/m²) q2 weeks. Liver surgery was performed according to q6wks multidisciplinary meetings. Results: There were 22F and 42 M, aged 33-76 years, with good PS (0/1/2: 40/22/2). Liver lesions were bilateral in 50 pts (78.1%), with a median of 10 metastases (1-50) and largest diameter of 52 mm (15-172). Five courses (1-13) were given to 61 pts (3 never treated), as 3-4th line for 35 pts (54.6%). Grade 3-4 toxicities per pt were neutropenia (40%), abdominal pain (26%), fatigue (18%) and diarrhea (16%). Tumor response (RECIST) was achieved in 28 pts (2 CR, 26 PR), with an objective response rate of 46% [95%CL, 33-58]. Disease control rate was 95% [90-100]. The main endpoint was met, with R0-R1 resections in 19/61 pts, for an overall complete macroscopic resection rate of 28% [17-39]. R0+R1 resections were performed in 33.3% Chrono pts and 25.6% Conv pts, with worse initial prognosis for the Chrono pts (>6 segments involved, 50% vs 9%; > 9 metastases, 75% vs 27%). Median progression-free and overall survival were 8.7 months [6.9-10.5], and 25.7 months [14.2-37.2] respectively. Conclusions: The combination of intravenous Cet and triplet hepatic artery infusion safely doubled the expected rate of complete resections of LM-CRC, despite prior failure of systemic chemo. This most effective treatment option now deserves upfront testing. Clinical trial information: NCT00852228.

247 Background: We demonstrated that DC Bead (Biocompatibles UK, Ltd) could be loaded with sunitinib and injected intra-arterially in the rabbit without unexpected toxicity. The purpose of this study is to evaluate the antitumoral effect of sunitinib eluting beads in the VX2 tumor model of liver cancer. Methods: VX2 tumors were implanted in the left liver lobe of New-Zealand white rabbits. Animals were assigned to 3 groups: Group 1 (n=6) received 1.5mg of sunitinib loaded in 0.05ml of 100-300um DC Bead, group 2 (n=5) received 0.05ml of 100-300um DC Bead, group 3 (n=5) received 0.05ml NaCl 0.9% in the left hepatic artery. One animal in each group was sacrificed at 24 hours and the others were followed for survival until day 15. Liver enzymes were measured daily. In group 1, plasmatic sunitinib concentration were measured daily by LC MS/MS tandem mass spectroscopy. At day 15 all living animals were sacrificed. After sacrifice, the livers were harvested for determination of the VEGF receptor tyrosine kinase activity by western blot and histopathological examination. Results: In group 1, no animals died during follow-up. In group 2, 2 animals died during follow-up on day x. In control group 3, 3 animals died during follow up on day x. In group 1 plasmatic sunitinib levels remained under therapeutic concentration throughout the experiment. Very high concentrations of sunitinib were measured in the liver tissue 24 and 15 days after embolization. Inhibition of the phosphorylation of the RTK was demonstrated at 24h and 15 days in groups 1. Sunitinib eluting beads seemed to penetrate in the tumor more effectively and there was more necrosis around the beads than their bland counterparts. Conclusions: Administration of sunitinib eluting beads in VX2 carrying rabbits resulted invery high drug concentrations at the site of embolization with minimal systemic passage. Despite the very high tissular sunitinib concentration we did not observe any additional toxicity with loaded beads. Sunitinib eluting beads inhibit the activation of RTK’s triggered by ischemia and seem to prolong survival of the treated animals. Therefore we consider that local treatment with sunitinib may provide a promising approach for the treatment of liver cancer.

3547 Background: HAI of chronomodulated (Chrono) irinotecan (I), 5-Fluorouracil (F) and oxaliplatin (O), or flat O combined with iv F-leucovorin allowed secondary metastases resections and prolonged survival in patients (pts) with CRC liver metastases despite prior chemotherapy failure (Bouchahda, Cancer 2009; Goere, Ann Surg 2010). Purpose: To prospectively evaluate safety and efficacy of combining iv Cet with HAI of IFO in pts with CRC liver metastases. Methods: This Phase II trial involved previously treated pts with unresectable CRC liver metastases. Pts received iv Cet (500 mg/m²) and Chrono or conventional HAI of I (180 mg/m²), F (2800 mg/m²), and O (85 mg/m²) q2 weeks. Liver metastases were resected if adequately downstaged. Results: Planned accrual of 60 pts was reached on 01/24/2012. 3 pts were not treated, 9 are ongoing. 48 consecutive treated pts (18F, 30 M; aged 32-76 years) are fully assessed and monitored. They had PS 0-1 (98%), bilobar liver lesions (69%), a median of 8 metastases (1-50; largest diameter, 57 mm – 15-172). Prior chemotherapy involved one (43%), or two or three (57%) lines. A median of 5 protocol courses (1-13) was given. Main grade 3-4 toxicities per pt were neutropenia (40%), abdominal pain (15%), fatigue (15%), nausea (15%), diarrhea (13%) and sensory neuropathy (4%). Objective response rate was 44% [30-58], with 6% radiological complete responses. Disease control rate was 85%. Secondary liver surgery was performed in 15 pts (31.3%).One pt with 25 metastases (1-6 cm) in all liver segments had pathologic complete response in 24 lesions removed through three-stage hepatectomy. She currently has an off treatment disease-free survival of 17+ months.With a follow up of 4 to 47 months, median progression-free survival is 14.2 months [9.7-18.8], 1- and 2-year survival rates are 92.8% and 59.2% respectively. Conclusions: OPTILIV offers a safe and unusually effective treatment option for patients with CRC liver metastases after failure of systemic chemotherapy within a coordinated medico-surgical strategy.

Acute lymphoblastic leukemia was diagnosed in a 7-year-old girl. Two months after insertion of a central venous catheter, she developed fever and complained of headache and abdominal pain. Physical examination revealed no focus of infection. A gram-negative nonfermenting bacillus was recurrently cultured from blood. Extensive biochemical testing and 16S ribosomal DNA sequencing led to the identification of Ralstonia gilardii .

Enterococcus cecorum was isolated as the etiologic agent of a continuous ambulatory peritoneal dialysis peritonitis episode in an alcoholic patient. To date, this is only the third infection due to this bacterium, found in the intestinal tract of many domestic animals, that has been reported in humans.

Clostridium tertium was isolated from two immunocompromised patients with septicemia, fever, and gastrointestinal symptoms. The strains were resistant to ceftazidime, cefepime, and clindamycin; intermediately resistant to penicillin; and susceptible to metronidazole, quinolones, and vancomycin.

Background Currently, most laboratories identify yeasts routinely on the basis of morphology and biochemical reactivity. This approach has quite often limited discriminatory power and may require long incubation periods. Due to the increase of fungal infections and due to specific antifungal resistence patterns for different species, accurate and rapid identification has become more important. Several molecular techniques have been described for fast and reliable identification of yeast isolates, but interlaboratory exchangeability of identification schemes of molecular techniques has hardly been studied. Here, we compared amplified ITS2 fragment length determination by an ABI Prism 310 (Applied Biosystems, Foster City, Ca.) capillary electrophoresis system with that obtained by a CEQ8000 (Beckman Coulter, Fullerton, Ca.) capillary electrophoresis system.