OASIS evaluation of the French surveillance network for antimicrobial resistance in diseased animals (RESAPATH): success factors underpinning a well-performing voluntary system.

Antimicrobial resistance is a One Health issue requiring the development of surveillance systems in the human, environmental and animal sectors. In the European Economic Area, the surveillance of antimicrobial resistance in zoonotic pathogens and indicator bacteria in healthy food-producing animals is required legally, while countries are also expected to extend their surveillance to diseased animals in the frame of national action plans. In this context, evaluating existing antimicrobial resistance surveillance systems in animal health is important to improve systems in place, but also to help other countries learn from these experiences, understand success factors and anticipate challenges. With this aim, the French surveillance network for antimicrobial resistance in bacteria from diseased animals (RESAPATH) was evaluated using the Outil d'Analyse des Systèmes d'Information en Santé (OASIS) assessment tool. Key performance factors included (i) a strong and inclusive central institutional organisation defining clear and well-accepted surveillance objectives, scope and procedures, (ii) strong skills in epidemiology and microbiology and (iii) a win-win approach enabling the voluntary participation of 71 field laboratories and where free annual proficiency testing plays a pivotal role. The main area for improvement of RESAPATH was its time-consuming data management system.

Applied precision cancer medicine in metastatic biliary tract cancer.

INTRODUCTION: Advanced therapy-refractory biliary tract cancer (BTC) has poor prognosis and constitutes a major challenge for adequate treatment strategies. By mapping the molecular profiles of advanced BTC patients, precision cancer medicine may provide targeted therapies for these patients. OBJECTIVE: In this analysis, we aimed to show the potential of PCM in metastatic BTC. METHODS: In this single-center, real-world retrospective analysis of our PCM platform, we describe the molecular profiling of 30 patients diagnosed with different types of metastatic BTC. Tumor samples of the patients were examined using a 161-gene next-generation sequencing panel, immunohistochemistry (IHC), and fluorescence in situ hybridization for chromosomal translocations. RESULTS: In total, we identified 35 molecular aberrations in 30 patients. The predominant mutations were KRAS (n = 8), TP53 (n = 7), IDH2 (n = 4), and IDH1 (n = 3) that accounted for the majority of all molecular alterations (62.86%). BRAF mutations were observed in two patients. Less frequent alterations were noted in ARID1A, CTNNB1, ESR1, FBXW7, FGFR2, MET, NOTCH2, PIK3CA, PTCH1, SMAD4, and SRC1, each in one case. FGFR fusion gene was detected in one patient. No mutations were detected in eight patients. IHC revealed EGFR and p-mTOR expression in 28 patients. Applying these results to our patients, targeted therapy was recommended for 60% of the patients (n = 18). One patient achieved stable disease. CONCLUSIONS: PCM is a feasible treatment approach and may provide molecular-guided therapy recommendations for metastatic BTC.

Abdominal fat distribution in diffuse idiopathic skeletal hyperostosis and ankylosing spondylitis patients compared to controls.

AIM: To evaluate abdominal fat distribution (subcutaneous adipose tissue [SAT] and visceral adipose tissue [VAT]) in two enthesopathy-related diseases with known correlation to metabolic syndrome (MS): diffuse idiopathic skeletal hyperostosis (DISH) and ankylosing spondylitis (AS) compared with controls. MATERIALS AND METHODS: Abdominal computed tomography (CT) examinations of 43 DISH (Resnick radiographic criteria) patients, 31 AS (Modified New York Criteria) patients and 42 age- and gender-matched (to DISH) controls (males: 29; 29; 27 and mean age: 71.7±7; 56.1±16; 72.7±8 years, respectively) were evaluated and compared for VAT and SAT surface areas on mid L3, L4, L5 levels. RESULTS: AS patients were significantly younger compared to DISH patients and controls. No significant differences were observed between VAT and SAT of DISH and AS patients or between SAT values in all groups even after correction for age. VAT was higher in DISH and AS patients compared to controls on all three levels, but reached significance (p<0.05) only for DISH patients (L3: 24.34/23.6/18.43; L4: 23.85/22.21/18.05; L5: 19.09/18.94/14.24 mm2, respectively). This did not change after correction for age. The VAT/SAT ratio was significantly larger in DISH and AS patients on all levels compared to controls. CONCLUSION: The higher VAT surface area, a known marker for MS, which by itself is associated with bone proliferation, in DISH and AS patients compared to controls substantiates its role as a potential surrogate marker for MS as well as suggests a potential shared pathogenic pathway for enthesopathic excessive bone production in DISH and AS.

Determination of Hydrogen Density by Swift Heavy Ions.

A novel method to determine the total hydrogen density and, accordingly, a precise plasma temperature in a lowly ionized hydrogen plasma is described. The key to the method is to analyze the energy loss of swift heavy ions interacting with the respective bound and free electrons of the plasma. A slowly developing and lowly ionized hydrogen theta-pinch plasma is prepared. A Boltzmann plot of the hydrogen Balmer series and the Stark broadening of the H_{ß} line preliminarily defines the plasma with a free electron density of (1.9±0.1)×10^{16} cm^{-3} and a free electron temperature of 0.8-1.3 eV. The temperature uncertainty results in a wide hydrogen density, ranging from 2.3×10^{16} to 7.8×10^{18} cm^{-3}. A 108 MHz pulsed beam of ^{48}Ca^{10+} with a velocity of 3.652 MeV/u is used as a probe to measure the total energy loss of the beam ions. Subtracting the calculated energy loss due to free electrons, the energy loss due to bound electrons is obtained, which linearly depends on the bound electron density. The total hydrogen density is thus determined as (1.9±0.7)×10^{17} cm^{-3}, and the free electron temperature can be precisely derived as 1.01±0.04 eV. This method should prove useful in many studies, e.g., inertial confinement fusion or warm dense matter.

Role of computer-assisted surgery in osteotomies around the knee.

PURPOSE: The goal of this article is to present our experience on navigation for osteotomies around the knee and especially osteotomies for coronal deformities. The first computer-assisted osteotomy was performed in March 2001 and since that time more than 1000 osteotomies have been performed in our department. METHODS: All the osteotomies were performed with the Orthopilot® device (B-Braun-Aesculap, Tuttlingen, Germany). The main indication was for genu varum deformities but several cases were operated for genu valgum. The surgical procedure as well as the indications and the rationale for each osteotomy (high tibial osteotomy-HTO, double-level osteotomy-DLO, femoral osteotomy-FO) are addressed in the article. RESULTS: The results are focused on several papers published by the authors since more than 10 years. Regarding HTO for genu varum, the preoperative goal (HKA angle: 184° ± 2°) was reached in 96 % of cases and the difference was statistically significant compared to the non-navigated series (71 %: p < 0.05). Regarding DLO for genu varum, the preoperative goal was reached in 92.7 % for the HKA angle and in 88.1 % for the medial proximal tibial mechanical angle (MPTMA). Regarding genu valgum deformity, the preoperative goal was achieved in 86.2 % of cases for the HKA angle and 100 % of cases for the MPTMA. CONCLUSION: According to these results, one can say that, regardless the type of osteotomy, the procedure is reliable, reproducible and accurate. Since 15 years, all the osteotomies around the knee are navigated in our department. Provided that one uses a reproducible radiograph protocol, navigation allows to perform double-level osteotomies, both for genu varum and genu valgum, with optimal accuracy in order to avoid oblique joint line, which will be difficult to revise to TKA. LEVEL OF EVIDENCE: IV.

Long-term outcomes of tension band wiring with a single K-wire in Rockwood type IV/V acute acromio-clavicular dislocations: 25 cases.

BACKGROUND: Our objective was to evaluate the long-term functional and radiological outcomes of tension band wiring with a single K-wire for acute Rockwood types IV and V acromio-clavicular dislocation (ACD). METHODS: Single-centre cross-sectional non-randomised observational cohort study of 25 shoulders treated surgically between January 2002 and December 2004, in 25 patients, 23 males and 2 females, with a mean age of 35±11years (24-46). The evaluation criteria were the absolute and weighted Constant scores, QuickDASH score, subjective shoulder value (SSV), visual analogue scale (VAS) pain score at rest and during activities, and radiographic features in clinically symptomatic patients. RESULTS: Mean values were as follows: follow-up, 150±17months (133-167); absolute Constant score, 88±17 (71-105); weighted Constant score, 92.5±12.5 (80-105); QuickDASH, 15.5±7 (8.5-22.5); SSV, 88±17% (71-105); VAS pain score at rest, 0.2±0.7 (0-0.9); and VAS pain score while active, 1.4±2.3 (0-3.7). The weighted Constant score was less than 70% in only 8% of patients. Of the 17 patients for whom radiographs were obtained, 8 had acromio-clavicular osteoarthritis. Mean coraco-clavicular distance was 12.3±4.3mm (8-16.6) and mean acromio-clavicular distance was 5±5mm (0-10). The recurrence rate was 8%. CONCLUSION: Tension band wiring with a single K-wire for acute acromio-clavicular dislocation reliably provides good long-term functional outcomes. Recurrences are uncommon and few patients experience symptoms (8%). LEVEL OF EVIDENCE: IV, retrospective study.

Prognostic impact of breast cancer subtypes in elderly patients.

We aimed to analyse the impact of breast cancer (BC) subtypes on the clinical course of disease with special emphasis on the occurrence of brain metastases (BM) and outcome in an elderly BC population. A total number of 706 patients ≥65 years receiving treatment for BC from 2007 to 2011 were identified from a BC database. 62 patients diagnosed with DCIS and 73 patients with incomplete datasets were excluded, leaving 571 patients for this analysis. Patient characteristics, biological tumour subtypes, and clinical outcome including overall survival (OS) were obtained by retrospective chart review. 380/571 (66, 5 %) patients aged 65-74 years were grouped among the young-old, 182/571 (31.9 %) patients aged 75-84 years among the old-old, and 29/571 (5.1 %) patients aged ≥85 years among the oldest-old. 392/571 (68.8 %) patients presented with luminal BC, 119/571 (20.8 %) with HER2-positive, and 59/571 (10.3 %) with triple-negative BC (TNBC). At 38 months median follow-up, 115/571 (20.1 %) patients presented with distant recurrence. A higher recurrence rate was observed in the HER2-positive subtype (43/119 (36.1 %)), as compared to TNBC (15/59 (25.4 %)) and luminal BC (57/392 (14.5 %); p < 0.001). BM were detected at a significantly higher rate in HER2-positive BC patients (9/119 (7.6 %)), as compared to TNBC (2/59 (3.4 %)) and luminal BC patients (6/392 (1.5 %); p = 0.003). Diagnosis of metastatic disease (HR 7.7; 95 % CI 5.2-11.4; p < 0.001) as well as development of BM (HR 3.5; 95 % CI 1.9-6.4; p < 0.001) had a significantly negative impact on OS in a time-dependent covariate cox regression model. In contrast to younger BC patients, outcome in this large cohort of elderly patients suggests that HER2-positive disease-not TNBC-featured the most aggressive clinical course with the highest rates of metastatic spread and BM. In-depth analysis regarding a potentially distinct biology of TNBC in elderly is therefore warranted.

Clinical and radiological outcomes with the Durom™ acetabular cup for large-diameter total hip arthroplasty: 177 implants after a mean of 80 months.

BACKGROUND: Large-diameter metal-on-metal hip prostheses are no longer used, but their outcomes after more than 5 years are unknown. We conducted a retrospective study with a 6.8-year mean follow-up to assess clinical outcomes after Durom™ cup implantation, including the dislocation rate, comparatively to the reference metal-on-polyethylene bearing. We determined the rate of failure ascribable to Durom™ cup use. We also looked for a sharp drop in the implant survival curve during the follow-up period and for factors associated with adverse reactions to metal debris (ARMDs). HYPOTHESIS: We hypothesised that clinical outcomes after Durom™ cup implantation were similar to those seen with a metal-on-polyethylene bearing, except for a lower rate of dislocation. PATIENTS AND METHODS: We included 177 consecutive THA procedures that were performed between 2005 and 2008 in 165 patients with a mean age of 57.6 ± 9.4 years (range, 31-76 years) and involved the implantation of a Durom™ cup, a femoral head greater than 36mm in diameter, and a PF(®) femoral stem (Zimmer, Etupes, France). The mini-posterior approach was used, with 2mm of acetabular overreaming in 82% of cases, a short femoral neck in 75% of cases, and a mean cup inclination of 34 ± 5° (range, 21-50°). RESULTS: Outcomes were assessed for 156 THA procedures in 146 patients after a mean follow-up of 6 years 8 months. The mean Postel-Merle d'Aubigné score improved from 9.7 ± 2.7 (range, 4-14) to 17.4 ± 1.7 (range, 15-18) and the mean Harris hip score from 45.2 ± 15.3 (range, 9-83) to 96.3 ± 7 (75-100). No episodes of dislocation were recorded. We identified 7 failures ascribable to the Durom™ cup including 6 due to ARMD and 1 to aseptic loosening. Implant survival after a mean of 80months was 95.5% (95% CI, 93.1-99.2), with no sharp drop in the survival curve. CONCLUSION: The Durom™ cup eliminates the risk of hip dislocation and produces similar functional outcomes to those seen with metal-on-polyethylene bearings after a mean follow-up of 80 months. Nevertheless, given the difficulty in predicting ARMD and hypersensitivity reactions, the Durom™ cup has been discarded and patients carrying it are monitored closely.

Are results of total knee arthroplasty for isolated patellofemoral OA as good as for medial compartment OA? A medium-term retrospective comparative study.

PURPOSE: The aim of this study was to compare the results of isolated patellofemoral arthritis (IPFA) treated using a total knee arthroplasty (TKA) compared to the results of medial tibiofemoral arthritis treated with a TKA. We hypothesised that there would be no difference between functional outcomes for the two groups. METHODS: Between 2003 and 2009, 32 TKAs were performed for IPFA (group I). Over this time period, a total of 813 primary TKAs were performed from which we identified a second group of patients who had undergone TKA for isolated medial tibiofemoral arthritis (group II: n = 32). These patients were matched based on age, sex, body mass index and average follow-up. The average age of patients in group I was 72.81 ± 6.6 years (59-83) and 71.97 ± 6.8 years in group II. The global International Knee Society (IKS) score was statistically significantly better in group I (114.72 ± 22 points) than in group II (84.9 ± 23.8). This difference was accounted for by better passive flexion, better walking distance and the absence of any coronal plane deformity. RESULTS: A total of 29 patients from group I were reviewed at an average follow-up of 64.58 ± 23.4 months, and 30 patients in group II were reviewed at an average follow-up of 66.13 ± 23.9 months (three were lost to follow-up in group I and two in group II). At final follow-up, there was statistically no difference between the two groups IKS score (175.34 ± 19.26 in group I vs. 170.13 ± 24.14 for group II) or Hospital for Special Surgery patella score (89.31 ± 9.98 points for group I vs. 89.16 ± 11.45 points for group II). We found no significant radiological difference between the two groups including patella height and orientation on axial views. CONCLUSIONS: The results of TKA for IPFA are as good as the results of TKA for isolated medial tibiofemoral arthritis with well-functioning prosthetic patellofemoral articulations. These results support our institutional preference for using TKA as treatment for IPFA in patients over 65-70 years old.

Port-a-Cath extravasation of vesicant cytotoxics: surgical options for a rare complication of cancer chemotherapy.

BACKGROUND: Although implantation of a central venous device such as a Port-a-Cath was initially considered safe, extravasation rates up to 4.7% have been reported. Therefore, the objective of this study was to propose a structured procedure for the management of extravasation of a cytotoxic treatment. METHODS: A total of eight patients were evaluated after port extravasation of epirubicin (n = 3), platinum compounds (n = 3), paclitaxel (n = 1), or trabectedin (n = 1) into the subcutaneous space. Immediate explantation of the port was performed in combination with a "Subcutaneous Wash-Out Procedure" (SWOP). When removal of the port was delayed, débridement and flap coverage were performed as necessary. Epirubicin concentrations present in the samples obtained during surgical intervention were subsequently analysed using high-performance liquid chromatography (HPLC). Patients were followed for at least six months and were examined for sequelae such as pain, induration, redness, and limited movement. RESULTS: All three patients whose extravasation event was detected during chemotherapy administration benefited from SWOP with acceptable side effects (e.g., erythema). The analysis of epirubicin concentrations demonstrated the active removal of relevant amounts of the compound by wound rinsing. In contrast, late detection of extravasation led to major débridement and flap coverage in four out of five patients. A high body mass index (BMI) value was associated with all of the patients that experienced port extravasation. CONCLUSION: Depending on when Port-a-Cath extravasations into subcutaneous tissue are detected, different treatments are appropriate. When extravasation is detected early, the SWOP was found to be beneficial.

Fulvestrant induces resistance by modulating GPER and CDK6 expression: implication of methyltransferases, deacetylases and the hSWI/SNF chromatin remodelling complex.

BACKGROUND: Breast cancer is the leading cause of cancer death in women living in the western hemisphere. Despite major advances in first-line endocrine therapy of advanced oestrogen receptor (ER)-positive breast cancer, the frequent recurrence of resistant cancer cells represents a serious obstacle to successful treatment. Understanding the mechanisms leading to acquired resistance, therefore, could pave the way to the development of second-line therapeutics. To this end, we generated an ER-positive breast cancer cell line (MCF-7) with resistance to the therapeutic anti-oestrogen fulvestrant (FUL) and studied the molecular changes involved in resistance. METHODS: Naive MCF-7 cells were treated with increasing FUL concentrations and the gene expression profile of the resulting FUL-resistant strain (FR.MCF-7) was compared with that of naive cells using GeneChip arrays. After validation by real-time PCR and/or western blotting, selected resistance-associated genes were functionally studied by siRNA-mediated silencing or pharmacological inhibition. Furthermore, general mechanisms causing aberrant gene expression were investigated. RESULTS: Fulvestrant resistance was associated with repression of GPER and the overexpression of CDK6, whereas ERBB2, ABCG2, ER and ER-related genes (GREB1, RERG) or genes expressed in resistant breast cancer (BCAR1, BCAR3) did not contribute to resistance. Aberrant GPER and CDK6 expression was most likely caused by modification of DNA methylation and histone acetylation, respectively. Therefore, part of the resistance mechanism was loss of RB1 control. The hSWI/SNF (human SWItch/Sucrose NonFermentable) chromatin remodelling complex, which is tightly linked to nucleosome acetylation and repositioning, was also affected, because as a stress response to FUL treatment-naive cells altered the expression of five subunits within a few hours (BRG1, BAF250A, BAF170, BAF155, BAF47). The aberrant constitutive expression of BAF250A, BAF170 and BAF155 and a deviant stress response of BRG1, BAF170 and BAF47 in FR.MCF-7 cells to FUL treatment accompanied acquired FUL resistance. The regular and aberrant expression profiles of BAF155 correlated directly with that of CDK6 in naive and in FR.MCF-7 cells corroborating the finding that CDK6 overexpression was due to nucleosome alterations. CONCLUSION: The study revealed that FUL resistance is associated with the dysregulation of GPER and CDK6. A mechanism leading to aberrant gene expression was most likely unscheduled chromatin remodelling by hSWI/SNF. Hence, three targets should be conceptually addressed in a second-line adjuvant therapy: the catalytic centre of SWI/SNF (BRG1) to delay the development of FUL resistance, GPER to increase sensitivity to FUL and the reconstitution of the RB1 pathway to overcome resistance.

Bone-patellar tendon-bone graft via a single minimally-invasive approach versus a classical approach in anterior cruciate ligament reconstruction: a prospective study.

UNLABELLED: Harvesting the patellar ligament for anterior cruciate ligament reconstructions can be a source of anterior knee pain and hypoesthesia of the lateral side of the knee. We analyzed the feasibility of a minimally-invasive technique via a single patellar approach and postulate that it reduces anterior pain and limits the hypoesthesia area. PATIENTS AND METHODS: A prospective, comparative, non-randomized, single-center study was conducted on two groups: one undergoing surgery with the classical anteromedial approach, the other with the minimally-invasive approach. Each group included 20 patients. Both series were reviewed between the 6th and 8th month after surgery. The revision was clinical, radiological, and ultrasonographic. RESULTS: The grafts harvested via the classical approach in all 20 cases presented good characteristics, versus eight out of 18 for the grafts harvested via the minimally-invasive approach. A prominent anterior tibial tuberosity improved the quality of the tibial bone block. A hypoesthesia zone was found in 16 cases out of 18 in the classical approach group, it measured a mean 10.3 ± 5.6 cm(2). A surface area of 3 cm(2) was noted in one case from the minimally-invasive group. No significant difference was found for the subjective and objective IKDC and Lille patellofemoral scores between the two groups. Anterior pain was present in four patients in the classical group and six in the minimally-invasive group. DISCUSSION: The minimally-invasive technique reduces the risk of cutaneous hypoesthesia. It does not prevent anterior pain related to harvesting the patellar tendon and a good-quality transplant can be obtained if the anterior tibial tuberosity is prominent. LEVEL OF EVIDENCE: Level III: case-control study.

Medial facet patelloplasty in patellar instability associated with patellofemoral dysplasia: a report of 26 cases.

INTRODUCTION: Numerous procedures may be used in the surgical treatment of patellar instability. We have noticed that certain types of patellar instability result in a bulge (or protrusion) of the distal medial facet of the patella which can hinder recentering in the trochlear groove. HYPOTHESIS: In certain patellofemoral dysplasias, trimming down of this bulge (patelloplasty) as long as it is associated with a "à la carte surgery" program, can improve stability, centering and lateral patellar tilt. PATIENTS AND METHODS: This retrospective series included 23 patients (26 knees), 13 women and 10 men, mean age 25.9±9.01 years old (15-52) operated between 1997-2008. Patellar dislocation had occurred at least once in all knees, and at least twice in 16 knees. The mean preoperative Kujala score was 79.1±6.1 points (68-91). There was a bulge on the medial facet of the patella in all cases, in particular on skyline views associated with the usual criteria for patellofemoral dysplasia. Patelloplasty was associated in all cases with resection of the lateral patellar retinaculum and anteromedialization of the tibial tubercle (7 were lowered) and four Albee trochleoplasties. RESULTS: Recurrent dislocation occurred in one case (4.7%). Nineteen patients (22 knees) were followed up for a mean 7.53±3.27 years (2-13 years). The mean postoperative Kujala score was 91.8±7.9 points (70-100) and the subjective results in 19/22 (86.5%) knees were satisfactory or very satisfactory. On skyline views, 21 patellae (95.5%) were well centered and one (4.5%) still presented with lateral tilt. There was no osteoarthritis in 15 knees (68%). CONCLUSION: In selected cases, patelloplasty of the medial facet of the patella has no particular morbidity at intermediate follow-up. Associated with other surgical procedures, the rate of satisfaction of patients is very high and especially encouraging. LEVEL OF EVIDENCE: Level IV, retrospective cohort study.

Brain metastases free survival differs between breast cancer subtypes.

BACKGROUND: Brain metastases (BM) are frequently diagnosed in patients with HER-2-positive metastatic breast cancer; in addition, an increasing incidence was reported for triple-negative tumours. We aimed to compare brain metastases free survival (BMFS) of breast cancer subtypes in patients treated between 1996 until 2010. METHODS: Brain metastases free survival was measured as the interval from diagnosis of extracranial breast cancer metastases until diagnosis of BM. HER-2 status was analysed by immunohistochemistry and reanalysed by fluorescent in situ hybridisation if a score of 2+ was gained. Oestrogen-receptor (ER) and progesterone-receptor (PgR) status was analysed by immunohistochemistry. Brain metastases free survival curves were estimated with the Kaplan-Meier method and compared with the log-rank test. RESULTS: Data of 213 patients (46 luminal/124 HER-2/43 triple-negative subtype) with BM from breast cancer were available for the analysis. Brain metastases free survival differed significantly between breast cancer subtypes. Median BMFS in triple-negative tumours was 14 months (95% CI: 11.34-16.66) compared with 18 months (95% CI: 14.46-21.54) in HER-2-positive tumours (P=0.001) and 34 months (95% CI: 23.71-44.29) in luminal tumours (P=0.001), respectively. In HER-2-positive patients, co-positivity for ER and HER-2 prolonged BMFS (26 vs 15 m; P=0.033); in luminal tumours, co-expression of ER and PgR was not significantly associated with BMFS. Brain metastases free survival in patients with lung metastases was significantly shorter (17 vs 21 months; P=0.014). CONCLUSION: Brain metastases free survival in triple-negative breast cancer, as well as in HER-2-positive/ER-negative, is significantly shorter compared with HER-2/ER co-positive or luminal tumours, mirroring the aggressiveness of these breast cancer subtypes.

Impact of anti-HER2 therapy on overall survival in HER2-overexpressing breast cancer patients with brain metastases.

BACKGROUND: Trastuzumab-based therapy after diagnosis of brain metastases (BM) may improve survival due to prolonged systemic disease control. We investigated whether lapatinib may yield additional survival benefit. METHODS: Eighty patients with BM from HER2-positive breast cancer were identified. Karnofsky Performance Score (KPS) of at least 70 was required. We included a control group of 37 patients treated before 2003, when continuation of trastuzumab after diagnosis of BM was not yet recommended. Remainders received either trastuzumab or lapatinib and trastuzumab (either concomitantly or sequentially) with or without chemotherapy. RESULTS: Median overall survival (OS) in patients receiving trastuzumab after diagnosis of BM was 13 months; corresponding numbers were 9 months in patients treated with chemotherapy, and 3 months with radiotherapy alone. Median OS was not reached in the lapatinib group. Addition of lapatinib prolonged OS over trastuzumab alone (P=0.002). After correction for potential confounders, lapatinib therapy remained an independent positive predictor for survival (HR 0.279; P=0.012). INTERPRETATION: This retrospective single-centre study suggests that the introduction of lapatinib improved survival in patients with BM from HER2-positive breast cancer. Patients with KPS ≥70 may benefit when treated with lapatinib in addition to trastuzumab after completion of local therapy.

NF-κB mediates the 12(S)-HETE-induced endothelial to mesenchymal transition of lymphendothelial cells during the intravasation of breast carcinoma cells.

BACKGROUND: The intravasation of breast cancer into the lymphendothelium is an early step of metastasis. Little is known about the mechanisms of bulky cancer invasion into lymph ducts. METHODS: To particularly address this issue, we developed a 3-dimensional co-culture model involving MCF-7 breast cancer cell spheroids and telomerase-immortalised human lymphendothelial cell (LEC) monolayers, which resembles intravasation in vivo and correlated the malignant phenotype with specific protein expression of LECs. RESULTS: We show that tumour spheroids generate 'circular chemorepellent-induced defects' (CCID) in LEC monolayers through retraction of LECs, which was induced by 12(S)-hydroxyeicosatetraenoic acid (HETE) secreted by MCF-7 spheroids. This 12(S)-HETE-regulated retraction of LECs during intravasation particularly allowed us to investigate the key regulators involved in the motility and plasticity of LECs. In all, 12(S)-HETE induced pro-metastatic protein expression patterns and showed NF-κB-dependent up-regulation of the mesenchymal marker protein S100A4 and of transcriptional repressor ZEB1 concomittant with down-regulation of the endothelial adherence junction component VE-cadherin. This was in accordance with ∼50% attenuation of CCID formation by treatment of cells with 10 µM Bay11-7082. Notably, 12(S)-HETE-induced VE-cadherin repression was regulated by either NF-κB or by ZEB1 since ZEB1 siRNA knockdown abrogated not only 12(S)-HETE-mediated VE-cadherin repression but inhibited VE-cadherin expression in general. INTERPRETATION: These data suggest an endothelial to mesenchymal transition-like process of LECs, which induces single cell motility during endothelial transmigration of breast carcinoma cells. In conclusion, this study demonstrates that the 12(S)-HETE-induced intravasation of MCF-7 spheroids through LECs require an NF-κB-dependent process of LECs triggering the disintegration of cell-cell contacts, migration, and the generation of CCID.

Long-term persistence of acquired resistance to 5-fluorouracil in the colon cancer cell line SW620.

Treatment resistance to antineoplastic drugs represents a major clinical problem. Here, we investigated the long-term stability of acquired resistance to 5-fluorouracil (FU) in an in vitro colon cancer model, using four sub-clones characterised by increasing FU-resistance derived from the cell line SW620. The resistance phenotype was preserved after FU withdrawal for 15weeks (~100 cell divisions) independent of the established level of drug resistance and of epigenetic silencing. Remarkably, resistant clones tolerated serum deprivation, adopted a CD133(+) CD44(-) phenotype, and further exhibited loss of membrane-bound E-cadherin together with predominant nuclear ß-catenin localisation. Thus, we provide evidence for a long-term memory of acquired drug resistance, driven by multiple cellular strategies (epithelial-mesenchymal transition and selective propagation of CD133(+) cells). These resistance phenomena, in turn, accentuate the malignant phenotype.