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Online ion range monitoring in hadron therapy can be performed via detection of secondary radiation, such as prompt γ-rays, emitted during treatment. The prompt γ emission profile is correlated with the ion depth-dose profile and can be reconstructed via Compton imaging. The line-cone reconstruction, using the intersection between the primary beam trajectory and the cone reconstructed via a Compton camera, requires negligible computation time compared to iterative algorithms. A recent report hypothesised that time of flight (TOF) based discrimination could improve the precision of the γ fall-off position measured via line-cone reconstruction, where TOF comprises both the proton transit time from the phantom entrance until γ emission, and the flight time of the γ-ray to the detector. The aim of this study was to implement such a method and investigate the influence of temporal resolution on the precision of the fall-off position. Monte Carlo simulations of a 160 MeV proton beam incident on a homogeneous PMMA phantom were performed using GATE. The Compton camera consisted of a silicon-based scatterer and CeBr3 scintillator absorber. The temporal resolution of the detection system (absorber + beam trigger) was varied between 0.1 and 1.3 ns RMS and a TOF-based discrimination method applied to eliminate unlikely solution(s) from the line-cone reconstruction. The fall-off position was obtained for varying temporal resolutions and its precision obtained from its shift across 100 independent γ emission profiles compared to a high statistics reference profile. The optimal temporal resolution for the given camera geometry and 108 primary protons was 0.2 ns where a precision of 2.30 ± 0.15 mm (1σ) on the fall-off position was found. This precision is comparable to current state of-the-art Compton imaging using iterative reconstruction methods or 1D imaging with mechanically collimated devices, and satisfies the requirement of being smaller than the clinical safety margins.

The aim of this work is to assess if cumulative dose (CD) and dose intensity (DI) of everolimus may affect survival of advanced pancreatic neuroendocrine tumors (PNETs) patients. One hundred and sixteen patients (62 males and 54 females, median age 55 years) with advanced PNETs were treated with everolimus for ≥3 months. According to a Receiver operating characteristics (ROC) analysis, patients were stratified into two groups, with CD ≤ 3000 mg (Group A; n = 68) and CD > 3000 mg (Group B; n = 48). The response rate and toxicity were comparable in the two groups. However, patients in group A experienced more dose modifications than patients in group B. Median OS was 24 months in Group A while in Group B it was not reached (HR: 26.9; 95% CI: 11.0–76.7; P

International audience; Large-size diamond detectors have been produced and tested with the aim of achieving reliable & efficient sensors for particle tracking or medical applications. Poly-and single-crystal CVD diamond samples were submitted to various ionizing particles. Their metallization was performed by using Distributed MicroWave Plasmas, a process developed by LPSC. Their detection performance was investigated using α and β radioactive sources, 95 MeV/u carbon beams from GANIL (Caen France) and short-bunched 8.5 keV photons from ESRF. This study is part of the ANR project MONODIAM-HE and of the CLaRyS collaboration for the on-line dose monitoring of hadrontherapy. The goal here is to provide large-area detectors with a high detection efficiency for carbon or proton beams, yielding time and position measurement at count rates greater than 100 MHz. A time resolution ranging from 20 ps up to 40 ps and an energy resolution varying from 7 % up to 10% were measured. It allowed us to conclude that poly-crystal CVD diamond detectors are good candidates for this application.

Recent anatomical and clinical studies have led to the hypothesis that in several cases of failure of response to renal denervation (RDN), the procedure has not been technically correct. To perform procedural reassessment in patients with true resistant hypertension who underwent RDN. We retrospectively reassessed the procedural technique of RDN in 10 true resistant hypertensive patients, comparing the sites of renal ablations with the knowledge of animal and human post mortem evidences. Procedural ablation technique was assessed in terms of number of ablations for each renal artery and site of ablation (quadrant and distance from renal ostium) by using the radiologic images of each RDN and the number of radiofrequency ablation attempts documented in the reports of each denervation session. 10 patients were studied, 9 denervated with Simplicity monoelectrode catheter, 1 with multielectrode balloon technique. Responders to the procedure underwent more ablations and particularly at least a quadrant ablation in one of the kidney arteries, >2 ablations in Dorsal plus Ventral quadrants and in 67% of then >10 ablations were done in superior inferior and ventral quadrants. This study confirms the importance of a well knowledge of renal artery anatomy and underlines the relevance of the choice of ablation sites in order to obtain a successful RDN procedure.

To assess the procedure for measuring blood pressure (BP) among hospital nurses and to assess if a training program would improve technique and accuracy.160 nurses from Molinette Hospital were included in the study. The program was based upon theoretical and practical lessons. It was one day long and it was held by trained nurses and physicians who have practice in the Hypertension Unit. An evaluation of nurses' measuring technique and accuracy was performed before and after the program, by using a 9-item checklist. Moreover we calculated the differences between measured and effective BP values before and after the training program.At baseline evaluation, we observed inadequate performance on some points of clinical BP measurement technique, specifically: only 10% of nurses inspected the arm diameter before placing the cuff, 4% measured BP in both arms, 80% placed the head of the stethoscope under the cuff, 43% did not remove all clothing that covered the location of cuff placement, did not have the patient seat comfortably with his legs uncrossed and with his back and arms supported. After the training we found a significant improvement in the technique for all items. We didn't observe any significant difference of measurement knowledge between nurses working in different settings such as medical or surgical departments.Periodical education in BP measurement may be required, and this may significantly improve the technique and consequently the accuracy.

Catalano V, Mellone P, d’Avino A, Shridhar V, Staccioli M P, Graziano F, Giordani P, Rossi D, Baldelli A M, Alessandroni P, Santini D, Lorenzon L, Testa E, D’Emidio S, De Nictolis M, Muretto P, Fedeli S L & Baldi A (2011) Histopathology58, 669–678 HtrA1, a potential predictor of response to cisplatin-based combination chemotherapy in gastric cancer Aims: HtrA1 is a member of the HtrA (high-temperature requirement factor A) family of serine proteases. HtrA1 plays a protective role in various malignancies due to its tumour suppressive properties. The aim of this study was to determine HtrA1 expression as a predictor of chemoresponse in patients with advanced gastric cancer. Methods and results: HtrA1 expression was determined by immunohistochemistry on specimens of primary gastric cancer from 80 patients treated consecutively with cisplatin-based combination chemotherapy. Response to chemotherapy was assessed according to Response Evaluation Criteria in Solid Tumours (RECIST) criteria. Our population consisted of males/females [51/29; median age 64 years (range 32–82)]. A complete or partial response was observed in 71.4% [95% confidence interval (CI) 54.7–88.2], 66.7% (95% CI 47.8–85.5) and 28.6% (95 CI 11.8–45.3) of tumours showing high, medium and low HtrA1 expression, respectively. A statistically significant association between HtrA1 expression and the clinical response was observed (P = 0.002). The median overall survival for patients with high/medium expression was 17 months compared to 9.5 months for patients with low HtrA1 expression (P = 0.037). Conclusions: Identification of HtrA1 in gastric cancer prior to chemotherapy indicates that levels of HtrA1 could be used to predict response to platinum-based combination therapies. Further assessment of HtrA1 expression is highly warranted in large, prospective studies.

A standardized approach to antihypertensive pharmacological therapy taking into account the pathophysiological mechanism of hypertension is not yet available. Laragh and colleagues have proposed a model based on the concept that the two final determinants of blood pressure are the volume status and the renin-angiotensin-aldosterone system. This model allows patients to be divided into two categories based on plasma renin activity ( 0.65 ng/mL) and with a different primary target for a specific therapy. In Laragh’s opinion, all antihypertensive drugs could be divided into two groups according to their mechanism of action: drugs that block the activity of the renin-angiotensin system (R drugs, such as β-adrenoceptor antagonists [β-blockers], ACE inhibitors, angiotensin II type 1 receptor antagonists [angiotensin receptor blockers] and centrally acting antihypertensive drugs) or drugs that increase renal sodium excretion (V drugs, such as diuretics, α-adrenoceptor antagonists [α-blockers] or calcium channel blockers). This article reviews the mechanisms underlying this antihypertensive drug classification.

Background/Aims: The vasculature has become an important target in the development of therapies for increasing numbers of human diseases, yet there are few reliable markers available to identify the endothelium in rodent models. We have characterized the expression, subcellular localization and accessibility of the rat pan-endothelial marker podocalyxin (podxl) using a newly developed monoclonal antibody (mAb), G278. Methods: podxl expression and accessibility to binding by G278 were determined in the rat by a variety of experimental approaches. Results: mAb G278 reliably immunostained blood vessels of all types and of every size in fresh-frozen, fixed-frozen and paraffin-embedded sections of all tissues, but did not stain lymphatic vessels. Western blotting, in vivo imaging and biodistribution analyses demonstrated that the highest levels of endothelial podxl were found in the lung and heart. We also determined that podxl is not enriched in caveolae and that its expression can be modulated in the tumor microenvironment. Conclusion: Our study shows that podxl is a better identifier of rat endothelia than are some of the more commonly used markers and that mAb G278 is a robust antibody for use not only in identifying rat blood vessels but also as a tool to elucidate podxl function.

How effectively and quickly endothelial caveolae can transcytose in vivo is unknown, yet critical for understanding their function and potential clinical utility. Here we use quantitative proteomics to identify aminopeptidase P (APP) concentrated in caveolae of lung endothelium. Electron microscopy confirms this and shows that APP antibody targets nanoparticles to caveolae. Dynamic intravital fluorescence microscopy reveals that targeted caveolae operate effectively as pumps, moving antibody within seconds from blood across endothelium into lung tissue, even against a concentration gradient. This active transcytosis requires normal caveolin-1 expression. Whole body gamma-scintigraphic imaging shows rapid, specific delivery into lung well beyond that achieved by standard vascular targeting. This caveolar trafficking in vivo may underscore a key physiological mechanism for selective transvascular exchange and may provide an enhanced delivery system for imaging agents, drugs, gene-therapy vectors and nanomedicines. 'In vivo proteomic imaging' as described here integrates organellar proteomics with multiple imaging techniques to identify an accessible target space that includes the transvascular pumping space of the caveola.

The purpose of this study was to experimentally assess the possibility to monitor carbon ion range variations--due to tumor shift and/or elongation or shrinking--using prompt-gamma (PG) emission with inhomogeneous phantoms. Such a study is related to the development of PG monitoring techniques to be used in a carbon ion therapy context.A 95 MeV/u carbon ion beam was used to irradiate phantoms with a variable density along the ion path to mimic the presence of bone and lung in homogeneous humanlike tissue. PG profiles were obtained after a longitudinal scan of the phantoms. A setup comprising a narrow single-slit collimator and two detectors placed at 90° with respect to the beam axis was used. The time of flight technique was applied to allow the selection between PG and background events.Using the positions at 50% entrance and 50% falloff of the PG profiles, a quantity called prompt-gamma profile length (PGPL) is defined. It is possible to observe shifts in the PGPL when there are absolute ion range shifts as small as 1-2 mm. Quantitatively, for an ion range shift of -1.33 ± 0.46 mm (insertion of a Teflon slab), a PGPL difference of -1.93 ± 0.58 mm and -1.84 ± 1.27 mm is obtained using a BaF2 and a NaI(Tl) detector, respectively. In turn, when an ion range shift of 4.59 ± 0.42 mm (insertion of a lung-equivalent material slab) is considered, the difference is of 4.10 ± 0.54 and 4.39 ± 0.80 mm for the same detectors.Herein, experimental evidence of the usefulness of employing PG to monitor carbon ion range using inhomogeneous phantoms is presented. Considering the homogeneous phantom as reference, the results show that the information provided by the PG emission allows for detecting ion range shifts as small as 1-2 mm. When considering the expected PG emission from an energy slice in a carbon ion therapy scenario, the experimental setup would allow to retrieve the same PGPL as the high statistics of the full experimental dataset in 58% of the times. However, this success rate increases to 93% when using a better optimized setup by means of Monte Carlo simulations.

The molecular diversity of the luminal endothelial cell surface arising in vivo from local variations in genetic expression and tissue microenvironment may create opportunities for achieving targeted molecular imaging and therapies. Here, we describe a strategy to identify probes and their cognate antigens for targeting vascular endothelia of specific organs in vivo . We differentially screen phage libraries to select organ-targeting antibodies by using luminal endothelial cell plasma membranes isolated directly from tissue and highly enriched in natively expressed proteins exposed to the bloodstream. To obviate liver uptake of intravenously injected phage, we convert the phage-displayed antibodies into scFv-Fc fusion proteins, which then are able to rapidly target select organ(s) in vivo as visualized directly by γ-scintigraphic whole-body imaging. Mass spectrometry helps identify the antigen targets. This comprehensive strategy provides new promise for harnessing the power of phage display for mapping vascular endothelia natively in tissue and for achieving vascular targeting of specific tissues in vivo .

Purpose A high interleukin-1β (IL-1B) and interleukin-1 receptor antagonist (IL-RN) ratio underlies an unfavorable proinflammatory status. Also, it seems to be involved in the mechanisms of cancer cachexia and tumor angiogenesis and metastasis. Two single nucleotide polymorphisms in IL-1B gene (IL-1B-511C/T,IL-1B-31T/C) and a variable number of tandem repeat polymorphisms in IL-RN gene (IL-1RNlong/2) enhance the circulating levels of the two cytokines. The prognostic role of IL-1B/IL-1RN genotypes was investigated in patients with relapsed and metastatic gastric cancer treated with palliative chemotherapy. Patients and Methods Before starting palliative chemotherapy, 123 prospectively enrolled patients supplied peripheral-blood samples for DNA extraction. Survival data were analyzed according to IL-1RN/IL-1B genotypes. Results Forty-two patients showed wild-type genotypes (IL-1RNlong/long, IL-1B-511C/C, and IL-1B-31T/T; group A). Forty-five patients showed the IL-1RN2 polymorphism, with wild-type IL-1B genotypes in seven patients and with IL-1B-511C/T and/or IL-1B-31T/C polymorphisms in 38 patients (group B). The remaining 36 patients demonstrated wild-type IL-1RN, with IL-1B-511C/T and/or IL-1B-31T/C polymorphisms (group C). In group A and B patients, the median progression-free survival (PFS) was 25 and 26 weeks, respectively, and median overall survival (OS) was 42 and 43 weeks, respectively. Group C patients showed worse PFS (median, 16 weeks) and OS (median, 28 weeks) than group A (P = .006 for PFS; P = .0001 for OS) and group B patients (P = .01 for PFS; P = .0001 for OS). The long/T/C haplotype was overrepresented in patients with shortened PFS (P = .001) and OS (P = .0005). Conclusion In patients with advanced gastric cancer, IL-1B polymorphisms showed adverse prognostic influence when coupled with wild-type IL-1RN genotype. These findings deserve further investigation for potential anticancer activity of recombinant IL-RN.

The aim of our analysis was to assess retrospectively the effect on local relapse, overall survival (OS) and disease-free survival (DFS) of a limited or an extended lymphadenectomy in radically resected gastric cancer patients. This study was performed in order to identify a subgroup of patients possibly not benefiting from a therapeutic approach such as chemoradiation therapy. We divided our patients into two groups according to lymphadenectomy type: group A for limited ( 25 resected lymph nodes) lymph nodes resection. A total of 418 patients were analysed: tumour stage at diagnosis was pT2–3 pN1–3 M0 in 339 patients and pT3 N0 M0 in 79 patients. Median age at diagnosis was 68 years (range 30–92 years). A total of 306 patients (73.2%) were in group A and 112 (26.8%) in group B. The median survival time (OS) for patients in groups A and B was 58.8 and 84.8 months, respectively (P=0.0371); median DFS was 28.8 months in group A and 59.9 months in group B (P=0.0027). At multivariate analysis, extension within the gastric wall, nodal involvement and the number of resected lymph nodes appeared to affect both OS and DFS. An inadequate lymph nodes resection can affect survival and result in a higher incidence of local relapse, making the latter group of patients optimal candidates for adjuvant chemoradiation.

Loss of E-cadherin expression has been related with an adverse outcome in patients with resected gastric cancer. More recently, experimental models with cancer cell lines showed that chemosensitivity may be affected by the E-cadherin expression status. We investigated whether E-cadherin expression is correlated with the response to chemotherapy and the survival of patients with advanced gastric cancer. Consecutive patients with advanced gastric cancer who underwent palliative chemotherapy were considered eligible for study entry. Measurable disease, complete follow-up information and availability of tumor specimens for immunohistochemistry were mandatory inclusion criteria. In 70 assessable patients, 30 patients had locoregional disease and 40 patients had visceral metastases. Chemotherapy consisted of cisplatin/fluorouracil/folinic acid in 33 patients and cisplatin/fluorouracil/epirubicin/folinic acid in 37 patients. There were 13 patients with complete response, 20 with partial response, 20 with stable disease and 17 patients progressed. Thirty-eight patients had80% E-cadherin-positive cancer cells (positive E-cadherin expression); 15 cases showed 25-70% (reduced E-cadherin expression), and in the remaining 17 cases E-cadherin expression was10% (negative E-cadherin expression). The response to chemotherapy was unrelated to the E-cadherin expression status. Conversely, survival in the 32 patients with reduced/negative E-cadherin expression (25 weeks) was significantly worse than that observed in the 38 patients with preserved E-cadherin expression (36 weeks) (p0.01). E-cadherin expression retained its independent prognostic role in the multivariate analysis. E-cadherin expression may give prognostic information in patients with advanced gastric cancer, but it does not seem to possess a predictive role in vivo. Some of the mechanisms inducing E-cadherin downregulation, like hypermethylation, may be potentially reversible, and they deserve further investigation as the target of novel therapeutic strategies.

The incidence of gastric cancer (GC) increases significantly after the fifth decade and palliative chemotherapy is the ultimate treatment in the majority of patients. We investigated safety and efficacy of a weekly regimen with cisplatin, fluorouracil and leucovorin as first-line chemotherapy for elderly patients with advanced GC. Chemotherapy-naive patients older than 65 years were considered eligible for study entry. Frail elderly patients were identified and excluded according to the following criteria: age85 years, dependence in one or more activities of daily living (activities of daily living and instrumental activities of daily living scales), three or more comorbid conditions, one or more geriatric syndromes. Chemotherapy consisted of 1-day per week administration of intravenous cisplatin 35 mg m(-2), 6S-stereoisomer leucovorin 250 mg m(-2) and fluorouracil 500 mg m(-2) (PLF). Patients were re-evaluated after eight weekly cycles and six additional weekly administrations were planned for patients without disease progression. A 5-day subcutaneous filgrastim (5 mug Kg(-1) day(-1), days +1-+5) was used after the first treatment delay for neutropenia and maintained thereafter. In the whole group, the best intention-to-treat overall response rate was 43% (95% CI: 30-56%). The time to disease progression and the median survival time were 5.3 and 8.6 months, respectively. Fatigue was the commonest nonhaematologic toxicity (71% of the patients). Filgrastim was used in 30 patients who showed grade II (20 patients) or grade III (10 patients) neutropenia. Neither grade IV toxicity nor toxic deaths were observed. The weekly PLF regimen resulted safe and effective in elderly patients with advanced GC. This outpatient regimen is based on old and low-cost drugs and it may represent an alternative to new and more expensive combinations.

Monoclonal antibody technology has generated invaluable tools for both the analytical and clinical sciences. However, standard immunization approaches frequently fail to provide monoclonal antibodies with the desired specificity. Subtractive immunization provides a powerful alternative to standard immunization and allows for the production of truly unique antibodies. With the intent of targeting specific epitopes within the proteome, subtractive immunization has been broadly and successfully implemented for the production of monoclonal antibodies otherwise unobtainable by standard immunization. Subtractive immunization utilizes a distinct immune tolerization approach that can substantially enhance the generation of monoclonal antibodies to desired antigens. The approach is based on tolerizing the host animal to immunodominant or otherwise undesired antigen(s) (tolerogen) that may be structurally or functionally related to the antigen of interest. Tolerization of the host animal can be achieved through one of three methods: High Zone, Neonatal, or Drug-induced tolerization. The tolerized animal is then inoculated with the desired antigen (immunogen) and antibodies generated by the subsequent immune response are screened for the desired antigenic reactivity. Over the past 15 years a large number of investigators have used the subtractive approach with cleverly chosen tolerogen–immunogen combinations and successfully generated uniquely reactive antibodies which are often neutralizing or function-blocking. This review will focus on the implementation of subtractive immunization for the production of antibodies otherwise unobtainable by standard immunization.

Current studies are investigating new E-cadherin gene (CDH1) mutations that may be responsible for diffuse gastric cancer susceptibility. Recently, a novel CDH1 germline variant presenting a G/A nucleotide change at cDNA position 2494 has been found in Japanese patients with familial diffuse gastric cancer. The consequent amino acid variation (Val/Met) may alter the binding activity to beta-catenin and the adhesive function of the E-cadherin protein. We have investigated its frequency in Italian cases of sporadic diffuse gastric cancer a well as in healthy controls. Peripheral blood samples were collected from consecutive patients with sporadic, diffuse gastric cancer and from healthy controls in the District of Urbino, Marche Region, Central Italy. After DNA extraction, standard techniques for molecular analyses were used to investigate the 2494 G/A germline nucleotide change in CDH1 cDNA. None of the 48 patients and 48 controls showed the G/A 2494 nucleotide change. Assuming a binomial distribution of the mutation among individuals and the absence of mutations in the 48 patients, the 95% upper bound for the underlying mutation frequency was 7.4%. The novel CDH1 nucleotide change is uncommon in Italian patients with sporadic diffuse gastric cancer. Given these results, further analyses in large population-based studies are not advisable.

The interactions of two cell lines having different metastatic properties, and the subsequent effects on dissemination were investigated using the chicken embryo metastasis assay. The highly aggressive human epidermoid cell line HEp-3 was tested alone or mixed with the mouse colon carcinoma cell line CL26 in this assay. When inoculated individually, each cell line forms experimental metastases in the chicken embryo, but only the HEp-3 cells give rise to spontaneous metastases. In embryos co-inoculated with both cell lines there was an overall reduction in metastatic burden in both the spontaneous and experimental metastasis assays. Furthers studies revealed that CL26 cells, when co-inoculated with HEp-3 cells did not acquire the ability to spontaneously metastasize. However, in the presence of CL26 cells, spontaneous HEp-3 metastasis was reduced. Intravenous co-inoculation of HEp-3 and CL26 cells also resulted in a reciprocal suppression of experimental metastasis by both cell lines. These studies demonstrate that the interactions of adjacent, phenotypically different tumor cells can have a suppressive effect on dissemination of one or both cell types.

Background Elderly patients have been often excluded from or underrepresented in the study populations of combination chemotherapy trials. The primary end point of this study was to determine the response rate and the toxicity of the weekly oxaliplatin, 5-fluorouracil and folinic acid (OXALF) regimen in elderly patients with advanced gastric cancer. The secondary objective was to measure the time to disease progression and the survival time. Methods Chemotherapy-naive patients with advanced gastric cancer aged 70 or older were considered eligible for study entry. Patients received weekly oxaliplatin 40 mg/m2, fluorouracil 500 mg/m2 and folinic acid 250 mg/m2. All drugs were given intravenously on a day-1 schedule. Results A total of 42 elderly patients were enrolled. Median age was 73 years and all patients had metastatic disease. The response rate according to RECIST criteria was 45.2% (95% CIs: 30%–56%) with two complete responses, 17 partial responses, 13 stable diseases and 10 progressions, for an overall tumor rate control of 76.2% (32 patients). Toxicity was generally mild and only three patients discontinued treatment because of treatment related adverse events. The most common treatment-related grade 3/4 adverse events were fatigue (7.1%), diarrhoea (4.8%), mucositis (2.4%), neurotoxicity (2.4%) and neutropenia (4.8%). The median response duration was 5.3 months (95% CIs: 2.13 – 7.34), the median time to disease progression was 5.0 months (95% CIs: 3.75 – 6.25) and the median survival time was 9.0 months (95% CIs: 6.18 – 11.82). Conclusion OXALF represents an active and well-tolerated treatment modality for elderly patients with locally advanced and metastatic gastric cancer.

Previously, we demonstrated the role of nitric oxide (NO) in transforming epithelial cells from a stationary to locomoting phenotype [E. Noiri, T. Peresleni, N. Srivastava, P. Weber, W. F. Bahou, N. Peunova, and M. S. Goligorsky. Am. J. Physiol. 270 ( Cell Physiol. 39): C794–C802, 1996] and its permissive function in endothelin-1-stimulated endothelial cell migration (E. Noiri, Y. Hu, W. F. Bahou, C. Keese, I. Giaever, and M. S. Goligorsky. J. Biol. Chem. 272: 1747–1753, 1997). In the present study, the role of functional NO synthase in executing the vascular endothelial growth factor (VEGF)-guided program of endothelial cell migration and angiogenesis was studied in two independent experimental settings. First, VEGF, shown to stimulate NO release from simian virus 40-immortalized microvascular endothelial cells, induced endothelial cell transwell migration, whereas NG-nitro-l-arginine methyl ester (l-NAME) or antisense oligonucleotides to endothelial NO synthase suppressed this effect of VEGF. Second, in a series of experiments on endothelial cell wound healing, the rate of VEGF-stimulated cell migration was significantly blunted by the inhibition of NO synthesis. To gain insight into the possible mode of NO action, we next addressed the possibility that NO modulates cell matrix adhesion by performing impedance analysis of endothelial cell monolayers subjected to NO. The data showed the presence of spontaneous fluctuations of the resistance in ostensibly stationary endothelial cells. Spontaneous oscillations were induced by NO, which also inhibited cell matrix adhesion. This process we propose to term “podokinesis” to emphasize a scalar form of micromotion that, in the presence of guidance cues, e.g., VEGF, is transformed to a vectorial movement. In conclusion, execution of the program for directional endothelial cell migration requires two coexisting messages: NO-induced podokinesis (scalar motion) and guidance cues, e.g., VEGF, which imparts a vectorial component to the movement. Such a requirement for the dual signaling may explain a mismatch in the demand and supply with newly formed vessels in different pathological states accompanied by the inhibition of NO synthase.

PURPOSE A multiinstitutional trial was performed to confirm the clinical activity, in terms of response rate and toxicity (primary objectives) and duration of responses and survival (secondary objectives), of an intensive weekly regimen in advanced gastric cancer. PATIENTS AND METHODS Patients with measurable unresectable and/or metastatic gastric carcinoma received 1-day per week administration of cisplatin (CDDP) 40 mg/m2, fluorouracil (5FU) 500 mg/m2, epi-doxorubicin (epi-ADR) 35 mg/m2, 6S-stereoisomer of leucovorin 250 mg/m2, and glutathione 1.5 g/m2. On the other days, filgrastim was administered by subcutaneous injection at a dose of 5 mg/kg. One cycle of therapy consisted of eight 1-week treatments. Patients who showed a response or stable disease received a further 6 weeks of therapy. RESULTS Of 105 enrolled patients, 11 had locally advanced unresectable disease only; 33 had primary nonresected and metastatic disease; 48 had metastatic disease and primary tumor resected; 10 had locoregional recurrence and metastatic disease; and three had locoregional recurrence only. After one cycle, 18 complete responses (CRs) and 47 partial responses (PRs) were achieved, for an overall response rate of 62% (95% confidence interval [CI], 53% to 71%). Twenty patients had stable disease and 20 progressed on therapy. The median survival duration of all 105 patients was 11 months, with 1- and 2-year survival rates of 42% and 5%, respectively. World Health Organization (WHO) grade III to IV toxicity, in terms of anemia, neutropenia, thrombocytopenia, and mucositis, was experienced by 40 patients (38%). There were no treatment-related deaths. CONCLUSION These data support the results of the pilot study and confirmed the high activity of the regimen, with acceptable toxicity. This schedule deserves evaluation in the adjuvant setting.

In this phase II trial we have evaluated the activity and toxicity of a combination regimen containing mitoxantrone, L-leucovorin, and fluorouracil in patients with advanced breast cancer pretreated with anthracyclines. Forty-six patients were included into the study; they received a total of 227 cycles of chemotherapy. Median age was 63 years (range 34-78), median performance status was 80 (range 60-100). Visceral metastases were present in 37 patients, 6 patients had bone involvement only, while 3 patients had soft tissue/lymph node disease. Median number of previous chemotherapy regimens for advanced disease was 2 (range 1-3). Ten patients had anthracycline primary resistance (progressive disease during treatment). Twenty-three patients received mitoxantrone 12 mg/sqm day 1; fluorouracil 370 mg/sqm and L-folinic acid 100 mg/sqm days 1-3 administered every three weeks. Another group of 23 patients were treated with the same regimen using a prolonged 5FU/L-FA schedule (5 days). Two complete responses and 6 partial responses were recorded with the 3-day schedule; 7 partial responses in the 5-day schedule (overall response rate 32.6%, 95% C.I. 19-46%). Two partial responses were observed in patients with anthracycline primary resistance. Median response duration was 9 months (range 3-16). Hematologic toxicity was mild: grade 3-4 leukopenia was recorded in 5 patients, grade 3-4 thrombocytopenia in 3 patients. Grade III-IV stomatitis and diarrhea was recorded in 4 and 5 patients respectively (all receiving the 5-day 5-FU/L-FA schedule). Cardiac toxicity was observed in two cases. This regimen proved active in advanced breast cancer following anthracycline-containing chemotherapy, and the 3-day schedule could be offered to such patients with acceptable toxicity.

Technologies are needed to map and image biological barriers in vivo that limit solid tumor delivery and, ultimately, the effectiveness of imaging and therapeutic agents. Here we integrate proteomic and imaging analyses of caveolae at the blood-tumor interface to discover an active transendothelial portal to infiltrate tumors. A post-translationally modified form of annexin A1 (AnnA1) is selectively concentrated in human and rodent tumor caveolae. To follow trafficking, we generated a specific AnnA1 antibody that targets caveolae in the tumor endothelium. Intravital microscopy of caveolae-immunotargeted fluorophores even at low intravenous doses showed rapid and robust pumping across the endothelium to enter mammary, prostate and lung tumors. Within 1 h, the fluorescence signal concentrated throughout tumors to exceed the peak levels in blood. This transvascular pumping required the expression of caveolin 1 and annexin A1. Tumor uptake with other antibodies were >100-fold less. This proteomic imaging strategy reveals a unique target, antibody and caveolae pumping system for solid tumor penetration.

submitted to conference record of IEEE NSS-MIC, Anaheim USA, 29 october-3 november 2012; International audience; A 8 channel Front End Electronics (FEE) circuit has been designed and fabricated in 0.35 μm CMOS process from Austria Micro System to be coupled with the Silicon Strip Detector (SSD) of the Compton Camera for quality control of hadrontherapy. Each channel includes a Charge Sensitive Amplifier (CSA) followed by two parallel CR-RC shapers. Slow and fast shapers, with 1 μs and 15 ns shaping time, are used to measure the energy and to time stamp all events respectively. The two sides of the SSD are read thanks to a configurable system for holes and electrons. The CSA presents an open loop gain of 67 dB and 90 degrees phase margin assuring a high stability. The circuit has been successfully tested. The test results are in good agreement with analytic and simulation calculations. Here, we describe the principles and present measured performances of the prototype. A high linearity over the range of 3E3 to 3E6 electrons is reached with a conversion gain of 3.6 mV/fC. The circuit achieves an ENC (Equivalent Noise Charge) of 412 electrons rms. 75% of the total noise is generated by the small value of the feedback resistor chosen to avoid pile up phenomenon due to the 1E5 hits/s occupancy rate. A cross-talk of 2 % was measured, 99% of which is due to the power supply disturbances. The power supply dissipation is 21 mW/channel for 3.3 V supply voltage. The area of this design is 2871×1881 μm2 including pads.

Summary In mammalian cultures, the activation and catalytic activity of both urokinase-type plasminogen activator (u-PA) and tissue-type PA (t-PA) is regulated in part by naturally occurring plasminogen activator inhibitors PAI-1 and PAI-2, members of the serpin family of serine protease inhibitors. Interaction of human PAIs with human u-PA (h-u-PA) and t-PA appears to be mediated by a putative PAI binding sequence in the enzymes. Comparison of the amino acid sequences of human, murine, porcine, bovine, simian and avian u-PA revealed specific sequences in the mammalian enzymes that are homologous to the recently discovered PAI-binding site in human u-PA and t-PA which is characterized by conserved basic amino acids. However, no homology was seen in the same region in chicken u-PA (c-u-PA), which is devoid of the consensus arginine and/or lysine residues. The chicken enzyme is a close structural and catalytic homolog of mammalian u-PA. The lack of the putative PAI binding sequence would predict that c-u-PA cannot interact with and be inhibited by human PAIs. To test this hypothesis, the effects of human PAI-1 and PAI-2 on avian u-PA were examined, using purified enzyme and inhibitors. Under conditions where h-u-PA activity is inhibited by human recombinant PAI-1 (rPAI-1), c-u-PA retains 95% activity. c-u-PA was also resistant to inhibition by human PAI-2. High molecular weight, SDS-stable complexes do not form readily when c-u-PA is incubated with PAI-1 or PAI-2, while human as well as murine u-PA formed complexes with both PAIs under the identical conditions. The c-u-PA molecule, thus, is a naturally occurring u-PA variant which lacks a PAI-binding region. The resistance of this enzyme to inhibition by human PAI-1 and PAI-2 supports the functional relevance of this sequence in mammalian u-PA and t-PA.

International audience; Purpose: Online in vivo control of the ion range in a patient during proton therapy is a major challenge for quality assurance of treatments. After measurements showed that prompt-γ emission is correlated to the ion range (Min et al 2006, Testa et al 2008), prompt-γ imaging emerged as apromising method (Verburg et al 2013). Fast methods are required to compute accurate prompt-γ emission maps to design and predict the camera response from treatment plans. An analytic computation method based on the structure of the dose calculation engines in treatment planning system has recently been proposed (Sterpin et al 2015). An alternative technique based on variance reduction in Monte Carlo (MC) calculations is developed here for computing prompt-γ emission maps in proton therapy.Materials/Methods: The track length estimator (TLE) method is a standard variance reduction technique in voxel-based dose computation in the kerma approximation (Williamson 1987), and similar approaches have also been developed for positron emitter distributions in proton therapy (Parodi et al 2007). A specific track length estimator has been developed here to design a continuous process along the proton track that locally deposits the expected value of the prompt-γ emission (induced by proton inelastic scattering) that would have occurred if a large number of protons with the same incident energy had followed the same step (i.e. track element). First an elemental database of prompt-γ emission spectra is established in the clinical energy range of incident protons for all elements in the composition of human tissues. This database of the prompt-γ spectra is built offline with high statistics. Regarding the implementation of the prompt-γ TLE MC tally, each proton deposits along its track the expectation of the prompt-γ spectra from the database according to the proton kinetic energy and the local material density and composition. All software developments have been carried out with the Gate/Geant4 toolkit.Results: A detailed statistical analysis is reported to characterize the dependency of the variance reduction on the geometrical (track length distribution) and physical (linear prompt-γ spectrum database) parameters. Benchmarking of the proposed technique with respect to an analogous MC technique is carried out. A large relative efficiency gain is reported, ca. 105. Such an efficiency gain could reduce the MC computing time of a full treatment from some weeks to less than one hour. Implementation issues are also addressed.Conclusions: This MC-based technique makes it possible to deal with complex situations such as heterogeneities for which proton straggling and secondary protons may have a decisive contribution. When considering translation to clinic, measurements for the prompt-γ s pectrum d atabase, or at least a sound calibration protocol of the simulated prompt-γ spectra, will have to be carried out.

The hypertensive patient is the most stable phenotype in psychosomatic medicine. Hypertensive patients represent a vulnerable population that deserves special attention from health care providers and systems, and psychosomatic medicine may be an important tool in the management of high blood pressure. Depression, anxiety disorders and personality features are often associated with elevated blood pressure (BP) and they may have a role in the development of mild high-renin essential hypertension. Besides, “white coat” hypertension and “masked” hypertension demonstrate how clinic blood pressure could be strongly related to trait anxiety. Hypertension is largely asymptomatic, and patients often have little understanding of the importance of achieving BP control. Medication adverse effects may become an important factor in poor adherence to the treatment and the antidepressant use increases the risk of hypertension. So, the challenge in the management of hypertensive patients is the adherence to non-pharmacological and behavioural treatments for hypertension.

International audience

Background Renal sympathetic denervation (RDN) is indicated for the treatment of resistant hypertension (RH). The RH is true when the causes of pseudo-hypertension as well as other modifiable causes, e. g secondary hypertension or poor adherence, are excluded. In the clinical practice there are patients who due to polyintolerances or allergies can't be treated with the maximal therapy and therefore are not considered as RH – patients in accordance with the current definition. Since they are also very difficult to treat with medical therapy we also considered them in this study as eligible for renal denervation. The goal of the study is to assess the percentage of eligibility to RDN and analyze the exclusion criteria in a group of resistant hypertensive patients. Methods Between 2011 and 2014 we retrospectively reviewed patients who came to our Center for an evaluation for RDN. We considered patients with office blood pressure ≥160 mmHg and patients with severe hypertension treated with fewer drugs because of poly-intolerances / allergies as eligible. At first a specialist optimized drug therapy and subsequently reassessed the blood pressure response. For each patient a diagnostic examination was conducted in order to rule out secondary hypertension. To exclude a white coat effect the patients each performed an arterial blood pressure monitoring (ABPM). In patients with true resistant hypertension, a CT angiography was performed in order to check the renal anatomic criteria of eligibility to the RDN. Results Out of the 35 participating patients, 27 were considered unsuitable for the RDN for at least one of these causes: blood pressure control with optimization of drug therapy (52%), evidence of white coat effect (22%), secondary hypertension (22%), lack of true resistant hypertension (18%), absence of consent to the procedure (29%). All eight eligible patients were sent by specialist centers while the major part of the unsuitable patients was sent by GPs. Conclusions Before giving the indication for RDN, it is necessary to carefully select patients with RH as most of the patients proved ineligible due to inadequacy of drug therapy and white coat hypertension.

The effect of the addition of 1,4-polyisoprene-1 ,4-polybutadiene (IBR) block copolymers on polymer compatibility and vulcanizate properties of a natural rubber/cis-1,4-polybutadiene rubber blend is studied. Differential scanning calorimetry and transmission electron microscopy are used to determine the degree of polymer compatibility and vulcamzate properties are measured in a sulfur cured, carbon-black filled rubber compound Block copolymer composition, concentration and molecular weight are found to be critical. Improvements in compatibility and vulcamzate properties are observed at low concentrations of IBR block copolymers.

Purpose: The utilization of protons and carbon ions for radiotherapy purposes provides high conformity and increased biological effectiveness. In order to fully benefit from the advantages, a precise beam range monitoring in real time is vital. The detection of prompt gammas with a collimated scintillating detector makes it possible to locate the Bragg peak position during proton or carbon ion irradiation. Accurate Monte Carlo simulations are essential both for the further development of an optimized detection array and for the accurate estimation of the applicability of the system in clinical conditions. Purpose of the current work is to assess and improve the performance of the Geant4 toolkit in the description of currently available prompt-gamma data.

This work presents some reflections on the issue of gender in mental health and occupational therapy practices of two historical situations showing two different but related practices: on one hand aspects linked to the performance of women in the public domain and on the other hand, stand out the implication of gender relations in such action. This paper intent to analyze the impact of those representations of gender in the field of mental health and its relationship with the current practices of occupational therapy. It presents the constituent elements of the concept of gender to identify care logic aimed at responding to the issue of mental health, reproducing the structure of power and domination of gender relations. Intents to include gender mainstreaming in occupational therapy practices wherever instrument interventions, strategies, or devices in health, bearing in mind that the tension between the practice of occupational therapy and the gender system constraints, is always in relation with the particular process of subjectivation of each of the subjects with which intervenes.

This study shows the rapid and differential production of the 40-43 kDa and the 70-90 kDa α1-acid glycoprotein (AGP) fucosylated glycoforms after treatment of the dorsal air pouch with bacterial lipopolysaccharide (LPS), HgCl2 or Freund's complete adjuvant (FCA). The 40-43 kDa and the 70-90 kDa AGP production is peaked 1-3 h post-LPS treatment. We observed that the responses to LPS and FCA are similar in that both AGP isoforms are induced whereas they differ in that the FCA exhibits a 6 h lag period. The response to HgCl2, however, exhibits the specific biphasic induction only of the 40-43 kDa AGP. The serum 40-43 kDa AGP glycoform gradually increases in response to all of the above stimulants and peaks by 24 h post- treatment. The increase of the 70-90 kDa AGP levels in the air pouch occurs in association with the accumulation of polymorphonuclear (PMN) cells while dexamethasone (DEX) increases only the 40-43 kDa AGP production in the absence of PMN accumulation. Macrophage-monocyte lineage cells forming the air pouch lining tissue may potentially be the cells that secrete the 40-43 kDa AGP while polymorphonuclear cells that infiltrate the air pouch secrete the 70-90 kDa AGP. The 40-43 kDa and 70-90 kDa AGP production induced by LPS in the air pouch precedes that of interleukin-1 (IL-1) or interleukin-6 (IL-6) while the 40-43 kDa AGP glycoform potentially increases IL-6 production by air pouch PMN exudate cells. These significant differences suggest a local pro-inflammatory role of AGP. Honeybee venom suppressed arthritis development and exhibited differential local or systemic regulation of AGP in serum vs. air pouch exudate or synovial fluid. This study with the air pouch model of facsimile synovium tissue suggests that local α1-acid glycoprotein (AGP) production may contribute to pro-inflammatory and anti-inflammatory activities during the local acute phase response or during chronic inflammatory stress as in arthritis. © 2010 Elsevier Inc.

We investigated the extent of blood pressure (BP) reduction and control (140/90 mmHg) in patients treated with appropriate or inappropriate drugs according to their plasma renin activity (PRA) level (natriuretic drugs and renin-angiotensin system blockers for low-renin and high-renin hypertension, respectively).One hundred and seventy Caucasian untreated hypertensive patients (61 females), aged 18-70 years, participated to the study. Patients with secondary hypertension, diabetes or established cardiovascular or renal disease were excluded. The physician prescribed an antihypertensive monotherapy chosen among all drug classes, unaware of patient's PRA levels. We compared effect of an inappropriate or appropriate drug, evaluating BP values after a month of treatment.Rate of BP control was not significantly higher in patients treated with an appropriate drug than the others (38% vs 29%, p=0.24). However, in a regression analysis, final diastolic BP (DBP) was lower in subjects treated with an appropriate drug (beta=-2.84, p=0.03).The present study does not clearly support the use of PRA in a general population of hypertensive patients to optimize BP control. However, the greater efficacy of a drug appropriate to PRA in reducing DBP may be clinically helpful in young hypertensive patients. Future studies are warranted to evaluate if PRA determination enhances the therapeutic success in patients with predominantly high values of DBP.

Mapping protein expression of endothelial cells (EC) in vivo is fundamental to understanding cellular function and may yield new tissue-selective targets. We have developed a monoclonal antibody, MAb J120, to a protein expressed primarily in rat lung and heart endothelium. The antigen was identified as CD34, a marker of hematopoietic stem cells and global marker of endothelial cells in human and mouse tissues. PCR-based cloning identified two CD34 variant proteins, full length and truncated, both of which are expressed on luminal endothelial cell plasma membranes (P) isolated from lung. Truncated CD34 predominated in heart P, and neither variant was detected in P from kidney or liver. CD34 in lung was readily accessible to125I-J120 inoculated intravenously, and immunohistochemistry showed strong CD34 expression in lung EC. Few microvessels stained in heart and kidney, and no CD34 was detected in vessels of other organs or in lymphatics. We present herein the first complete sequence of a rat CD34 variant and show for the first time that the encoded truncated variant is endogenously expressed on EC in vivo. We also demonstrate that CD34 expression in rat EC, unlike mouse and human, is restricted in its distribution enabling quite specific lung targeting in vivo.

The level of blood pressure, the type of antihypertensive treatment and the prevalence of resistant hypertension at the first examination were evaluated in 6254 patients referred to a hospital Hypertension Unit from 1989 to 2003. From 1989-1993 to 1999-2003, we observed a reduced prevalence of grade 2 and grade 3 hypertension, and an increase in the prevalence of grade 1 hypertension, the proportion of treated subjects, the average number of antihypertensive drugs per patient and the prevalence of resistant hypertension.

No established second-line chemotherapy is available for patients with advanced gastric cancer failing to respond or progressing to first-line chemotherapy. However, 20-40% of these patients commonly receive second-line chemotherapy. We evaluated the influence of clinico-pathologic factors on the survival of 175 advanced gastric cancer patients, who received second-line chemotherapy at three oncology departments. Univariate and multivariate analyses found five factors which were independently associated with poor overall survival: performance status 2 (hazard ratio (HR), 1.79; 95% CI, 1.16-2.77; P=0.008), haemoglobin/=11.5 g l(-1) (HR, 1.48; 95% CI, 1.06-2.05; P=0.019), CEA level50 ng ml(-1) (HR, 1.86; 95% CI, 1.21-2.88; P=0.004), the presence of greater than or equal to three metastatic sites of disease (HR, 1.72; 95% CI, 1.16-2.53; P=0.006), and time-to-progression under first-line chemotherapy/=6 months (HR, 1.97; 95% CI, 1.39-2.80; P0.0001). A prognostic index was constructed dividing patients into low- (no risk factor), intermediate- (one to two risk factors), or high- (three to five risk factors) risk groups, and median survival times for each group were 12.7 months, 7.1 months, and 3.3 months, respectively (P0.001). In the absence of data deriving from randomised trials, this analysis suggests that some easily available clinical factors may help to select patients with advanced gastric cancer who could derive more benefit from second-line chemotherapy.