Jean-Yves Bansard

Expression of nine tumour antigens in a series of human glioblastoma multiforme: interest of EGFRvIII, IL-13Rα2, gp100 and TRP-2 for immunotherapy

In this study, we investigated the mRNA and protein expression of nine tumour antigens in human glioblastoma multiforme with a view to their possible use in dendritic cell-based immunotherapy. Expression of ALK, EGFRvIII, GALT3, gp100, IL-13Rα2, MAGE-A3, NA17-A, TRP-2 and tyrosinase were studied by real-time RT-PCR on frozen tissues using a series of 47 tumour samples from patients with glioblastoma. Results were compared with non-neoplastic brain expression or glioblastoma samples with very low levels of expression near the limits of detection for EGFRvIII and MAGE-A3, as these latter two antigens were not detected in non-neoplastic brain. Tumour antigens showing a 5-fold increase in mRNA expression were considered as positive, and only antigens displaying an mRNA over-expression in a significant number of cases were analysed by immunohistochemistry on paraffin-embedded sections. Using real time RT-PCR, we found EGFRvIII, gp100, IL-13Rα2 and TRP-2 to be positive in 64, 38, 32 and 21% of cases, respectively. While we observed no over-expression for ALK, GALT3 and tyrosinase, 3 samples out of 47 were positive for MAGE-3 and 1 sample for NA17-A. More than 25% of tumour cells showed strong protein expression in 13, 34, 85 and 96% of GBM samples for gp100, TRP-2, EGFRvIII and IL-13Rα2, respectively. Interestingly, protein expression of at least 3 antigens was observed in 38% of cases. These results point out the importance of EGFRvIII, IL-13Rα2 and, to a less extent gp100 and TRP-2, for developing an immunotherapy strategy against glioblastoma.

Value of immunohistochemical Ki-67 and p53 determinations as predictive factors of outcome in renal cell carcinoma☆

OBJECTIVES Nuclear grade and tumor stage are important prognostic factors in renal cell carcinoma, but tumors of similar stage and grade can exhibit a wide variation in biologic behavior and clinical outcome. In this retrospective study, we evaluated the immunologic markers, Ki-67 (MIB1) and p53, in 73 cases of conventional (clear cell) renal cell carcinoma and compared these markers with the accepted prognostic features of grade, stage, and tumor size in predicting outcome. METHODS Specimens of 73 renal cell carcinomas of different nuclear grade (20 Furhman I/II, 32 Fuhrman III, and 21 Fuhrman IV) and different stage (10 pT1, 23 pT2, 36 pT3, and 4 pT4) were immunostained with monoclonal antibodies against Ki-67 and p53. RESULTS Univariate statistical analysis showed that tumor size (P <0. 001), nuclear grade (P <0.01), tumor stage (P <0.01), Ki-67 index (P <0.001), and p53 immunostaining (P <0.03) correlated significantly with a poor prognosis. A Ki-67 index of 20% was a powerful predictor of survival in all patients (P <0.00001), with strong predictive values. On multivariate analysis, the Ki-67 index and metastases were significant independent prognostic factors (P <0.02 and <0.01, respectively). CONCLUSIONS Ki-67 immunostaining appeared to be an additional prognostic indicator of biologic aggressiveness in renal cell carcinoma. Immunohistochemical assessment of tumor antigens could be used to identify patients at high risk of tumor progression in addition to conventional prognostic factors.

Effect of remifentanil with and without atropine on heart rate variability and RR interval in children.

Remifentanil can cause bradycardia either by parasympathetic activation or by other negative chronotropic effects. The high frequency (HF) component of heart rate variability (HRV) is a marker of parasympathetic activity. This study aimed to evaluate the effect of remifentanil on RR interval and on HRV in children. Forty children ASA I or II were studied after approval by the human studies committee and informed parental consent was obtained. After stabilisation at sevoflurane 1 MAC, they were randomly divided into two groups: one received a 20 μg.kg−1 atropine injection (AT + REMI) and the other ringer lactate solution (REMI). Three minutes later, a 1 μg.kg−1 bolus of remifentanil was administered over 1 min, followed by a continual infusion at 0.25 μg.kg−1.min−1 for 10 min increased to 0.5 μg.kg−1.min−1 for a further 10 min. A time varying, autoregressive analysis of RR sequences was used to estimate classical spectral parameters: low (0.04–0.15 Hz; LF) and high (0.15–0.45 Hz; HF) frequency, whereas the root mean square of successive differences of RR intervals (rmssd) was derived directly from the temporal sequence. Statistical analyses were conducted by means of the multiple correspondence analysis and with non parametrical tests. Remifentanil induced an RR interval lengthening, i.e. bradycardia, in both groups compared to pretreatment values and was associated with an increase of HF and rmssd only for the REMI group. The parasympathetic inhibition by atropine did not totally prevent remifentanils negative chronotropic effect. A direct negative chronotropic effect of remifentanil is proposed.

Heterogeneous distribution of polyamines in temporal lobe epilepsy.

Polyamine contents were determined in human temporal lobe epilepsy. In the seven patients studied, stereoelectroencephalography (SEEG) located the epileptogenic focus in Ammons horn and neuropathological findings were limited to hippocampal gliosis and sclerosis. Each polyamine exhibited a specific regional distribution. The most important variations were observed for spermidine and spermine while putrescine levels varied less. The regional variation was predominant in middle > posterior > anterior parts of the temporal lobe. Spermine contents and the spermidine/spermine (SPD/SPM) index varied especially in the middle and posterior parts of the hippocampus. Metabolic SPD/SPM index and spermidine levels were found to be drastically increased in almost all limbic parts when compared to neocortical regions. The opposite was observed for spermine. The heterogeneous distribution of polyamines was compared to abnormal electrical activities recorded by SEEG: SPD/SPM index and spermidine levels were sharply increased in seizure onset areas and high levels of spermine were detected in temporal cortex propagation areas. The presently reported heterogeneity of polyamine contents might contribute to modulate differentially the local control of excitability in human temporal epilepsy.

Erythrocyte polyamines and prognosis in stage D2 prostatic carcinoma patients.

We studied 43 patients with newly diagnosed, untreated, stage D2 prostatic carcinoma, and correlated the initial performance status, hemoglobin, prostate specific antigen levels, tumor Gleason grade, extent of disease on the bone scan, and erythrocyte spermidine and spermine levels with progression. Three patients died of unrelated causes and were excluded from the study, 16 remained in remission with a mean 28 +/- 11 months of followup and 24 had progression (18, or 75%, of whom died of the cancer) with a mean 12 +/- 9 months of followup (p < 0.05 for followup) after initiation of hormonal therapy. Pretreatment performance status, hemoglobin, and erythrocyte spermidine and spermine levels were correlated with progression, hemoglobin and spermine being the most significant independent variables (p = 0.006 and p = 0.001, respectively). Concerning cause-specific survival, only hemoglobin and spermine erythrocyte levels were significant independent variables (p = 0.02 and p = 0.0025, respectively). If confirmed, polyamine erythrocyte levels obtained by a simple blood sample could discriminate at diagnosis patients with a high risk of rapid hormonal relapse who may benefit from a more aggressive primary management.

Polyamine deprivation provokes an antalgic effect

It is well established that inhibition of putrescine formation using D,L-2-(difluoromethyl)ornithine and feeding a polyamine-deficient diet together with non-absorbable antibiotics (neomycin and metronidazole), prevent almost completely the growth of tumors in rats. A similar regimen given to patients with prostate cancer not only reduced the titer of prostate specific antigen in serum, but surprisingly provoked at the same time an antalgic effect. This observation led us to study the potentiation effect of polyamine deprivation on pain threshold in healthy rats. Animals were fed for 2 weeks with an artificial diet of known polyamine content, in combination with antibiotics and 2-(difluoromethyl)ornithine, and were then submitted to pain stimuli using two models, the Randall-Selitto test and the Tail-Flick test. Polyamine deprivation produced in these models an increase in the latency of the response, even under conditions which did not produce significant changes of the polyamine concentrations in blood and brain. From these observations, we may conclude that the polyamines play a role in the perception of nociceptive stimuli under physiological conditions.

In Vivo Antitumor Activity of Clitocine, an Exocyclic Amino Nucleoside Isolated from Lepista inversa

A biologically guided fractionation from Lepista inversa (Scop.: Fr.) led to the isolation of clitocine, an exocyclic amino nucleoside. This compound and two mixtures of β/α anomers (mixture A, 40:60 and mixture B, 80:20) were synthesized or isolated depending on the purification procedure. The β anomer and clitocine mixtures A and B showed similar cytotoxic activities with IC50 values ranging from 20.5 to 42 nM in murine cancer cell lines (3LL and L1210) and from 185 to 578 nM in human cancer cell lines (DU145, K‐562, MCF7, and U251). An in vivo study of mixture B was carried out on 3LL‐ and L1210‐tumor‐bearing mice. Clitocine solubilized in β‐hydroxypropylcyclodextrin and injected at concentrations of 0.5, 3, and 5 mg kg−1 did not significantly increase the survival rate and lifespan of 3LL‐tumor‐bearing mice. In contrast, clitocine showed antitumor activity on L1210‐tumor‐bearing mice with a significant increase in lifespan and a decrease in the development of ascites observed at 3 mg kg−1. The induction of apoptosis may be the basis of the antitumor activity of clitocine against L1210 as suggested by flow‐cytometry analysis of cells treated in vitro.

Electrocardiographic and hemodynamic effects of intravenous infusion of bupivacaine, ropivacaine, levobupivacaine, and lidocaine in anesthetized ewes.

Background and Objectives: Neural blockade techniques are associated with a risk of acute cardiac toxicity after accidental intravenous (IV) injection of local anesthetics. The aim of this study was to compare electrocardiographic (ECG) and hemodynamic (HEM) effects induced by IV infusion of local anesthetics in an anesthetized ewe model. Methods: Thirty-two anesthetized ewes received IV bupivacaine (BUPI), ropivacaine (ROPI), or levobupivacaine (S-BUPI) at an equimolar dose, or lidocaine (LIDO) at a 3-fold higher rate (n = 8 in each group). RR, PR, QRS, and QT intervals (QTc), changes (&Dgr;) in systolic and diastolic arterial pressure (SAP and DAP), and in myocardial contractility (dP/dt), were assessed every 30 seconds for 7 minutes. From main ECG variables (RR, PR, QRS, QT), we proposed to use multiple correspondence analysis and hierarchical ascending classification to explore the structure of statistical dependencies among those measurements, and to determine the different patterns of ECG and HEM changes induced by infusion of BUPI, ROPI, S-BUPI, and LIDO. Results: Graphic representation of multiple correspondence analysis associated BUPI with the most pronounced modifications in ECG and HEM variables, followed by in decreasing order ROPI, S-BUPI, and LIDO. Comparisons of clusters identified by hierarchical ascending classification confirmed this classification for ECG variables. Ventricular tachycardia occurred only in the BUPI group. Conclusions: In our anesthetized ewe model, high dose IV infusion of BUPI induced the most marked changes in RR, PR, QRS, QT, QTc intervals, &Dgr;SAP, and &Dgr;dP/dt. ROPI altered ECG variables less than BUPI but more than S-BUPI. LIDO was associated with the smallest changes.

Erythrocyte spermine levels: a prognostic parameter in childhood common acute lymphoblastic leukemia.

Polyamines have been implicated to play a role in cell proliferation and in cancer development. Ninety percent of the circulating spermidine (Spd) and spermine (Spm) are transported by red blood cells (RBC). RBC Spd and Spm levels were prospectively determined in 63 unselected children with common acute lymphoblastic leukemia. The Spm and Spd levels were not correlated with white blood cell (WBC) count. On the basis of the polyamine levels it was possible to discriminate four groups with P < 10−3. In C1, C2, C3 and C4 group the Spm level was respectively 90 (39–597), 3.75 (1–7.45), 9.95 (2.9–12.6) and 17(6.3–33.8). The probability of relapse-free survival (RFS) of the 58 children who entered complete remission was 55% ± 9. For the groups C1 (n = 6), C2 (n = 16), C3 (n = 21) and C4 (n = 15) groups, the RFS was 25% ± 20, 73% ± 12, 73% ± 13 and 32% ± 13 respectively. For children with Spm levels <13/⩾13 nmol/8 × 109 RBC, event-free survival (EFS) was 54% ± 11/33% ± 10 and RFS was 64% ± 12/38% ± 11 respectively (P < 0.03, P < 0.005). our clinical study shows clearly that an rbc spermine level could be used as parameter of prognosis at the time of diagnosis, particularly for patients with intermediary wbc count.

An analysis of IEEE publications

This study reports on a few elements of the scientific production of the IEEE. Other features could be displayed that would be interesting for a better understanding of the trajectories of the societies, journals, etc. The possibility of projecting new data onto the current spaces allows researchers to see if journals are static (the concepts and methods remaining stable) or dynamic (evolutions, ruptures can be tracked). In other words, this type of analysis can be used as a strategic tool to follow the impact and trends in engineering sciences. The fact that the authors concentrate on the IEEE publications prohibits any comparison with other societies publishing engineering papers. Such insights are feasible through the analysis of the INSPEC database. This could bring other clues on the coverage, the competition, and the reaction to new areas.