Sensitization of ovarian carcinoma cells with zoledronate restores the cytotoxic capacity of Vγ9Vδ2 T cells impaired by the prostaglandin E2 immunosuppressive factor: implications for immunotherapy.

Epithelial ovarian cancer (EOC) usually spreads into the peritoneal cavity, thereby providing an opportunity for intraperitoneal adoptive immunotherapy with Vγ9Vδ2 T lymphocytes, a T cell subpopulation endowed with high lytic properties against tumor cells. However, previous studies have reported that Vγ9Vδ2 T cells fail to expand from peripheral blood mononuclear cells in one-third of patients with cancer. Here, from a cohort of 37 patients with EOC, a multiple correspondence analysis identified three populations, one of which was not suitable for Vγ9Vδ2 T-cell adoptive therapy. Interestingly, the ineligible patients were identified based on the frequency of Vγ9Vδ2 T cells in their peripheral blood and the patients' age. The average time to tumor recurrence was also found to be significantly different between the three populations, suggesting that the innate immune response is involved in EOC prognosis. A dramatic decrease in the lytic properties of Vγ9Vδ2 T cells occurred following incubation with ascitic supernatant and was found to be associated with reduced perforin/granzyme degranulation. Prostaglandin E2, but not IL-6, IL-10, VEGF or TGF-ß, showed immunosuppressive effects in Vγ9Vδ2 T cells. Interestingly, our results emphasize that pretreating ovarian tumor cells with zoledronate partially reverses the immunosuppressive effects of ovarian cancer-associated ascites and restores a high level of lytic activity. These data sustain that optimal Vγ9Vδ2 T-cell adoptive immunotherapy previously requires counteracting the tumor immunosuppressive microenvironment. Altogether, our findings provide a rationale for clinically evaluating Vγ9Vδ2 T-cell adoptive immunotherapy with intraperitoneal carcinomatosis presensitization by zoledronate in patients with EOC.

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 (Delta) 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, DeltaSAP, and DeltadP/dt. ROPI altered ECG variables less than BUPI but more than S-BUPI. LIDO was associated with the smallest changes.

Uncorrelated randomness of the heart rate is associated with sepsis in sick premature infants.

BACKGROUND: Late-onset sepsis in the premature infant is frequently revealed by severe, unusual and recurrent bradycardias. In view of the high morbidity and mortality associated with infection, reliable markers are needed. OBJECTIVES: It was the aim of this study to determine if heart rate (HR) behavior may help the diagnosis of infection in premature infants with such cardiac decelerations. METHODS: Electrocardiogram recordings were collected in 51 premature infants with a postmenstrual age <33 weeks with frequent bradycardias. Newborns in the sepsis group (C-reactive protein increase and positive blood culture) were compared with a no-sepsis group (C-reactive protein <5 mg/l before and 24 h after recording and negative blood cultures) for their HR characteristics, i.e. RR series distribution (mean, median, skewness, kurtosis, sample asymmetry), magnitude of variability in time and frequency domain, fractal exponents (alpha(1), alpha(2)) and complexity measurements (approximate and sample entropy). Results are presented as the median (25%, 75%). RESULTS: Gestational, chronological and postmenstrual age and gender were similar in the sepsis (n = 10) and no-sepsis group (n = 38). Three infants had an increase in C-reactive protein but negative cultures. Low entropy measurements [approximate entropy 0.4 (0.3, 0.5) vs. 0.8 (0.6, 1); p < 0.001] and long-range fractal exponent [alpha(2) 0.78 (0.71, 0.83) vs. 0.92 (0.8, 1.1); p < 0.05] were significantly associated with sepsis. No other HR characteristic was associated with sepsis. The decrease in 0.1 units of approximate entropy was associated with an over 2-fold increase in the odds of sepsis. CONCLUSION: Late-onset sepsis is associated with uncorrelated randomness of the HR. This abnormal HR behavior may help to monitor premature infants presenting with frequent and severe bradycardias.

WISE-2005: effect of aerobic and resistive exercises on orthostatic tolerance during 60 days bed rest in women.

Cardiovascular deconditioning after long duration spaceflight is especially challenging in women who have a lower orthostatic tolerance (OT) compared with men. We hypothesized that an exercise prescription, combining supine aerobic treadmill exercise in a lower body negative pressure (LBNP) chamber followed by 10 min of resting LBNP, three to four times a week, and flywheel resistive training every third day would maintain orthostatic tolerance (OT) in women during a 60-day head-down-tilt bed rest (HDBR). Sixteen women were assigned to two groups (exercise, control). Pre and post HDBR OT was assessed with a tilt/LBNP test until presyncope. OT time (mean +/- SE) decreased from 17.5 +/- 1.0 min to 9.1 +/- 1.5 min (-50 +/- 6%) in control group (P < 0.001) and from 19.3 +/- 1.3 min to 13.0 +/- 1.9 min (-35 +/- 7%) in exercise group (P < 0.001), with no significant difference in OT time between the two groups after HDBR (P = 0.13). Nevertheless, compared with controls post HDBR, exercisers had a lower heart rate during supine rest (mean +/- SE, 71 +/- 3 vs. 85 +/- 4, P < 0.01), a slower increase in heart rate and a slower decrease in stroke volume over the course of tilt/LBNP test (P < 0.05). Blood volume (mean +/- SE) decreased in controls (-9 +/- 2%, P < 0.01) but was maintained in exercisers (-4 +/- 3%, P = 0.17).Our results suggest that the combined exercise countermeasure did not significantly improve OT but protected blood volume and cardiovascular response to sub tolerance levels of orthostatic stress.

Reproducibility of CT-based bone texture parameters of cancellous calf bone samples: influence of slice thickness.

Bone microarchitecture is an important determinant of the fracture risk, independently of bone mineral density. At present, bone biopsy is required for microarchitecture assessment, and accessible non-invasive techniques are needed. In this study, we tested the short-term reproducibility and parameter changes of a non-invasive method for microarchitecture assessment with a medical computed tomography. Texture parameters (run lengths and co-occurrence) were extracted from bone sample images. Reproducibility and the influence of slice thickness (1, 3, 5 and 8mm) were also studied. After five repositionings, short-term reproducibility was found to be good. All run length parameters but one fell significantly with increasing slice thickness. Co-occurrence parameters showed different patterns of change. Short-term coefficients of variation of texture parameters used to assess bone microarchitecture were similar to those obtained elsewhere with other techniques. The results were influenced by slice thicknesses, emphasizing the importance of the conditions of acquisition.

Expression of nine tumour antigens in a series of human glioblastoma multiforme: interest of EGFRvIII, IL-13Ralpha2, 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-13Ralpha2, 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-13Ralpha2 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-13Ralpha2, respectively. Interestingly, protein expression of at least 3 antigens was observed in 38% of cases. These results point out the importance of EGFRvIII, IL-13Ralpha2 and, to a less extent gp100 and TRP-2, for developing an immunotherapy strategy against glioblastoma.

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 beta/alpha anomers (mixture A, 40:60 and mixture B, 80:20) were synthesized or isolated depending on the purification procedure. The beta 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 beta-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.

From anatomic standardization analysis of perfusion SPECT data to perfusion pattern modeling: evidence of functional networks in healthy subjects and temporal lobe epilepsy patients.

RATIONALE AND OBJECTIVES: In the general context of perfusion pattern modeling from single-photon emission computed tomographic (SPECT) data, the purpose of this study is to characterize interindividual functional variability and functional connectivity between anatomic structures in a set of SPECT data acquired from a homogeneous population of subjects. MATERIALS AND METHODS: From volume of interest (VOI)-perfusion measurements performed on anatomically standardized SPECT data, we proposed to use correspondence analysis (CA) and hierarchical clustering (HC) to explore the structure of statistical dependencies among these measurements. The method was applied to study the perfusion pattern in two populations of subjects; namely, SPECT data from 27 healthy subjects and ictal SPECT data from 10 patients with mesio-temporal lobe epilepsy (MTLE). RESULTS: For healthy subjects, anatomic structures showing statistically dependent perfusion patterns were classified into four groups; namely, temporomesial structures, internal structures, posterior structures, and remaining cortex. For patients with MTLE, they were classified as temporomesial structures, surrounding temporal structures, internal structures, and remaining cortex. Anatomic structures of each group showed similar perfusion behavior so that they may be functionally connected and may belong to the same network. Our main result is that the temporal pole and lenticular nucleus seemed to be highly relevant to characterize ictal perfusion in patients with MTLE. This exploratory analysis suggests that a network involving temporal structures, lenticular nucleus, brainstem, and cerebellum seems to be involved during MTLE seizures. CONCLUSION: CA followed by HC is a promising approach to explore brain perfusion patterns from SPECT VOI measurements.

Immunohistochemical analysis of tumor polyamines discriminates high-risk patients undergoing nephrectomy for renal cell carcinoma.

Renal cell carcinoma (RCC) is known to display a wide variation in biological behavior and clinical outcome. Although usual bioclinical prognostic parameters (eg, nuclear grade, tumor stage) are to a certain extent useful in predicting the outcome of RCC after radical nephrectomy, they now appear to be insufficient. The polyamines (spermidine, spermine, and putrescine) are ubiquitous polycations that are essential for cell proliferation. To support their excessive proliferation, cancer cells have high rates of polyamine metabolism. Indeed, malignant cells typically have higher polyamine levels than their normal counterparts. Before this report, antipolyamine antibodies that are potentially valuable tools for the in situ observation of polyamines had not been exploited in clinical conditions. In the present study, tumor tissues obtained from radical nephrectomy performed for RCC (n = 73) were immunostained with the anti-spermine monoclonal antibody Spm8-2, and the immunoreactivity was evaluated as a prognostic tool. RCC cells displayed various reactivity to the antibody Spm8-2 that translated into a heterogeneous cytoplasmic staining. The prognostic value of the labeling index (LI) on clinical outcome was correlated with the usual clinicopathologic parameters, and the cell proliferation rate was evaluated using Ki-67 labeling. Multiple correspondence analysis and ascending hierarchical classification were performed to determine significant prognostic factors. Univariate statistical survival analysis demonstrated that tumor size (P < .001), nuclear grade (P < .01), necrosis (P < .007), tumor stage (P < .004), metastasis (P < .001), Ki-67 LI (P < .0003), and Spm8-2 immunoreactivity (P < .0001) were predictors of tumor-related death. A positive correlation was found between Ki-67 LI and Spm8-2 immunoreactivity (r' = .53). Multivariate analysis revealed that only Ki-67 LI and Spm8-2 immunoreactivity were significant independent factors in patients with metastases (P < .04 and <.001, respectively) and in patients without metastases (P < .006 and <.001, respectively). Moreover, 100% of the patients with Spm8-2 immunoreactivity <10% were alive at the end of the follow-up. In terms of predictive values, Spm8-2 immunoreactivity had the highest predictive values (sensitivity, 89; specificity, 75; risk ratio, 11) of all clinicopathologic parameters. This study demonstrates that the anti-spermine monoclonal antibody Spm8-2 may be used at the time of radical nephrectomy as a reliable prognostic marker for defining RCC patients at high risk for progression.

Comparison of heart rate response to an epinephrine test dose and painful stimulus in children during sevoflurane anesthesia: heart rate variability and beat-to-beat analysis.

BACKGROUND AND OBJECTIVES: During regional anesthesia, various stimuli leading to an adrenergic response can occur. However, simulation of an epidural test dose by using intravenous administration of epinephrine (EPI) has always been compared with an intravenous saline infusion as the control. The aim of this study was to evaluate the possibility of distinguishing in children the effect on HR by an intravascular epinephrine infusion and a painful stimulus, using heart rate variability (HRV) and beat-to-beat analysis of HR. METHODS: Thirty American Society of Anesthesiologists physical status P I children who required elective surgery were studied. At 1 minimum alveolar concentration (MAC) of sevoflurane, electrocardiogram was recorded continuously. Systolic blood pressure (SBP) was measured every minute. Measurements were performed after an intravenous administration of 0.5 microg/kg of epinephrine and during a small skin surgical incision (SI). Time-varying auto-regressive modeling of the interpolated RR sequences was performed for estimating power spectrum (msec(2)). The HF bands were defined by (0.15-0.4 Hz). RESULTS: Median (range) age and weight of all children were 3.5 (1-10) years and 16 (9-30) kg. EPI produced a lower increase in HR than did SI. SBP increased significantly more than did after SI. T-wave amplitude increased significantly after EPI but not after SI. Sixty seconds after the first change in HR, a secondary decrease (in comparison to control value) can be detected with EPI in contrast to SI. HF spectral power increased significantly after EPI administration but decreased after SI. The sensitivity, specificity, and positive and negative predictive value were respectively for DeltaHR >10 beats per minuteof 56%, 26%, 43%, and 38%; for DeltaSBP >15 mm Hg of 60%, 86%, 81%, and 67%; and for DeltaT-wave amplitude >25% of 86%, 73%, 76%, and 84%. Using detection of the secondary decrease of HR, 60 seconds after the first change in HR, sensitivity, specificity, and positive and a negative predictive value were respectively 96%, 100%, 100%, and 96%. CONCLUSIONS: Detection of the secondary HR decrease, 60 seconds after the first change in HR, allows us to distinguish the effects of a painful stimulus from those related to the epinephrine test dose at 1 MAC of sevoflurane. This secondary HR decrease induced by epinephrine appears primarily because of a compensatory increase in parasympathetic tone.

Impedance cardiographic waveforms in children: Involvement in left ventricular ejection time determination during anesthesia.

OBJECTIVE: The physiologic basis of the impedance cardiographic (ICG) signal is still a matter of debate. An accurate determination of left ventricular ejection time (LVET) is needed to calculate stroke volume. In children, several shapes of the ICG signal are observed during anesthesia or in critical care. The aim of this study was to determine the relationship between the shape of the ICG signal and various morphologic and hemodynamic determinants obtained with Doppler echocardiography to highlight the effect of the various shapes in the accuracy of LVET determination. DESIGN: Prospective study. SETTING: Pediatric surgery in a university hospital. PATIENTS: 103 children, ASA physical status I or II. INTERVENTIONS: General anesthesia for elective surgery. Measurements: Electrocardiography, ICG, and Doppler echocardiography were recorded simultaneously. Classic hemodynamic variables, such as heart rate, LVET, stroke volume, or systemic vascular resistance, were measured or calculated. A mathematical model of the ICG signal was used to determine the shape of the ICG signal and its potential relationship to various morphologic and hemodynamic determinants. The ICG signal shape was then modeled with the normalized amplitudes (NHA) and phase (NHP) of the three harmonics of the fundamental signal. Analysis was performed with a multiple correspondence analysis. RESULTS: ICG method LVET did not significantly correlate with LVET determined with Doppler or with heart rate. NHA and NHP of the ICG signal were not significantly related to LVET determined with Doppler or to stroke volume. NHA were linked to heart rate and the relative length of the cardiac diastolic period. CONCLUSION: The absence of relationship between LVET determined with Doppler and the main variables defining the shape of the ICG signal highlight the relative limit of stroke volume determination with ICG method. The involvement of the arterial compliance in the ICG signal shape can be discussed in regard to the links between NHA and both heart rate and the relative length of the cardiac diastolic period, respectively.