Gene expression profiling in peripheral blood cells of patients with rheumatoid arthritis in response to anti-TNF-alpha treatments.

The efficacy of anti-TNF-α therapies highlights the role of TNF-α in the pathogenesis of rheumatoid arthritis (RA). However, the mechanism of action of these agents is poorly understood at the molecular level. The aim of this study was to characterize the effects of anti-TNF-α treatment on the global gene expression profile in peripheral blood mononuclear cells (PBMCs) of responder RA patients. Changes in gene expression were determined using oligonucleotide microarrays (25,341 genes) in PBMCs obtained before and after 12 wk of treatment with either etanercept or adalimumab from responder RA patients. Two hundred fifty-one genes displayed significant changes (false discovery rate < 0.1%) in expression level (178 upregulations with mean fold change = 1.5 and 73 downregulations with mean fold change = -1.50) after 12 wk of treatment. Importantly, the expression of several genes, including those coding for the calcium binding proteins S100A12 and A8, CD14 antigen, Selectin P, or ribosomal protein L39, reported to be upregulated in RA patients, were found to be decreased after anti-TNF-α treatment. Globally, inflammation, immune response, apoptosis, protein synthesis, and mitochondrial oxido-reduction were the most affected pathways in response to anti-TNF-α treatment. The obtained gene expression signature in PBMCs provides new information to better understand the mechanisms of action of anti-TNF-α treatment in RA patients.

The Fourier transform spectrometer of the Université Pierre et Marie Curie QualAir platform.

A Bruker Optics IFS 125HR Fourier transform spectrometer (FTS) and the Laboratoire de Physique Moléculaire pour l'Atmosphère et l'Astrophysique retrieval algorithm were adapted for ground based atmospheric measurements. As one of the major instruments of the experimental research platform QualAir, this FTS is dedicated to study the urban air composition of large megacity such as Paris. The precise concentration measurements of the most important atmospheric pollutants are a key to improve the understanding and modeling of urban air pollution processes. Located in the center of Paris, this remote sensing spectrometer enables to monitor many pollutants. Examples for NO(2) and CO are demonstrating the performances of this new experimental setup.

Peripheral blood lymphocytes from patients with rheumatoid arthritis are differentially sensitive to apoptosis induced by anti-tumour necrosis factor-alpha therapy.

OBJECTIVE: The efficacy of anti-tumour necrosis factor-alpha (TNF-alpha) therapies in rheumatoid arthritis (RA) has been mainly attributed to TNF-alpha neutralisation. Other mechanism as immune cell apoptosis, which is impaired in RA, may also be induced by anti-TNF-alpha therapies. The aim of our study was to investigate whether TNF-alpha inhibitors could induce apoptosis in vitro of the peripheral blood lymphocytes of RA patients. METHODS: Peripheral blood mononuclear cells (PBMC) isolated from 24 patients with RA and 18 healthy donors were incubated with anti-TNF-alpha agents, infliximab or etanercept, in comparison with no agent and including an isotypic control, for 48 hours. Apoptosis was detected and quantified by annexin V labelling of phosphatidylserine externalization using cytofluorometric analysis and compared with PBMC production TNF-alpha in vitro. RESULTS: In healthy donors, induced apoptosis was observed in 0.3% to 3.8% of lymphocytes with both therapies. In RA patients the treatment induced lymphocyte apoptosis in 17 of 24 patients with a percentage of annexin V-positive lymphocytes ranging from 0.1% to 25%. Among these 17 RA patients, a significant in vitro lymphocyte apoptosis (> 4%) was observed in 11 patients (46%) compared with healthy donors (p < 0.01). The variability of the response to anti-TNF-alpha within the RA population was not dependent on TNF-alpha synthesis or disease activity. CONCLUSION: In vitro induction of lymphocyte apoptosis by anti-TNF-alpha was observed in a subgroup of RA patients. Based on these data, it would be of interest to further study the interindividual variations of sensitivity to apoptosis induced by TNF alpha inhibitors in relation to treatment efficacy or resistance observed in RA patients.

Development of a stabilized low temperature infrared absorption cell for use in low temperature and collisional cooling experiments.

We have constructed a stabilized low temperature infrared absorption cell cooled by an open cycle refrigerator, which can run with liquid nitrogen from 250 to 80K or with liquid helium from 80K to a few kelvin. Several CO infrared spectra were recorded at low temperature using a tunable diode laser spectrometer. These spectra were analyzed taking into account the detailed effects of collisions on the line profile when the pressure increases. We also recorded spectra at very low pressure to accurately model the diode laser emission. Spectra of the R(2) line in the fundamental band of 13CO cooled by collisions with helium buffer gas at 10.5K and at pressures near 1 Torr have been recorded. The He-pressure broadening parameter (gamma(0) = 0.3 cm(-1) atm(-1)) has been derived from the simultaneous analysis of four spectra at different pressures.

Diode laser emission linewidth determination: application to H2O line profile studies in the 5 and 1.4 microm regions.

In order to study absorption line profiles using the stabilized diode laser spectrometer of Laboratoire de Physique Moléculaire et Applications (LPMA), a reliable determination of the emission line shape of different diodes laser is needed. In the near infrared region (1.39 and 1.66 microm) we used Distributed Feed Back diode lasers which operate around room temperature and in the middle infrared (5 and 8 microm) we used lead salt diode lasers cooled in a helium closed cycle cryostat or in a liquid nitrogen dewar. Some results obtained in H2O line profile studies in the 1.39 and 5 microm regions are presented as examples demonstrating how absorption line profile measurements can lead to erroneous values of the spectroscopic parameters when the contribution of the diode laser emission line width is neglected.

Pressure broadening study at low temperature: application to methane.

We studied the R(0) line profile in the CH4 v4 band from room temperature to 188 K with N2 as a perturber, to 100 K with O2 as perturber, and from room temperature to 15 K using He as a perturber. The N2 and O2 measurements were performed over a total pressure range of 15-110 mbar, and for the He measurements the maximum sample pressure at 15 K was 1.10 mbar. Broadening parameters were determined, taking into account the confinement narrowing (Dicke effect), and narrowing parameters, deduced from the soft or hard collision model, are compared with the dynamic friction coefficient calculated values. We also obtained preliminary values for the temperature dependence of the N2, O2 and He line broadening parameters for this transition.

High-Lying Rotational Levels of Water: An Analysis of the Energy Levels of the Five First Vibrational States.

As a continuation of the work carried out on the ground and (010) vibrational states of water (R. Lanquetin, L. H. Coudert, and C. Camy-Peyret, 1999, J. Mol. Spectrosc. 195, 54-57), rotational energy levels for these two states are revisited here and new accurate rotational energy levels are considered for the three next vibrational states, that is, the (020), (100), and (001) states. Experimental rotational energies, along with their uncertainties, are retrieved through analyses of already published data sets and of discharge and flame emission spectra. The maximum value of J for the obtained levels is 25 for the ground state, 21 for the (010) state, and 20 for the three next states. Based on the bending-rotation Hamiltonian approach (L. H. Coudert, 1997, J. Mol. Spectrosc. 181, 246-273), a new theoretical approach is proposed to calculate rotational energies in the five interacting vibrational states under consideration and is used to carry out an analysis of the experimental energies. Comparisons with other existing energy level data sets are also presented. Copyright 2001 Academic Press.

Absolute Intensities of the nu(1) and nu(3) Bands of (16)O(3).

New experimental data on the nu(1) and nu(3) bands of (16)O(3) improving the value of absolute line intensities have been obtained. The intensities of 295 lines have been measured with an average accuracy between 2.5% and 3% and the rotational expansion of the transition moment operators for the nu(1) and nu(3) bands has been deduced. Finally, a complete listing of line intensities has been computed with an intensity cutoff of 1x10(-25) cm(-1)/molecule cm(-2). Copyright 2001 Academic Press.

High-Order Resonances in the Water Molecule.

The water-vapor spectra in the near-infrared and visible region were reanalyzed with the purpose of finding experimental evidences of unusual high-order resonance between "dark" high-bending and "bright" stretch vibration states. About 70 transitions to the (050), (060), (070), (080), (160), (061), (170), (071), and, even (0 10 0) bending states, and their resonating partners were assigned in the spectra that gives the experimental energy levels lying near or above the potential energy barrier to linearity. The assignments were confirmed by combination differences and simultaneous observation of both perturbed and perturbing levels. It was found that the high-order resonances with large changing of vibration quantum numbers are typical for the water molecule and they are caused by the strong centrifugal distortion near the linear configuration. These resonances destroy the usual polyad scheme originating from well-known Coriolis, Darling-Dennison, and Fermi resonances in H(2)O molecule. Copyright 2001 Academic Press.

Labile conjugation of a hydrophilic drug to PLA oligomers to modify a drug delivery system: cephradin in a PLAGA matrix.

The physical entrapment of a hydrophilic drug within degradable microspheres is generally difficult because of poor entrapment yield and/or fast release, depending on the microsphere fabrication method. In order to counter the effects of drug hydrophilicity, it is proposed to covalently attach the drug to lactic acid oligomers, with the aim of achieving temporary hydrophobization and slower release controlled by the separation of the drug from the degradable link within the polymer matrix. This strategy was tested on microspheres of the antibiotic cephradin. As the prodrug form, the entrapment of the drug was almost quantitative. The prodrug did degrade in an aqueous medium, modelling body fluids, but cleavage did not occur at the drug-oligomer junction and drug molecules bearing two lactyl residual units were released. When the prodrug is entrapped within a PLAGA matrix, no release was observed within the experimental time period. However, data suggest that conjugation via a bond more sensitive to hydrolysis than the main chain PLA ester bonds should make the system work as desired.

The nu(1) and nu(3) Bands of the (17)O(16)O(17)O Isotopomer of Ozone.

Using 0.002 cm(-1) resolution Fourier transform absorption spectra of an (17)O-enriched ozone sample, an extensive analysis of the nu(3) band together with a partial identification of the nu(1) band of the (17)O(16)O(17)O isotopomer of ozone has been performed for the first time. As for other C(2v)-type ozone isotopomers [J.-M. Flaud and R. Bacis, Spectrochim. Acta, Part A 54, 3-16 (1998)], the (001) rotational levels are involved in a Coriolis-type resonance with the levels of the (100) vibrational state. The experimental rotational levels of the (001) and (100) vibrational states have been satisfactorily reproduced using a Hamiltonian matrix which takes into account the observed rovibrational resonances. In this way precise vibrational energies and rotational and coupling constants were deduced and the following band centers nu(0)(nu(3)) = 1030.0946 cm(-1) and nu(0)(nu(1)) = 1086.7490 cm(-1) were obtained for the nu(3) and nu(1) bands, respectively. Copyright 2000 Academic Press.

High-Lying Rotational Levels of Water: Comparison of Calculated and Experimental Energy Levels for (000) and (010) up to J = 25 and 21.

Emission spectra of methane-oxygen low-pressure flames have been recorded at a resolution of 0.02 cm-1 with an infrared Fourier transform spectrometer in the spectral ranges 780-1370 and 1800-5000 cm-1. The flame temperature was about 1850 K and a large number of transitions involving J values as high as 34 for an extended set of vibrational states could be assigned. Combined with already published data sets on H2O, our line position analysis yielded rotational energy levels for many of these states, but only the results relevant to the ground and the (010) states are presented here. The experimental energies for these two states have been fitted with the help of the bending-rotation Hamiltonian approach [L. H. Coudert, J. Mol. Spectrosc. 181, 246-273 (1997)], and for each rotational level, the calculated energy along with its uncertainty is reported and compared with the observed value. Comparisons with other available energy level data sets for the ground and (010) states are also presented. Copyright 1999 Academic Press.

Cephradin-plaga microspheres for sustained delivery to cattle.

In the field of controlled drug delivery, most of the reported work is aimed at introducing new systems, or at providing basic information on the critical parameters which affect release profiles in vitro and occasionally in vivo. The situation is totally different when one wants to fulfil the specific requirements imposed by the marketing of a sustained release device to be used in humans or in animals eaten by human beings. The control of the release characteristics is then a difficult challenge. In this work, attempts were made to combine cephradin, a hydrophilic beta-lactam antibiotic, and bioresorbable polymeric matrices of a poly(alpha-hydroxy acid) in the form of microspheres with the aim of delivering the antibiotic to cattle at a dose rate of 4-5 mg/kg/day over a 3-4 days period after i.m. injection. PLAGA aliphatic polyesters were selected because they are already FDA approved as matrices. The solvent evaporation technique using PVA as the emulsion stabilizer was selected because it is efficient and can be extended to an industrial scale. Various experimental conditions were used in order to obtain the highest encapsulation yields compatible with the desired specifications. Decreasing the volume of the aqueous phase and adding a water-miscible organic solvent/non-solvent of cephradin failed. In contrast, microspheres containing up to 30% cephradin were prepared after addition of sodium chloride to the aqueous dispersing phase. The amount of entrapped drug was raised to 40% by decreasing the temperature and the pressure. Preliminary investigations using dogs showed that 20% cephradin microspheres prepared under these conditions extended the presence of cephradin in the blood circulation up to 48 h. Increasing the load led to higher blood concentrations but shorter sustained release. The fact that the microspheres were for cattle limited the volume of the injection and thus the amount of microspheres to be administered. The other limiting factors were related to microsphere morphology.

Detector Nonlinearity Correction Scheme for the LPMA Balloonborne Fourier Transform Spectrometer.

The Limb Profile Monitor of the Atmosphere (LPMA) instrument is a Fourier transform spectrometer designed to record stratospheric (and in some cases tropospheric) absorption spectra from a balloon gondola. This spectrometer operates with two-detector output optics (photoconductive HgCdTe and photovoltaic InSb, liquid-nitrogen cooled). The response of the HgCdTe detector becomes nonlinear for high photon fluxes, which is the case for solar occultation. We have designed a processing scheme, based on the minimization of out-of-optical-band spectral artifacts, to correct for the effect of nonlinearity in the useful spectral range. The method is explained, and sample results are presented for spectra recorded in different balloon flight conditions and with two different HgCdTe detectors.

The Water Vapor Linestrengths between 11 600 and 12 750 cm-1

The water vapor linestrengths in the region of the 3nu + delta resonance polyad of interacting vibrational states (the corresponding upper states are (310), (211), (112), (013), (131), (230), (032), and (051)) have been analyzed leading to accurate dipole moment transition parameters. The effective rotational Hamiltonian constants used to calculate the vibration-rotation wavefunctions (J.-M. Flaud, C. Camy-Peyret, A. Bykov, O. Naumenko, T. Petrova, A. Scherbakov, L. Sinitsa, 1994. J. Mol. Spectrosc. 183, 300-309) take into account both strong centrifugal distortion effects and dark states presence. These effects are known to be important for the highly excited vibrational states of water-like molecules. The input data set included the line intensities measured by Toth (R. Toth, 1994. J. Mol. Spectrosc. 166, 176-183) and the line intensities of the weak bands 2nu1 + 3nu2, 3nu2 + 2nu3, and 3nu1 + nu2 derived from peak absorptions of a spectrum recorded at a pressure of 17.0 Torr and a path length of 434 m. The parameters of the effective dipole moment operator determined by least square fitting give a very satisfactory agreement with experimental values since the mean error for the 876 experimental linestrengths is only 3.9%. It is worth noticing that such an agreement could be reached only because high-order resonance couplings with dark states were explicitly taken into account. Copyright 1997 Academic Press. Copyright 1997Academic Press

The High-Resolution Spectrum of Water Vapor between 11 600 and 12 750 cm-1

The absorption spectrum of water vapor has been recorded between 11 600 and 12 750 cm-1 with a Fourier transform spectrometer (Kitt Peak, Az) at a resolution of 0.012 cm-1 and with a path length of 434 m. The line assignment has led to the determination of 506 accurate energy levels of the (310) (211), (112), (013), (230), (131), (032), and (051) vibrational states which belong to the so-called 3nu + delta resonance polyad. The rotational energy levels obtained are on the average in agreement with those reported recently by R. Toth (J. Mol. Spectrosc. 166, 176-183 (1994)) for the strong bands, but there are differences for high J levels or weak bands levels (about 15% of all levels). The experimental rotational energy levels have been fitted using Pade-Borel approximants and a set of 104 vibrational energies and rotational, resonance, and centrifugal distortion constants for the (310), (211), (112), (013), (230), (131), (032), and (051) vibrational states have been determined.

[Limitations of ultrasonography in the detection of small cancers of the left kidney]. / Limites de l'échographie dans la détection des petits cancers du rein gauche.

Renal ultrasound seems to be more difficult on the left than on the right side. In a retrospective analysis of 343 patients in whom a radical nephrectomy for renal tumor diagnosed by abdominal ultrasonography had been carried out, we analyzed the characteristics of these tumors. TNM classification was used. Right kidney tumors (n = 170) were smaller than the left ones (n = 172), 58 +/- 31.7 mm and 66.8 +/- 31.5 mm respectively, the difference being statistically significant (p = 0.01). Right kidney tumors of a less advanced stage (p < 0.05). Asymptomatic renal tumors were found more frequently on the right side (58.7%) than on the left (41.3%) (p = 0.03) and were smaller on the right (44 mm +/- 19.6 vs 54.5 mm +/- 27.2) (p = 0.04). In conclusion, Ultrasound exploration of the left kidney seems to be more difficult, mostly because the exploration of the right kidney is facilitated by the acoustic window of the liver.

[CD4 TIL (Tumor Infiltrating Lymphocytes) induce complete response in patients treated with IL-2 (Interleukin-2). Preliminary study]. / Les TIL (Tumor Infiltrating Lymphocytes) CD4 induisent des résponses complètes chez des patients traités par IL-2 (Interleukin-2). Etude préliminaire.

This study analyzed clinical response induced by TIL in patients with renal cell carcinoma previously treated by interleukin-2. Six patients (4 men, 2 women, mean age 44.3 years) with measurable metastatic localizations have been treated by TIL reinjection (0.02 to 7.6 x 10(10) cells). TIL phenotype was a combination of CD4 and CD8 in 3 patients, predominantly CD4 in two patients and predominantly CD8 in one patient. Previous treatment by interleukin-2 induced one partial response, 2 stabilizations of the disease and 3 tumoral progressions. TIL led to an amelioration for 4 patients: 2 were in complete response, 2 were stabilized and 2 had tumoral progression and decreased. This study shows that CD4 TIL may improve an initially response induced by IL-2 therapy.