Efficacy of early cognitive-linguistic treatment and communicative treatment in aphasia after stroke: a randomised controlled trial (RATS-2).

BACKGROUND: The two main approaches in aphasia treatment are cognitive-linguistic treatment (CLT), aimed at restoring the linguistic levels affected, semantics, phonology or syntax, and communicative treatment, aimed at optimising information transfer by training compensatory strategies and use of residual language skills. The hypothesis that CLT is more effective than communicative treatment in the early stages after stroke was tested in this study. METHODS: In this multicentre, randomised, parallel group trial with blinded outcome assessment, 80 patients with aphasia after stroke were included within 3 weeks post-stroke. Patients received 6 months of CLT, comprising semantic and/or phonological training, or communicative treatment for at least 2 h per week. They were assessed before treatment and at 3 and 6 months with the Amsterdam-Nijmegen Everyday Language Test (ANELT-A, primary outcome) and semantic and phonological tests (secondary outcomes). The intervention effect was evaluated by means of analysis of covariance, with adjustment for baseline scores. RESULTS: There was no difference between the mean ANELT-A score of the CLT group (n=38) and the communicative treatment group (n=42), at 3 months (adjusted difference 1.5, 95% CI -2.6 to 5.6) or at 6 months (adjusted difference 1.6, 95% CI -2.3 to 5.6) post-stroke. On two of six specific semantic and phonological tests, the mean scores differed significantly, both in favour of CLT. CONCLUSION: This study does not confirm the hypothesis that patients with aphasia after stroke benefit more from CLT, aimed at activation of the underlying semantic and phonologic processes, than from general, non-specific communicative treatment (ISRCTN67723958 Current Controlled Trials).

Oxytocin- and vasopressin-induced growth of human small-cell lung cancer is mediated by the mitogen-activated protein kinase pathway.

Malignant growth of small-cell lung carcinoma is promoted by various neuroendocrine autocrine/paracrine loops. Therefore, to interfere with this mitogenic process, it is crucial to elucidate the mechanisms involved. It is known that the oxytocin (OT) and vasopressin (VP) genes, normally transcriptionally restricted in their expression, are activated in small-cell lung cancer (SCLC), concomitantly with expression of their receptors (OTR, V1aR, V1bR/V3R and V2R). The aim of the present study was to characterize, in concentrations close to physiological and pharmacological conditions, intracellular signalling events triggered by OT and VP binding to their specific receptors in SCLC cells and to identify factors mediating OT- and VP-induced mitogenic effects on SCLC. Known agonists for OTR ([Thr4,Gly7]OT) and V1aR (F180), in addition to OT and VP, were able to elicit increases in cytosolic Ca2+ levels and this effect could be blocked using an OTR antagonist (OVTA) or a V1aR antagonist (SR49059) respectively. There was no activation of the cAMP pathway detected after VP, dDAVP (a V2R agonist), or OT treatment. Stimulation of SCLC cells with OT and VP led to an increase of extracellular signal-regulated kinase (ERK) 1/2 phosphorylation, maximal at 5 min, and the subsequent phosphorylation of its downstream target p90 ribosomal S6 kinase (p90RSK). Pre-incubation with OVTA and SR49059, and with inhibitors of phospholipase C (PLC), protein kinase C (PKC), mitogen-activated protein kinase/ERK kinase (MEK) 1/2 and a Ca2+ chelator significantly reduced OT- and VP-induced ERK1/2 phosphorylations. OVTA, SR49059 as well as MEK1/2 and PKC inhibitors also downregulated OT- and VP-induced p90RSK phosphorylation. In [3H]thymidine-uptake experiments, we subsequently observed that PLC, Ca2+, PKC and ERK1/2 are absolutely required for the OT- and VP-stimulated SCLC cellular growth process. In conclusion, the results presented here indicate that OT- and VP-induced mitogenic effects on SCLC are respectively mediated by OTR and V1aR signalling and that this mitogenic signalling passes through the phosphorylation of ERK1/2 and p90RSK in a PLC-, Ca2+-, PKC- and MEK1/2-dependent pathway.

Human chorionic gonadotropin and growth factors at the embryonic-endometrial interface control leukemia inhibitory factor (LIF) and interleukin 6 (IL-6) secretion by human endometrial epithelium.

BACKGROUND: The elucidation of the molecular mechanisms by which the embryo contributes to its implantation is an area of extensive research. The main objective of this study was to investigate the pattern of leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) secretion by human endometrial epithelium, and their regulation by human chorionic gonadotropin (hCG) and other growth factors present at the embryonic-endometrial interface. METHODS: Endometrial epithelial cells (EEC) were isolated from biopsies collected at both proliferative and secretory phases of fertile women. RESULTS: HCG (1-50 IU/ml) increased LIF secretion by EEC cultures derived from follicular phase (up to 285+/-75%) or from secretory phase (up to 212+/-16%). In contrast, hCG reduced IL-6 secretion by EEC in both phases. The hCG/LH receptor gene was transcribed by EEC as evidenced by RT-PCR. Insulin-like growth factors 1 and 2 increased LIF secretion by EEC. Transforming growth factor beta1 stimulated LIF and reduced IL-6 secretion. CONCLUSIONS: Through hCG, the blastocyst may be involved in the control of its implantation (via an increase of proimplantatory LIF) and tolerance (via an inhibition of proinflammatory IL-6). Other growth factors present at the embryonic-endometrial interface are also involved in the control of LIF and IL-6 endometrial secretion.

[Urinary excretion of 6-sulphatoxymelatonin in normal subjects: statistical approach to the influence of age and sex]. / Approche statistique de l'influence de l'âge et du sexe sur l'excrétion de 6-sulfatoxymélatonine urinaire (a-MT6s) chez l'individu normal.

A radioimmunoassay of urinary 6-sulphatoxymelatonin (a-MT6s) was performed in 90 normal subjects: 44 males and 46 females (17-67 years). Patients treated with betablokers or antidepressants were not included in this study. Urine samples were collected over three periods of time: 7 to 11 p.m., 11 p.m. to 7 a.m., and 7 to 11 a.m. Between 11 p.m. and 7 a.m., the subjects slept in their normal environment and had not ingested alcohol for 24 hours. We searched for a possible relation between urinary a-MT6s excretion (expressed in ng/l/h) and age. From 7 to 11 p.m. and from 7 to 11 a.m. no significant relation could be found. On the contrary, between 11 p.m. and 7 a.m. there was a significant relation indicating decrease of a-MT6s secretion with increasing age. Several linear or non-linear curve patters were tested: Boltzmann sigmoid (1(st), 2(nd), and 3(rd) degree), polynomial curves. The Boltzmann sigmoid showed the best fit judging by the r-squared value (0.152) and the runs test (p=0.64). On this curve the inflection point was located at 53 4 years (SDM, standard deviation of the mean). From 19 to 45 years, the upper sigmoid plateau was located at 1381 91 ng/l/h (SDM). The decrease was found between 45 and 60 years and the lower sigmoid plateau then stabilized at 467 370 ng/l/h (\SDM). In the study group, there was no significant difference between men and women according to the Mann-Withney test. Finally, use of oral contraceptives did not affect urinary a-MT6s (Mann-Withney).

Novel plasma extraction procedure and development of a specific enzyme-immunoassay of oxytocin: application to clinical and biological investigations of small cell carcinoma of the lung.

Paraneoplastic secretion of the lactation-inducing hormone oxytocin (OT) has been reported in about 30% of cases of small cell carcinoma of the lung (SCCL). In order to investigate the role of OT in the biology of SCCL tumours, a specific enzyme-immunoassay (EIA) for OT, which can be applied to both human plasma and culture medium, has been developed. OT EIA is performed on 96-well microtiter plates coated with a rabbit polyclonal antibody (Ab) anti-OT (04). This antibody does not exhibit any significant cross-reactivity either with vasopressin (VP) or with vasotocin (VT). The immunological reaction involving Ab anti-OT is a competition between the tracer (biotinylated OT) and synthetic OT (standard curve) or OT present in biological samples. In order to limit interference induced by plasma proteins, plasma samples are filtrated by a one-step centrifugation on centricon YM-3 (cut-off 3000 Da). After plasma filtration, 90.7 +/- 5.1 (SD) % (n = 22) immunoreactive (IR) OT is recovered. The sensitivity of OT EIA is 1 pmol/L, while intra- and inter-assay coefficients of variation (CV) are around 3.41% and 2.84%, respectively. In healthy volunteers, plasma IR OT is 7.28 +/- 4.49 (SD) pmol/L (n = 32) with no gender difference. As shown by the data both from plasma of SCCL patients and from supernatants and cell contents of SCCL cell lines, this EIA procedure offers a novel, reproducible, specific and sensitive method for the measurement of IR OT.

XAS investigation of Al2O3-coated nano-composite ZrO2.

In order to complement the structural characterisation by high resolution electron microscopy and perturbed angular correlation spectroscopy (PAC) of Al2O3 coated nano-composite ZrO2, XAFS spectra have been acquired and analysed. The electron micrographs showed fringes of a well ordered lattice and well defined crystal faces of the as-produced powder whereas the PAC spectra indicated a strongly distorted short range order. On the basis of the XAFS data, a structural model for the ZrO2 core of the nano-composite ZrO2/Al2O3 with a well ordered Zr lattice and a sevenfold, strongly distorted nearest neighbour oxygen shell is proposed. A smooth temperature dependence without an indication for a phase transformation up to a temperature of 700K has been revealed.

Tumor necrosis factor alpha decreases, and interleukin-10 increases, the sensitivity of human monocytes to dexamethasone: potential regulation of the glucocorticoid receptor.

Resistance to glucocorticoid therapy has been observed in patients with autoimmune/inflammatory diseases and may be related to the inflammatory process itself. The aim of this study was to examine the ability of tumor necrosis factor alpha (TNFalpha, a proinflammatory cytokine) and interleukin (IL)-10 (an anti-inflammatory cytokine) to differentially regulate the sensitivity of human monocytes/macrophages to glucocorticoids. To accomplish this, we first analyzed the pattern of TNFalpha and IL-10 inhibition by dexamethasone in LPS-stimulated whole-blood cell cultures. Second, we studied the modulation of the sensitivity of these cells to dexamethasone by preincubation with TNFalpha or IL-10 and measurement of LPS-stimulated IL-6 secretion. In addition, we evaluated the effect of dexamethasone on phorbolmyristate-acetate-stimulated IL-1 receptor antagonist secretion by the human monocytic cell line U937. Finally, we investigated whether the modulation of corticosensitivity in TNFalpha- and IL-10-pretreated U937 cells was related to a change of the glucocorticoid receptor concentration and affinity. Dexamethasone had different effects on LPS-induced TNFalpha and IL-10 secretion; whereas it suppressed TNFalpha in a dose-dependent fashion, its effect on IL-10 secretion was biphasic, producing stimulation at lower, and inhibition at higher doses. The concentration of LPS employed influenced the effect of dexamethasone on IL-10 secretion (P < 0.001). Pretreatment with TNFalpha diminished, and with IL-10 improved, the ability of dexamethasone to suppress IL-6 secretion in whole-blood cell cultures (P < 0.01 for both) and to enhance IL-1 receptor antagonist secretion by U937 cells (P < 0.05 for both). TNFalpha decreased (P < 0.001), while IL-10 increased (P < 0.001), the concentration of dexamethasone binding sites in these cells, with no discernible effect on their binding affinity. We conclude that glucocorticoids differentially modulate TNFalpha and IL-10 secretion by human monocytes in a LPS dose-dependent fashion and that the sensitivity of these cells to glucocorticoids is altered by TNFalpha or IL-10 pretreatment; TNFalpha blocks their effects, whereas IL-10 acts synergistically with glucocorticoids. This is accompanied by opposite glucocorticoid receptor changes, respectively opposing and favoring glucocorticoid actions. This study suggests that the pattern of pro-/antiinflammatory cytokine secretion may alter the response of patients to glucocorticoid therapy.

Characterization of the insulin-like growth factor axis in the human thymus.

The components of the insulin-like growth factor (IGF) axis have been investigated in the normal human thymus. Using ribonuclease protection assays (RPA), IGF-II transcripts were detected in the normal human thymus. By reverse transcriptase polymerase chain reaction (RT-PCR) analyses, promoters P3 and P4 were found to be active in the transcription of IGF2 gene within human thymic epithelial cells (TEC). No IGF-II mRNA could be detected in human lymphoid Jurkat T cells with 30 cycles of RT-PCR. By Northern blot analyses, IGFBP-2 to -6 (but not IGFBP-1) were found to be expressed in TEC with a predominance of IGFBP-4. Interestingly, Jurkat T cells only express IGFBP-2 but at high levels. The type 1 IGF receptor was detected in Jurkat T cells but not in human TEC. The identification of the components of the IGF axis within separate compartments of the human thymus adds further evidence for a role of this axis in the control of T-cell development. The precise influence of thymic IGF axis upon T-cell differentiation and immunological self-tolerance however needs to be further investigated.

The solution structure of a superpotent B-chain-shortened single-replacement insulin analogue.

This paper reports on an insulin analogue with 12.5-fold receptor affinity, the highest increase observed for a single replacement, and on its solution structure, determined by NMR spectroscopy. The analogue is [D-AlaB26]des-(B27-B30)-tetrapeptide-insulin-B26-amide. C-terminal truncation of the B-chain by four (or five) residues is known not to affect the functional properties of insulin, provided the new carboxylate charge is neutralized. As opposed to the dramatic increase in receptor affinity caused by the substitution of D-Ala for the wild-type residue TyrB26 in the truncated molecule, this very substitution reduces it to only 18% of that of the wild-type hormone when the B-chain is present in full length. The insulin molecule in solution is visualized as an ensemble of conformers interrelated by a dynamic equilibrium. The question is whether the "active" conformation of the hormone, sought after in innumerable structure/function studies, is or is not included in the accessible conformational space, so that it could be adopted also in the absence of the receptor. If there were any chance for the active conformation, or at least a predisposed state to be populated to a detectable extent, this chance should be best in the case of a superpotent analogue. This was the motivation for the determination of the three-dimensional structure of [D-AlaB26]des-(B27-B30)-tetrapeptide-insulin-B26-amide. However, neither the NMR data nor CD spectroscopic comparison of a number of related analogues provided a clue concerning structural features predisposing insulin to high receptor affinity. After the present study it seems more likely than before that insulin will adopt its active conformation only when exposed to the force field of the receptor surface.

Nanosecond-resolved XMCD on ID24 at the ESRF to investigate the element-selective dynamics of magnetization switching of Gd-Co amorphous thin film.

The one-bunch filling mode of the ESRF is combined with a microcoil to generate a pulsed-magnetic-field pump phased with respect to the probe that is given by the bunch of photons emitted each turn (357 kHz). Nanosecond-resolved X-ray magnetic circular dichroism (XMCD) is carried out. Besides the microcoil, the two other key-elements are the energy-dispersive XAS spectrometer, which yields parallel data acquisition, and the diamond-based quarter-wave plate, which tunes the helicity of the photon alternatively left and right.

A scattering filter for energy-dispersive optics.

A filtering technique to remove parasitic scattering from X-ray absorption spectra that are acquired in energy-dispersive mode has been developed and tested at the European Synchrotron Radiation Facility. The improved set-up removes small-angle scattering of the sample or the windows of sample cells which may spoil the energy resolution or reduce the intensity of prominent features in the absorption spectrum, such as the white line at the Pt L(III) edge. The sample is placed behind the curved monochromator and between two plane perfect crystals in the Bonse-Hart configuration. The dispersion of the Bonse-Hart double-crystal camera is matched to the dispersion of the curved monochromator by inclining the scattering planes of the two optical elements against each other.

Quarter-wave plates and X-ray magnetic circular dichroism on ID24 at the ESRF.

The first XMCD measurements carried out on the ID24 energy-dispersive XAS beamline at the ESRF are reported. Circular-polarized X-rays are obtained using perfect diamond crystals as quarter-wave plates. The very small source divergence allows circular polarizations close to unity to be obtained.

An exchangeable Bragg/Laue polychromator for energy-dispersive XAFS.

An original design for a cooled exchangeable polychromator for energy-dispersive XAFS (X-ray absorption fine structure) working either in the transmission configuration (Laue case) or in the reflection configuration (Bragg case) is presented. It enables the acquisition of X-ray absorption spectra between 5 and 25 keV with a spot size on the sample that can reach to less than 20 microm FWHM for some energies. Only 1 h is needed to exchange both benders in operative mode. Parallel transmission spectra with a bandpass between 5 and 10% can be obtained in the full energy range. The dispersive optics and mechanics of ID24 (ESRF, Grenoble, France) have been designed to obtain XAFS spectra in less than 1 s and, in some cases, in the millisecond range.

Curved Crystal Transmission Optics for Energy-Dispersive X-ray Absorption Spectroscopy.

The development of a curved crystal monochromator of the Laue type for energy-dispersive X-ray absorption spectroscopy is presented. The quality of the X-ray absorption spectra at high photon energies is compared with spectra measured with silicon crystals in the more frequently used Bragg geometry. In the Bragg case, an asymmetric broadening of the reflectivity profile leads to strong distortions of the near-edge fine structure and to a reduction in spectral resolution. The reflectivity profiles of fiat and curved crystals for Laue and Bragg geometry have been calculated using dynamical theory and are compared with experimental data. The new optics have been used for in situ time-resolved X-ray absorption spectroscopy. An example of the application of the technique for the characterization of a Pd catalyst is given. The X-ray absorption fine structure at the Pd K-edge has been measured during the activation and during the heterogeneous catalytic oxidation of carbon monoxide.

Immunoreactive somatostatin in the rat ovary.

Immunoreactive SRIH was detected in the rat ovary (15.6 pg/mg wet weight, 520 pg/mg protein) and was localized to the granulosa cells (168 +/- 6 pg/10(6) cells). Serial dilution studies showed parallelism of the inhibition curve for synthetic SRIH-14 and those of extracts of whole ovary and media conditioned by granulosa cells. The quantity of immunoreactive SRIH released into granulosa cell conditioned media decreased with time, while the intracellular content remained relatively constant. Gel chromatography showed peaks of immunoreactivity co-eluting with SRIH-14 (38%), SRIH-28 (31%) and a high molecular weight component. The addition of synthetic SRIH-14 stimulated meiotic maturation in cumulus-enclosed rat oocytes, with dose dependency being observed at SRIH-14 concentrations between 1 and 1000 pmol/l. No evidence of pre-pro-SRIH gene expression could be demonstrated in either rat ovary or testis using both Northern analysis and reverse transcriptase/polymerase chain reaction amplification of polyadenylated RNA. SRIH may be produced in the ovary during a specific stage of ontogeny or by an alternative gene. It is also possible that SRIH is actively taken up and stored by granulosa cells without being produced locally.

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. II. Inhibin and aromatase inhibitor activity.

The inhibin content and aromatase inhibitor activity (AIA) of 72 follicular fluids (FF) obtained from 42 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) were studied as a function of IVF ET outcome. Inhibin levels were determined by bioassay (BA) and RIA; AIA was measured by BA. The inhibin content of follicles characterized as immature by their estradiol (E2) levels and E2/progesterone (P) ratios was significantly lower (P less than 0.05) than that of mature follicles (i.e. leading to pregnancy). The mean AIA for mature follicles were significantly lower than AIA in groups where pregnancy was not obtained. AIA for follicles from which a pregnancy was obtained for each ET was also significantly lower than that in FF characterized as immature of hypermature. The highest E2/AIA and inhibin BA/AIA ratios were associated with the highest incidence of successful IVF ET outcome. No correlation was found between AIA and inhibin, on the one hand, and E2, delta 4-androstenedione, E2/P, and PRL, on the other. However, a positive correlation was found between inhibin (RIA and BA) and P, reflecting the production of inhibin by granulosa cells during luteinization. These studies allowed us to conclude that FF inhibin levels do not differ according to IVF ET outcome, but are an index of follicular maturation. AIA not only constitutes an index of follicular maturation and granulosa cell luteinization, but is of predictive value for IVF ET outcome as E2/AIA and inhibin BA/AIA ratios.

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. III. Proteoglycans.

Using a specific proteoglycan (PG) radioimmunoassay (RIA) in which human cartilage antiserum was directed against the PG protein core, the PG content of follicular fluid (FF) obtained from 42 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) was studied as a function of IVF-ET outcome. Inhibition curves of purified PG cartilage preparations were parallel to those of large and small nonstimulated follicles and follicles that had been stimulated by a luteinizing hormone-releasing hormone (LHRH) agonist, d-tryptophan-6 (Decapeptyl: D-Trp6 analogue, Beaufour Laboratories, IPSEN Biotech, Paris, France), and human menopausal gonadotropin (hMG). While FF levels of immunoreactive PG-like material (Ir-PG) did not differ according to IVF-ET outcome, highly significant negative correlations were obtained between FF 17 beta-estradiol levels and FF Ir-PG levels in oocyte groups where pregnancy was obtained, i.e., oocytes were fertilized and cleaved, and pregnancy followed either for each ET or for one of two embryos reimplanted. The correlation persisted but weakened when all groups were pooled together. No correlation was observed between FF Ir-PG and progesterone. RIA or bioassay showed a positive correlation between FF inhibin and Ir-PG for the group in which each ET led to a pregnancy. Ir-PG concentrations were significantly greater in smaller than in larger follicles collected from untreated women. Upon induction of ovulation with either pure follicle-stimulating hormone (FSH), FSH + human chorionic gonadotropin (hCG), or D-Trp6/hMG + hCG, this difference no longer appeared. These results indicate that the reduction of Ir-PG concentrations constitutes an index of follicular maturation.(ABSTRACT TRUNCATED AT 250 WORDS)

Correlation between follicular fluid content and the results of in vitro fertilization and embryo transfer. I. Sex steroids.

The steroid content of 72 follicular fluids (FF) obtained from 42 women subjected to ovulation induction with the luteinizing hormone-releasing hormone analogue D-Trp6 and human menopausal gonadotropin was studied in terms of the evolution of in vitro fertilization (IVF) and embryo transfer (ET) results. The FF were classified into several categories based on oocyte evolution. Individual values of FF estrone and estradiol (E2), as well as androstenedione and testosterone could not be correlated with ET outcome. However, FF progesterone (P) levels for follicles leading to pregnancy were significantly lower when compared with those in the other categories. The correlation between the E2/P ratio and E2 permitted the definition of a band wherein IVF-ET outcome was successful and enabled the characterization of different functional follicular maturational states.

[Endocrine, paracrine and autocrine mechanisms of aromatase activity]. / Mécanismes endocrines, paracrines et autocrines de l'activité aromatasique.

The endocrine, paracrine and autocrine mechanisms involved in aromatase activity. Development of a single follicle during the menstrual cycle is under control of hormones stimulating follicular maturation, ovulation and luteogenesis. Several factors intervene locally to avoid other follicles developing at the same time as the dominant follicle. These other follicles remain quiescent or go on to atresia. Atresia results from the action of several endocrine, paracrine and autocrine mechanisms which synergistically inhibit aromatase activity. The subsequent lack of estrogens reduces granulosa cell multiplication.