Expression of interleukin-3 and tumor necrosis factor-beta mRNAs in cultured microglia.

The function of interleukin-3 (or multi-CSF) in the hemopoietic system has been studied in great detail. Although its growth promoting activity on brain microglial cells has been confirmed both in vitro and in vivo, its presence in the brain and even in cultured brain cells has repeatedly been questioned. We have shown recently that isolated rat microglia express mRNA(IL-3) and synthesize IL-3 polypeptide. It is shown here by use of the PCR method, that mRNA(IL-3) is found also in C6 glioblastoma, in rat aggregate cultures, and in newborn and adult rat brain. Quantitation of amplified cDNA(IL-3) was achieved by non-competitive RT-PCR using an elongated internal standard. IL-3 messenger RNA was almost undetectable in vivo and low in (serum-free) aggregate cultures. In isolated microglia, mRNA(IL-3) was increased upon treatment with LPS, PHA, with the cytokines IL-1 or TNF-alpha, with retinoic acid, dbcAMP or the phorbol ester TPA. Effects of LPS were inhibited by dexamethasone, while the glucocorticoid by itself had no effect on basal IL-3 expression. LPS increased mRNA(IL-3) in a concentration-dependent manner beginning with 10 pg/ml and reaching plateau levels at 10 ng/ml. LPS also increased mRNAs of TNF-alpha and TNF-beta. TNF-alpha mRNA was already detectable in untreated microglia and LPS-increased levels were sustained for a few days. In contrast, TNF-beta mRNA was observed only between 4 and 16 h of LPS incubation. It was absent in LPS-free microglia, and after 24 h of LPS-treatment or later.(ABSTRACT TRUNCATED AT 250 WORDS)

Long-range heterogeneity at the 3' ends of human mRNAs.

The publication of a draft of the human genome and of large collections of transcribed sequences has made it possible to study the complex relationship between the transcriptome and the genome. In the work presented here, we have focused on mapping mRNA 3' ends onto the genome by use of the raw data generated by the expressed sequence tag (EST) sequencing projects. We find that at least half of the human genes encode multiple transcripts whose polyadenylation is driven by multiple signals. The corresponding transcript 3' ends are spread over distances in the kilobase range. This finding has profound implications for our understanding of gene expression regulation and of the diversity of human transcripts, for the design of cDNA microarray probes, and for the interpretation of gene expression profiling experiments.

Coronary MR imaging using free-breathing 3D steady-state free precession with radial k-space sampling.

PURPOSE: To investigate the potential of free-breathing 3D steady-state free precession (SSFP) imaging with radial k-space sampling for coronary MR-angiography (MRA), coronary projection MR-angiography and coronary vessel wall imaging. MATERIALS AND METHODS: A navigator-gated free-breathing T2-prepared 3D SSFP sequence (TR = 6.1 ms, TE = 3.0 ms, flip angle = 120 degrees, field-of-view = 360 mm(2)) with radial k-space sampling (384 radials) was implemented for coronary MRA. For projection coronary MRA, this sequence was combined with a 2D selective aortic spin tagging pulse. Coronary vessel wall imaging was performed using a high-resolution inversion-recovery black-blood 3D radial SSFP sequence (384 radials, TR = 5.3 ms, TE = 2.7 ms, flip angle = 55 degrees, reconstructed resolution 0.35 x 0.35 x 1.2 mm(3)) and a local re-inversion pulse. Six healthy volunteers (two for each sequence) were investigated. Motion artifact level was assessed by two radiologists. Results: In coronary MRA, the coronary lumen was displayed with a high signal and high contrast to the surrounding lumen. Projection coronary MRA demonstrated selective visualization of the coronary lumen while surrounding tissue was almost completely suppressed. In coronary vessel wall imaging, the vessel wall was displayed with a high signal when compared to the blood pool and the surrounding tissue. No visible motion artifacts were seen. Conclusion: 3D radial SSFP imaging enables coronary MRA, coronary projection MRA and coronary vessel wall imaging with a low motion artifact level.

Development and maintenance of the neuronal cytoskeleton in aggregated cell cultures of fetal rat telencephalon and influence of elevated K+ concentrations.

Serum-free aggregating cell cultures of fetal rat telencephalon were examined by biochemical and immunocytochemical methods for their development-dependent expression of several cytoskeletal proteins, including the heavy- and medium-sized neurofilament subunits (H-NF and M-NF, respectively); brain spectrin; synapsin I; beta-tubulin; and the microtubule-associated proteins (MAPs) 1, 2, and 5 and tau protein. It was found that with time in culture the levels of most of these cytoskeletal proteins increased greatly, with the exceptions of the particular beta-tubulin form studied, which remained unchanged, and MAP 5, which greatly decreased. Among the neurofilament proteins, expression of M-NF preceded that of H-NF, with the latter being detectable only after approximately 3 weeks in culture. Furthermore, MAP 2 and tau protein showed a development-dependent change in expression from the juvenile toward the adult form. The comparison of these developmental changes in cytoskeletal protein levels with those observed in rat brain tissue revealed that protein expression in aggregate cultures is nearly identical to that in vivo during maturation of the neuronal cytoskeleton. Aggregate cultures deprived of glial cells, i.e., neuron-enriched cultures prepared by treating early cultures with the antimitotic drug cytosine arabinoside, exhibited pronounced deficits in M-NF, H-NF, MAP 2, MAP 1, synapsin I, and brain spectrin, with increased levels of a 145-kDa brain spectrin breakdown product. These adverse effects of glial cell deprivation could be reversed by the maintenance of neuron-enriched cultures at elevated concentrations of KCl (30 mM). This chronic treatment had to be started at an early developmental stage to be effective, a finding suggesting that sustained depolarization by KCl is able to enhance the developmental expression and maturation of the neuronal cytoskeleton.

Genotype 3 is associated with accelerated fibrosis progression in chronic hepatitis C.

BACKGROUND/AIMS: While several risk factors for the histological progression of chronic hepatitis C have been identified, the contribution of HCV genotypes to liver fibrosis evolution remains controversial. The aim of this study was to assess independent predictors for fibrosis progression. METHODS: We identified 1189 patients from the Swiss Hepatitis C Cohort database with at least one biopsy prior to antiviral treatment and assessable date of infection. Stage-constant fibrosis progression rate was assessed using the ratio of fibrosis Metavir score to duration of infection. Stage-specific fibrosis progression rates were obtained using a Markov model. Risk factors were assessed by univariate and multivariate regression models. RESULTS: Independent risk factors for accelerated stage-constant fibrosis progression (>0.083 fibrosis units/year) included male sex (OR=1.60, [95% CI 1.21-2.12], P<0.001), age at infection (OR=1.08, [1.06-1.09], P<0.001), histological activity (OR=2.03, [1.54-2.68], P<0.001) and genotype 3 (OR=1.89, [1.37-2.61], P<0.001). Slower progression rates were observed in patients infected by blood transfusion (P=0.02) and invasive procedures or needle stick (P=0.03), compared to those infected by intravenous drug use. Maximum likelihood estimates (95% CI) of stage-specific progression rates (fibrosis units/year) for genotype 3 versus the other genotypes were: F0-->F1: 0.126 (0.106-0.145) versus 0.091 (0.083-0.100), F1-->F2: 0.099 (0.080-0.117) versus 0.065 (0.058-0.073), F2-->F3: 0.077 (0.058-0.096) versus 0.068 (0.057-0.080) and F3-->F4: 0.171 (0.106-0.236) versus 0.112 (0.083-0.142, overall P<0.001). CONCLUSIONS: This study shows a significant association of genotype 3 with accelerated fibrosis using both stage-constant and stage-specific estimates of fibrosis progression rates. This observation may have important consequences for the management of patients infected with this genotype.

Affymetrix Whole-Transcript Human Gene 1.0 ST array is highly concordant with standard 3' expression arrays.

The recently released Affymetrix Human Gene 1.0 ST array has two major differences compared with standard 3' based arrays: (i) it interrogates the entire mRNA transcript, and (ii) it uses DNA targets. To assess the impact of these differences on array performance, we performed a series of comparative hybridizations between the Human Gene 1.0 ST and the Affymetrix HG-U133 Plus 2.0 and the Illumina HumanRef-8 BeadChip arrays. Additionally, both RNA and DNA targets were hybridized on HG-U133 Plus 2.0 arrays. The results show that the overall reproducibility of the Gene 1.0 ST array is best. When looking only at the high intensity probes, the reproducibility of the Gene 1.0 ST array and the Illumina BeadChip array is equally good. Concordance of array results was assessed using different inter-platform mappings. Agreements are best between the two labeling protocols using HG-U133 Plus 2.0 array. The Gene 1.0 ST array is most concordant with the HG-U133 array hybridized with cDNA targets. This may reflect the impact of the target type. Overall, the high degree of correspondence provides strong evidence for the reliability of the Gene 1.0 ST array.

Outcomes of patients on dual-boosted PI regimens : experience of the Swiss HIV cohort study

Rapport de SynthèseLa thérapie antirétrovirale a progressée de manière significative depuis te début de l'épidémie du syndrome d'immunodéficience acquise (SIDA). Durant les 20 dernières années, plusieurs combinaisons de traitements ont été utilisées avec succès menant à une réduction de la mortalité associée. Par contre, le traitement a aussi engendré des cas de résistances multiples avec comme résultat, le besoin d'utiliser plusieurs molécules en combinaison, et une augmentation des cas de toxicité. Une stratégie souvent employée fût la combinaison de deux molécules inhibitrices de la protéase en même temps en combinaison avec une troisième molécule, le ritonavir. (DBPI).La cohorte Suisse sur le VIH existe depuis 1987 et permet d'étudier de façon longitudinale les patients qui y sont inscrits. Pour ce travail de thèse, nous avons étudié les patients inscrits à la cohorte suisse de 1996 à 2007 qui ont reçu une combinaison DBPI.Pendant la période étudiée, un total de 405 patients ont reçu un traitement DBPI, dont 295 patients ont reçu le DBPI pour plus de 6 mois. La durée médiane du traitement était de 2.2 ans. Sur les 287 patients qui étaient en échec viral au début du traitement (défini comme HIV RNA>400 copies/ml), 64.1% ont réussi à supprimer la virémie et 54.4% ont eu une suppression dans les 24 semaines qui ont suivi le début de la thérapie. Les patients avaient reçu en moyenne 6 combinaisons de traitement différentes avant le début de la thérapie DBPi. Pour les patients qui ont arrêté le traitement DBPI, la cause principale de l'arrêt était due au souhait du patient (48.3%), à l'échec virologique (22.5%) et à la toxicité (15.8%). Les patients ayant reçu le traitement après 1999, ou ayant été traités avec une combinaison de Lopinavir-ritonvir/saquinavir ou lopinavir-ritonavir/atazanavir arrivaient à supprimer leur virémie plus souvent que ceux qui avaient reçu d'autres combinaisons.Cette étude constitue la plus grande étude publiée sur le sujet de l'utilisation des DBPI pour les patients à résistances multiples. Malgré le fait que c'est une étude observationnelle, nous pouvons attester que le taux de succès était de 64.4%, le taux de toxicité était relativement bas (15.8%) et que la plus part des patients ont toléré ces combinaisons, malgré le taux élevé d'effets secondaires souvent rapportés. En somme, cette approche pourrait être envisagée dans des situations ou les nouveaux traitements tels que les inhibiteurs de l'intégrase et du CCR5 ne sont pas encore disponibles.

Thy-1-mediated cell-cell contact induces astrocyte migration through the engagement of αVβ3 integrin and syndecan-4.

Cell adhesion to the extracellular matrix proteins occurs through interactions with integrins that bind to Arg-Gly-Asp (RGD) tripeptides, and syndecan-4, which recognizes the heparin-binding domain of other proteins. Both receptors trigger signaling pathways, including those that activate RhoGTPases such as RhoA and Rac1. This sequence of events modulates cell adhesion to the ECM and cell migration. Using a neuron-astrocyte model, we have reported that the neuronal protein Thy-1 engages αVβ3 integrin and syndecan-4 to induce RhoA activation and strong astrocyte adhesion to their underlying substrate. Thus, because cell-cell interactions and strong cell attachment to the matrix are considered antagonistic to cell migration, we hypothesized that Thy-1 stimulation of astrocytes should preclude cell migration. Here, we studied the effect of Thy-1 expressing neurons on astrocyte polarization and migration using a wound-healing assay and immunofluorescence analysis. Signaling molecules involved were studied by affinity precipitation, western blotting and the usage of specific antibodies. Intriguingly, Thy-1 interaction with its two receptors was found to increase astrocyte polarization and migration. The latter events required interactions of these receptors with both the RGD-like sequence and the heparin-binding domain of Thy-1. Additionally, prolonged Thy-1-receptor interactions inhibited RhoA activation while activating FAK, PI3K and Rac1. Therefore, sustained engagement of integrin and syndecan-4 with the neuronal surface protein Thy-1 induces astrocyte migration. Interestingly we identify here, a cell-cell interaction that despite initially inducing strong cell attachment, favors cell migration upon persistent stimulation by engaging the same signaling receptors and molecules as those utilized by the extracellular matrix proteins to stimulate cell movement.

Right coronary MR angiography at 7 T: a direct quantitative and qualitative comparison with 3 T in young healthy volunteers.

PURPOSE: To objectively compare quantitative parameters related to image quality attained at coronary magnetic resonance (MR) angiography of the right coronary artery (RCA) performed at 7 T and 3 T. MATERIALS AND METHODS: Institutional review board approval was obtained, and volunteers provided signed informed consent. Ten healthy adult volunteers (mean age ± standard deviation, 25 years ± 4; seven men, three women) underwent navigator-gated three-dimensional MR angiography of the RCA at 7 T and 3 T. For 7 T, a custom-built quadrature radiofrequency transmit-receive surface coil was used. At 3 T, a commercial body radiofrequency transmit coil and a cardiac coil array for signal reception were used. Segmented k-space gradient-echo imaging with spectrally selective adiabatic fat suppression was performed, and imaging parameters were similar at both field strengths. Contrast-to-noise ratio between blood and epicardial fat; signal-to-noise ratio of the blood pool; RCA vessel sharpness, diameter, and length; and navigator efficiency were quantified at both field strengths and compared by using a Mann-Whitney U test. RESULTS: The contrast-to-noise ratio between blood and epicardial fat was significantly improved at 7 T when compared with that at 3 T (87 ± 34 versus 52 ± 13; P = .01). Signal-to-noise ratio of the blood pool was increased at 7 T (109 ± 47 versus 67 ± 19; P = .02). Vessel sharpness obtained at 7 T was also higher (58% ± 9 versus 50% ± 5; P = .04). At the same time, RCA vessel diameter and length and navigator efficiency showed no significant field strength-dependent difference. CONCLUSION: In our quantitative and qualitative study comparing in vivo human imaging of the RCA at 7 T and 3 T in young healthy volunteers, parameters related to image quality attained at 7 T equal or surpass those from 3 T.

Mejoras de funcionalidades en la gestión de usuariosde la plataforma K-PAX

Esta memoria trata de las mejoras propuestas en la plataforma educativa kPax de la UOC referentes a la gestión de usuarios.

Motivational effects of cannabinoids are mediated by μ-opioid and k-opioid receptor

Repeated THC administration produces motivational and somaticadaptive changes leading to dependence in rodents. Toinvestigate the molecular basis for cannabinoid dependenceand its possible relationship with the endogenous opioid system,we explored Δ9-tetrahydrocannabinol (THC) activity in mice lacking μ-, δ- or κ-opioid receptor genes. Acute THCinduced hypothermia, antinociception, and ypolocomotion remained unaffected in these mice, whereas THC tolerance and withdrawal were minimally modified in mutant animals. In contrast, profound phenotypic changes are observed in several place conditioning protocols that reveal both THC rewarding and aversive properties. Absence of μ receptors abolishes THC place preference. Deletion of κ receptors ablates THC place aversion and furthermore unmasks THC place preference. Thus, an opposing activity of μ- and κ-opioid receptors in modulating reward pathways forms the basis for the dual euphoric–dysphoric activity of THC.

[Archives de la Parole]. , Nadar dadā and Dalang dadā : two songs in Kōrwā from Sarguja (C.P.) / [Sir George Grierson], éd. ; Linguistic survey of India ; [Nathu Kōrwā], voix

[Traditions. Asie. Inde. Chhattisgarh. Surguja]