Two major goals of regenerative medicine are to reproducibly transform adult somatic cells into a pluripotent state and to control their differentiation into specific cell fates. Progress toward these goals would be greatly helped by obtaining a complete picture of the RNA isoforms produced by these cells due to alternative splicing (AS) and alternative promoter selection (APS). To investigate the roles of AS and APS, reciprocal exon-exon junctions were interrogated on a genome-wide scale in differentiating mouse embryonic stem (ES) cells with a prototype Affymetrix microarray. Using a recently released open-source software package named AltAnalyze, we identified 144 genes for 170 putative isoform variants, the majority (67%) of which were predicted to alter protein sequence and domain composition. Verified alternative exons were largely associated with pathways of Wnt signaling and cell-cycle control, and most were conserved between mouse and human. To examine the functional impact of AS, we characterized isoforms for two genes. As predicted by AltAnalyze, we found that alternative isoforms of the gene Serca2 were targeted by distinct microRNAs (miRNA-200b, miRNA-214), suggesting a critical role for AS in cardiac development. Analysis of the Wnt transcription factor Tcf3, using selective knockdown of an ES cell-enriched and characterized isoform, revealed several distinct targets for transcriptional repression (Stmn2, Ccnd2, Atf3, Klf4, Nodal, and Jun) as well as distinct differentiation outcomes in ES cells. The findings herein illustrate a critical role for AS in the specification of ES cells with differentiation, and highlight the utility of global functional analyses of AS.

Eye movements were examined to determine how readers with Down syndrome process sentences online. Participants were 9 individuals with Down syndrome ranging in reading level from Grades 1 to 3 and a reading-level-matched control group. For syntactically simple sentences, the pattern of reading times was similar for the two groups, with longer reading times found at sentence end. This "wrap-up" effect was also found in the first reading of more complex sentences for the control group, whereas it only emerged later for the readers with Down syndrome. Our results provide evidence that eye movements can be used to investigate reading in individuals with Down syndrome and underline the need for future studies.

This study deals with the determinants of prosodic phrasing in French schoolchildren's narratives. Children (aged 7 to 11) told picture stories to a silent same-age peer. The establishment of temporal and/or causal relations between the events was more or less guided by the drawings (ordered vs. arbitrary sequences). The comprehension of the referential links was more or less supported by the way the frames were displayed (simultaneous vs. consecutive display mode). Four storytelling conditions that differed by the constraints imposed on inference-resolving and memory-searching were thus defined. Naïve French listeners were asked to segment tape-recorded narrations using prosodic variation as a criterion, and to decide whether each prosodic segment was "conclusive" or "continuative." The comparison of the listeners' segmentation labels to those of an expert (functional and formal annotation) showed that more than 91% of the labels corresponded to prosodic boundaries and more than 78% of the non-terminal labels corresponded to non-terminal boundaries, but only 55% of the terminal labels corresponded to terminal boundaries. The storytellers' boundaries were then analyzed as a function of age and storytelling conditions. Non-terminal and terminal boundaries varied with the picture-display mode. Terminal boundaries also depended on the type of event sequence, and non-terminal boundaries on the improvement of the linguistic and communicative skills of the narrators. Terminal judgments of non-terminal boundaries mainly occurred in texts where each event was told in a single proposition either without anaphoric references to the main character or with anaphoric pronouns.

Although FGFs are known to affect mesoderm patterning, their influence on intermediate mesoderm specification during gastrulation is ignored. Here, we show that pronephros precursors are exposed to FGF, but a strict control of FGF signals is necessary to allow pronephros development. We provide evidence that this control is mediated by the paired-like homeobox genes Mix.1 and Mix.2. Morpholino-based Mix.1/2 knockdown, or repression of Mix.1 target genes with an enRMix.1 construct, causes an expansion of FGF4 and FGF8 expression in the lateral marginal zone at gastrula stage, together with an inhibition of pronephros development at neurula and tailbud stages. Expression of the nephrogenic mesoderm markers Xlim-1 and XPax-8 can be rescued in Mix.1/2 morphants by intrablastocoelic injections of the FGFR inhibitor SU5402 at mid-gastrula stage, showing that inhibition of pronephros development results from an increase of FGF signalling. We further show that Mix.1 overexpression results in the down-regulation of FGF3, 4, 8 and XmyoD, in addition to Xbra. However, cells overexpressing Mix.1 can normally populate somites, indicating that Mix.1 does not affect their fate cell autonomously. These data support the idea that Mix.1/2 regulates levels and/or duration of FGF signals to which pronephros precursors are exposed during gastrulation.

PURPOSE: The regulator of angiogenesis most extensively studied is VEGF. VEGF mRNA in plasma from patients with colorectal cancer was analyzed as a possible surrogate marker of tumor angiogenesis. METHODS: VEGF mRNA was measured by quantitative PCR in plasma, tumors and circulating tumor cells from colorectal cancer patients. Circulating VEGF protein was analyzed by ELISA. Microvessel density was determined. RESULTS: Levels of VEGF mRNA and protein in plasma were higher in patients than in controls. VEGF mRNA was overexpressed in tumors with respect to normal tissues. Levels of VEGF protein were associated with VEGF mRNA in plasma, but no associations with tumor samples were found. A trend to statistical significance was shown between high VEGF mRNA and vascular invasion. MVD was not related to VEGF mRNA in plasma. CONCLUSIONS: Thus, VEGF mRNA could be a marker similar to VEGF protein in plasma.

OBJECTIVE: There is great controversy concerning treatment for menopausal symptoms. We evaluated homeopathic treatments for hot flushes and their effect on quality of life in menopausal women. METHODS: Open, multi-national prospective, pragmatic and non-comparative observational study of homeopathic treatments prescribed and their effectiveness, observing their impact on quality of life. RESULTS: Ninety-nine physicians in 8 countries took part in this study and included 438 patients with an average age of 55. Homeopathic medicines were prescribed to all patients; 98% of the prescription lines were for homeopathic medicines. Lachesis mutus, Belladonna, Sepia officinalis, Sulphur and Sanguinaria canadensis were the most prescribed. A non-homeopathic treatment and/or food supplement prescribed for 5% of the patients. This observational study revealed a significant reduction (p<0.001) in the frequency of hot flushes by day and night and a significant reduction in the daily discomfort they caused (mean fall of 3.6 and 3.8 points respectively, on a 10cm visual analogue scale; p<0.001). Ninety percent of the women reported disappearance or lessening of their symptoms, these changes mostly taking place within 15 days of starting homeopathic treatment. CONCLUSIONS: The results of this observational study suggest that homeopathic treatment for hot flushes in menopausal women is effective. Further studies including randomized controlled trials should be conducted.

The effects and the mode of action of hypericin (1) were studied, in the dark, on the action potential (AP) and the L-type Ca2+ channel of frog atrial heart muscle, using intracellular microelectrode and patch-clamp techniques, respectively. In the presence of Ca2+ in Ringer solution, hypericin (1 to 4 muM) did not markedly modify the AP. Total replacement of Ca2+ by Sr2+ in the solution (Ringer Sr2+) revealed that hypericin (4 muM) prolonged the AP duration (APD). Hypericin dose-dependently increased the magnitude of the Sr2+current, which develops through L-type Ca2+ channels in the Ringer solution containing tetrodotoxin (0.7 muM) and tetraethylammonium (10 mM), but did not modify the kinetics of activation and inactivation. This revealed that hypericin increased L-type Ca2+ channel conductance, which accounted for the APD lengthening. The hypericin-induced APD lengthening recorded in the Ringer Sr2+ was not prevented by (i) a blockade of alpha- and beta-adrenoceptors by yohimbine (1 muM), urapidil (1 muM), and propanolol (50 muM), respectively, and (ii) PKC blockade by staurosporine (1 muM). The hypericin-induced APD lengthening recorded in the Ringer Sr2+ was prevented by blocking soluble guanylate cyclase (sGC) activity by 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one (13 muM), which mimicked the effects of hypericin. Hypericin decreased the cellular cGMP level by 69% in atrial myocytes. The compound also decreased the cellular cGMP level by inhibiting sGC, thus cancelling the nucleotide inhibitory effect on the cardiac L-type Ca2+ channel.