The Natural Chemopreventive Agent Sulforaphane Inhibits STAT5 Activity

Signal transducer and activator of transcription STAT5 is an essential mediator of cytokine, growth factor and hormone signaling. While its activity is tightly regulated in normal cells, its constitutive activation directly contributes to oncogenesis and is associated to a number of hematological and solid tumor cancers. We previously showed that deacetylase inhibitors can inhibit STAT5 transcriptional activity. We now investigated whether the dietary chemopreventive agent sulforaphane, known for its activity as deacetylase inhibitor, might also inhibit STAT5 activity and thus could act as a chemopreventive agent in STAT5-associated cancers. We describe here sulforaphane (SFN) as a novel STAT5 inhibitor. We showed that SFN, like the deacetylase inhibitor trichostatin A (TSA), can inhibit expression of STAT5 target genes in the B cell line Ba/F3, as well as in its transformed counterpart Ba/F3-1*6 and in the human leukemic cell line K562 both of which express a constitutively active form of STAT5. Similarly to TSA, SFN does not alter STAT5 initial activation by phosphorylation or binding to the promoter of specific target genes, in favor of a downstream transcriptional inhibitory effect. Chromatin immunoprecipitation assays revealed that, in contrast to TSA however, SFN only partially impaired the recruitment of RNA polymerase II at STAT5 target genes and did not alter histone H3 and H4 acetylation, suggesting an inhibitory mechanism distinct from that of TSA. Altogether, our data revealed that the natural compound sulforaphane can inhibit STAT5 downstream activity, and as such represents an attractive cancer chemoprotective agent targeting the STAT5 signaling pathway

Stadtgestalt und Stadtgestaltung. Design und die creative city

Müller A-L. Stadtgestalt und Stadtgestaltung. Design und die creative city. In: Moebius S, Prinz S, eds. Das Design der Gesellschaft. Zur Kultursoziologie des Designs. Sozialtheorie. Bielefeld: Transcript-Verlag; 2012: 303-326

Thalamic deep brain stimulation for Tourette Syndrome: A naturalistic trial with brief randomized, double-blinded sham-controlled periods

Background: There is still a lack of controlled studies to prove efficacy of thalamic deep brain stimulation for Tourette's Syndrome. Objectives: In this controlled trial, we investigated the course of tic severity, comorbidities and quality of life during thalamic stimulation and whether changes in tic severity can be assigned to ongoing compared to sham stimulation. Methods: We included eight adult patients with medically refractory Tourette's syndrome. Bilateral electrodes were implanted in the centromedian-parafascicular-complex and the nucleus ventro-oralis internus. Tic severity, quality of life and comorbidities were assessed before surgery as well as six and twelve months after. Short randomized, double-blinded sham-controlled crossover sequences with either active or sham stimulation were implemented at both six-and twelve-months' assessments. The primary outcome measurement was the difference in the Yale Global Tic Severity Scale tic score between active and sham stimulation. Adverse events were systematically surveyed for all patients to evaluate safety. Results: Active stimulation resulted in significantly higher tic reductions than sham stimulation (F = 79.5; p = 0.001). Overall quality of life and comorbidities improved significantly in the open-label phase. Over the course of the trial two severe adverse events occurred that were resolved without sequelae. Conclusion: Our results provide evidence that thalamic stimulation is effective in improving tic severity and overall quality of life. Crucially, the reduction of tic severity was primarily driven by active stimulation. Further research may focus on improving stimulation protocols and refining patient selection to improve efficacy and safety of deep brain stimulation for Tourette's Syndrome. (c) 2021 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

A Regulatory Potential of the <em>Xist</em> Gene Promoter in Vole <em>M. rossiaemeridionalis</em>

<div><p>X chromosome inactivation takes place in the early development of female mammals and depends on the <em>Xist</em> gene expression. The mechanisms of <em>Xist</em> expression regulation have not been well understood so far. In this work, we compared <em>Xist</em> promoter region of vole <em>Microtus rossiaemeridionalis</em> and other mammalian species. We observed three conserved regions which were characterized by computational analysis, DNaseI <em>in vitro</em> footprinting, and reporter construct assay. Regulatory factors potentially involved in <em>Xist</em> activation and repression in voles were determined. The role of CpG methylation in vole <em>Xist</em> expression regulation was established. A CTCF binding site was found in the 5′ flanking region of the <em>Xist</em> promoter on the active X chromosome in both males and females. We suggest that CTCF acts as an insulator which defines an inactive <em>Xist</em> domain on the active X chromosome in voles.</p> </div

Analysis of the effect of −43G/A substitution on the activity of reporter constructs.

<p>Schemes of the constructs pCx14G/A and pCx5G/A are shown to the left; green and red rectangles denote the conserved regions CNS1 and CNS2, respectively. Relative luciferase activity of constructs in the fibroblast culture Sd10 is shown to the right. Arrow shows the <i>Xist</i> transcription start site; R.U., relative luciferase activity units.</p

CTCF interaction with the vole <i>Xist</i> regulatory region.

<p>(A) ChIP performed using anti-CTCF monoclonal antibodies (CTCF Ab) followed by PCR analysis. The fourth <i>Xist</i> exon and Igf2/H19 ICR were used as negative and positive controls of CTCF binding, respectively. (B) Real-time PCR analysis of ChIP experiments. Each bar indicates the average of two independent PCR analyses with the standard deviation. Percent of input is calculated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033994#s4" target="_blank">Materials and Methods</a>.</p

Figure 2

<p>(A) Multiple alignment of CNS3 sequences of <i>Xist</i> 5′ regulatory regions in different mammalian lineages. (Bt) <i>Bos taurus</i>; (Cf) <i>Canis familiaris</i>; (Dn) <i>Dasypus novemcinctus</i> (armadillo); (Ec) <i>Equus caballus</i> (horse); (Hs) <i>Homo sapiens</i>, (La) <i>Loxodonta africana</i> (elephant); (Oc) O<i>ryctolagus cuniculus</i> (rabbit); (Sa) <i>Sorex araneus</i> (shrew); (Ss) <i>Sus scrofa</i> (pig); (Cp) <i>Cavia porcellus</i> (guinea pig); (Mm) <i>Mus musculus</i>; (Rn) <i>Rattus norvegicus</i>; (Mr) <i>M. rossiaemeridionalis</i> (vole). Putative CTCF binding sites are shown with yellow frames. Consensus of CTCF binding site is present <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033994#pone.0033994-Kim1" target="_blank">[60]</a>. (B) Alignment of <i>Xist</i> minimal promoter. Human CTCF is shown with red frames <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033994#pone.0033994-Pugacheva1" target="_blank">[14]</a>. Vole AP2 binding site is shown with blue frame.</p

CTCF interacts with the <i>Xist</i> allele on the active X chromosome in voles.

<p>Sequencing of the vole <i>Xist</i> minimal promoter using as a template Sad4 or Sa006 genomic DNA (A), CTCF-bound fraction from line Sad4 (B), CTCF-bound fraction from line Sa006 (C).</p

Methylation profile of the <i>Xist</i> 5′ regulatory region in the Sa006 and Sad4 vole fibroblast lines.

<p>White circles denote unmethylated CpG sites; black, methylated; the <i>Xist</i> transcription start site is indicated by arrow; Xa, active X chromosome; Xi, inactive X chromosome; <i>M. r.</i>, <i>Microtus rossiaemeridionalis</i>.</p