SMARCA4/Brg1 coordinates genetic and epigenetic networks underlying Shh-type medulloblastoma development.

Recent large-scale genomic studies have classified medulloblastoma into four subtypes: Wnt, Shh, Group 3 and Group 4. Each is characterized by specific mutations and distinct epigenetic states. Previously, we showed that a chromatin regulator SMARCA4/Brg1 is required for Gli-mediated transcription activation in Sonic hedgehog (Shh) signaling. We report here that Brg1 controls a transcriptional program that specifically regulates Shh-type medulloblastoma growth. Using a mouse model of Shh-type medulloblastoma, we deleted Brg1 in precancerous progenitors and primary or transplanted tumors. Brg1 deletion significantly inhibited tumor formation and progression. Genome-wide expression analyses and binding experiments indicate that Brg1 specifically coordinates with key transcription factors including Gli1, Atoh1 and REST to regulate the expression of both oncogenes and tumor suppressors that are required for medulloblastoma identity and proliferation. Shh-type medulloblastoma displays distinct H3K27me3 properties. We demonstrate that Brg1 modulates activities of H3K27me3 modifiers to regulate the expression of medulloblastoma genes. Brg1-regulated pathways are conserved in human Shh-type medulloblastoma, and Brg1 is important for the growth of a human medulloblastoma cell line. Thus, Brg1 coordinates a genetic and epigenetic network that regulates the transcriptional program underlying the Shh-type medulloblastoma development.

IR spectroscopy of α- and ß-protonated pyrrole via argon complex photodissociation.

Protonated pyrrole cations are produced in a pulsed discharge/supersonic expansion source, mass-selected in a time-of-flight spectrometer, and studied with infrared photodissociation spectroscopy. Vibrational spectra in both the fingerprint and C-H/N-H stretching regions are obtained using the method of tagging with argon. Sharp vibrational structure is compared to IR spectra predicted by theory for the possible α-, ß-, and N-protonated structures. The spectral differences among these isomers are much larger than the frequency shifts due to argon attachment at alternative sites. Though α-protonation predominates thermodynamically, the kinetically favored ß-protonated species is also observed for the first time (in 3-4 times lower abundance under the conditions employed here). Theoretical investigations attribute the greater stability of α-protonated pyrrole to topological charge stabilization, rather than merely to the greater number of resonance contributors. The far-IR pattern of protonated pyrrole does not match the interstellar UIR bands.

Prevention of postoperative nausea and vomiting after intrathecal morphine for Cesarean section: a randomized comparison of dexamethasone, droperidol, and a combination.

BACKGROUND: Intrathecal morphine provides good analgesia after cesarean delivery but the side effects include nausea and vomiting. Low-dose droperidol (0.625 mg) combined with dexamethasone 4 mg is postulated to have an additive antiemetic effect with less side effects. We therefore compared single doses of dexamethasone and droperidol alone with a low-dose combination of the two, to prevent spinal morphine-induced nausea and vomiting after cesarean section. METHODS: In a double-blind study, 120 women undergoing elective cesarean section under spinal anesthesia (using 0.5% bupivacaine 10 mg and morphine 0.2 mg) were allocated randomly to receive dexamethasone 8 mg, droperidol 1.25 mg, dexamethasone 4 mg and droperidol 0.625 mg, or placebo, before the end of surgery. The incidences of nausea and vomiting, sedative score, pain score, and side effects were recorded. RESULTS: The incidence of nausea and vomiting within 6 h postoperatively was lower and incidence of no nausea and vomiting for 24 h postoperatively was significantly higher for the combination group compared to the placebo group and the dexamethasone only group. Sedation scores within 3 h postoperatively and incidence of restlessness for the combination group were significantly lower than in the droperidol only group. CONCLUSION: An additive antiemetic effect and no significant side effects were shown for the combination of dexamethasone 4 mg and droperidol 0.625 mg. This combination was more effective than either dexamethasone 8 mg or droperidol 1.25 mg alone in preventing nausea and vomiting after spinal anesthesia using 0.5% bupivacaine and morphine 0.2 mg.

Three regulatory systems control expression of glutamine synthetase in Saccharomyces cerevisiae at the level of transcription.

The GLN1 gene of Saccharomyces cerevisiae was cloned by complementation of a gln1 auxotroph. A GLN1-lacZ fusion was constructed to assay GLN1 promoter activity. beta-Galactosidase and glutamine synthetase expression in chromosomally integrated GLN1-lacZ fusion strains were co-regulated in response to a shift from glutamine to glutamate as the nitrogen source, purine limitation, and 3-aminotriazole-induced histidine starvation. Regulation of GLN1 expression by each of the three pathways occurred at the transcriptional level. Increased accumulation of GLN1 mRNA was observed within 5 min after a shift from glutamine to glutamate as the nitrogen source. After 5 min, GLN1 mRNA levels were constant. The level of GLN1 transcript was reduced by approximately 75% within 5 min following glutamine addition to the cells growing with glutamate as nitrogen source. This indicates that the GLN1 message is unstable and has a half-life of approximately 3 min. Deletion analysis indicated that the sequences required for GLN1 expression are located within approximately 350 bp upstream from the transcriptional initiation site.