BKM120 induces apoptosis and inhibits tumor growth in medulloblastoma.

Medulloblastoma (MB) is the most common malignant brain tumor in children, accounting for nearly 20 percent of all childhood brain tumors. New treatment strategies are needed to improve patient survival outcomes and to reduce adverse effects of current therapy. The phosphatidylinositol-3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) intracellular signaling pathway plays a key role in cellular metabolism, proliferation, survival and angiogenesis, and is often constitutively activated in human cancers, providing unique opportunities for anticancer therapeutic intervention. The aim of this study was to evaluate the pre-clinical activity of BKM120, a selective pan-class I PI3K inhibitor, on MB cell lines and primary samples. IC50 values of BKM120 in the twelve MB cell lines tested ranged from 0.279 to 4.38 µM as determined by cell viability assay. IncuCyte ZOOM Live-Cell Imaging system was used for kinetic monitoring of cytotoxicity of BKM120 and apoptosis in MB cells. BKM120 exhibited cytotoxicity in MB cells in a dose and time-dependent manner by inhibiting activation of downstream signaling molecules AKT and mTOR, and activating caspase-mediated apoptotic pathways. Furthermore, BKM120 decreased cellular glycolytic metabolic activity in MB cell lines in a dose-dependent manner demonstrated by ATP level per cell. In MB xenograft mouse study, DAOY cells were implanted in the flank of nude mice and treated with vehicle, BKM120 at 30 mg/kg and 60 mg/kg via oral gavage daily. BKM120 significantly suppressed tumor growth and prolonged mouse survival. These findings help to establish a basis for clinical trials of BKM120, which could be a novel therapy for the treatment of medulloblastoma patients.

MRI injury patterns in surgically confirmed and reconstructed posterolateral corner knee injuries.

PURPOSE: The posterolateral corner (PLC) of the knee is anatomically complex with similarly complex MR imaging findings in acutely injured knees. The purpose of this study was to define the MRI pattern of injury in cases of PLC disruption requiring surgery because of clinical instability. METHODS: The knee MRIs of 22 patients who underwent surgical repair and/or reconstruction of PLC injury were retrospectively reviewed. The fibular collateral ligament (FCL), popliteus tendon (PT), biceps femoris (BF), popliteofibular ligament (PFL), arcuate ligament (AL), and fabellofibular ligament (FFL) were evaluated and graded as follows: complete tear, high-grade partial tear, low-grade partial tear, and normal. RESULTS: In the 22 cases of PLC injury that necessitated surgery, a constellation of findings involving the larger structures of the PLC was identified. Of the FCL, PT, and BF (considered larger structures), at least two were abnormal in all 22 injury cases. Of the PFL, AL, and FFL (considered smaller structures), the PFL appeared abnormal in 19 cases, yet neither the AL nor FFL were confidently characterized in the injury group. CONCLUSION: The larger structures of the PLC are easily evaluated using standard MRI techniques. This study identified a predictable pattern of imaging findings involving these more easily assessed structures in those patients who were felt to be clinically unstable and underwent surgical reconstruction, as at least two were abnormal in all 22 cases. The smaller structures of the PLC are difficult to assess with MRI; however, direct visualization of their involvement on MRI is not necessary to report a clinically unstable PLC injury. Emphasis of this simplified but critical analysis of the FCL, BF and PT on MRI scans reviewed by radiologists and orthopaedic surgeons may help to prevent delayed diagnosis of unstable PLC injuries. LEVEL OF EVIDENCE: III.

Photoluminescence imaging of suspended single-walled carbon nanotubes.

Single-walled carbon nanotubes (SWNTs) suspended in air over trenches are imaged using their intrinsic near-infrared (NIR) photoluminescence (1.0-1.6 microm). Far-field emission from extended suspended lengths (approximately 50 microm) is both spatially and spectrally resolved, and SWNTs are classified based on the spatial uniformity of their emission intensity and emission wavelength. In a few cases, emission assigned to different (n,m) species is observed along the same suspended segment. Most SWNTs imaged on millisecond time scales show steady emission, but a few fluctuate and suffer a reduction of intensity. The quantum efficiency is dramatically higher than that in previous reports and is estimated at 7%, a value that is precise but subject to corrections because of assumptions about absorption and coherence.

Ca2+ source-dependent transcription of CRE-containing genes in vascular smooth muscle.

Altered Ca2+ handling has immediate physiological and long-term genomic effects on vascular smooth muscle function. Previously we showed that Ca2+ entry through voltage-dependent Ca2+ channels (VDCCs) or store-operated Ca2+ channels (SOCCs) results in phosphorylation of the Ca2+/cAMP response element (CRE)-binding protein in cerebral arteries. Here, oligonucleotide array analysis was used to determine gene transcription profiles resulting from these two Ca2+ entry pathways in human cerebrovascular smooth muscle cell cultures. Results were confirmed and expanded using quantitative RT-PCR, Western blot, and immunofluorescence. A distinct, yet overlapping, set of CRE-regulated genes was induced by VDCC activation using K+ membrane depolarization vs. SOCC activation by thapsigargin (TG). Membrane depolarization selectively induced a sustained increase in early growth response-1 (Egr-1) mRNA and protein, which were inhibited by the VDCC blocker nimodipine and the SOCC inhibitor 2-aminoethoxydiphenylborate (2-APB). TG selectively induced a sustained increase in MAPK phosphatase-1 (MKP-1) mRNA and protein, and these effects were decreased by 2-APB, but not by nimodipine. The physiological agonist ANG II also stimulated expression of Egr-1 and MKP-1. Coadministration of 2-APB prevented expression of Egr-1 and MKP-1, whereas nimodipine blocked only Egr-1 expression. TG and ANG II induced phosphorylation of ERK, which was sensitive to 2-APB and was selectively required for CRE-binding protein phosphorylation. Our findings thus indicate that Ca2+ entry through VDCCs and store-operated Ca2+ entry can differentially regulate CRE-containing genes in vascular smooth muscle and also imply that agonist-induced signals involved in modulation of gene transcription can be controlled by multiple sources of Ca2+.