Translational control analysis by translationally active RNA capture/microarray analysis (TrIP–Chip)

We have developed a new approach to systematically study post-transcriptional regulation in a small number of cells. Actively translating mRNAs are associated with polysomes and the newly synthesized peptide chains are closely associated with molecular chaperones such as hsp70s, which assist in the proper folding of nascent polypeptides into higher ordered structures. These chaperones provide an anchor with which to separate actively translating mRNAs associated with polysomes from free mRNAs. Affinity capture beads were developed to capture hsp70 chaperones associated with the polysome complexes. The isolated actively translating mRNAs were used for high-throughput expression profiling analysis. Feasibility was demonstrated using an in vitro translation system with known translationally regulated mRNA transcript thymidylate synthase (TS). We further developed the approach using HCT-116 colon cancer cells with both TS and p53 as positive controls. The steady-state levels of TS and p53 mRNAs were unaltered after 5-fluorouracil treatment as assessed by real-time qRT-PCR analysis. In contrast, the protein expression and polysome-associated mRNA levels of both genes were increased. These differences in translational rate were revealed with our new approach from 500 cells. This technology has the potential to make investigation of translational control feasible with limited quantities of clinical specimens

Deciphering the Non-Equivalence of Serine and Threonine O-Glycosylation Points: Implications for Molecular Recognition of the Tn Antigen by an anti-MUC1 Antibody.

The structural features of MUC1-like glycopeptides bearing the Tn antigen (α-O-GalNAc-Ser/Thr) in complex with an anti MUC-1 antibody are reported at atomic resolution. For the α-O-GalNAc-Ser derivative, the glycosidic linkage adopts a high-energy conformation, barely populated in the free state. This unusual structure (also observed in an α-S-GalNAc-Cys mimic) is stabilized by hydrogen bonds between the peptidic fragment and the sugar. The selection of a particular peptide structure by the antibody is thus propagated to the carbohydrate through carbohydrate/peptide contacts, which force a change in the orientation of the sugar moiety. This seems to be unfeasible in the α-O-GalNAc-Thr glycopeptide owing to the more limited flexibility of the side chain imposed by the methyl group. Our data demonstrate the non-equivalence of Ser and Thr O-glycosylation points in molecular recognition processes. These features provide insight into the occurrence in nature of the APDTRP epitope for anti-MUC1 antibodies.We thank the Ministerio de Econom a y Competitividad/FEDER (project CTQ2012-36365, CTQ2012-32065, BFU2010-19504, CTQ2013-44367-C2-2-P, UNLR13-4E-1931 and grant I.C.) and DGA (B89) for financial support. N.M.-S. and D.M. thank Universidad de La Rioja for FPI grants. We thank Katherine Stott (Department of Biochemistry, Cambridge University) for technical help with the BLI experiments. G.J.L.B. thanks financial support from the EPSRC. G.J.L.B. is a Royal Society University Research Fellow. M.S. thanks the Generalitat de Catalunya and Universitat Rovira i Virgili for financial support.This is the published version. It first appeared at http://onlinelibrary.wiley.com/doi/10.1002/anie.201502813/abstract

Simultaneous Regeneration of Two 160 Gbit/s WDM Channels in a Single Highly Nonlinear Fiber

We experimentally demonstrate simultaneous all-optical regeneration of two 160-Gbit/s wavelength-division multiplexed (WDM) channels in a single highly nonlinear fiber (HNLF). The multi-channel regeneration performance is confirmed by bit-error rate (BER) measurements. The receiver powers at a BER of 10-9 are improved by about 4.9 dB and 2.1 dB for the two channels, respectively. The BER performance is not degraded by the presence of a second channel. Mitigation of the inter-channel nonlinearities is achieved through bidirectional propagation.</p

4 × 160-Gbit/s multi-channel regeneration in a single fiber

Simultaneous regeneration of four high-speed (160 Gbit/s) wavelength-division multiplexed (WDM) and polarization-division multiplexed (PDM) signals in a single highly nonlinear fiber (HNLF) is demonstrated. The regeneration operation is based on four-wave mixing in HNLF, where the degraded data signals are applied as the pump. As a result, the noise on both '0' and '1' levels can be suppressed simultaneously in our scheme. The stimulated Brillouin scattering (SBS) from the continuous wave (CW) is suppressed by cross-phase modulation (XPM) from the data pump, relieving the requirement of external phase modulation of the CW light. Mitigation of the inter-channel nonlinearities is achieved mainly through an inter-channel 0.5 bit slot time delay. Bidirectional propagation is also applied to relieve the inter-channel four-wave mixing. The multi-channel regeneration performance is validated by bit-error rate (BER) measurements. The receiver powers at the BER of 10-9 are improved by 1.9 dB, 1.8 dB, 1.6 dB and 1.5 dB for the four data channels, respectively.</p

Skin Aging and Photoaging Alter Fatty Acids Composition, Including 11,14,17-eicosatrienoic Acid, in the Epidermis of Human Skin

We investigated the alterations of major fatty acid components in epidermis by natural aging and photoaging processes, and by acute ultraviolet (UV) irradiation in human skin. Interestingly, we found that 11,14,17-eicosatrienoic acid (ETA), which is one of the omega-3 polyunsaturated acids, was significantly increased in photoaged human epidermis in vivo and also in the acutely UV-irradiated human skin in vivo, while it was significantly decreased in intrinsically aged human epidermis. The increased ETA content in the epidermis of photoaged human skin and acute UV-irradiated human skin is associated with enhanced expression of human elongase 1 and calcium-independent phophodiesterase A2. We demonstrated that ETA inhibited matrix metalloproteinase (MMP)-1 expression after UV-irradiation, and that inhibition of ETA synthesis using EPTC and NA-TCA, which are elongase inhibitors, increased MMP-1 expression. Therefore, our results suggest that the UV increases the ETA levels, which may have a photoprotective effect in the human skin

Exogenous dopamine reduces GABA receptor availability in the human brain

Background: While it has recently been shown that dopamine release stimulates conscious self‐monitoring through the generation of gamma oscillations in medial prefrontal/anterior cingulate cortex, and that the GABAergic system is effective in producing such oscillations, interaction of the two transmitter systems has not been demonstrated in humans. We here hypothesize that dopamine challenge stimulates the GABA system directly in the medial prefrontal/anterior cingulate region in the human brain. // Methods: Positron emission tomography (PET) with the GABA receptor α1/α5 subtype ligand [11C] Ro15‐4513 was used to detect changes in GABA receptor availability after clinical oral doses of levodopa in a double blind controlled study. // Results: We here provide the first direct evidence for such coupling in the cerebral cortex, in particular in the medial prefrontal anterior cingulate region, by showing that exogenous dopamine decreases [11C] Ro15‐4513 binding widely in the human brain compatible with a fall in α1 subtype availability in GABA complexes due to increased GABA activity

Monitoring conterminous United States (CONUS) land cover change with Web-Enabled Landsat Data (WELD)

Forest cover loss and bare ground gain from 2006 to 2010 for the conterminous United States (CONUS) were quantified at a 30 m spatial resolution using Web-Enabled Landsat Data available from the USGS Center for Earth Resources Observation and Science (EROS) (http://landsat.usgs.gov/WELD.php). The approach related multi-temporal WELD metrics and expert-derived training data for forest cover loss and bare ground gain through a decision tree classification algorithm. Forest cover loss was reported at state and ecoregional scales, and the identification of core forests\u27 absent of change was made and verified using LiDAR data from the GLAS (Geoscience Laser Altimetry System) instrument. Bare ground gain correlated with population change for large metropolitan statistical areas (MSAs) outside of desert or semi-desert environments. Google Earth™ time series images were used to validate the products. Mapped forest cover loss totaled 53,084 km2 and was found to be depicted conservatively, with a user\u27s accuracy of 78% and a producer\u27s accuracy of 68%. Excluding errors of adjacency, user\u27s and producer\u27s accuracies rose to 93% and 89%, respectively. Mapped bare ground gain equaled 5974 km2 and nearly matched the estimated area from the reference (Google Earth™) classification; however, user\u27s (42%) and producer\u27s (49%) accuracies were much less than those of the forest cover loss product. Excluding errors of adjacency, user\u27s and producer\u27s accuracies rose to 62% and 75%, respectively. Compared to recent 2001–2006 USGS National Land Cover Database validation data for forest loss (82% and 30% for respective user\u27s and producer\u27s accuracies) and urban gain (72% and 18% for respective user\u27s and producer\u27s accuracies), results using a single CONUS-scale model with WELD data are promising and point to the potential for national scale operational mapping of key land cover transitions. However, validation results highlighted limitations, some of which can be addressed by improving training data, creating a more robust image feature space, adding contemporaneous Landsat 5 data to the inputs, and modifying definition sets to account for differences in temporal and spatial observational scales. The presented land cover extent and change data are available via the official WELD website (ftp://weldftp.cr.usgs.gov/CONUS_5Y_LandCover/ftp://weldftp.cr.usgs. gov/CONUS_5Y_LandCover/)

Sodium glucose co-transporter 2 inhibition increases epidermal growth factor expression and improves outcomes in patients with type 2 diabetes

Underlying molecular mechanisms of the kidney protective effects of sodium glucose co-transporter 2 (SGLT2) inhibitors are not fully elucidated. Therefore, we studied the association between urinary epidermal growth factor (uEGF), a mitogenic factor involved in kidney repair, and kidney outcomes in patients with type 2 diabetes (T2D). The underlying molecular mechanisms of the SGLT2 inhibitor canagliflozin on EGF using single-cell RNA sequencing from kidney tissue were examined. Urinary EGF-to-creatinine ratio (uEGF/Cr) was measured in 3521 CANagliflozin cardioVascular Assessment Study (CANVAS) participants at baseline and week 52. Associations of uEGF/Cr with kidney outcome were assessed using multivariable-adjusted Cox regression models. Single-cell RNA sequencing was performed using protocol kidney biopsy tissue from ten young patients with T2D on SGLT2i, six patients with T2D on standard care only, and six healthy controls (HCs). In CANVAS, each doubling in baseline uEGF/Cr was associated with a 12% (95% confidence interval 1-22) decreased risk of kidney outcome. uEGF/Cr decreased after 52 weeks with placebo and remained stable with canagliflozin (between-group difference +7.3% (2.0-12.8). In young persons with T2D, EGF mRNA was primarily expressed in the thick ascending loop of Henle. Expression in biopsies from T2D without SGLT2i was significantly lower compared to HCs, whereas treatment with SGLT2i increased EGF levels closer to the healthy state. In young persons with T2D without SGLT2i, endothelin-1 emerged as a key regulator of the EGF co-expression network. SGLT2i treatment was associated with a shift towards normal EGF expression. Thus, decreased uEGF represents increased risk of kidney disease progression in patients with T2D. Canagliflozin increased kidney tissue expression of EGF and was associated with a downstream signaling cascade linked to tubular repair and reversal of tubular injury.</p