Rapid Total Synthesis of DARPin pE59 and Barnase

We report the convergent total synthesis of two proteins: DARPin pE59 and Bacillus amyloliquefaciens RNase (Barnase). Leveraging our recently developed fast-flow peptide-synthesis platform, we rapidly explored numerous conditions for the assembly of long polypeptides, and were able to mitigate common side reactions, including deletion and aspartimide products. We report general strategies for improving the synthetic quality of difficult peptide sequences with our system. High-quality protein fragments produced under optimal synthetic conditions were subjected to convergent native chemical ligation, which afforded native full-length proteins after a final desulfurization step. Both DARPin and Barnase were folded and found to be as active as their recombinant analogues.MIT Faculty Start-up FundMassachusetts Institute of Technology (Charles E. Reed Faculty Initiative Fund)Deshpande Center for Technological InnovationDamon Runyon-Rachleff (Innovation Award)Sontag Foundation (Distinguished Scientist Award)AstraZeneca (Firm) (Distinguished Graduate Student Fellowship)Daniel S. Kemp Summer FellowshipNational Institutes of Health (U.S.). Biotechnology Training Program (5T32GM008334-25

Nanostructured optical waveguide with a highly confined mode

We propose a transmission line working at telecom wavelengths with cross section as small as ${\lambda}^2/39$, which is 1.6 times smaller than that of optimized silicon waveguide. The proposed line can be implemented as a subwavelength fiber with plasmonic cladding. This considerable decrease in the line cross section is achieved by utilizing a plasmonic quasi-antisymmetric mode. The required plasmonic cladding is rather thin, therefore, losses are moderate and could be compensated by using amplifying core materials. Such a transmission line can find applications in densely integrated optical systems

Influence of nanoparticles and metal vapors on the color of laboratory and atmospheric discharges

Currently, electrical discharges occurring at altitudes of tens to hundreds of kilometers from the Earth’s surface attract considerable attention from researchers from all over the world. A significant number of (nano)particles coming from outer space burn up at these altitudes. As a result, vapors of various substances, including metals, are formed at different altitudes. This paper deals with the influence of vapors and particles released from metal electrodes on the color and shape of pulse-periodic discharge in air, nitrogen, argon, and hydrogen. It presents the results of experimental studies. The discharge was implemented under an inhomogeneous electric field and was accompanied by the generation of runaway electrons and the formation of mini-jets. It was established that regardless of the voltage pulse polarity, the electrode material significantly affects the color of spherical- and cylindrical-shaped mini jets formed when bright spots appear on electrodes. Similar jets are observed when the discharge is transformed into a spark. It was shown that the color of the plasma of mini-jets is similar to that of atmospheric discharges (red sprites, blue jets, and ghosts) at altitudes of dozens of kilometers and differs from the color of plasma of pulsed diffuse discharges in air and nitrogen at the same pressure. It was revealed that to observe the red, blue and green mini-jets, it is necessary to use aluminum, iron, and copper electrodes, respectivel

Effect of working gas pressure on interlayer mixing in magnetron-deposited Mo/Si multilayers

By methods of cross-sectional transmission electron microscopy and small-angle x-ray scattering (λ = 0.154 nm) the influence of Ar gas pressure (1 to 4 mTorr) on the growth of amorphous interfaces in Mo/Si multilayers (MLs) deposited by DC magnetron sputtering is studied. The significant reduction in the ML period, which is evident as a volumetric contraction, is observed in MLs deposited at Ar pressure where the mean-free path for the sputtered atoms is comparable with the magnetronsubstrate distance. Some reduction in the thickness of the amorphous interlayers with Ar pressure increase is found, where the composition of the interlayers is enriched with molybdenum. The interface modification resulted in an increase in EUV reflectance of the Mo/Si ML

Rapid Flow-Based Peptide Synthesis

A flow-based solid-phase peptide synthesis methodology that enables the incorporation of an amino acid residue every 1.8 min under automatic control or every 3 min under manual control is described. This is accomplished by passing a stream of reagent through a heat exchanger into a low volume, low backpressure reaction vessel, and through a UV detector. These features enable continuous delivery of heated solvents and reagents to the solid support at high flow rate, thereby maintaining maximal concentration of reagents in the reaction vessel, quickly exchanging reagents, and eliminating the need to rapidly heat reagents after they have been added to the vessel. The UV detector enables continuous monitoring of the process. To demonstrate the broad applicability and reliability of this method, it was employed in the total synthesis of a small protein, as well as dozens of peptides. The quality of the material obtained with this method is comparable to that for traditional batch methods, and, in all cases, the desired material was readily purifiable by RP-HPLC. The application of this method to the synthesis of the 113-residue Bacillus amyloliquefaciens RNase and the 130-residue DARPin pE59 is described in the accompanying manuscript.MIT Faculty Start-up FundMassachusetts Institute of Technology (Charles E. Reed Faculty Initiative Fund)Deshpande Center for Technological InnovationDamon Runyon-Rachleff (Innovation Award)Sontag Foundation (Distinguished Scientist Award)C. P. Chu and Y. Lai FellowshipDaniel S. Kemp Summer FellowshipNational Institute of General Medical Sciences (U.S.). Biotechnology Training Program (Grant 5T32GM008334-25)National Institutes of Health (U.S.) (Fellowship F32GM101762

Isolation and Characterization of Human Colon Adenocarcinoma Stem-Like Cells Based on the Endogenous Expression of the Stem Markers.

BACKGROUND: Cancer stem cells\u27 (CSCs) self-maintenance is regulated via the pluripotency pathways promoting the most aggressive tumor phenotype. This study aimed to use the activity of these pathways for the CSCs\u27 subpopulation enrichment and separating cells characterized by the OCT4 and SOX2 expression. METHODS: To select and analyze CSCs, we used the SORE6x lentiviral reporter plasmid for viral transduction of colon adenocarcinoma cells. Additionally, we assessed cell chemoresistance, clonogenic, invasive and migratory activity and the data of mRNA-seq and intrinsic disorder predisposition protein analysis (IDPPA). RESULTS: We obtained the line of CSC-like cells selected on the basis of the expression of the OCT4 and SOX2 stem cell factors. The enriched CSC-like subpopulation had increased chemoresistance as well as clonogenic and migration activities. The bioinformatic analysis of mRNA seq data identified the up-regulation of pluripotency, development, drug resistance and phototransduction pathways, and the downregulation of pathways related to proliferation, cell cycle, aging, and differentiation. IDPPA indicated that CSC-like cells are predisposed to increased intrinsic protein disorder. CONCLUSION: The use of the SORE6x reporter construct for CSCs enrichment allows us to obtain CSC-like population that can be used as a model to search for the new prognostic factors and potential therapeutic targets for colon cancer treatment

Cross sections and double-helicity asymmetries of midrapidity inclusive charged hadrons in p+p collisions at sqrt(s)=62.4 GeV

Unpolarized cross sections and double-helicity asymmetries of single-inclusive positive and negative charged hadrons at midrapidity from p+p collisions at sqrt(s)=62.4 GeV are presented. The PHENIX measurements for 1.0 < p_T < 4.5 GeV/c are consistent with perturbative QCD calculations at next-to-leading order in the strong coupling constant, alpha_s. Resummed pQCD calculations including terms with next-to-leading-log accuracy, yielding reduced theoretical uncertainties, also agree with the data. The double-helicity asymmetry, sensitive at leading order to the gluon polarization in a momentum-fraction range of 0.05 ~< x_gluon ~< 0.2, is consistent with recent global parameterizations disfavoring large gluon polarization.Comment: PHENIX Collaboration. 447 authors, 12 pages, 5 figures, 5 tables. Submitted to Physical Review

Inclusive cross section and double helicity asymmetry for pi^0 production in p+p collisions at sqrt(s) = 62.4 GeV

The PHENIX experiment presents results from the RHIC 2006 run with polarized proton collisions at sqrt(s) = 62.4 GeV for inclusive pi^0 production at mid-rapidity. Unpolarized cross section results are measured for transverse momenta p_T = 0.5 to 7 GeV/c. Next-to-leading order perturbative quantum chromodynamics calculations are compared with the data, and while the calculations are consistent with the measurements, next-to-leading logarithmic corrections improve the agreement. Double helicity asymmetries A_LL are presented for p_T = 1 to 4 GeV/c and probe the higher range of Bjorken_x of the gluon (x_g) with better statistical precision than our previous measurements at sqrt(s)=200 GeV. These measurements are sensitive to the gluon polarization in the proton for 0.06 < x_g < 0.4.Comment: 387 authors from 63 institutions, 10 pages, 6 figures, 1 table. Submitted to Physical Review D. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Inclusive cross section and single-transverse-spin asymmetry for very forward neutron production in polarized p+p collisions at sqrt(s)=200 GeV

The energy dependence of the single-transverse-spin asymmetry, A_N, and the cross section for neutron production at very forward angles were measured in the PHENIX experiment at RHIC for polarized p+p collisions at sqrt(s)=200 GeV. The neutrons were observed in forward detectors covering an angular range of up to 2.2 mrad. We report results for neutrons with momentum fraction of x_F=0.45 to 1.0. The energy dependence of the measured cross sections were consistent with x_F scaling, compared to measurements by an ISR experiment which measured neutron production in unpolarized p+p collisions at sqrt(s)=30.6--62.7 GeV. The cross sections for large x_F neutron production for p+p collisions, as well as those in e+p collisions measured at HERA, are described by a pion exchange mechanism. The observed forward neutron asymmetries were large, reaching A_N=-0.08+/-0.02 for x_F=0.8; the measured backward asymmetries, for negative x_F, were consistent with zero. The observed asymmetry for forward neutron production is discussed within the pion exchange framework, with interference between the spin-flip amplitude due to the pion exchange and nonflip amplitudes from all Reggeon exchanges. Within the pion exchange description, the measured neutron asymmetry is sensitive to the contribution of other Reggeon exchanges even for small amplitudes.Comment: 383 authors, 16 pages, 18 figures, 6 tables. Submitted to Phys. Rev. D. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm