Toward a Synthetic Yeast Endosymbiont with a Minimal Genome

Based on the endosymbiotic theory, one of the key events that occurred during mitochondrial evolution was an extensive loss of nonessential genes from the protomitochondrial endosymbiont genome and transfer of some of the essential endosymbiont genes to the host nucleus. We have developed an approach to recapitulate various aspects of endosymbiont genome minimization using a synthetic system consisting of Escherichia coli endosymbionts within host yeast cells. As a first step, we identified a number of E. coli auxotrophs of central metabolites that can form viable endosymbionts within yeast cells. These studies provide a platform to identify nonessential biosynthetic pathways that can be deleted in the E. coli endosymbionts to investigate the evolutionary adaptations in the host and endosymbiont during the evolution of mitochondria

Additional file 1: of Efficacy and safety profile of nab-paclitaxel plus gemcitabine in patients with metastatic pancreatic cancer treated to disease progression: a subanalysis from a phase 3 trial (MPACT)

List of Independent Ethics Committees and Institutional Review Boards for MPACT. (DOCX 39 kb

Aromatic Aldehyde and Hydrazine Activated Peptide Coated Quantum Dots for Easy Bioconjugation and Live Cell Imaging

We present a robust scheme for preparation of semiconductor quantum dots (QDs) and cognate partners in a conjugation ready format. Our approach is based on bis-aryl hydrazone bond formation mediated by aromatic aldehyde and hydrazinonicotinate acetone hydrazone (HyNic) activated peptide coated quantum dots. We demonstrate controlled preparation of antibody–QD bioconjugates for specific targeting of endogenous epidermal growth factor receptors in breast cancer cells and for single QD tracking of transmembrane proteins via an extracellular epitope. The same approach was also used for optical mapping of RNA polymerases bound to combed genomic DNA in vitro

Aromatic Aldehyde and Hydrazine Activated Peptide Coated Quantum Dots for Easy Bioconjugation and Live Cell Imaging

We present a robust scheme for preparation of semiconductor quantum dots (QDs) and cognate partners in a conjugation ready format. Our approach is based on bis-aryl hydrazone bond formation mediated by aromatic aldehyde and hydrazinonicotinate acetone hydrazone (HyNic) activated peptide coated quantum dots. We demonstrate controlled preparation of antibody–QD bioconjugates for specific targeting of endogenous epidermal growth factor receptors in breast cancer cells and for single QD tracking of transmembrane proteins via an extracellular epitope. The same approach was also used for optical mapping of RNA polymerases bound to combed genomic DNA in vitro

Aromatic Aldehyde and Hydrazine Activated Peptide Coated Quantum Dots for Easy Bioconjugation and Live Cell Imaging

We present a robust scheme for preparation of semiconductor quantum dots (QDs) and cognate partners in a conjugation ready format. Our approach is based on bis-aryl hydrazone bond formation mediated by aromatic aldehyde and hydrazinonicotinate acetone hydrazone (HyNic) activated peptide coated quantum dots. We demonstrate controlled preparation of antibody–QD bioconjugates for specific targeting of endogenous epidermal growth factor receptors in breast cancer cells and for single QD tracking of transmembrane proteins via an extracellular epitope. The same approach was also used for optical mapping of RNA polymerases bound to combed genomic DNA in vitro

Characterizing the Quantum Confined Stark Effect in Semiconductor Quantum Dots and Nanorods for Single-Molecule Electrophysiology

We optimized the performance of quantum confined Stark effect QCSE based voltage nanosensors. A high throughput approach for single particle QCSE characterization was developed and utilized to screen a library of such nanosensors. Type II ZnSe CdS seeded nanorods were found to have the best performance among the different nanosensors evaluated in this work. The degree of correlation between intensity changes and spectral changes of the excitons emission under applied field was characterized. An upper limit for the temporal response of individual ZnSe CdS nanorods to voltage modulation was characterized by high throughput, high temporal resolution intensity measurements using a novel photon counting camera. The measured 3.5 us response time is limited by the voltage modulation electronics and represents about 30 times higher bandwidth than needed for recording an action potential in a neuron

Expression of CHOP and GADD34 Correlates with Cell Fate

<div><p>(A) MEFs were treated with increasing concentrations of TG or TM for the indicated times, followed by cell lysis and immunoblot for CHOP, or α-actin or Sec61β to judge loading. The concentrations outlined by the gray box are those that allow for survival. For every time-point, the TM and TG panels were taken from the same blot, and the same exposure time.</p> <p>(B) MEFs were treated for up to 5 d in the continuous presence of 25- or 50-ng/ml TM, with the media and stressor refreshed each day; and expression of CHOP, BiP, and α-actin was probed by immunoblot.</p> <p>(C) Same as in (A), probing instead for GADD34, or α-actin or TRAPα as loading controls.</p> <p>(D) Cells treated continuously in TG at the indicated concentrations were probed for expression of BiP, GADD34, and α-actin by immunoblot.</p></div

UPR Activation Can Permit Cell Survival and Proliferation

<div><p>(A) MEFs were cultured for 24 h in the presence of increasing concentrations of TM or TG. Cell lysates were then probed by immunoblot with antibodies specific for BiP, or α-actin as a loading control.</p> <p>(B) MEFs were cultured in the presence of 25-ng/ml TM or vehicle. The ER fraction was isolated from each by differential centrifugation, and was then separated by two-dimensional SDS-PAGE. The gels were stained for total protein with SYPRO Ruby. The series of spots corresponding to the ER-resident sentinel glycoprotein HSP47, identified by mass spectrometry, is indicated, with fully glycosylated (HSP47-CHO, filled arrow), underglycosylated (HSP47-CHO, open arrow) and unglycosylated (HSP47, open arrow) species visible. There are four isoforms of HSP47 that differ in isolectric point. Treatment of cells in a higher concentration of TM confirmed that the faster-migrating species are under- or non-glycosylated forms of the protein.</p> <p>(C) MEFs were treated overnight in the indicated concentration of TM. Cells were fixed in 2.5% glutaraldehyde, then prepared for transmission electron microscopic analysis. Nuclei (N), mitochondria (M), and rough ER (RER) are indicated. All images are at 25,000× magnification. Scale bar represents 500 nm.</p> <p>(D) MEFs were plated in replicate and exposed to varying concentrations of TM or TG, with the media and stressor refreshed daily. At each day, cells from individual plates were washed and trypsinized, and the cell concentration in the disrupted cell suspension was determined using an automated cell counter. Each suspension was counted twice, and error bars represent means ± SDM from triplicate plates. The numbers given on the <i>y</i>-axis represent cell number per milliliter in the diluted cell suspension. The gray bar is extended from the starting cell count plus the standard deviation, to allow net growth of the culture, or lack thereof, to be more readily assessed.</p> <p>(E) Cells were cultured for 6 d in the presence of 25-ng/ml TM, refreshed daily, and analyzed by electron microscopy for ER structure as in (C).</p></div

RSV characterised transcripts at 16 hours post infection.

The first datasheet shows the list of transcript groups identified by the pipeline. For each transcript group it is noted how many sequence reads belong to the group, the average poly A length, the features present on the transcript (as well as the predicted splice acceptor/donor pair usage if there is an apparent intron—in the case of RSV this is clearly not genuine splicing) and what ORFS are found that are present in the features file that accompanied the analysis. (XLSX)</p