Site-specific incorporation of phosphotyrosine using an expanded genetic code.

Access to phosphoproteins with stoichiometric and site-specific phosphorylation status is key to understanding the role of protein phosphorylation. Here we report an efficient method to generate pure, active phosphotyrosine-containing proteins by genetically encoding a stable phosphotyrosine analog that is convertible to native phosphotyrosine. We demonstrate its general compatibility with proteins of various sizes, phosphotyrosine sites and functions, and reveal a possible role of tyrosine phosphorylation in negative regulation of ubiquitination

Internal and external cooling methods and their effect on body temperature, thermal perception and dexterity

© 2018 The Authors. Published by PLOS. This is an open access article available under a Creative Commons licence. The published version can be accessed at the following link on the publisher’s website: https://doi.org/10.1371/journal.pone.0191416© 2018 Maley et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Objective The present study aimed to compare a range of cooling methods possibly utilised by occupational workers, focusing on their effect on body temperature, perception and manual dexterity. Methods Ten male participants completed eight trials involving 30 min of seated rest followed by 30 min of cooling or control of no cooling (CON) (34C, 58% relative humidity). The cooling methods utilised were: ice cooling vest (CV0), phase change cooling vest melting at 14C (CV14), evaporative cooling vest (CVEV), arm immersion in 10C water (AI), portable water-perfused suit (WPS), heliox inhalation (HE) and ice slushy ingestion (SL). Immediately before and after cooling, participants were assessed for fine (Purdue pegboard task) and gross (grip and pinch strength) manual dexterity. Rectal and skin temperature, as well as thermal sensation and comfort, were monitored throughout. Results Compared with CON, SL was the only method to reduce rectal temperature (P = 0.012). All externally applied cooling methods reduced skin temperature (P0.05). Conclusion The present study observed that ice ingestion or ice applied to the skin produced the greatest effect on rectal and skin temperature, respectively. AI should not be utilised if workers require subsequent fine manual dexterity. These results will help inform future studies investigating appropriate pre-cooling methods for the occupational worker.This project is financially supported by the US Government through the Technical Support Working Group within the Combating Terrorism Technical Support Office.Published versio

Profiling of Genes Related to Cross Protection and Competition for NbTOM1 by HLSV and TMV

10.1371/journal.pone.0073725PLoS ONE89-POLN

New chemolysis for urological calcium phosphate calculi – a study in vitro

BACKGROUND: Advances in techniques have left very few indications for open surgical extraction of urinary stones currently. These advances notwithstanding, the search continues for medical approaches to urinary stone management. In this study, we perform an in vitro study analyzing the efficiency and prospect of two new complex solutions in urological calcium phosphate calculi dissolution. METHODS: Eighteen stones composed mainly of calcium phosphates were taken from patients who underwent kidney stone surgery. These stones were large enough (weight range 0.514–0.928 g) to be fragmented and matched equally into six groups. Chemolysis of phosphate stones was done with six different solvents and was repeated 3 times with 6 stones for each solution. At 24, 48 and 72 h, reduction in weight, percentage weight change, and dissolution rate; the dissolution rates at pH 5.0, 7.0 and 8.5 for each solution, using different cations (Na(+), K(+ )or Ca(2+)), according to different dilutions (1:1, 1:2, 1:3, 1:4) of S1 and S2 were simultaneously determined. RESULTS: Calcium phosphate calculi were poorly dissolved by Phys and Art, and they had a low dissolution rate in pH 8.5 EDTA. The most effective solutions were S1, S2 and R, with 72 h mean dissolution rates: 5.75 ± 0.44 mg/hr (S1), 5.2 ± 0.63 mg/hr (S2), 4.55 ± 0.46 mg/hr (R) ([Image: see text] ± s, p < 0.01 R, S1 and S2 vs Phys, Art and EDTA; p < 0.05, S1 vs R, LSD-test). The mean percentage weight loss at 72 h was: 52.1 ± 15.75 % (S1), 44.4 ± 7.37 % (S2) and 40.5 ± 3.67 % (R) ([Image: see text] ± s, p < 0.01 R, S1 and S2 vs Phys, Art and EDTA, LSD-test). Diluted twice, S1 and S2 had even better effectiveness than their initial solution. The additive of Na(+), K(+ )or Ca(2+ )greatly reduced the dissolution rates of S1, S2. CONCLUSION: Our data indicate that test solutions S1 and S2 are effective solvents in the chemolysis of calcium phosphate stones. At twice dilutions, these solutions are even more useful in the treatment of stone disease

Evaluating Statistical Methods Using Plasmode Data Sets in the Age of Massive Public Databases: An Illustration Using False Discovery Rates

Plasmode is a term coined several years ago to describe data sets that are derived from real data but for which some truth is known. Omic techniques, most especially microarray and genomewide association studies, have catalyzed a new zeitgeist of data sharing that is making data and data sets publicly available on an unprecedented scale. Coupling such data resources with a science of plasmode use would allow statistical methodologists to vet proposed techniques empirically (as opposed to only theoretically) and with data that are by definition realistic and representative. We illustrate the technique of empirical statistics by consideration of a common task when analyzing high dimensional data: the simultaneous testing of hundreds or thousands of hypotheses to determine which, if any, show statistical significance warranting follow-on research. The now-common practice of multiple testing in high dimensional experiment (HDE) settings has generated new methods for detecting statistically significant results. Although such methods have heretofore been subject to comparative performance analysis using simulated data, simulating data that realistically reflect data from an actual HDE remains a challenge. We describe a simulation procedure using actual data from an HDE where some truth regarding parameters of interest is known. We use the procedure to compare estimates for the proportion of true null hypotheses, the false discovery rate (FDR), and a local version of FDR obtained from 15 different statistical methods

Over-Expression of LSD1 Promotes Proliferation, Migration and Invasion in Non-Small Cell Lung Cancer

Background: Lysine specific demethylase 1 (LSD1) has been identified and biochemically characterized in epigenetics, but the pathological roles of its dysfunction in lung cancer remain to be elucidated. The aim of this study was to evaluate the prognostic significance of LSD1 expression in patients with non-small cell lung cancer (NSCLC) and to define its exact role in lung cancer proliferation, migration and invasion. Methods: The protein levels of LSD1 in surgically resected samples from NSCLC patients were detected by immunohistochemistry or Western blotting. The mRNA levels of LSD1 were detected by qRT-PCR. The correlation of LSD1 expression with clinical characteristics and prognosis was determined by statistical analysis. Cell proliferation rate was assessed by MTS assay and immunofluorescence. Cell migration and invasion were detected by scratch test, matrigel assay and transwell invasion assay. Results: LSD1 expression was higher in lung cancer tissue more than in normal lung tissue. Our results showed that overexpression of LSD1 protein were associated with shorter overall survival of NSCLC patients. LSD1 was localized mainly to the cancer cell nucleus. Interruption of LSD1 using siRNA or a chemical inhibitor, pargyline, suppressed proliferation, migration and invasion of A549, H460 and 293T cells. Meanwhile, over-expression of LSD1 enhanced cell growth. Finally, LSD1 was shown to regulate epithelial-to-mesenchymal transition in lung cancer cells

Binary Willshaw learning yields high synaptic capacity for long-term familiarity memory

We investigate from a computational perspective the efficiency of the Willshaw synaptic update rule in the context of familiarity discrimination, a binary-answer, memory-related task that has been linked through psychophysical experiments with modified neural activity patterns in the prefrontal and perirhinal cortex regions. Our motivation for recovering this well-known learning prescription is two-fold: first, the switch-like nature of the induced synaptic bonds, as there is evidence that biological synaptic transitions might occur in a discrete stepwise fashion. Second, the possibility that in the mammalian brain, unused, silent synapses might be pruned in the long-term. Besides the usual pattern and network capacities, we calculate the synaptic capacity of the model, a recently proposed measure where only the functional subset of synapses is taken into account. We find that in terms of network capacity, Willshaw learning is strongly affected by the pattern coding rates, which have to be kept fixed and very low at any time to achieve a non-zero capacity in the large network limit. The information carried per functional synapse, however, diverges and is comparable to that of the pattern association case, even for more realistic moderately low activity levels that are a function of network size.Comment: 20 pages, 4 figure

ZnSe/ZnSeTe Superlattice Nanotips

The authors report the growth of ZnSe/ZnSeTe superlattice nanotips on oxidized Si(100) substrate. It was found the nanotips exhibit mixture of cubic zinc-blende and hexagonal wurtzite structures. It was also found that photoluminescence intensities observed from the ZnSe/ZnSeTe superlattice nanotips were much larger than that observed from the homogeneous ZnSeTe nanotips. Furthermore, it was found that activation energies for the ZnSe/ZnSeTe superlattice nanotips with well widths of 16, 20, and 24 nm were 76, 46, and 19 meV, respectively

A Selectable and Excisable Marker System for the Rapid Creation of Recombinant Poxviruses

Genetic manipulation of poxvirus genomes through attenuation, or insertion of therapeutic genes has led to a number of vector candidates for the treatment of a variety of human diseases. The development of recombinant poxviruses often involves the genomic insertion of a selectable marker for purification and selection purposes. The use of marker genes however inevitably results in a vector that contains unwanted genetic information of no therapeutic value.Here we describe an improved strategy that allows for the creation of marker-free recombinant poxviruses of any species. The Selectable and Excisable Marker (SEM) system incorporates a unique fusion marker gene for the efficient selection of poxvirus recombinants and the Cre/loxP system to facilitate the subsequent removal of the marker. We have defined and characterized this new methodological tool by insertion of a foreign gene into vaccinia virus, with the subsequent removal of the selectable marker. We then analyzed the importance of loxP orientation during Cre recombination, and show that the SEM system can be used to introduce site-specific deletions or inversions into the viral genome. Finally, we demonstrate that the SEM strategy is amenable to other poxviruses, as demonstrated here with the creation of an ectromelia virus recombinant lacking the EVM002 gene.The system described here thus provides a faster, simpler and more efficient means to create clinic-ready recombinant poxviruses for therapeutic gene therapy applications

Overexpression of Nrdp1 in the Heart Exacerbates Doxorubicin-Induced Cardiac Dysfunction in Mice

BACKGROUND: Cardiac cell death and generation of oxidative stress contribute to doxorubicin (DOX)-induced cardiac dysfunction. E3 ligase Nrdp1 plays a critical role in the regulation of cell apoptosis, inflammation and production of reactive oxygen species (ROS), which may contribute to heart failure. However, the role of Nrdp1 in DOX-induced cardiac injury remains to be determined. METHODS AND RESULTS: We examined the effect of Nrdp1 overexpression with DOX treatment in rat neonatal cardiomyocytes and mouse heart tissue. Cardiomyocytes were infected with adenovirus containing GFP (Ad-GFP), Nrdp1 wild-type (Ad-Nrdp1) or the dominant-negative form of Nrdp1 (Ad-Dn-Nrdp1), then treated with DOX for 24 hr. DOX treatment increased cell death and apoptosis, with Ad-Nrdp1 infection enhancing these actions but Ad-Dn-Nrdp1 infection attenuating these effects. Furthermore, 5 days after a single injection of DOX (20 mg/kg, intraperitoneally), Nrdp1 transgenic mice (TG) showed decreased cardiac function and increased apoptosis, autophagy and oxidative stress as compared with wild-type (WT) mice (P<0.01). Survival rate was significantly lower in Nrdp1 TG mice than in WT mice 10 days after DOX injection (P<0.01). CONCLUSIONS/SIGNIFICANCE: These results were associated with decreased activation of Akt, extracellular signal-regulated kinase 1/2 (ERK1/2) and signal transducer and activator of transcription 3 (STAT3) signaling pathways. Nrdp1 may be a key mediator in the development of cardiac dysfunction after DOX treatment and associated with inhibition of Akt, ERK1/2 and STAT3. Nrdp1 may be a new therapeutic target in protecting against the cardiotoxic effects of DOX