Protein Expression and Purification of Human Hormone Sensitive Lipase

The purpose of this project was to develop a new product, purified human hormone sensitive lipase (HSL), for Blue Sky Biotech\u27s product line. This was accomplished by cloning the HSL gene by gene synthesis, amplifying the annealed templates by PCR, cloning the amplicons into a plasmid in E. coli, packaging the gene into baculovirus, expressing the gene in cultured insect cells, and purifying the protein by affinity chromatography using histidine and biotin tags contained in the engineered vector. The results indicate that HSL has been purified and remains biologically active. Blue Sky Biotech will be able to sell this product to laboratories currently researching lipolysis pathways involved in diseases such as obesity, diabetes, and hyperlipidemia

Stem emissions of monoterpenes, acetaldehyde, and methanol from Scots pine (Pinus sylvestris L.) affected by tree water relations and cambial growth

Abstract Tree stems are an overlooked source of volatile organic compounds (VOCs). Their contribution to ecosystem processes and total VOC fluxes is not well studied, and assessing it requires better understanding of stem emission dynamics and their driving processes. To gain more mechanistic insight into stem emission patterns, we measured monoterpene, methanol, and acetaldehyde emissions from the stems of mature Scots pines (Pinus sylvestris L.) in a boreal forest over three summers. We analysed the effects of temperature, soil water content, tree water status, transpiration, and growth on the VOC emissions, and used generalized linear models to test their relative importance in explaining the emissions. We show that Scots pine stems are considerable sources of monoterpenes, methanol, and acetaldehyde, and their emissions are strongly regulated by temperature. However, even small changes in water availability affected the emission potentials: increased soil water content increased the monoterpene emissions within a day, whereas acetaldehyde and methanol emissions responded within two to four days. This lag corresponded to their transport time in the xylem sap from the roots to the stem. Moreover, the emissions of monoterpenes, methanol, and acetaldehyde were influenced by the cambial growth rate of the stem with six- to ten-day lags. This article is protected by copyright. All rights reserved.Peer reviewe

Effect of strain rate on tensile mechanical properties of high-purity niobium single crystals for SRF applications

An investigation of the mechanical properties of high-purity niobium single crystals is presented. Specimens were cut with different crystallographic orientations from a large grain niobium disk and uniaxial tensile tests were conducted at strain rates between 10-4 and 103 s-1. The logarithmic strain rate sensitivity for crystals oriented close to the center of a tensile axis inverse pole figure (IPF) is ~0.14 for all strain rates. The strain at failure (ranging from 0.4 to 0.9) is very sensitive to crystal orientation and maximal at ~10-2 s-1 for crystals oriented close to the center of an IPF. The high anisotropy observed at quasi-static strain rates decreased with increasing strain rate. The activation of multiple slip systems in the dynamic tests could account for this reduction in anisotropy. A transition from strain hardening to softening in the plastic domain was observed at strain rates greater than approximately 6 × 10-2 s-1 for crystals oriented close to the center of a tensile axis IPF. Shear bands were observed in specimens with orientations having similarly high Schmid factors on both {110} and {112} slip families, and they are correlated with reduced ductility. Crystal rotations at fracture are compared for the different orientations using scanning electron microscopy images and EBSD orientation maps. A rotation toward the terminal stable [101] orientation was measured for the majority of specimens (with tensile axes more than ~17° from the [001] direction) at strain rates between 1.28 × 10-2 and 1000 s-1.The authors would like to acknowledge the work of CERN's Materials, Metrology and Non-Destructive Testing (EN-MME-MM) section for granting access to their equipment for specimen preparation and scanning electron microscope (SEM) analyses. The authors would also like to thank Mr. Larry Vladic of Elite Motion LLC for lending us the high-speed camera during the high strain rate tests performed ASU. This Marie Sklodowska-Curie Action (MSCA) Innovative Training Network (ITN) receives funding from the European Union's H2020 Framework Programme under grant agreement no. 764879. T.R. Bieler, D. Kang, E. Pai Kulyadi, P. Eisenlohr, C. Kale, and K.N. Solanki acknowledge support from DOE/OHEP grant DE-SC0009962

Biological, clinical and population relevance of 95 loci for blood lipids.

Plasma concentrations of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol and triglycerides are among the most important risk factors for coronary artery disease (CAD) and are targets for therapeutic intervention. We screened the genome for common variants associated with plasma lipids in >100,000 individuals of European ancestry. Here we report 95 significantly associated loci (P < 5 x 10(-8)), with 59 showing genome-wide significant association with lipid traits for the first time. The newly reported associations include single nucleotide polymorphisms (SNPs) near known lipid regulators (for example, CYP7A1, NPC1L1 and SCARB1) as well as in scores of loci not previously implicated in lipoprotein metabolism. The 95 loci contribute not only to normal variation in lipid traits but also to extreme lipid phenotypes and have an impact on lipid traits in three non-European populations (East Asians, South Asians and African Americans). Our results identify several novel loci associated with plasma lipids that are also associated with CAD. Finally, we validated three of the novel genes-GALNT2, PPP1R3B and TTC39B-with experiments in mouse models. Taken together, our findings provide the foundation to develop a broader biological understanding of lipoprotein metabolism and to identify new therapeutic opportunities for the prevention of CAD

Cytotoxicity of ZnO Nanoparticles Can Be Tailored by Modifying Their Surface Structure: A Green Chemistry Approach for Safer Nanomaterials

ZnO nanoparticles (NP) are extensively used in numerous nanotechnology applications; however, they also happen to be one of the most toxic nanomaterials. This raises significant environmental and health concerns and calls for the need to develop new synthetic approaches to produce safer ZnO NP, while preserving their attractive optical, electronic, and structural properties. In this work, we demonstrate that the cytotoxicity of ZnO NP can be tailored by modifying their surface-bound chemical groups, while maintaining the core ZnO structure and related properties. Two equally sized (9.26 ± 0.11 nm) ZnO NP samples were synthesized from the same zinc acetate precursor using a forced hydrolysis process, and their surface chemical structures were modified by using different reaction solvents. X-ray diffraction and optical studies showed that the lattice parameters, optical properties, and band gap (3.44 eV) of the two ZnO NP samples were similar. However, FTIR spectroscopy showed significant differences in the surface structures and surface-bound chemical groups. This led to major differences in the zeta potential, hydrodynamic size, photocatalytic rate constant, and more importantly, their cytotoxic effects on Hut-78 cancer cells. The ZnO NP sample with the higher zeta potential and catalytic activity displayed a 1.5-fold stronger cytotoxic effect on cancer cells. These results suggest that by modifying the synthesis parameters/conditions and the surface chemical structures of the nanocrystals, their surface charge density, catalytic activity, and cytotoxicity can be tailored. This provides a green chemistry approach to produce safer ZnO NP

Nondisjunction and transmission ratio distortion ofChromosome 2 in a (2.8) Robertsonian translocation mouse strain

Aneuploidy results from nondisjunction of chromosomes in meiosis and is the leading cause of developmental disabilities and mental retardation in humans. Therefore, understanding aspects of chromosome segregation in a genetic model is of value. Mice heterozygous for a (2.8) Robertsonian translocation were intercrossed with chromosomally normal mice and Chromosome 2 was genotyped for number and parental origin in 836 individuals at 8.5 dpc. The frequency of nondisjunction of this Robertsonian chromosome is 1.58%. Trisomy of Chromosome 2 with two maternally derived chromosomes is the most developmentally successful aneuploid karyotype at 8.5 dpc. Trisomy of Chromosome 2 with two paternally derived chromosomes is developmentally delayed and less frequent than the converse. Individuals with maternal or paternal uniparental disomy of Chromosome 2 were not detected at 8.5 dpc. Nondisjunction events were distributed randomly across litters, i.e., no evidence for clustering was found. Transmission ratio distortion is frequently observed in Robertsonian chromosomes and a bias against the transmission of the (2.8) Chromosome was detected. Interestingly, this was observed for female and male transmitting parents

Does Prehabilitation modify muscle mass in patients with rectal cancer undergoing neoadjuvant therapy?:A subanalysis from the REx Randomised Controlled Trial

Background: Patients with rectal cancer who present with sarcopenia (low muscle mass) are at significantly greater risk of postoperative complications and reduction in disease-free survival. We performed a subanalysis of a randomised controlled study [the REx trial; www.isrctn.com; 62859294] to assess the potential of prehabilitation to modify muscle mass in patients having neoadjuvant chemoradiotherapy (NACRT). Methods: Patients scheduled for NACRT, then potentially curative surgery (August 2014–March 2016) had baseline physical assessment and psoas muscle mass measurement (total psoas index using computed tomography-based measurements). Participants were randomised to either the intervention (13–17-week telephone-guided graduated walking programme) or control group (standard care). Follow-up testing was performed 1–2 weeks before surgery. Results: The 44 patients had a mean age of 66.8 years (SD 9.6) and were male (64%); white (98%); American Society of Anesthesiologists class 2 (66%); co-morbid (58%); overweight (72%) (body mass index ≥ 25 kg/m2). At baseline, 14% were sarcopenic. At follow-up, 13 (65%) of patients in the prehabilitation group had increased muscle mass versus 7 (35%) that experienced a decrease. Conversely, 16 (67%) controls experienced a decrease in muscle mass and 8 (33%) showed an increase. An adjusted linear regression model estimated a mean treatment difference in Total Psoas Index of 40.2mm2/m2 (95% CI − 3.4 to 83.7) between groups in change from baseline (p = 0.07). Conclusions: Prehabilitation improved muscle mass in patients with rectal cancer who had NACRT. These results need to be explored in a larger trial to determine if the poorer short- and long-term patient outcomes associated with low muscle mass can be minimised by prehabilitation

Evaluation of the chemical composition of gas- And particle-phase products of aromatic oxidation

Aromatic volatile organic compounds (VOCs) are key anthropogenic pollutants emitted to the atmosphere and are important for both ozone and secondary organic aerosol (SOA) formation in urban areas. Recent studies have indicated that aromatic hydrocarbons may follow previously unknown xidation chemistry pathways, including autoxidation that can lead to the formation of highly oxidised products. In this study we evaluate the gas- and particle-phase ions measured by online mass spectrometry during the hydroxyl radical oxidation of substituted C9-aromatic isomers (1,3,5-trimethylbenzene, 1,2,4-trimethylbenzene, propylbenzene and isopropylbenzene) and a substituted polyaromatic hydrocarbon (1-methylnaphthalene) under low- and medium-NOx conditions. A time-of-flight chemical ionisation mass spectrometer (ToF-CIMS) with iodide anion ionisation was used with a filter inlet for gases and aerosols (FIGAERO) for the detection of products in the particle phase, while a Vocus protontransfer- reaction mass spectrometer (Vocus-PTR-MS) was sed for the detection of products in the gas phase. The signal of product ions observed in the mass spectra were compared or the different precursors and experimental conditions. The majority of mass spectral product signal in both the gas and particle phases comes from ions which are common to all precursors, though signal distributions are distinct for different VOCs. Gas- and particle-phase composition are distinct from one another. Ions corresponding to products contained in the near-explicit gas phase Master Chemical Mechanism (MCM version 3.3.1) are utilised as a benchmark of current scientific understanding, and a comparison of these with observations shows that the MCM is missing a range of highly oxidised products from its mechanism. In the particle phase, the bulk of the product signal from all precursors comes from ring scission ions, a large proportion of which are more oxidised than previously reported and ave undergone further oxidation to form highly oxygenated organic molecules (HOMs). Under the perturbation of OH oxidation with increased NOx , the contribution of HOM-ion signals to the particle-phase signal remains elevated for more substituted aromatic precursors. Up to 43%of product signal comes from ring-retaining ions including HOMs; this is most mportant for the more substituted aromatics. Unique products are a minor component in these systems, and many of the dominant ions have ion formulae concurrent with other systems, highlighting the challenges in utilising marker ions for SOA