scholarly journals An engineered IL-2 reprogrammed for anti-tumor therapy using a semi-synthetic organism

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jerod L. Ptacin ◽  
Carolina E. Caffaro ◽  
Lina Ma ◽  
Kristine M. San Jose Gall ◽  
Hans R. Aerni ◽  
...  
Keyword(s):  
Nk Cells ◽  
Large Scale ◽  
Tumor Therapy ◽  
Covalent Modification ◽  
Synthetic Dna ◽  
Expansion Characteristic ◽  
Synthetic Organisms ◽  
Dose Dependent ◽  
Receptor Beta

AbstractThe implementation of applied engineering principles to create synthetic biological systems promises to revolutionize medicine, but application of fundamentally redesigned organisms has thus far not impacted practical drug development. Here we utilize an engineered microbial organism with a six-letter semi-synthetic DNA code to generate a library of site-specific, click chemistry compatible amino acid substitutions in the human cytokine IL-2. Targeted covalent modification of IL-2 variants with PEG polymers and screening identifies compounds with distinct IL-2 receptor specificities and improved pharmacological properties. One variant, termed THOR-707, selectively engages the IL-2 receptor beta/gamma complex without engagement of the IL-2 receptor alpha. In mice, administration of THOR-707 results in large-scale activation and amplification of CD8+ T cells and NK cells, without Treg expansion characteristic of IL-2. In syngeneic B16-F10 tumor-bearing mice, THOR-707 enhances drug accumulation in the tumor tissue, stimulates tumor-infiltrating CD8+ T and NK cells, and leads to a dose-dependent reduction of tumor growth. These results support further characterization of the immune modulatory, anti-tumor properties of THOR-707 and represent a fundamental advance in the application of synthetic biology to medicine, leveraging engineered semi-synthetic organisms as cellular factories to facilitate discovery and production of differentiated classes of chemically modified biologics.

scholarly journals Purification and molecular characterization of recombinant rat betacellulin

2001 ◽  
Vol 27 (2) ◽  
pp. 239-247 ◽  
Author(s):  
AJ Dunbar ◽  
IK Priebe ◽  
MP Sanderson ◽  
C Goddard
Keyword(s):  
Fusion Protein ◽  
Large Scale ◽  
Gel Filtration ◽  
Factor Xa ◽  
Dependent Manner ◽  
3T3 Fibroblasts ◽  
Reducing Conditions ◽  
Dose Dependent ◽  
Heparin Affinity

A method for the large scale expression and purification of rat betacellulin (BTC) from Escherichia coli has been developed using a cleavable fusion protein strategy. Insoluble fusion protein collected as inclusion bodies was dissolved in urea under reducing conditions, re-folded, and purified by gel filtration chromatography and C(4) RP-HPLC. Authentic rat BTC was obtained after proteolytic cleavage of the fusion protein with Factor Xa. Factor Xa cleaved an additional site within the BTC protein, generating a truncated isoform separable from full-length BTC by heparin-affinity chromatography. Recombinant rat BTC stimulated the proliferation of mouse Balb/c 3T3 fibroblasts and competed for binding to the ErbB1 receptor in a dose-dependent manner analogous to that of BTC purified from natural sources.

scholarly journals Characterization of the bovine brain cytosolic phosphatidylinositol 3-kinase complex

1993 ◽  
Vol 290 (2) ◽  
pp. 609-616 ◽  
Author(s):  
F Ruiz-Larrea ◽  
P Vicendo ◽  
P Yaish ◽  
P End ◽  
G Panayotou ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Binding Site ◽  
Large Scale ◽  
Kinase Activity ◽  
Bovine Brain ◽  
Covalent Modification ◽  
Phosphatidylinositol 3 Kinase ◽  
Protein Subunits ◽  
Atp Binding Site

Receptor-linked phosphatidylinositol (PtdIns) 3-kinase may generate a second-messenger signal. Here a large-scale purification of the bovine brain enzyme, based on methods developed by Morgan, Smith and Parker [(1990) Eur. J. Biochem. 191, 761-767] and Fry, Panayotou, Dhand, Ruiz-Larrea, Gout, Nguyen, Courtneidge and Waterfield [(1992) Biochem. J. 288, 383-393] is described. The purified enzyme is shown to be a heterodimer of 85 kDa and 110 kDa protein subunits (p85 and p110). Labelling with 5′-p-fluorosulphonylbenzoyladenosine shows that p110 contains an ATP-binding site and confers catalytic activity to the complex. The purified complex is known to be highly phosphorylated on both p85 alpha and p110 subunits, and dephosphorylation generates a deactivated complex, indicating that phosphorylation is an important covalent modification of the complex and may modulate PtdIns 3-kinase activity.

Some applications of electron beam microanalysis techniques in VLSI process evaluation

1983 ◽  
Vol 41 ◽  
pp. 160-161
Author(s):  
Simon Thomas
Keyword(s):  
Integrated Circuits ◽  
Process Evaluation ◽  
Large Scale ◽  
Technology Development ◽  
Analytical Techniques ◽  
Successful Implementation ◽  
Analysis Of Failures ◽  
Characterization Of Materials ◽  
Circuits Vlsi

Trends in the technology development of very large scale integrated circuits (VLSI) have been in the direction of higher density of components with smaller dimensions. The scaling down of device dimensions has been not only laterally but also in depth. Such efforts in miniaturization bring with them new developments in materials and processing. Successful implementation of these efforts is, to a large extent, dependent on the proper understanding of the material properties, process technologies and reliability issues, through adequate analytical studies. The analytical instrumentation technology has, fortunately, kept pace with the basic requirements of devices with lateral dimensions in the micron/ submicron range and depths of the order of nonometers. Often, newer analytical techniques have emerged or the more conventional techniques have been adapted to meet the more stringent requirements. As such, a variety of analytical techniques are available today to aid an analyst in the efforts of VLSI process evaluation. Generally such analytical efforts are divided into the characterization of materials, evaluation of processing steps and the analysis of failures.

Characterization of seawater reverse osmosis fouled membranes from large scale commercial desalination plant

2019 ◽  
Author(s):  
Chem Int
Keyword(s):  
Reverse Osmosis ◽  
Large Scale ◽  
Retention Rate ◽  
Local Level ◽  
Microscopic Analysis ◽  
Transmembrane Pressure ◽  
Hydraulic Permeability ◽  
Seawater Reverse Osmosis ◽  
A Chain

The objective of this work is to study the ageing state of a used reverse osmosis (RO) membrane taken in Algeria from the Benisaf Water Company seawater desalination unit. The study consists of an autopsy procedure used to perform a chain of analyses on a membrane sheet. Wear of the membrane is characterized by a degradation of its performance due to a significant increase in hydraulic permeability (25%) and pressure drop as well as a decrease in salt retention (10% to 30%). In most cases the effects of ageing are little or poorly known at the local level and global measurements such as (flux, transmembrane pressure, permeate flow, retention rate, etc.) do not allow characterization. Therefore, a used RO (reverse osmosis) membrane was selected at the site to perform the membrane autopsy tests. These tests make it possible to analyze and identify the cause as well as to understand the links between performance degradation observed at the macroscopic scale and at the scale at which ageing takes place. External and internal visual observations allow seeing the state of degradation. Microscopic analysis of the used membranes surface shows the importance of fouling. In addition, quantification and identification analyses determine a high fouling rate in the used membrane whose foulants is of inorganic and organic nature. Moreover, the analyses proved the presence of a biofilm composed of protein.

Characterization of Interconnect Defects Using Scanning Thermal Conductivity Microscopy

2004 ◽  
Author(s):  
H.W. Ho ◽  
J.C.H. Phang ◽  
A. Altes ◽  
L.J. Balk
Keyword(s):  
Thermal Conductivity ◽  
Integrated Circuits ◽  
Large Scale

Abstract In this paper, scanning thermal conductivity microscopy is used to characterize interconnect defects due to electromigration. Similar features are observed both in the temperature and thermal conductivity micrographs. The key advantage of the thermal conductivity mode is that specimen bias is not required. This is an important advantage for the characterization of defects in large scale integrated circuits. The thermal conductivity micrographs of extrusion, exposed and subsurface voids are presented and compared with the corresponding topography and temperature micrographs.

Implantable neural interfaces

2018 ◽  
Author(s):  
Stefano Vassanelli
Keyword(s):  
Brain Function ◽  
Large Scale ◽  
Ionic Channels ◽  
Patch Clamp Technique ◽  
Advantages And Disadvantages ◽  
Optogenetic Stimulation ◽  
Physical Interfaces ◽  
The Brain

Establishing direct communication with the brain through physical interfaces is a fundamental strategy to investigate brain function. Starting with the patch-clamp technique in the seventies, neuroscience has moved from detailed characterization of ionic channels to the analysis of single neurons and, more recently, microcircuits in brain neuronal networks. Development of new biohybrid probes with electrodes for recording and stimulating neurons in the living animal is a natural consequence of this trend. The recent introduction of optogenetic stimulation and advanced high-resolution large-scale electrical recording approaches demonstrates this need. Brain implants for real-time neurophysiology are also opening new avenues for neuroprosthetics to restore brain function after injury or in neurological disorders. This chapter provides an overview on existing and emergent neurophysiology technologies with particular focus on those intended to interface neuronal microcircuits in vivo. Chemical, electrical, and optogenetic-based interfaces are presented, with an analysis of advantages and disadvantages of the different technical approaches.

scholarly journals Effects of D-allulose on glucose tolerance and insulin response to a standard oral sucrose load: results of a prospective, randomized, crossover study

2021 ◽  
Vol 9 (1) ◽  
pp. e001939
Author(s):  
Francesco Franchi ◽  
Dmitry M Yaranov ◽  
Fabiana Rollini ◽  
Andrea Rivas ◽  
Jose Rivas Rios ◽  
...  
Keyword(s):  
Crossover Study ◽  
Plasma Glucose ◽  
Large Scale ◽  
Insulin Response ◽  
Mean Difference ◽  
Dietary Guidelines ◽  
Sugar Intake ◽  
Time Points ◽  
Insulin Levels ◽  
Dose Dependent

IntroductionCurrent dietary guidelines recommend limiting sugar intake for the prevention of diabetes mellitus (DM). Reduction in sugar intake may require sugar substitutes. Among these, D-allulose is a non-calorie rare monosaccharide with 70% sweetness of sucrose, which has shown anti-DM effects in Asian populations. However, there is limited data on the effects of D-allulose in other populations, including Westerners.Research design and methodsThis was a prospective, randomized, double-blind, placebo-controlled, crossover study conducted in 30 subjects without DM. Study participants were given a standard oral (50 g) sucrose load and randomized to placebo or escalating doses of D-allulose (2.5, 5.0, 7.5, 10.0 g). Subjects crossed-over to the alternate study treatment after 7–14 days of wash out. Plasma glucose and insulin levels were measured at five time points: before and at 30, 60, 90 and 120 min after ingestion.ResultsD-allulose was associated with a dose-dependent reduction of plasma glucose at 30 min compared with placebo. In particular, glucose was significantly lower with the 7.5 g (mean difference: 11; 95% CI 3 to 19; p=0.005) and 10 g (mean difference: 12; 95% CI 4 to 20; p=0.002) doses. Although glucose was not reduced at the other time points, there was a dose-dependent reduction in glucose excursion compared with placebo, which was significant with the 10 g dose (p=0.023). Accordingly, at 30 min D-allulose was associated with a trend towards lower insulin levels compared with placebo, which was significant with the 10 g dose (mean difference: 14; 95% CI 4 to 25; p=0.006). D-allulose did not reduce insulin at any other time point, but there was a significant dose-dependent reduction in insulin excursion compared with placebo (p=0.028), which was significant with the 10 g dose (p=0.002).ConclusionsThis is the largest study assessing the effects of D-allulose in Westerners demonstrating an early dose-dependent reduction in plasma glucose and insulin levels as well as decreased postprandial glucose and insulin excursion in subjects without DM. These pilot observations set the basis for large-scale investigations to support the anti-DM effects of D-allulose.Trial registration numberNCT02714413.

scholarly journals Remote Sensing Methods for the Biophysical Characterization of Protected Areas Globally: Challenges and Opportunities

2021 ◽  
Vol 10 (6) ◽  
pp. 384
Author(s):  
Javier Martínez-López ◽  
Bastian Bertzky ◽  
Simon Willcock ◽  
Marine Robuchon ◽  
María Almagro ◽  
...  
Keyword(s):  
Remote Sensing ◽  
Protected Areas ◽  
Large Scale ◽  
Anthropogenic Activities ◽  
Global Scale ◽  
Area Network ◽  
Biophysical Characterization ◽  
Challenges And Opportunities ◽  
Scale Characterization

Protected areas (PAs) are a key strategy to reverse global biodiversity declines, but they are under increasing pressure from anthropogenic activities and concomitant effects. Thus, the heterogeneous landscapes within PAs, containing a number of different habitats and ecosystem types, are in various degrees of disturbance. Characterizing habitats and ecosystems within the global protected area network requires large-scale monitoring over long time scales. This study reviews methods for the biophysical characterization of terrestrial PAs at a global scale by means of remote sensing (RS) and provides further recommendations. To this end, we first discuss the importance of taking into account the structural and functional attributes, as well as integrating a broad spectrum of variables, to account for the different ecosystem and habitat types within PAs, considering examples at local and regional scales. We then discuss potential variables, challenges and limitations of existing global environmental stratifications, as well as the biophysical characterization of PAs, and finally offer some recommendations. Computational and interoperability issues are also discussed, as well as the potential of cloud-based platforms linked to earth observations to support large-scale characterization of PAs. Using RS to characterize PAs globally is a crucial approach to help ensure sustainable development, but it requires further work before such studies are able to inform large-scale conservation actions. This study proposes 14 recommendations in order to improve existing initiatives to biophysically characterize PAs at a global scale.

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