The Role of Cholesterol on Triterpenoid Saponin-Induced Endolysosomal Escape of a Saporin-Based Immunotoxin

Cholesterol seems to play a central role in the augmentation of saporin-based immunotoxin (IT) cytotoxicity by triterpenoid saponins. Endolysosomal escape has been proposed as one mechanism for the saponin-mediated enhancement of targeted toxins. We investigated the effects of lipid depletion followed by repletion on Saponinum album (SA)-induced endolysosomal escape of Alexa Fluor labelled saporin and the saporin-based immunotoxin OKT10-SAP, directed against CD38, in Daudi lymphoma cells. Lipid deprived cells showed reduced SA-induced endolysosomal escape at two concentrations of SA, as determined by a flow cytometric method. The repletion of membrane cholesterol by low density lipoprotein (LDL) restored SA-induced endolysosomal escape at a concentration of 5 µg/mL SA but not at 1 µg/mL SA. When LDL was used to restore the cholesterol levels in lipid deprived cells, the SA augmentation of OKT10-SAP cytotoxicity was partially restored at 1 µg/mL SA and fully restored at 5 µg/mL SA. These results suggest that different mechanisms of action might be involved for the two different concentrations of SA and that endosomal escape may not be the main mechanism for the augmentation of saporin IT cytotoxicity by SA at the sub-lytic concentration of 1 µg/mL SA.     

Modeling of Droplet Generation in a Top Blowing Steelmaking Process

Metallurgical and Materials Transactions B - Quantification of metal droplets ejected due to impinging gas jet on the surface of liquid metal is an important parameter for the understanding and for...     

Buying Power

The global push toward finalizing and implementing the next generation of connected smart machinery—the Industry 4.0 protocols—begs the question: how will these new standards influence equipment purchasing decisions for plastics manufacturers?     

How verified (or tested) is my code? Falsification-driven verification and testing

Formal verification has advanced to the point that developers can verify the correctness of small, critical modules. Unfortunately, despite considerable efforts, determining if a “verification” verifies what the author intends is still difficult. Previous approaches are difficult to understand and often limited in applicability. Developers need verification coverage in terms of the software they are verifying, not model checking diagnostics. We propose a methodology to allow developers to determine (and correct) what it is that they have verified, and tools to support that methodology. Our basic approach is based on a novel variation of mutation analysis and the idea of verification driven by falsification. We use the CBMC model checker to show that this approach is applicable not only to simple data structures and sorting routines, and verification of a routine in Mozilla’s JavaScript engine, but to understanding an ongoing effort to verify the Linux kernel read-copy-update mechanism. Moreover, we show that despite the probabilistic nature of random testing and the tendency to incompleteness of testing as opposed to verification, the same techniques, with suitable modifications, apply to automated test generation as well as to formal verification. In essence, it is the number of surviving mutants that drives the scalability of our methods, not the underlying method for detecting faults in a program. From the point of view of a Popperian analysis where an unkilled mutant is a weakness (in terms of its falsifiability) in a “scientific theory” of program behavior, it is only the number of weaknesses to be examined by a user that is important.     

TSTL: the template scripting testing language

International Journal on Software Tools for Technology Transfer - A test harness, in automated test generation, defines the set of valid tests for a system, as well as their correctness properties....     

Microbial growth,communities and sensory characteristics of vacuum and modified atmosphere packaged lamb shoulders

Packaging fresh lamb in a vacuum (VAC) versus a 100% CO₂ modified atmosphere (MAP) may influence product shelf-life and the bacterial communities. While VAC is a common packing method and 100% CO₂ MAP is used in some countries, there is little information about how these different techniques affect the growth of spoilage bacteria and sensory attributes of lamb. The aim of this study was to assess changes in microbiological and organoleptic properties, and determine differences in microbial communities by terminal restriction fragment length polymorphism (TRFLP) and 454 pyrosequencing, in bone-in (BI) and bone-out (BO) MAP- and VAC-packed lamb shoulders stored at −0.3 °C over 12 wk. VAC and MAP lamb shoulders were acceptable in sensory test scores over 12 wk of storage at −0.3 °C, despite total viable count (TVC) and lactic acid bacteria (LAB) levels increasing to 8 log₁₀ CFU/cm² for VAC lamb and 4–6 log₁₀ CFU/cm² for MAP lamb. Similar to the sensory results, there were no significant differences in microbial communities between BI and BO product. However, types of bacteria were different between VAC and MAP packaging. Specifically, while VAC shoulder became dominated by Carnobacterium spp. in the middle of the storage period, the MAP shoulder microbial population remained similar from the start until later storage times.