Blocking Muscarinic Receptor 3 Attenuates Tumor Growth and Decreases Immunosuppressive and Cholinergic Markers in an Orthotopic Mouse Model of Colorectal Cancer

Tumor cells have evolved to express immunosuppressive molecules allowing their evasion from the host’s immune system. These molecules include programmed death ligands 1 and 2 (PD-L1 and PD-L2). Cancer cells can also produce acetylcholine (ACh), which plays a role in tumor development. Moreover, tumor innervation can stimulate vascularization leading to tumor growth and metastasis. The effects of atropine and muscarinic receptor 3 (M3R) blocker, 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide (4-DAMP), on cancer growth and spread were evaluated in vitro using murine colon cancer cell line, CT-26, and in vivo in an orthotopic mouse model of colorectal cancer. In the in vitro model, atropine and 4-DAMP significantly inhibited CT-26 cell proliferation in a dose dependent manner and induced apoptosis. Atropine attenuated immunosuppressive markers and M3R via inhibition of EGFR/AKT/ERK signaling pathways. However, 4-DAMP showed no effect on the expression of PD-L1, PD-L2, and choline acetyltransferase (ChAT) on CT-26 cells but attenuated M3R by suppressing the phosphorylation of AKT and ERK. Blocking of M3R in vivo decreased tumor growth and expression of immunosuppressive, cholinergic, and angiogenic markers through inhibition of AKT and ERK, leading to an improved immune response against cancer. The expression of immunosuppressive and cholinergic markers may hold potential in determining prognosis and treatment regimens for colorectal cancer patients. This study’s results demonstrate that blocking M3R has pronounced antitumor effects via several mechanisms, including inhibition of immunosuppressive molecules, enhancement of antitumor immune response, and suppression of tumor angiogenesis via suppression of the AKT/ERK signaling pathway. These findings suggest a crosstalk between the cholinergic and immune systems during cancer development. In addition, the cholinergic system influences cancer evasion from the host’s immunity.     

Scalable Distributed Sensor Fault Diagnosis for Smart Buildings

The enormous energy use of the building sector and the requirements for indoor living quality that aim to improve occupants’ productivity and health, prioritize Smart Buildings as an emerging technology. The Heating, Ventilation and Air-Conditioning (HVAC) system is considered one of the most critical and essential parts in buildings since it consumes the largest amount of energy and is responsible for humans comfort. Due to the intermittent operation of HVAC systems, faults are more likely to occur, possibly increasing eventually building’s energy consumption and/or downgrading indoor living quality. The complexity and large scale nature of HVAC systems complicate the diagnosis of faults in a centralized framework. This paper presents a distributed intelligent fault diagnosis algorithm for detecting and isolating multiple sensor faults in large-scale HVAC systems. Modeling the HVAC system as a network of interconnected subsystems allows the design of a set of distributed sensor fault diagnosis agents capable of isolating multiple sensor faults by applying a combinatorial decision logic and diagnostic reasoning. The performance of the proposed method is investigated with respect to robustness, fault detectability and scalability. Simulations are used to illustrate the effectiveness of the proposed method in the presence of multiple sensor faults applied to a 83-zone HVAC system and to evaluate the sensitivity of the method with respect to sensor noise variance.      

Unveiling the Essential Role of Arkadia’s Non-RING Elements in the Ubiquitination Process

Arkadia is a positive regulator of the TGFβ-SMAD2/3 pathway, acting through its C-terminal RING-H2 domain and targeting for degradation of its negative regulators. Here we explore the role of regions outside the RING domain (non-RING elements) of Arkadia on the E2-E3 interaction. The contribution of the non-RING elements was addressed using Arkadia RING 68 aa and Arkadia 119 aa polypeptides. The highly conserved NRGA (asparagine-arginine-glycine-alanine) and TIER (threonine-isoleucine-glutamine-arginine) motifs within the 119 aa Arkadia polypeptide, have been shown to be required for pSMAD2/3 substrate recognition and ubiquitination in vivo. However, the role of the NRGA and TIER motifs in the enzymatic activity of Arkadia has not been addressed. Here, nuclear magnetic resonance interaction studies with the E2 enzyme, UBCH5B, C85S UBCH5B-Ub oxyester hydrolysis, and auto-ubiquitination assays were used to address the role of the non-RING elements in E2-E3 interaction and in the enzymatic activity of the RING. The results support that the non-RING elements including the NRGA and TIER motifs are required for E2-E3 recognition and interaction and for efficient auto-ubiquitination. Furthermore, while Arkadia isoform-2 and its close homologue Arkadia 2C are known to interact with free ubiquitin, the results here showed that Arkadia isoform-1 does not interact with free ubiquitin.     

INFLUENCE OF SOLUTE TEMPERATURE AND CONCENTRATION ON THE COMBINED OSMOTIC AND AIR DRYING

The effect of solute concentration (15-45 % sugars) and temperature (10-40 °C) on the osmotic dehydration of apple was investigated. Cylindrical samples of apple were immersed in glucose or sucrose solutions and the water loss, the volume of solids and the porosity were measured as a function of time. Water loss was proportional to the square root of time, while the volume of solids decreased and the porosity increased with time. Sugar gain and water loss decreased the compression stress of the apple samples. Osmotic pre-treatment reduced the shrinkage and the porosity of apple solids during air-drying, compared to the no-treated samples. The results of this investigation are useful in the design of efficient osmotic dehydration processes, and in the evaluation of texture of dehydration of products.     

The effect of alternative sweeteners on batter rheology and cake properties

BACKGROUND: The aim of this research was to investigate whether certain polyols (mannitol, maltitol, sorbitol, lactitol), fructose, oligofructose and polydextrose can replace sugar (by an equal amount of each substitute) in cake formulations. The rheological behaviour of the cake batter and the physical characteristics of the cakes containing sugar substitutes were compared with the respective attributes of the control cake. Differential scanning calorimetry was used to investigate the effect of sugar substitutes on starch gelatinisation. Furthermore sensorial characteristics were evaluated by instrumental measurements and sensory evaluation. The correlation of the batter characteristics with the textural attributes of the final product was also attempted. RESULTS: The best results were obtained by using oligofructose, lactitol or maltitol as sugar replacers, which exhibited similar behaviour to sucrose in terms of batter rheology and increased starch gelatinisation temperature. Fructose and mannitol led to cakes of poor quality characteristics, as was demonstrated by instrumental measurements and sensory evaluation. CONCLUSIONS: Batter rheological behaviour as well as the ability of sugar substitutes to increase starch gelatinisation temperature proved to be controlling factors of the textural properties and volume of the cakes. The sensory evaluation indicated that overall acceptance followed closely the scores of tenderness and taste. Copyright © 2011 Society of Chemical Industry     

Rapid enrichment and analysis of yeast phosphoproteins using affinity chromatography, 2D-PAGE and peptide mass fingerprinting

A combination of affinity purification, 2D-PAGE and peptide mass fingerprinting was employed to study the phosphoprotein complement of Saccharomyces cerevisiae. Protein extracts were first passed through a phosphoprotein affinity column, and the phosphoprotein-enriched eluate fractions were then separated on 2D gels and visualized by staining with SYPRO Ruby. Proteins were excised from the gels and identified by peptide mass fingerprinting; 11/13 protein spots identified from a gel of the phosphoprotein-enriched fraction had prior published evidence indicating that they were phosphoproteins. Additional experiments using a specific stain for phosphoproteins, prior incubation of the protein extract with alkaline phosphatase and blotting with monoclonal antibodies to phosphothreonine, phosphoserine and phosphotyrosine demonstrated that the phosphoprotein affinity column was an effective method for enriching phosphoproteins. Further validating the method, growth of yeast in the presence of sorbic acid resulted in altered phosphorylation of 17 proteins, 13 of which had prior published evidence that they were phosphoproteins or had ATP binding activity.     

Robustifying Dynamic Positioning of Crane Vessels for Heavy Lifting Operation

Construction crane vessels make use of dynamic positioning(DP)systems during the installation and removal of offshore structures to maintain the vessel’s position.Studies have reported cases of instability of DP systems during offshore operation caused by uncertainties,such as mooring forces.DP"robustification"for heavy lift operations,i.e.,handling such uncertainties systematically and with stability guarantees,is a long-standing challenge in DP design.A new DP method,composed by an observer and a controller,is proposed to address this challenge,with stability guarantees in the presence of uncertainties.We test the proposed method on an integrated cranevessel simulation environment,where the integration of several subsystems(winch dynamics,crane forces,thruster dynamics,fuel injection system etc.)allow a realistic validation under a wide set of uncertainties.     

Cooperative Multi-Agent Control for Autonomous Ship Towing Under Environmental Disturbances

Among the promising application of autonomous surface vessels (ASVs) is the utilization of multiple autonomous tugs for manipulating a floating object such as an oil platform, a broken ship, or a ship in port areas. Considering the real conditions and operations of maritime practice, this paper proposes a multi-agent control algorithm to manipulate a ship to a desired position with a desired heading and velocity under the environmental disturbances. The control architecture consists of a supervisory controller in the higher layer and tug controllers in the lower layer. The supervisory controller allocates the towing forces and angles between the tugs and the ship by minimizing the error in the position and velocity of the ship. The weight coefficients in the cost function are designed to be adaptive to guarantee that the towing system functions well under environmental disturbances, and to enhance the efficiency of the towing system. The tug controller provides the forces to tow the ship and tracks the reference trajectory that is computed online based on the towing angles calculated by the supervisory controller. Simulation results show that the proposed algorithm can make the two autonomous tugs cooperatively tow a ship to a desired position with a desired heading and velocity under the (even harsh) environmental disturbances.      

Electrophoretic characterization of protein interactions suggesting limited feasibility of accelerated shelf-life testing of ultra-high temperature milk

Accelerated shelf-life testing is applied to a variety of products to estimate keeping quality over a short period of time. The industry has not been successful in applying this approach to ultra-high temperature (UHT) milk because of chemical and physical changes in the milk proteins that take place during processing and storage. We investigated these protein changes, applying accelerated shelf-life principles to UHT milk samples with different fat levels and using native- and sodium dodecyl sulfate-PAGE. Samples of UHT skim and whole milk were stored at 20, 30, 40, and 50°C for 28 d. Irrespective of fat content, UHT treatment had a similar effect on the electrophoretic patterns of milk proteins. At the start of testing, proteins were bonded mainly through disulfide and noncovalent interactions. However, storage at and above 30°C enhanced protein aggregation via covalent interactions. The extent of aggregation appeared to be influenced by fat content; whole milk contained more fat than skim milk, implying aggregation via melted or oxidized fat, or both. Based on reduction in loss in absolute quantity of individual proteins, covalent crosslinking in whole milk was facilitated mainly by products of lipid oxidation and increased access to caseins for crosslinking reactions. Maillard and dehydroalanine products were the main contributors involved in protein changes in skim milk. Protein crosslinking appeared to follow a different pathway at higher temperatures (≥40°C) than at lower temperatures, making it very difficult to extrapolate these changes to protein interactions at lower temperatures.     

INFLUENCE OF SOLUTE TEMPERATURE AND CONCENTRATION ON THE COMBINED OSMOTIC AND AIR DRYING

ABSTRACT

The effect of solute concentration (15–45 % sugars) and temperature (10–40 °C) on the osmotic dehydration of apple was investigated. Cylindrical samples of apple were immersed in glucose or sucrose solutions and the water loss, the volume of solids and the porosity were measured as a function of time. Water loss was proportional to the square root of time, while the volume of solids decreased and the porosity increased with time. Sugar gain and water loss decreased the compression stress of the apple samples. Osmotic pre-treatment reduced the shrinkage and the porosity of apple solids during air-drying, compared to the no-treated samples. The results of this investigation are useful in the design of efficient osmotic dehydration processes, and in the evaluation of texture of dehydration of products.