Transfection of the human heme oxygenase gene into rabbit coronary microvessel endothelial cells: protective effect against heme and hemoglobin toxicity.

Heme oxygenase (HO) is a stress protein and has been suggested to participate in defense mechanisms against agents that may induce oxidative injury such as metals, endotoxin, heme/hemoglobin, and various cytokines. Overexpression of HO in cells might therefore protect against oxidative stress produced by certain of these agents, specifically heme and hemoglobin, by catalyzing their degradation to bilirubin, which itself has antioxidant properties. We report here the successful in vitro transfection of rabbit coronary microvessel endothelial cells with a functioning gene encoding the human HO enzyme. A plasmid containing the cytomegalovirus promoter and the human HO cDNA complexed to cationic liposomes (Lipofectin) was used to transfect rabbit endothelial cells. Cells transfected with human HO exhibited an approximately 3.0-fold increase in enzyme activity and expressed a severalfold induction of human HO mRNA as compared with endogenous rabbit HO mRNA. Transfected and nontransfected cells expressed factor VIII antigen and exhibited similar acetylated low-density lipoprotein uptake (two important features that characterize endothelial cells) with > 85% of cells staining positive for each marker. Moreover, cells transfected with the human HO gene acquired substantial resistance to toxicity produced by exposure to recombinant hemoglobin and heme as compared with nontransfected cells. The protective effect of HO overexpression against heme/hemoglobin toxicity in endothelial cells shown in these studies provides direct evidence that the inductive response of human HO to such injurious stimuli represents an important tissue adaptive mechanism for moderating the severity of cell damage produced by these blood components.

The effect of protein concentration on ion binding.

The concentration of protein in a solution has been found to have a significant effect on ion binding affinity. It is well known that an increase in ionic strength of the solvent medium by addition of salt modulates the ion-binding affinity of a charged protein due to electrostatic screening. In recent Monte Carlo simulations, a similar screening has been detected to arise from an increase in the concentration of the protein itself. Experimental results are presented here that verify the theoretical predictions; high concentrations of the negatively charged proteins calbindin D9k and calmodulin are found to reduce their affinity for divalent cations. The Ca(2+)-binding constant of the C-terminal site in the Asn-56 --> Ala mutant of calbindin D9k has been measured at seven different protein concentrations ranging from 27 microM to 7.35 mM by using 1H NMR. A 94% reduction in affinity is observed when going from the lowest to the highest protein concentration. For calmodulin, we have measured the average Mg(2+)-binding constant of sites I and II at 0.325, 1.08, and 3.25 mM protein and find a 13-fold difference between the two extremes. Monte Carlo calculations have been performed for the two cases described above to provide a direct comparison of the experimental and simulated effects of protein concentration on metal ion affinities. The overall agreement between theory and experiment is good. The results have important implications for all biological systems involving interactions between charged species.

Effect of lactose solid-state on the aerosolization performance of drug-carrier mixtures

Lactose, in particular α-lactose monohydrate, is the most used carrier for inhalation. Its surface and solid-state properties are of paramount importance in determining drug aerosolization performance. However, these properties may be altered by processing, such as micronization, thus affecting the product performance and stability. The present research project focused on the study of the effect of lactose solid-state on the aerosolization performance of drug-carrier mixtures, giving particular attention to the impact of micronization on lactose physico-chemical properties. The formation of a fraction of hygroscopic anhydrous α-lactose, rather than amorphous lactose, as a consequence of the mechanical stress stemming from micronization was evidenced by 1H NMR, XRPD and DSC analyses performed on samples of micronized lactose. The development of a new DVS method capable to identify and quantify different forms of α-lactose (hygroscopic anhydrous, stable anhydrous and amorphous), even simultaneously present in the same sample, confirmed the results obtained with the above-mentioned techniques. The influence of lactose solid-state on drug respirability was then evaluated through the preparation and in vitro aerodynamic assessment of ternary and binary mixtures containing two different drugs. In particular, the use, as carriers, of anhydrous forms of α-lactose in place of the conventional α-lactose monohydrate resulted in significantly improved respirability in the case of salbutamol sulphate and poorer performance in the case of budesonide. In an attempt to rationalize the obtained results, IGC was selected as a tool to investigate possible variations in the surface energy of the studied lactose carriers and APIs. A direct correlation between the total surface free energy of lactose carriers and drug respirability was not found. However, salbutamol sulphate and budesonide exhibited different specific surface free energy, to which the difference in the aerosolization performance may be, at least in part, ascribed.

Effect of water vapour condensation on the radon content in subsurface air in a hypogeal inactive-volcanic environment in Galdar cave, Spain

Fluctuations of trace gas activity as a response to variations in weather and microclimate conditions were monitored over a year in a shallow volcanic cave (Painted Cave, Galdar, Canary Islands, Spain). 222Rn concentration was used due to its greater sensitivity to hygrothermal variations than CO2 concentration. Radon concentration in the cave increases as effective vapour condensation within the porous system of the rock surfaces inside the cave increases due to humidity levels of more than 70%. Condensed water content in pores was assessed and linked to a reduction in the direct passage of trace gases. Fluctuations in radon activity as a response to variations in weather and microclimate conditions were statistically identified by clustering entropy changes on the radon signal and parameterised to predict radon concentration anomalies. This raises important implications for other research fields, including the surveillance of shallow volcanic and seismic activity, preventive conservation of cultural heritage in indoor spaces, indoor air quality control and studies to improve understanding of the role of subterranean terrestrial ecosystems as reservoirs and/or temporary sources of trace gases.

Direct versus indirect channels: Differentiated loss aversion in a high‐involvement, non‐frequently purchased hedonic product

Purpose – This article aims to investigate whether intermediaries reduce loss aversion in the context of a high-involvement non-frequently purchased hedonic product (tourism packages). Design/methodology/approach – The study incorporates the reference-dependent model into a multinomial logit model with random parameters, which controls for heterogeneity and allows representation of different correlation patterns between non-independent alternatives. Findings – Differentiated loss aversion is found: consumers buying high-involvement non-frequently purchased hedonic products are less loss averse when using an intermediary than when dealing with each provider separately and booking their services independently. This result can be taken as identifying consumer-based added value provided by the intermediaries. Practical implications – Knowing the effect of an increase in their prices is crucial for tourism collective brands (e.g. “sun and sea”, “inland”, “green destinations”, “World Heritage destinations”). This is especially applicable nowadays on account of the fact that many destinations have lowered prices to attract tourists (although, in the future, they will have to put prices back up to their normal levels). The negative effect of raising prices can be absorbed more easily via indirect channels when compared to individual providers, as the influence of loss aversion is lower for the former than the latter. The key implication is that intermediaries can – and should – add value in competition with direct e-tailing. Originality/value – Research on loss aversion in retailing has been prolific, exclusively focused on low-involvement and frequently purchased products without distinguishing the direct or indirect character of the distribution channel. However, less is known about other types of products such as high-involvement non-frequently purchased hedonic products. This article focuses on the latter and analyzes different patterns of loss aversion in direct and indirect channels.

Law and Foreign Direct Investment. IES WORKING PAPER 4/2011

In this paper we examine the effect of law on foreign direct investment outflows with a specific interest in the relationship between international investment law and domestic private property laws. Our results indicate that FDI investor is indifferent to host country property rights, hence shareholder protection by law is not a significant determinant of FDI outflows. We argue that FDI, in contrast with other types of capital flows, can effectively mitigate the agency problem through majority ownership and control, hence reduce exposure to ex-post expropriation by the affiliate. On the other hand, FDI investor remains exposed to risk of expropriation by the host government and is strongly sensitive to the enforcement of law in the host country. In contrast with recent literature we conclude that there are no causal relationship between bilateral investment treaties and FDI.

Direct and general support maintenance manual : truck tractor, 10-ton, 6x6, M123 (2320-395-1875, M123C (2320-294-9552), M1231C (2320-226-6081), M123E2 (2320-879-6177), and truck cargo, 10-ton, 6x6, M125 (2320-219-7340).

Includes index.

Direct support and general support maintenance manual : truck, utility, 1/4 ton, 4x4, M151 (2320-542-4783) ... truck, utility, 1/4 ton, 4x4, M151A1C (2320-763-1091) ... truck, ambulance, frontline, 1/4 ton, 4x4, M718 (2310-782-6056), M718A1 (2310-177-9256).

Includes index.

Direct and general support and depot maintenance repair parts and special tools lists : tractor, full tracked, low speed : diesel engine, medium drawbar pull, oscillating track, 74-in. minimum gage, power shift transmission, w/bulldozer, hydraulic-tilting, w/backrip scarifier, w/winch, reversible, rear, 1 drum (Allis-Chalmers model HD16M) ....

"April 1972."

Direct support and general support maintenance manual : tractor, full tracked, low speed, DED medium drawbar pull, oscillating track, 78-in. gage (Caterpillar model D7F), FSN 2410-177-7283 w/ripper, FSN 2410-177-7284 w/winch.

Includes index.

Direct and general support and depot maintenance manual : loader scoop type, pneu tired, diesel engine driven, hinged frame steer ...

Includes index.

Operator, organizational, direct and general support, and depot maintenance manual : loader, scoop type, DED, 4-wheel drive, pneumatic tires, hinged frame steer, w/forklift, front mounted, hydraulic operated, 10,500-lb capacity (Eaton, Yale & Towne Trojan model 3000M) FSN 3805-074-6378.

Includes index.

Operator's, organizational, direct support, and general support maintenance manual : bridge, armored-vehicle-launched, scissoring type, aluminum, 60-ft. span, for use with M-48 and M-60 launcher (all makes and models), FSN 5420-522-9599).

"October 1972."

Direct and general support and depot maintenance repair parts and special tools lists : theodolite, directional, 1 sec graduation, 5.9 in. length telescope w/tripod, carrying case, and accessories (Wild Heerbrugg instruments), model T-2, serial numbers 5602 through 17897 and 27235 through 30594 (FSN 6675-232-8972), model T-2 56 ....

"September 1968."

Operator, organizational, direct support and general support maintenance manual : tent, frame type, universal field maintenance, expandable, cotton duck, fire, mildrew [sic], water, and weather resistant, w/o floor, floor area 27 ft w, 22 ft h, 64 ft lg, length expandable in 8-foot increments, w/covers, frames, ground anchor kit, liners, FSN 8340-935-6372.

"January 1970."