Bradley A. Saville

Finite element modeling of drug distribution in the vitreous humor of the rabbit eye

Direct intravitreal injection of drug is a common method for treating diseases of the retina or vitreous. The stagnant nature of the vitreous humor and surrounding tissue barriers creates concentration gradients within the vitreous that must be accounted for when developing drug therapy. The objective of this research was to study drug distribution in the vitreous humor of the rabbit eye after an intravitreal injection, using a finite element model. Fluorescein and fluorescein glucuronide were selected as model compounds due to available experimental data. All required model parameters were known except for the permeability of these compounds through the retina, which was determined by fitting model predictions to experimental data. The location of the intravitreal injection in the experimental studies was not precisely known; therefore, several injection locations were considered, and best-fit retinal permeability was determined for each case. Retinal permeability of fluorescein and fluorescein glucuronide estimated by the model ranged from 1.94×10−5 to 3.5×10−5 cm s−1 and from 0 to 7.62×10−7 cm s−1, respectively, depending on the assumed site of the injection. These permeability values were compared with values previously calculated from other models, and the limitations of the models are discussed. Intravitreal injection position was found to be an important variable that must be controlled in both experimental and clinical settings.

Production of L-DOPA from tyrosinase immobilized on nylon 6,6 : enzyme stability and scaleup

Abstract The production of l -3,4 dihydroxyphenylalanine ( l -DOPA) from mushroom tyrosinase immobilized on chemically modified nylon 6,6 membranes was investigated in a batch reactor. The effects of product inhibition, oxygen partial pressure, and scaleup upon l -DOPA production rates and tyrosinase activity were studied. l -DOPA production rates were strongly influenced by oxygen and l -DOPA concentrations. Kinetic modeling revealed that l -DOPA concentrations as low as 1 m m would reduce the l -DOPA production rate by approximately 50% due to competitive inhibition. The average l -DOPA production rate increased by 10% when the oxygen partial pressure was increased from 21 kPa and 100 kPa; however, at higher oxygen partial pressures (50 kPa and 100 kPa), the rate of oxidative inactivation of tyrosinase increased, causing the enzyme half-life to decrease from 46 h under 21 kPa oxygen to 7.7 h under 100 kPa oxygen. The turnover number was approximately 1,170 under 21 kPa oxygen, but dropped to approximately 150 under 100 kPa oxygen. Scaleup from a 500-ml batch reactor to 1-l and 2-l reactors proved to be straightforward. Essentially identical production rates were obtained as long as the enzyme concentration within the reactor was held constant. If the enzyme concentration was varied, a proportional change in the l -DOPA production rate was not observed due to the effects of product inhibition.

Degradation of phenol using tyrosinase immobilized on siliceous supports.

The degradation of phenol by tyrosinase immobilized on chemically modified sodium aluminosilicate (NaA), calcium aluminosilicate (CaA), and silica gel was studied. Phenol conversion by immobilized tyrosinase ranged between approximately 15% and 60%, depending upon the initial phenol concentration, pH, and enzyme loading. Tyrosinase immobilized on CaA and on NaA could be re-used repeatedly without any decrease in performance. However, in studies at pH 8.0, significant enzyme inhibition was observed, since phenol conversion was rapid for approximately 20 min, then reached a plateau. The inhibition was reversible; activity was restored upon placing the immobilized enzyme in fresh substrate. Reducing the pH to 6.8 from 8.0 led to higher conversion of phenol, and decreased the inhibition of the immobilized enzyme.

L-DOPA production from tyrosinase immobilized on nylon 6,6.

The production of L‐DOPA immobilized on chemically modified nylon 6,6 membranes was studied in a batch reactor. Tyrosinase was immobilized on nylon using glutaraldehyde as a crosslinking agent. The effects of membrane pore size and glutaraldehyde concentration upon enzyme uptake and L‐DOPA production were investigated. Enzyme uptake was unaffected by glutaraldehyde concentration; approximately 70% uptake was observed when 25% w/v (group 1), 5% (group 2), and 3% (group 3) glutaraldehyde were used, indicating that glutaraldehyde was in excess. Similarly, uptake was the same for membranes with 0.20 and 10 μm pore sizes.

A subconjunctival degradable implant for cyclosporine delivery in corneal transplant therapy

The effect of local cyclosporine therapy upon corneal transplant survival was investigated. A high risk rabbit model with vascularized corneas was used to assess the efficacy of subconjunctivally implanted degradable devices for cyclosporine therapy. Animals were divided into four groups, receiving either no therapy, a placebo PLGA device, or drug containing devices implanted either at the time of transplantation or two weeks previous. The mean survival times of animals in the control and placebo groups were statistically equivalent (21 +/- 4 days vs 18 +/- 4 days). Devices containing CsA improved the survival time of grafts. Predosing the animals with CsA improved the survival time to 28 +/- 7 days, and CsA devices implanted at the time of transplantation increased the survival time to 35 +/- 7 days. The improvement in survival times was consistent with the in vitro drug release profiles. No systemic CsA was detected, suggesting that the effect may have been local. Histological assessment indicated that devices were well tolerated.

Electricity Production from Anaerobic Digestion of Household Organic Waste in Ontario: Techno-Economic and GHG Emission Analyses

The first Feed-in-Tariff (FiT) program in North America was recently implemented in Ontario, Canada to stimulate the generation of electricity from renewable sources. The life cycle greenhouse gas (GHG) emissions and economics of electricity generation through anaerobic digestion (AD) of household source-separated organic waste (HSSOW) are investigated within the FiT program. AD can potentially provide considerable GHG emission reductions (up to 1 t CO(2)eq/t HSSOW) at relatively low to moderate cost (-

Models of Hepatic Drug Elimination

35 to 160/t CO(2)eq) by displacing fossil electricity and preventing the emission of landfill gas. It is a cost-effective GHG mitigation option compared to some other FiT technologies (e.g., wind, solar photovoltaic) and provides unique additional benefits (waste diversion, nutrient recycling). The combination of electricity sales at a premium rate, savings in waste management costs, and economies of scale allow AD facilities processing >30,000 t/yr to be cost-competitive against landfilling. However, the FiT does not sufficiently support smaller-scale facilities that are needed as a transition to larger, more economically viable facilities. Refocusing of the FiT program and waste policies are needed to support the adoption of AD of HSSOW, which has not yet been developed in the Province, while more costly technologies (e.g., photovoltaic) have been deployed.

Characterization and Performance of Immobilized Amylase and Cellulase

The liver is, by nature, heterogeneous. It contains a complex vascular network for blood flow and a stationary phase consisting of enzymes within parenchymal cells. Several physiological processes, therefore, may combine to give observed ranges in drug elimination. Net changes in concentration are a consequence of a series of steps: uptake of substrate into liver cells, enzymatic reactions within the cells, release of metabolites and unconverted substrate from the cells into the sinusoids, and the net flow of the perfusing medium in the vasculature. In addition, substrate binding to proteins in the blood and in the liver can influence hepatic elimination. An understanding of each of these processes is necessary to fully comprehend the overall process of drug elimination, and these processes must be accounted for, either individually or by grouping and approximation, if a model for drug elimination is to be developed. Existing models of hepatic elimination may be classified according to their treatment of mixing within the vasculature and whether or not the model explicitly accounts for mass transfer between the heterogeneous phases of the liver. Four major classes may be defined: 1. Nonparametric homogeneous models, which assume that either complete mixing or no mixing occurs within the vasculature of the organ. 2. Homogeneous mixing models, which allow for a range of mixing phenomena. 3. Heterogeneous micromixing models, which allow for mass transport between the cells and vasculature and describe mixing within the vasculature on a microscopic level. 4. Heterogeneous compartmental models, which also describe interphase mass transfer but assume complete mixing on a microscopic level, and therefore use a time and spatially averaged approach to model mixing. The utility of these models of hepatic elimination will be critically assessed based upon (1) their ability to account for the influence of the aforementioned physiological processes upon elimination; (2) the data requirements of the model, in addition to its mathematical complexity and ease of use; and (3) the range of compounds and metabolites which may be described using the model.

A compartmental model for the ocular pharmacokinetics of cyclosporine in rabbits.

The performance of cellulase and amylase immobilized on siliceous supports was investigated. Enzyme uptake onto the support depended on the enzyme source and immobilization conditions. For amylase, the uptake ranged between 20 and 60%, and for cellulase, 7–10%. Immobilized amylase performance was assessed by batch kinetics in 100–300 g/L of corn flour at 65°C. Depending on the substrate and enzyme loading, between 40 and 60% starch conversion was obtained. Immobilized amylase was more stable than soluble amylase. Enzyme samples were preincubated in a water bath at various temperatures, then tested for activity. At 105°C, soluble amylase lost ∼55% of its activity, compared with ∼30% loss for immobilized amylase. The performance of immobilized cellulase was evaluated from batch kinetics in 10 g/L of substrate (shredded wastepaper) at 55°C. Significant hydrolysis of the wastepaper was also observed, indicating that immobilization does not preclude access to and hydrolysis of insoluble cellulose.

Life cycle greenhouse gas impacts of ethanol, biomethane and limonene production from citrus waste

Studies were conducted in rabbits to determine the ocular distribution and elimination of cyclosporine, with the objective of developing a comprehensive pharmacokinetic model. Following a bolus dose into the anterior chamber, drug levels were measured in the aqueous humor, cornea, iris/ciliary body, lens, sclera, and conjunctiva. Cyclosporine was rapidly eliminated from the aqueous, but drug levels in ocular tissues persisted for in excess of 48 hours, particularly in the cornea and iris/ciliary body. The terminal elimination half life from these tissues was 45 hr and 30 hr, respectively, providing evidence that these tissues could act as a reservoir for the drug. It was found that a compartmental model accurately described the experimental data. A single compartment was used for each of the tissues and fluids sampled, except for the cornea, which was subdivided into two compartments, representing its tissue and aqueous regions.