Heat and mass transfer scale-up issues during freeze-drying,I: Atypical radiation and the edge vial effect

The aim of this study is to determine whether radiation heat transfer is responsible for the position dependence of heat transfer known as the edge vial effect. Freeze drying was performed on a laboratory-scale freeze dryer using pure water with vials that were fully stoppered but had precision cut metal tubes inserted in them to ensure uniformity in resistance to vapor flow. Sublimation rates were determined gravimetrically. Vials were sputter-coated with gold and placed at selected positions on the shelf. Average sublimation rates were determined for vials located at the front, side, and center of an array of vials. Sublimation rates were also determined with and without the use of aluminum foil as a radiation shield. The effect of the guardrail material and its contribution to the edge vial effect by conduction heat transfer was studied by replacing the stainless steel band with a low-thermal conductivity material (styrofoam). The emissivities (ε) of relevant surfaces were measured using an infrared thermometer. Sublimation rate experiments were also conducted with vials suspended off the shelf to study the role of convection heat transfer. It was found that sublimation rates were significantly higher for vials located in the front compared to vials in the center. Additional radiation shields in the form of aluminum foil on the inside door resulted in a decrease in sublimation rates for the front vials and to a lesser extent, the center vials. There was a significant decrease in sublimation rate for goldcoated vials (ε≈0.4) placed at the front of an array when compared to that of clear vials (ε≈0.9). In the case of experiments with vials suspended off the shelf, the heat transfer coefficient was found to be independent of chamber pressure, indicating that pure convection plays no significant role in heat transfer. Higher sublimation rates were observed when the steel band was used instead of Styrofoam while the highest sublimation rates were obtained in the absence of the guardrail, indicating that the metal band can act as a thermal shield but also transmits some heat from the shelf via conduction and radiation. Atypical radiation heat transfer is responsible for higher sublimation rates for vials located at the front and side of an array. However, the guardrail contributes a little to heat transfer by conduction.     

Compartmentalized energy transfer in cardiomyocytes: use of mathematical modeling for analysis of in vivo regulation of respiration.

The mathematical model of the compartmentalized energy transfer system in cardiac myocytes presented includes mitochondrial synthesis of ATP by ATP synthase, phosphocreatine production in the coupled mitochondrial creatine kinase reaction, the myofibrillar and cytoplasmic creatine kinase reactions, ATP utilization by actomyosin ATPase during the contraction cycle, and diffusional exchange of metabolites between different compartments. The model was used to calculate the changes in metabolite profiles during the cardiac cycle, metabolite and energy fluxes in different cellular compartments at high workload (corresponding to the rate of oxygen consumption of 46 mu atoms of O.(g wet mass)-1.min-1) under varying conditions of restricted ADP diffusion across mitochondrial outer membrane and creatine kinase isoenzyme "switchoff." In the complete system, restricted diffusion of ADP across the outer mitochondrial membrane stabilizes phosphocreatine production in cardiac mitochondria and increases the role of the phosphocreatine shuttle in energy transport and respiration regulation. Selective inhibition of myoplasmic or mitochondrial creatine kinase (modeling the experiments with transgenic animals) results in "takeover" of their function by another, active creatine kinase isoenzyme. This mathematical modeling also shows that assumption of the creatine kinase equilibrium in the cell may only be a very rough approximation to the reality at increased workload. The mathematical model developed can be used as a basis for further quantitative analyses of energy fluxes in the cell and their regulation, particularly by adding modules for adenylate kinase, the glycolytic system, and other reactions of energy metabolism of the cell.     

Differential transform semi-numerical analysis of biofluid-particle suspension flow and heat transfer in non-Darcian porous media

The differential transform method (DTM) is semi-numerical method which is used to study the steady, laminar buoyancy-driven convection heat transfer of a particulate biofluid suspension in a channel containing a porous material. A two-phase continuum model is used. A set of variables is implemented to reduce the ordinary differential equations for momentum and energy conservation (for both phases) to a dimensionless system. DTM solutions are obtained for the dimensionless system under appropriate boundary conditions. We examine the influence of momentum inverse Stokes number (Sk_m), Darcy number (Da), Forchheimer number (Fs), particle loading parameter (p_L), particle-phase wall slip parameter (Ω) and buoyancy parameter (B) on the fluid-phase velocity (U) and particle-phase velocity (U_p). Padé approximants are also employed to achieve satisfaction of boundary conditions. Excellent correlation is obtained between the DTM and numerical quadrature solutions. The results indicate that there is a strong decrease in fluid-phase velocities with increasing Darcian (first-order) drag and the second-order Forchheimer drag, and a weaker reduction in particle-phase velocity field. Fluid and particle-phase velocities are also strongly affected with inverse momentum Stokes number. DTM is shown to be a powerful tool providing engineers with an alternative simulation approach to other traditional methods for multi-phase computational biofluid mechanics. The model finds applications in haemotological separation and biotechnological processing.     

Contribution to the mathematical theory of mass behavior: I. The propagation of single acts

The propagation of a single act in a large population is supposed to depend on some external circumstance and on an “imitation component”, where encounters with individuals who are performing or have already performed the act contribute to the tendency of an individual to perform it. The “tendency” to perform is supposed to be measured by the average frequency of stimuli, randomly distributed in time, impinging on the individual. The deduced equation is a relation between the fraction of the population who have performed the act and time, provided the time course of the “external circumstance” and the way in which the imitation component contributes are known. Several special cases are studied, in particular, cases without the imitation component, cases with imitation only, and various mixed cases. Examples are given of social situations in which such factors may operate and general suggestions are made for the systematization of observations and/or experiments to test the assumptions of the theory.     

Computational fluid dynamics modeling of mass transfer behavior in a bioreactor for hairy root culture. I. Model development and experimental validation

A two-dimensional axisymmetric computational fluid dynamics (CFD) model based on a porous media model and a discrete population balance model was established to investigate the hydrodynamics and mass transfer behavior in an airlift bioreactor for hairy root culture.During the hairy root culture of Echinacea purpurea, liquid and gas velocity, gas holdup, mass transfer rate, as well as oxygen concentration distribution in the airlift bioreactor were simulated by this CFD model. Simulative results indicated that liquid flow and turbulence played a dominant role in oxygen mass transfer in the growth domain of the hairy root culture. The dissolved oxygen concentration in the hairy root clump increased from the bottom to the top of the bioreactor cultured with the hairy roots, which was verified by the experimental detection of dissolved oxygen concentration in the hairy root clump. This methodology provided insight understanding on the complex system of hairy root culture and will help to eventually guide the bioreactor design and process intensification of large-scale hairy root culture.     

A mathematical analysis of rebound in a target-mediated drug disposition model: I.Without feedback

We consider the possibility of free receptor (antigen/cytokine) levels rebounding to higher than the baseline level after one or more applications of an antibody drug using a target-mediated drug disposition model. Using geometry and dynamical systems analysis, we show that rebound will occur if and only if the elimination rate of the drug–receptor product is slower than the elimination rates of the drug and of the receptor. We also analyse the magnitude of rebound through approximations and simulations and demonstrate that it increases if the drug dose increases or if the difference between the elimination rate of the drug–receptor product and the minimum of the elimination rates of the drug and of the receptor increases.     

The rotorfermentor. I. Description of the apparatus,power requirements,and mass transfer characteristics

A novel fermentation device, the rotorfermentor, is described and some experimental results are presented on power requirements and oxygen mass transfer characteristics of the rotorfermentor. This fermentation device is designed to achieve high cell concentrations in batch and continuous cultures. Basically, the rotorfermentor consists of a rotating microporous membrane which is enclosed within a stationary fermentor vessel. The metabolic products in the broth are continuously removed by filtration through the rotating microporous membrane while the growing cells can be retained inside the fermentor. This dual function of cell growth and concentration with the simultaneous removal of metabolic products is the essential characteristic of the rotorfermentor.     

A reduced mathematical model of the acute inflammatory response: I. Derivation of model and analysis of anti-inflammation

The acute inflammatory response, triggered by a variety of biological or physical stresses on an organism, is a delicate system of checks and balances that, although aimed at promoting healing and restoring homeostasis, can result in undesired and occasionally lethal physiological responses. In this work, we derive a reduced conceptual model for the acute inflammatory response to infection, built up from consideration of direct interactions of fundamental effectors. We harness this model to explore the importance of dynamic anti-inflammation in promoting resolution of infection and homeostasis. Further, we offer a clinical correlation between model predictions and potential therapeutic interventions based on modulation of immunity by anti-inflammatory agents.     

Biodegradation of Methyl Orange by alginate-immobilized Aeromonas sp. in a packed bed reactor: external mass transfer modeling

Azo dyes are recalcitrant and xenobiotic nature makes these compounds a challenging task for continuous biodegradation up to satisfactorily levels in large-scale. In the present report, the biodegradation efficiency of alginate immobilized indigenous Aeromonas sp. MNK1 on Methyl Orange (MO) in a packed bed reactor was explored. The experimental results were used to determine the external mass transfer model. Complete MO degradation and COD removal were observed at 0.20 cm bead size and 120 ml/h flow rate at 300 mg/l of initial dye concentration. The degradation of MO decreased with increasing bead sizes and flow rates, which may be attributed to the decrease in surface of the beads and higher flux of MO, respectively. The experimental rate constants (k _ps) for various beads sizes and flow rates were calculated and compared with theoretically obtained rate constants using external film diffusion models. From the experimental data, the external mass transfer effect was correlated with a model J _D = K Re ^?(1 ? n). The model was tested with K value (5.7) and the Colburn factor correlation model for 0.20, 0.40 and 0.60 bead sizes were J _D = 5.7 Re ^?0.15, J _D = 5.7 Re ^?0.36 and J _D = 5.7 Re ^?0.48, respectively. Based on the results, the Colburn factor correlation models were found to predict the experimental data accurately. The proposed model was constructive to design and direct industrial applications in packed bed reactors within acceptable limits.     

Mixing and modes of mass transfer in the third cerebral ventricle: a computational analysis

Anatomic, velocimetric, and brain motion MRI scans were combined with a computational fluid dynamics model to investigate cerebrospinal fluid (CSF) mixing in the third cerebral ventricle of a healthy male adult. It was found that advection dominates over diffusion in most of the third ventricle. Three zones where diffusion plays an important role in the mixing process were identified. One of these zones, consisting of recessus infundibulus, recessus opticus and the adjacent regions up to commissura anterior, is likely to exist in the general population. We hypothesize that this zone may act as a buffer to flatten concentration peaks of pituitary gland hormones released into the CSF of the third ventricle. We further hypothesize that this zone may facilitate the communication between hypothalamus and the pituitary gland through the third ventricle cerebrospinal fluid by prolonging residence times of the communicated hormones.     

Differences in the processes of beta-lactoglobulin cold and heat denaturations.

The changes in beta-lactoglobulin upon cold and heat denaturation were studied by scanning calorimetry, CD, and NMR spectroscopy. It is shown that, in the presence of urea, these processes of beta-lactoglobulin denaturation below and above 308 K are accompanied by different structural and thermodynamic changes. Analysis of the NOE spectra of beta-lactoglobulin shows that changes in the spin diffusion of beta-lactoglobulin after disruption of the unique tertiary structure upon cold denaturation are much more substantial than those upon heat denaturation. In cold denatured beta-lactoglobulin, the network of residual interactions in hydrophobic and hydrophilic regions of the molecules is more extensive than after heat denaturation. This suggests that upon cold- and heat-induced unfolding, the molecule undergoes different structural rearrangements, passing through different denaturation intermediates. From this point of view, cold denaturation can be considered to be a two stage process with a stable intermediate. A similar equilibrium intermediate can be obtained at 35 degrees C in 6.0 M urea solution, where the molecule has no tertiary structure. Cooling or heating of the solution from this temperature leads to unfolding of the intermediate. However, these processes differ in cooperativity, showing noncommensurate sigmoidal-like changes in efficiency of spin diffusion, ellipticity at 222 nm, and partial heat capacity. The disruption with cooling is accompanied by cooperative changes in heat capacity, whereas with heating the heat capacity changes only gradually. Considering the sigmoidal shape of the heat capacity change an extended heat absorption peak, we propose that the intermediate state is stabilized by enthalpic interactions.     

Structure and phospholipid transfer activity of human PLTP: analysis by molecular modeling and site-directed mutagenesis.

The plasma phospholipid transfer protein (PLTP) is an important regulator of high density lipoprotein (HDL) metabolism. We have here, based on sequence alignments of the plasma LPS-binding/lipid transfer protein family and the X-ray structure of the bactericidal/permeability increasing protein (BPI), modeled the structure of PLTP. The model predicts a two-domain architecture with conserved lipid-binding pockets consisting of apolar residues in each domain. By site-directed mutagenesis of selected amino acid residues and transient expression of the protein variants in HeLa cells, the pockets are shown to be essential for PLTP-mediated phospholipid transfer. A solid phase ligand binding assay was used to determine the HDL-binding ability of the mutants. The results suggest that the observed decreases in phospholipid transfer activity of the N-terminal pocket mutants cannot be attributed to altered HDL-binding, but the C-terminal lipid-binding pocket may be involved in the association of PLTP with HDL. Further, the essential structural role of a disulfide bridge between cysteine residues 146 and 185 is demonstrated. The structural model and the mutants characterized here provide powerful tools for the detailed analysis of the mechanisms of PLTP function.     

Quantitative analysis of metabolic processes. II. A model-system for the synthesis of the dissociable enzyme and mathematical formulation of the process

Schemes are presented for induced synthesis of the dissociable enzyme in which repeated use of the template is made. The role of the inducer is to release the repression. A mathematical analysis is carried out and expressions are obtained to describe the kinetics of enzyme formation. A practical case (penicillinase synthesis) is compared with theoretically derived equations by using an analogue computer to simulate an enzyme forming system. A good correlation between theoretical and experimental data is obtained.     

Thermodynamic analysis of thermal transitions in globular proteins. I. Calorimetric study of chymotrypsinogen, ribonuclease and myoglobin

Thermal confonnalional transformations of globular proteins, chymotrypsinogen, ribo-nudcase and myoglobin in solutions at different pH values were studied microcalorimetrically. It was shown that the heat effects observed in the process of heating have a complicated form and their explanation necessitates an assumption on the existence of two stages of the process separated by temperature: (I) a pre-denaturational stage where the protein partial heat capacity changes are probably connected with a labilization of the globule structure and (II) a denaturational stage representing a single-step transition of the protein into a state with a higher enthalpy. Transitions without enthalpy change, the existence of which is shown for the first time in globular proteins, necessitates a more cautions approach to the thennudynamical analysis of different physical parameters on the basis of the van't Hoff equation.     

Quantitative analysis of steroid profiles from urine by capillary gas chromatography : I. Accuracy and reproducibility of the sample preparation

A method is described for the determination of steroid profiles from urine by means of gas chromatography using high-efficiency glass capillary columns. The accuracy and reproducibility of essential steps in the sample preparation (extraction of steroids and steroid conjugates by means of XAD-2, enzymatic hydrolysis with Helix pomatia juice, solvolysis in acidified ethyl acetate and alkali wash) are established using different endogenously labelled urine samples, obtained from normal subjects to whom labelled steroids had been administered. Preliminary results are given on the reproducibility of the derivatization procedure (formation of methoxime-trimethylsilyl (MO-TMS) ethers), the gas chromatographic analysis and the whole method. Two procedures for the purification of MO-TMS steroid derivatives are compared. Application of the method to urine samples of patients with various endocrine disorders is included.     

Characterization of the heat shock response in cultured sugarcane cells : I. Physiology of the heat shock response and heat shock protein synthesis

Effect of heat shock on the growth of cultured sugarcane cells (Saccharum officinarum L.) was measured. Heat shock (HS) treatment at 36 to 38°C (2 hours) induced the development of maximum thermotolerance to otherwise nonpermissive heat stress at 54°C (7 minutes). Optimum thermotolerance was observed 8 hours after heat shock. Development of thermotolerance was initiated by treatments as short as 30 minutes at 36°C. Temperatures below 36°C or above 40°C failed to induce maximum thermotolerance. In vivo labeling revealed that HS at 32 to 34°C induced several high molecular mass heat shock proteins (HSPs). A complex of 18 kilodalton HSPs required at least 36°C treatment for induction. The majority of the HSPs began to accumulate within 10 minutes, whereas the synthesis of low molecular mass peptides in the 18 kilodalton range became evident 30 minutes after initiation of HS. HS above 38°C resulted in progressively decreased HSP synthesis with inhibition first observed for HSPs larger than 50 kilodaltons. Analysis of two-dimensional gels revealed a complex pattern of label incorporation including the synthesis of four major HSPs in the 18 kilodalton range and continued synthesis of constitutive proteins during HS.     

Quantitative analysis of metabolic processes III. A model-system for enzyme synthesis by sequential induction and mathematical formulation of the process

A postulated scheme for sequential enzyme synthesis coupled with linear chain reaction is proposed. Mathematical calculations are carried out for the transient-state formation of enzyme coupled with enzyme-substrate reactions. Examples are presented to demonstrate the relations between various operational parameters during enzyme synthesis as a function of time. Significance of the phenomenon of sequential synthesis is discussed in relation to abnormal growth and cellular injury.