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Simulation and optimization of a supercritical extraction process for recovering provitamin A.

In this work, a simulation procedure of a supercritical extraction process was developed through the use of the commercial simulator HYSYS (Hyprotech Ltd.), adapting the existing units to the operating conditions typical of the supercritical extraction process. The objective is to recover provitamin A (beta-carotene) from palm oil (esterified) using carbon dioxide/ethanol as the supercritical mixed solvent. This example characterizes the problem for recovering high added value product from natural sources, as the palm oil, which is desired by the market. Owing to the fact that esterified palm oil is a complex mixture, made by several components, in order to characterize this system in the simulator, it was necessary to create hypothetical components using the UNIFAC (universal function-group activity coefficients model) group contribution, because they are not present in a conventional database and, then, their physical properties must be estimated and/or predicted before the simulation. The optimization was carried out in each simulation for each equipment, in terms of operating conditions (temperature and pressure), in order to obtain the maximum recovery of carotenes. According to the results, it was possible to concentrate carotenes through two cycles of supercritical extraction with high yield. Furthermore, ethyl esters (biodiesel) were also obtained, as a byproduct of the proposed process, which can also be used as an alternative fuel, with the important characteristic that it is renewable.

Carbon Dioxide↗

The effect of processing conditions on the properties of poly(methyl methacrylate) fibers.

Brittle failure of bone cement remains a problem for the long-term stability of hip prostheses. Fibers have been developed from poly(methyl methacrylate) (PMMA) that retain the chemistry of bone cement, but improve the mechanical properties greatly. To fabricate the fibers, a polymer melt is extruded out of a small die (spinneret) and pulled onto a take-up wheel. Varying the speed of extrusion and temperature of the melt can control the viscosity of the molten polymer. This study examines the effect of melt viscosity on the resultant properties of fibers fabricated from PMMA. The goals are to optimize fiber processing and determine processing-structure--property relationships. Resultant fibers had moderate to high levels of retained molecular orientation, and ultimate tensile strengths (UTS) ranging from 60 to 225 MPa, moduli from 1.5 to 3.5 GPa, and strain to failure from 10 to 40%. Fibers fabricated at a constant viscosity and draw velocity had identical properties, whereas decreasing the viscosity generally increased the mechanical properties and retained orientation. Linear regression models were constructed to predict how the processing variables affect the structure (orientation) of the fiber and how the structure affects the UTS. This can be used to design efficient processing methods for PMMA fibers.

Anisotropy↗

Optimization of a heterogeneous reaction system for the production of optically active D-amino acids using thermostable D-hydantoinase.

A thermostable D-hydantoinase from Bacillus stearothermophilus SD-1 was previously mass-produced by batch cultivation of the recombinant E. coli harboring the gene encoding the enzyme (Lee et al., 1997). In this work, we attempted to optimize the process for the production of N-carbamoyl-D-p-hydroxyphenylglycine, which is readily hydrolyzed to D-p-hydroxyphenylglycine under acidic conditions, from 5-(4-hydroxyphenyl)hydantoin using the mass-produced D-hydantoinase. In an effort to overcome the low solubility of the substrate, enzyme reaction was carried out in a heterogeneous system consisting of a high substrate concentration up to 300 g/L. In this reaction system, most of substrate is present in suspended particles. Optimal temperature and pH were determined to be 45 degrees C and 8.5, respectively, by taking into account the reaction rate and conversion yield. When the free enzyme was employed as a biocatalyst, enzyme loading higher than 300 unit/g-substrate was required to achieve maximum conversion. Use of whole cell enzyme resulted in maximum conversion even at lower enzyme loadings than the free enzyme, showing 96% conversion yield at 300 g/L substrate. The heterogeneous reaction system used in this work might be applied to the enzymatic production of other valuable compounds from a rarely water-soluble substrate.

Amidohydrolases↗

Poly(dimethylsiloxane) electrospray devices fabricated with diamond-like carbon-poly(dimethylsiloxane) coated SU-8 masters.

This study presents coupling of a poly(dimethylsiloxane) (PDMS) micro-chip with electrospray ionization-mass spectrometry (ESI-MS). Stable electrospray is generated directly from a PDMS micro-channel without pressure assistance. Hydrophobic PDMS aids the formation of a small Taylor cone in the ESI process and facilitates straightforward and low-cost batch production of the ESI-MS chips. PDMS chips were replicated with masters fabricated from SU-8 negative photoresist. A novel coating, an amorphous diamond-like carbon-poly(dimethylsiloxane) hybrid, deposited on the masters by the filtered pulsed plasma arc discharge technique, improved significantly the lifetime of the masters in PDMS replications. PDMS chip fabrication conditions were observed to affect the amount of background peaks in the MS spectra. With an optimized fabrication process (PDMS curing agent/silicone elastomer base ratio of 1/8 (w/w), curing at 70 degree C for 48 h) low background spectra were recorded for the analytes. The performance of PDMS devices was examined in the ESI-MS analysis of some pharmaceutical compounds and amino acids.

Arginine↗

Biological network design strategies: discovery through dynamic optimization.

An important challenge in systems biology is the inherent complexity of biological network models, which complicates the task of relating network structure to function and of understanding the conceptual design principles by which a given network operates. Here we investigate an approach to analyze the relationship between a network structure and its function using the framework of optimization. A common feature found in a variety of biochemical networks involves the opposition of a pair of enzymatic chemical modification reactions such as phosphorylation-dephosphorylation or methylation-demethylation. The modification pair frequently adjusts biochemical properties of its target, such as activating and deactivating function. We applied optimization methodology to study a reversible modification network unit commonly found in signal transduction systems, and we explored the use of this methodology to discover design principles. The results demonstrate that different sets of rate constants used to parameterize the same network topology represent different compromises made in the resulting network operating characteristics. Moreover, the same topology can be used to encode different strategies for achieving performance goals. The ability to adopt multiple strategies may lead to significantly improved performance across a range of conditions through rate modulation or evolutionary processes. The optimization framework explored here is a practical approach to support the discovery of design principles in biological networks.

Computer Simulation↗

[Study on the dynamic model of supercritical fluid extraction of effective ingredients from natural plant].

In order to obtain the theoretical mass transfer model in the process of extracting effective ingredients from natural plant by Supercritical Fluid, the mass transfer mechanism was analyzed and the analytical solution of the dynamic model of Supercritical Fluid Extraction was deduced through rational simplifying the differential mass balance equations. The complete extraction time, extraction yield as a function of time and the influence of the diameter of solid material on the extraction yield were calculated by the model and the results of computation fit well with the experimental data of literature. This model can be used to analyze, predict, design and optimize the process of extracting effective ingredients from natural plant by Supercritical Fluid.

Chromatography, Supercritical Fluid↗

Modulation of the digestive-lysosomal system in Paramecium caudatum. I. Effects of temperature.

The heterophagic pathway of the digestive-lysosomal system in axenically grown Paramecium caudatum is divisible into vacuole formation, vacuole acidification-condensation, lysosomal fusion-digestion and defecation. These four processes can be separated in time, thus permitting the study of the effects of temperature on each process. The optimal growth temperature for this cell was 27 degrees C. The rate of digestive vacuole (DV) formation at varying temperatures was represented by a skewed bell-shaped curve having an optimum between 28 and 30 degrees C. The time course for the acidification-condensation step was lengthened below 26 degrees C, but was not accelerated above this temperature. The rate but not the extent of vacuole condensation was decreased at 19 and 22 degrees C. Temperature increase above 22 degrees C shortened, slightly, the duration of the lysosomal fusion-digestion process, whereas below 22 degrees C small temperature decreases greatly extended this period. Within a given experiment the rates of defecation were proportional to temperatures above 17 degrees C. However, these rates varied widely among different experiments. Interestingly, the activation energies for both the formation and defecation processes averaged 19 kcal/mol. Furthermore, Paramecium appeared to readily adapt to environmental temperature changes, since the length of the processing periods and the rates of defecation were similar in cells with or without a 24 h acclimation. These results indicated that the four processes in the digestive cycle in P. caudatum are distinct but each is energy-dependent.

Animals↗

Optimization of error-correction processes with respect to time. Comparison to free energy aspects.

Time aspects of selection processes with the possibility of error correction through proofreading branches are studied by mathematical modelling of the kinetics of the reaction network. The methods are similar to those previously developed for free energy aspects. The minimum time delay that is necessary for achieving a certain accuracy level can then be calculated. The main difference to previous results lies in the initial association-dissociation step. In the free energy picture, this shall be essentially equilibrated, but that would yield a too large time delay in the time picture. Characteristic features that are indicative for the optimization strategy of the cell are discussed.

Energy Metabolism↗

Denaturation and reactivation of dimeric human glutathione reductase--an assay for folding inhibitors.

Human glutathione reductase (GR; which catalyzes the reaction NADPH + GSSG + H+ --> 2 GSH + NADP+) is an obligatory FAD-containing homodimer of known geometry. Native human GR, a potential target of antimalarial and cytostatic agents, cannot be dissociated by dilution or by means of subunit-interface mimetics, similarly to well-studied viral dimeric proteins. However, ab initio folding and/or dimerization of human GR can be inhibited by point mutations or by peptides corresponding to subunit-interface areas, for example synthetic peptide P11, which represents the intersubunit-contact helix H11. The structure of this peptide, which might assist inhibitor design, was solved by high-resolution NMR spectroscopy. Residues 440-453, were found to be alpha helical in the isolated peptide. To quantitate the efficacy of inhibitors such as P11, we developed the following unfolding/reactivation assay. The effects of various guanidine hydrochloride (Gdn/HCl) concentrations were studied by analytical ultracentrifugation. It was shown that human GR denatured by greater than 3 M Gdn/HCl is monomeric and free of FAD. Circular-dichroism experiments at 223 nm indicated a half-life of approximately 20 s at 20 degrees C for the unfolding process. To optimize the reactivation yield, four parameters [protein concentration (x) in the range 0.3-10 microg/ml, cofactor supplementation, temperature (y: 0-32 degrees C), and time (0-72 h)] were varied systematically, and a reactivation score z was given to each constellation of parameters. This type of analysis might be useful to optimize refolding and activation yields for other proteins. For human GR, the highest recovery was found not to occur at one of the corners of the x,y plane, but close to its center. Consequently, the optimal assay conditions for folding and dimerization inhibitors are as follows. The enzyme (at 300 microg/ml) is denatured by 5 M guanidine hydrochloride/5 mM dithiothreitol, then reactivated by dilution to 1 microg/ml at pH 6.9 and 20 degrees C. In the absence of inhibitors, this procedure leads to 70% of the control activity within 8 h. Peptides representing the upper subunit interface (for instance residues 436-478) of human GR were found to inhibit refolding with EC50% values in the micromolar range, whereas fragments from other regions of the protein had no influence on this process. For peptide P11, the EC50% value was 20 microM. In conclusion, hGR, enzyme with a tight intersubunit contact area of 21 nm2, appears to be suitable for studying protein folding, dimerization, and prosthetic-group complexation in the absence and presence of compounds that inhibit these processes. There is a shortage, at least for oligomeric enzymes of eukaryotes, of published systematic studies on protein (re)activation.

Dimerization↗

Paramedian forehead flap reconstruction for nasal defects.

BACKGROUND: The paramedian forehead flap is the ultimate reconstructive method for repair of extensive nasal defects. Changes in technique have resulted in the evolution of the modern-day forehead flap, which is a streamlined, efficient, reliable flap that can be counted on to provide superior function and cosmesis in the reconstruction of large nasal defects. OBJECTIVE: Consistent success in the execution of a forehead flap hinges on a careful stepwise approach to the defect, the patient, and the surgical technique. Characterization of these steps was undertaken to assist the surgeon in achieving consistent post- operative results. METHODS: The process of executing a paramedian forehead flap beginning with preoperative assessment through the intraoperative procedure and culminating in the postoperative care is elucidated and discussed. RESULTS: Through thoughtful planning and correct execution of technique, very large nasal defects are reconstructed, with excellent functional and esthetic results. Specific examples illustrate the range of approaches that can be used to address a variety of nasal tissue loss. CONCLUSION: With careful attention to the reconstruction of all components of a nasal defect, a forehead flap can restore virtually any large nasal defect with excellent functional and cosmetic results. The skill sets that help optimize the process of nasal reconstruction are important to acquire. With careful planning and surgical finesse, forehead flaps can often result in nearly imperceptible restoration of the nose.

Forehead↗

An oscillatory hierarchy controlling neuronal excitability and stimulus processing in the auditory cortex.

EEG oscillations are hypothesized to reflect cyclical variations in the neuronal excitability, with particular frequency bands reflecting differing spatial scales of brain operation. However, despite decades of clinical and scientific investigation, there is no unifying theory of EEG organization, and the role of ongoing activity in sensory processing remains controversial. This study analyzed laminar profiles of synaptic activity [current source density CSD] and multiunit activity (MUA), both spontaneous and stimulus-driven, in primary auditory cortex of awake macaque monkeys. Our results reveal that the EEG is hierarchically organized; delta (1-4 Hz) phase modulates theta (4-10 Hz) amplitude, and theta phase modulates gamma (30-50 Hz) amplitude. This oscillatory hierarchy controls baseline excitability and thus stimulus-related responses in a neuronal ensemble. We propose that the hierarchical organization of ambient oscillatory activity allows auditory cortex to structure its temporal activity pattern so as to optimize the processing of rhythmic inputs.

Acoustic Stimulation↗

Optimal control of multiphoton ionization processes in aligned I2 molecules with time-dependent polarization pulses.

Multiphoton ionization processes in aligned I2 molecules are actively controlled by the homemade pulse shaping system, with which a time-dependent polarization pulse can be generated and controlled. We find a correlation between a femtosecond time-dependent polarization pulse and the production efficiency of evenly or oddly charged molecular ions. We achieve much better controllability of the correlation with a time-dependent polarization pulse than with a pulse having a fixed ellipticity. The results suggest the existence of an unknown tunnel ionization mechanism which is characteristic of a time-dependent polarization pulse. Our experiments point to new directions in optimal control studies with molecular systems, as discussed in the text.

Journal Article↗

Development of 5-iodo-2'-deoxyuridine milling process to reduce initial burst release from PLGA microparticles.

The aim of this study was to prepare 5-iodo-2'-deoxyuridine (IdUrd) loaded poly(d,l-lactide-co-glycolide) (PLGA) microspheres with a reduced initial burst in the in vitro release profile, by modifying the drug grinding conditions. IdUrd particle size reduction has been performed using spray-drying or ball milling. Spray-drying significantly reduced drug particle size with a change of the initial crystalline form to an amorphous one and led to a high initial burst. Conversely, ball milling did not affect the initial IdUrd crystallinity. Therefore, the grinding process was optimized to emphasize the initial burst reduction. A first step allowed us to set qualitative parameters such as ball number (7) and cooling with liquid nitrogen to obtain a mean size reduction and a narrow distribution. In a second step, three parameters including milling speed, drug amount and time were studied by a response surface analysis. The interrelationship between drug amount and milling speed was the most significant factor. To reduce particle size it should be necessary to use a moderate speed associated with a sufficient drug amount (400-500 mg). IdUrd release from microparticles prepared by the o/w emulsion/extraction solvent evaporation process with the lowest crystalline particle size (15.3 microns) was studied. Burst effect could be reduced significantly. Concerning the first phase of drug release, the burst was 8.7% for 15.3 microns compared to 19% for 19.5 microns milled drug particles.

Chemistry, Pharmaceutical↗

Optimizing candidate selection--a vision in business limited conference. 1-2 December 1998, Basel, Switzerland.

The pharmaceutical industry is faced with filtering hundreds of thousands of compounds to identify successful drug candidates. Given these numbers, how does the pharmaceutical industry identify optimal therapeutic agents rapidly, efficiently, economically and successfully, with the ultimate result of the patient receiving the best drug? The conference summarized the present and future requirements for evaluating emerging technologies, integrating that technology into a filter for large and growing numbers of compounds, building and linking diverse knowledge bases, and establishing predictive foundations that will optimize and accelerate drug discovery and development. Specific conference topics focused on organizational and management approaches as well as some of the major technologies and emerging techniques for supporting drug candidate selection and optimization. It is predicted that the pharmaceutical industry will be synthesizing and screening a million or more compounds for multiple therapeutic targets in the near future. Pulling together the resources of current and emerging technology, knowledge, and multidisciplinary teamwork, so that discovery and selection of successful drug candidates from this large pool of compounds can take place rapidly, is a significant challenge. This conference focused on the organizational issues and experimental tools that can provide for a shortening of discovery time, identification of current and future selection techniques and criteria, the linking of technologies and business strategies to reduce risk, and novel processes for optimizing candidates more quickly and efficiently. The conference was directed at industrial scientists involved in all stages along the drug discovery and development interface. This conference was well-attended, with approximately 100 participants.

Journal Article↗

Electronic data processing in the Danish cytogenetic central register and EDP problems of registers in general.

A brief introduction to the Danish Cytogenetic Central Register (DCCR) is given, and possibilities, principles and problems concerning the establishment and maintenance of a national cytogenetic register are presented. Various data carrier media for registers in general are discussed, of which the magnetic disc is considered most appropriate. General principles for programs capable of performing insertions, deletions and other modifications in the data base are outlined as well as the principles for the programs in the DCCR. The individual records should preferably be identified by aid of a central person registration number (CPR) rather than by name. The data should be stored and sorted by this identification in order to facilitate retrieval of a desired record. The structure of the records is discussed with regard to prevention of the occurrence of certain errors as well as the optimization of processing. Flexibility and economy of space are achieved by using programs able to handle records of unequal length, and problems occurring in connection with this are discussed. The question of how to protect sensitive data is dealt with, and two different methods used in the DCCR are outlined. Programs capable of analyzing karyotypes with the purpose of recognizing various cytogenetic syndromes have been developed for use in the DCCR. Various examples of computing times of typical program runs are presented.

Chromosome Aberrations↗

Analysis of a mammography teaching program based on an affordance design model.

RATIONALE AND OBJECTIVES: The wide use of computer technology in education, particularly in mammogram reading, asks for e-learning evaluation. The existing media comparative studies, learner attitude evaluations, and performance tests are problematic. Based on an affordance design model, this study examined an existing e-learning program on mammogram reading. MATERIALS AND METHODS: The selection criteria include content relatedness, representativeness, e-learning orientation, image quality, program completeness, and accessibility. A case study was conducted to examine the affordance features, functions, and presentations of the selected software. Data collection and analysis methods include interviews, protocol-based document analysis, and usability tests and inspection. Also some statistics were calculated. RESULTS: The examination of PBE identified that this educational software designed and programmed some tools. The learner can use these tools in the process of optimizing displays, scanning images, comparing different projections, marking the region of interests, constructing a descriptive report, assessing one's learning outcomes, and comparing one's decisions with the experts' decisions. Further, PBE provides some resources for the learner to construct one's knowledge and skills, including a categorized image library, a term-searching function, and some teaching links. Besides, users found it easy to navigate and carry out tasks. The users also reacted positively toward PBE's navigation system, instructional aids, layout, pace and flow of information, graphics, and other presentation design. CONCLUSION: The software provides learners with some cognitive tools, supporting their perceptual problem-solving processes and extending their capabilities. Learners can internalize the mental models in mammogram reading through multiple perceptual triangulations, sensitization of related features, semantic description of mammogram findings, and expert-guided semantic report construction. The design of these cognitive tools and the software interface matches the findings and principles in human learning and instructional design. Working with PBE's case-based simulations and categorized gallery, learners can enrich and transfer their experience to their jobs.

Breast Neoplasms↗

Orthographic processing of polysyllabic words by native and nonnative English speakers.

How polysyllabic English words are analyzed in silent reading was examined in three experiments by comparing lexical decision responses to words physically split on the screen. The gap was compatible either with the Maximal Onset Principle or the Maximal Coda Principle. The former corresponds to the spoken syllable (e.g., ca det), except when the word has a stressed short first vowel (e.g., ra dish), while the reverse is true for the latter (giving cad et and rad ish). Native English speakers demonstrated a general preference for the Max Coda analysis and a correlation with reading ability when such an analysis did not correspond with the spoken syllable. Native Japanese speakers, on the other hand, showed a Max Onset preference regardless of the type of word, while native Mandarin Chinese speakers showed no preference at all. It is concluded that a maximization of the coda is the optimal representation of polysyllabic words in English and that poorer native readers are more influenced by phonology than are better readers. The way that nonnative readers mentally represent polysyllabic English words is affected by the way such words are structured in their native language, which may not lead to optimal English processing.

Adult↗

[Free radical mechanisms of memory disorders of toxic origin and experimental therapy of the condition].

Influence of the pesticides deltametrin and dichlophos on memory processes and activity of the antioxidative enzymes superoxide dismutase (SOD) and catalase in the brain and blood of female rats was studied. Xenobiotics disturbed processes of formation and storage of signs of memory, which were tested in the conditional reaction of passive avoidance and in the conditional reaction of active avoidance; they also lowered SOD activity (but not catalase) in the brain (deltametrin by 25%, dichlophos by 21%) 3 h after their injection to the animals. Indices of SOD activity and blood catalase didn't change. SOD inhibition is suggested to be a result of the action of biotransformation product of deltametrin and dichlophos molecules on it, since in in vitro experiments pesticides don't exert influence on the inhibition kinetics of epinephrine autooxidation by brain homogenates. Prophylactic use of alpha-tocopherol or unithiol optimizes the processes of learning and memory in rats, intoxicated with these toxicants; a more marked antiamnestic, effect is observed in the combined injection of these preparations.

Animals↗