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Chemical and biological characteristics of a west African weaning food supplemented with compea (Vigna unguiculata).

The effects of cowpea and amino acid supplementation on the protein quality and chemical characteristics of a maize-based West African traditional weaning food were studied. Process optimization for improved nutritive value was also determined. Supplementation of the traditional weaning food with cowpea increased the lysine, tryptophan and threonine content while the sulphur-amino acids decreased with increasing levels of cowpea. Further supplementation of 70:30 maize/cowpea blends with lysine, threonine or methionine did not significantly improve (p > 0.05) protein quality in terms of the biological value (BV) and net protein utilization (NPU), although significant increases in the protein scores were noted. However, considerable improvements in the BV and NPU were recorded in blends fortified with either tryptophan alone or a combination of lysine, tryptophan, methionine and threonine. Cooking whole cowpea seeds for 45 min before incorporating in the blend formulation also significantly improved (p < 0.05) the protein quality of maize/cowpea blends. The BV and NPU of blends containing 30% pre-cooked cowpea increased from 52 to 76% and 50 to 71% respectively compared to pure maize porridge. The protein content increased from 10 to 14% and the utilizable proteins more than doubled. The weight increase of experimental rats fed with these blends was comparable to that of rats on casein diet. A 30% supplementation of the maize-based weaning food with cowpea therefore greatly enhances the nutritive value especially when the cowpea is pre-cooked for 45 min. Use of chemical scores alone for such blends cannot be a reliable index of blend quality.

Africa, Western↗

Some factors influencing the proportion of periplasmic hepatitis B virus pre-S2 antigen in the recombinant yeast Hansenula polymorpha.

A central composite design (CCD) was used to evaluate, for the purpose of future process optimization, the influence of pH, yeast extract and ammonium chloride concentrations on the proportion of periplasmic hepatitis B pre-S2 antigen in the recombinant yeast Hansenula polymorpha. Each factor was tested at five levels, and a second order polynomial model for the proportion of periplasmic antigen was fitted to the treatment combinations. pH showed the greatest effect: the proportion of periplasmic antigen was greatly increased at the higher pH levels. At the higher pH levels used, the proportion of periplasmic antigen was enhanced by a high concentration of ammonium chloride. Additional experiments have confirmed both the validity of the selected model and the optimal conditions found. A significant correlation was found between the proportion of periplasmic antigen and the total yield of antigen. These results indicated that it should be possible to modulate the distribution of the pre-S2 antigen between the periplasm and the cytoplasm of the yeast.

Ammonium Chloride↗

The use of particle concentration fluorescence immunoassay technology for the analysis of rDNA products.

The electrophoretic and immunological techniques typically used to detect potentially useful biopharmaceutical proteins are sensitive with detection limits in the nanogram range. However, quantitation of a recombinant protein can be cumbersome, and involve large numbers of samples throughout process optimization schemes. Although electrophoretic methods (i.e., SDS-PAGE and Western blots) now avail themselves to quantitation by densitometry, these techniques are time consuming because of the lack of appropriate automated systems. Biological activity assays, when available, often require relatively pure material and are not suitable for analyzing and quantitating impure or semi-purified samples, typical of the fermentation milieu. The optimization of several rDNA-derived protein systems from both prokaryotic and eukaryotic hosts has been completed using PCFIA, a rapid, sensitive system with high throughput. The development of Particle Concentration Fluorescence Immunoassay (CFIA) procedures for several of these rDNA-derived proteins of interest as potential biopharmaceuticals (e.g., alpha-1-antitrypsin, tPA, soluble CD4, and a malaria vaccine candidate) are discussed.

Binding, Competitive↗

Optimization of cultivation and nutrition conditions and substrate pretreatment for solid-substrate fermentation of wheat straw by Coriolus versicolor.

Bioconversion of wheat straw by solid-substrate fermentation (SSF) with Coriolus versicolor was optimized by varying its physiological parameters. Selective delignification (more lignin than holocellulose degradation) and increases in crude protein (CP) content and in vitro dry matter digestibility (IVDMD) were taken as the criteria to select optimum levels of these parameters. The fungus behaved optimally under the following set of cultural and nutritional conditions: pH 5.5, moisture level 55%, temperature 30 degrees C, duration of fermentation 21 d, form of inoculum--grain culture, turning frequency--once at mid-incubation, urea (nitrogen source) 1.5% (sterile) or 3.0% (nonsterile), single superphosphate (phosphorus + sulfur source) 1.0%, no addition of free polysaccharides (as whey or molasses). A maximum of 17.5% increase in IVDMD involving 4.3% degradation of lignin, was attained in the optimized SSF under laboratory conditions. The digestibility improvement could be further increased by using a substrate pretreatment (physical/chemical/biological) in the following order of preference: NaOH treatment, urea or urine treatment, ensiling, steaming, grinding. For practical farm applications, urea treatment and ensiling appeared most feasible. The laboratory optimized process was also scaled up to 4 kg (sterile and unsterile) and 50 kg (unsterile) fermentations.

Animal Feed↗

Synthesis of cyclodextrin glucosyl transferase by Bacillus cereus for the production of cyclodextrins.

A potent indigenous bacillus isolate identified as Bacillus cereus (RJ-30) was found to produce Cyclodextrin Glucosyl Transferase (CGTase) extracellularly. Process optimization of various fermentation parameters has been established for optimal growth of bacillus and the maximum enzyme synthesis. The organism had the highest specific growth rate (0.7 mu) with a generation time of 1 h in glucose containing medium at the conditions of pH 7.0, 37 degrees C at 300 rpm, 1.5 vvm of agitation, and aeration. At these conditions, it exhibited the maximum activity of 54 U/mL at the synthesis rate of 2.7 U/L/h. CGTase was produced from the early exponential growth and peaked during the midsporulating stage of about 16 h thereafter maintained at the same level of 50 U/mL. Saccharides containing media were better inducers than starch, and the influence of carbohydrate substrates has shown that enzyme synthesis is promoted by xylose (65 U/mL) and, more remarkably, by the supplementation of wheat bran extract in glucose medium (106 U/mL). This organism produced CGTase stably in a chemostat culturing over a period of 400 h with a maximum productivity of 5.4 kU/L/h (threefold higher than obtained in batch culturing [1.75 kU/L/h]). Comparatively, CGTase was produced by immobilized cells in a continuous fluidized bed reactor for over approx 360 h, at a relatively high dilution rate of 0.88 h-1 resulting in the productivity of 23.0 kU/L/h.

Bacillus cereus↗

Optimization study on the formulation of roxithromycin dispersible tablet using experimental design.

This study set out to improve the physical and pharmaceutical characteristics of the present formulation using an appropriate experimental design. The work described here concerns the formulation of the dispersible tablet applying direct compression method containing roxithromycin in the form of coated granules. In this study 2(3) factorial design was used as screening test model and Central Composite Design (CCC) associated with response surface methodology was used as optimization study model to develop and to optimize the proper formulation of roxithromycin dispersible tablet. The three independent variables investigated were functional excipients like binder (X1), disintegrant (X2) and lubricant (X3). The effects of these variables were investigated on the following responses: hardness (Y1), friability (Y2) and disintegration time (Y3) of tablet. Three replicates at the center levels of the each design were used to independently calculate the experimental error and to detect any curvature in the response surface. This enabled the best formulations to be selected objectively. The effect order of each term to all response variable was X3> X2> X1> X1*X2> X2*X2> X2*X3> X3*X3> X1*X3> X1*X1 and model equations on each response variables were generated. Optimized compositions of formula were accordingly computed using those model equations and confirmed by following demonstration study. As a result, this study has demonstrated the efficiency and effectiveness of using a systematic formulation optimization process to develop the tablet formulation of roxithromycin dispersible tablet with limited experiment.

Anti-Bacterial Agents↗

Development of an interactive graphical user interface for therapy simulation.

Because of the ongoing development to more complex non-coplanar techniques in radiotherapy, the use of modem computer graphics while designing a dose plan becomes increasingly essential and more important. In this paper, we describe a concept to simulate 3D conformal treatment techniques on the computer. All important components of the treatment device and relevant patient structures are mapped to an internal model, which allows simulation of motion sequences as well as the interactive adjustment of treatment parameters. The intention of this user interface is to save time by using mainly graphical modules in the optimization process instead of running through the dose calculation every time.

Algorithms↗

On-line monitoring of opaque liquids by photoacoustic spectroscopy.

A new photoacoustic sensor system for on-line monitoring of highly concentrated and optical opaque liquid samples is presented. The dyeing of textiles is performed with highly concentrated dye solutions with concentrations ranging from 50 mg L(-1) up to 40 g L(-1). For process optimization and control of the wastewater, an on-line monitoring of the dye concentration is needed. Optical transmission measurements allow the determination of the dye concentration in a relatively small range. Samples with concentrations in the upper mg L(-1) and g L(-1) range have to be diluted before the measurement due to their optical opacity. Additionally, light-scattering particles have a strong effect on the transmitted light intensity. By photoacoustic spectroscopy, concentrations in condensed matter can be determined over several orders of magnitude. Furthermore, scattering particles do not generate any photoacoustic signal.

Journal Article↗

Fed-batch production of D-ribose from sugar mixtures by transketolase-deficient Bacillus subtilis SPK1.

D-ribose, a five-carbon sugar, is used as a key intermediate for the production of various biomaterials, such as riboflavin and inosine monophosphate. A high D-ribose-producing Bacillus subtilis SPK1 strain was constructed by the chemical mutation of the transketolase-deficient strain, B. subtilis JY1. Batch fermentation of B. subtilis SPK1 with 20 g l(-1) xylose and 20 g l(-1) glucose resulted in 4.78 g l(-1) dry cell mass, 23.0 g l(-1) D-ribose concentration, and 0.72 g l(-1) h(-1) productivity, corresponding to a 1.5- to 1.7-fold increase when compared with values for the parental strain. A late-exponential phase was chosen as the best point for switching to a fed-batch process. Optimized fed-batch fermentation of B. subtilis SPK1, feeding a mixture of 200 g l(-1) xylose and 50 g l(-1) glucose after the late-exponential phase reduced the residual xylose and glucose concentrations to less than 7.0 g l(-1) and gave the best results of 46.6 g l(-1) D-ribose concentration and 0.88 g l(-1) h(-1) productivity which were 2.0- and 1.2-fold higher than the corresponding values in a simple batch fermentation.

Bacillus subtilis↗

Improvement of microbial strains and fermentation processes.

Improvement of microbial strains for the overproduction of industrial products has been the hallmark of all commercial fermentation processes. Conventionally, strain improvement has been achieved through mutation, selection, or genetic recombination. Overproduction of primary or secondary metabolites is a complex process, and successful development of improved strains requires a knowledge of physiology, pathway regulation and control, and the design of creative screening procedures. In addition, it requires mastery of the fermentation process for each new strain, as well as sound engineering know-how for mediaoptimization and the fine-tuning of process conditions. This review focuses on the various options that may be employed to improve microbial strains and addresses the complex problems of screening, the tools and technology behind the selection of targeted organisms, and the importance of process optimization. Furthermore, this review discusses new and emerging technologies and designing optimized media for tracking mutants with enhanced productivity or other desired attributes.

Bacteria↗

Prehension synergies during nonvertical grasping, I: experimental observations.

The mechanical complexities of rotating an object through the gravity field present a formidable challenge to the human central nervous system (CNS). The current study documents the finger force patterns selected by the CNS when performing one-, two-, and four-finger grasping while holding an object statically at various orientations with respect to vertical. Numerous mechanically "unnecessary" behaviors were observed. These included: nonzero tangential forces for horizontal handle orientations, large internal forces (i.e., those in excess of equilibrium requirements) for all orientations, and safety margins between 50 and 90%. Additionally, none of the investigated measures were constant across orientations or could be represented as a simple trigonometric function of orientation. Nonetheless, all measures varied in systematic (and sometimes symmetric) ways with orientation. The results suggest that the CNS selects force patterns that are based on mechanical principles but also that are not simply related to object orientation. This study is complemented by a second paper that provides an in-depth analysis of the mechanics of nonvertical grasping and accounts for many of the observed results with numerical optimization (see Part II - current issue). Together, the papers demonstrate that the CNS is likely to utilize optimization processes when controlling prehensile actions.

Adaptation, Physiological↗

Impedance characteristics of a neuromusculoskeletal model of the human arm II. Movement control.

The modulation of neuromusculoskeletal impedance during movements is analysed using a motor control model of the human arm. The motor control system combines feedback and feedforward control and both control modes are determined in one optimization process. In the model, the stiffness varies at the double movement frequency for 2-Hz oscillatory elbow movements and has high values at the movement reversals. During goal-directed two-degrees-of-freedom arm movements, the stiffness is decreased during the movement and may be increased in the initial and final phases, depending on the movement velocity. The stiffness has a considerable curl during the movement, as was also observed in experimental data. The dynamic stiffness patterns of the model can be explained basically by the alpha-gamma coactivation scheme where feedback gains covary with motor control signals. In addition to the modulation of the gain factors, it is argued that the variation of the intrinsic stiffness has a considerable effect on movement control, especially during fast movements.

Arm↗

Measurement of bubble and pellet size distributions: past and current image analysis technology.

Measurements of bubble and pellet size distributions are useful for biochemical process optimizations. The accuracy, representation, and simplicity of these measurements improve when the measurement is performed on-line and in situ rather than off-line using a sample. Historical and currently available measurement systems for photographic methods are summarized for bubble and pellet (morphology) measurement applications. Applications to cells, mycelia, and pellets measurements have driven key technological developments that have been applied for bubble measurements. Measurement trade-offs exist to maximize accuracy, extend range, and attain reasonable cycle times. Mathematical characterization of distributions using standard statistical techniques is straightforward, facilitating data presentation and analysis. For the specific application of bubble size distributions, selected bioreactor operating parameters and physicochemical conditions alter distributions. Empirical relationships have been established in some cases where sufficient data have been collected. In addition, parameters and conditions with substantial effects on bubble size distributions were identified and their relative effects quantified. This information was used to guide required accuracy and precision targets for bubble size distribution measurements from newly developed novel on-line and in situ bubble measurement devices.

Forecasting↗

A biomechanical model of mammographic compressions.

A number of biomechanical models have been proposed to improve nonrigid registration techniques for multimodal breast image alignment. A deformable breast model may also be useful for overcoming difficulties in interpreting 2D X-ray projections (mammograms) of 3D volumes (breast tissues). If a deformable model could accurately predict the shape changes that breasts undergo during mammography, then the model could serve to localize suspicious masses (visible in mammograms) in the unloaded state, or in any other deformed state required for further investigations (such as biopsy or other medical imaging modalities). In this paper, we present a validation study that was conducted in order to develop a biomechanical model based on the well-established theory of continuum mechanics (finite elasticity theory with contact mechanics) and demonstrate its use for this application. Experimental studies using gel phantoms were conducted to test the accuracy in predicting mammographic-like deformations. The material properties of the gel phantom were estimated using a nonlinear optimization process, which minimized the errors between the experimental and the model-predicted surface data by adjusting the parameter associated with the neo-Hookean constitutive relation. Two compressions (the equivalent of cranio-caudal and medio-lateral mammograms) were performed on the phantom, and the corresponding deformations were recorded using a MRI scanner. Finite element simulations were performed to mimic the experiments using the estimated material properties with appropriate boundary conditions. The simulation results matched the experimental recordings of the deformed phantom, with a sub-millimeter root-mean-square error for each compression state. Having now validated our finite element model of breast compression, the next stage is to apply the model to clinical images.

Biomechanical Phenomena↗

A growth kinetic model of Kluyveromyces marxianus cultures on cheese whey as substrate.

This work presents a multi-route, non-structured kinetic model for determination of microbial growth and substrate consumption in an experimental batch bioreactor in which beta-galactosidase is produced by Kluyveromyces marxianus growing on cheese whey. The main metabolic routes for lactose, and oxygen consumption, cell growth, and ethanol production are derived based on experimental data. When these individual rates are combined into a single growth rate, by rewriting the model equations, the model re-interpretation has a complexity similar to that of the usual variations of the Monod kinetic model, available in the literature. Furthermore, the proposed model is in good agreement with the experimental data for different growth temperatures, being acceptable for dynamic simulations, processes optimization, and implementations of model-based control technologies.

Bioreactors↗

Application of the method of fundamental solutions to potential-based inverse electrocardiography.

Potential-based inverse electrocardiography is a method for the noninvasive computation of epicardial potentials from measured body surface electrocardiographic data. From the computed epicardial potentials, epicardial electrograms and isochrones (activation sequences), as well as repolarization patterns can be constructed. We term this noninvasive procedure Electrocardiographic Imaging (ECGI). The method of choice for computing epicardial potentials has been the Boundary Element Method (BEM) which requires meshing the heart and torso surfaces and optimizing the mesh, a very time-consuming operation that requires manual editing. Moreover, it can introduce mesh-related artifacts in the reconstructed epicardial images. Here we introduce the application of a meshless method, the Method of Fundamental Solutions (MFS) to ECGI. This new approach that does not require meshing is evaluated on data from animal experiments and human studies, and compared to BEM. Results demonstrate similar accuracy, with the following advantages: 1. Elimination of meshing and manual mesh optimization processes, thereby enhancing automation and speeding the ECGI procedure. 2. Elimination of mesh-induced artifacts. 3. Elimination of complex singular integrals that must be carefully computed in BEM. 4. Simpler implementation. These properties of MFS enhance the practical application of ECGI as a clinical diagnostic tool.

Action Potentials↗

On-column refolding and purification of recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) expressed as inclusion body in Escherichia coli.

Recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) was produced in E. coli as an inclusion body. rHuIL-1ra was purified to Over 98% purity by anion exchange chromatography after on-column refolding. The optimized processes produced more than 2 g pure refolded rHuIL-1ra per 1 l culture, corresponding to a 44% recovery, without an intermediate dialysis step. Refolded rHuIL-1ra had full biological activity with the MTT assay. An intramolecular disulfide linkage in the oxidized recombinant protein was suggested by data from HPLC and non-reducing SDS-PAGE.

Chromatography, Ion Exchange↗

Simultaneous saccharification and L-(+)-lactic acid fermentation of protease-treated wheat bran using mixed culture of lactobacilli.

Protease-treated wheat bran (20% w/v) of particle size less than 300 microm containing 65% (w/w) starch was used for the simultaneous saccharification and L-(+)-lactic acid fermentation by the mixed cultures of Lactobacillus casei and Lactobacillus delbrueckii. Maximum lactate yield after various process optimizations was 123 g l(-1) with a productivity of 2.3 g l(-1) h(-1) corresponding to a conversion of 0.95 g lactic acid per gram starch after 54 h at 37 degrees C. By using protease-treated wheat bran around tenfold decrease in supplementation of the costly medium component, like yeast extract, was achieved together with a considerable increase in the production level.

Bioreactors↗