Optimization of feed rate profile for the monoclonal antibody production

The optimal control algorithm to calculate the optimal feed rate profile of nutrient solution containing two limiting nutrients was proposed. Different from other conventional optimization methods, the proposed algorithm calculated the optimal control profiles for different initial and feed conditions. The singular optimal control algorithm, dynamic programming, and nonsingular transformation algorithm were used for the optimization of simple problems of the 4th order and the performances were compared. With the proposed transformation algorithm, the final MAb concentration increased and the CPU time decreased. For the different initial glucose and glutamine conditions, the optimal control profiles were calculated with the proposed transformation algorithm. As the initial glutamine concentration increased, the final MAb concentration also increased due to the cell viability increase. This was also applied to the different feed compositions. When the glutamine concentration was increased in the feed stream, the final MAb concentration also increased.     

Interplay between Constraints,Objectives, and Optimality for Genome-Scale Stoichiometric Models

High-throughput data generation and genome-scale stoichiometric models have greatly facilitated the comprehensive study of metabolic networks. The computation of all feasible metabolic routes with these models, given stoichiometric, thermodynamic, and steady-state constraints, provides important insights into the metabolic capacities of a cell. How the feasible metabolic routes emerge from the interplay between flux constraints, optimality objectives, and the entire metabolic network of a cell is, however, only partially understood. We show how optimal metabolic routes, resulting from flux balance analysis computations, arise out of elementary flux modes, constraints, and optimization objectives. We illustrate our findings with a genome-scale stoichiometric model of Escherichia coli metabolism. In the case of one flux constraint, all feasible optimal flux routes can be derived from elementary flux modes alone. We found up to 120 million of such optimal elementary flux modes. We introduce a new computational method to compute the corner points of the optimal solution space fast and efficiently. Optimal flux routes no longer depend exclusively on elementary flux modes when we impose additional constraints; new optimal metabolic routes arise out of combinations of elementary flux modes. The solution space of feasible metabolic routes shrinks enormously when additional objectives---e.g. those related to pathway expression costs or pathway length---are introduced. In many cases, only a single metabolic route remains that is both feasible and optimal. This paper contributes to reaching a complete topological understanding of the metabolic capacity of organisms in terms of metabolic flux routes, one that is most natural to biochemists and biotechnologists studying and engineering metabolism.     

Optimum operating mode for a class of fermentation

This paper is concerned with optimization of the operating mode of a fermentor. Combining the various modes of operation—batch, semibatch, and continuous—the operating pattern which maximizes the desired metabolic product in a single fermentor is determined by using Kelley's transformation method with Pontryagin's maximum principle. Kelley's transformation method is a device which avoids the singular situation which occurs when the usual procedure of selecting the optimal control function by the maximum principle breaks down. This is the case in the problem considered in this paper. For lysine fermentation, the best operating mode depends on the fermentor capacity and operating time. The results of this study are summarized thus: (i) when the operating time is “long enough,” optimal conditions require that continuous operation follows either semibatch and/or batch operation, and (ii) when the fermentor capacity becomes “large enough,” semibatch operation becomes important.     

CONGENITAL HEART DISEASE—Changing Concepts in the Surgical Treatment

Probably the most important continuing advance in the treatment of congenital heart disease is the ever-diminishing risk of operations on the open heart. The uncomplicated septal defect or valvular stenosis is now corrected under direct vision with essentially the same risk as that which attends the routine operation for patent ductus arteriosus. Perfusion systems, and corrective heart operations, are now available for any patient who weighs 10 kilograms or more; palliative operations are often prescribed for critically ill patients weighing less than 10 kilograms.With respect to the future, successful removal and replantation of the heart in dogs opens the door for imaginative approaches to many states now considered inoperable. Still more inspiring is the realization that cardiac homotransplantation is surgically feasible and immunologically possible, if specific transplantation antigens can be isolated.     

Computer-assisted stereotactic brain operations by means including computerized axial tomography.

With a computerized program system for stereotactic brain operations it becomes possible for the first time to react even before running a possible risk, e.g., in case of punctures in the midbrain, the brain stem, or in the hypothalamus, by simulating the operative procedure even before starting the operation itself. This is effected by the ability to change the penetration angle of the electrode or by choosing a different point of trepanation. The inclusion of computerized axial tomography, especially through the presentation of the CT scan, made to measure with the help of the linear transformation, and of the input of the cranial and ventricular coordinates through a digitizer, together with the coordinates resulting from the X-ray picture, brings the definition of the target point to a still greater optimum. Thus the safety and the precision of the stereotactic operation have been improved even further.     

Phenotypic characterization of <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis> using elementary modes towards synthesis of amino acids

Elementary flux mode (EFM) analysis is a powerful tool to represent the metabolic network structure and can be further utilized for flux analysis. The method enables characterization and quantification of feasible phenotypes in microbes. EFM analysis was employed to characterize the phenotype of Corynebacterium glutamicum to yield various amino acids. The metabolic network of C. glutamicum yielded 62 elementary modes by incorporating the accumulation of amino acids namely, lysine, alanine, valine, glutamine and glutamate. The analysis also allowed us to compute the maximum theoretical yield for the synthesis of various amino acids. These 62 elementary modes were further used to obtain optimal phenotypic space towards accumulation of biomass and lysine. The study indicated that the optimal solution space from 62 elementary modes forms a super space which incorporates various mutants including lysine producing strain of C. glutamicum. The analysis was also extended to obtain sensitivity of the network to variation in the stoichiometry of NADP in the definition of biomass.     

Testing Methods: The Evaluation of Discovery Operations in Evolutionary Biology

Scientific discovery requires both abstract, theoretically defined concepts and discovery operations formed by sets of rules that permit the empirical detection of instances of those concepts. In this paper, I examine the ontological status of discovery operations and the tests employed to evaluate them in evolutionary biology. Attention is drawn to the distinction between nomothetic (universal, predictive) and ideographic (historical, retrodictive) discovery operations, and between complementary and exclusive discovery operations. Three types of tests of discovery operations are commonly employed in evolutionary biology. Theoretical tests aim to show that a discovery operation is inconsistent with accepted, well-corroborated, empirical theories. Empirical tests evaluate the performance of competing discovery operations in terms of their results when applied to the same empirical data sets. Philosophical tests aim to show that an operation is inconsistent with logical and epistemological principles. Appropriately designed theoretical and philosophical tests of ideographic discovery operations may be scientifically valid. Empirical tests, however, are incapable of evaluating the scientific merits of competing discovery operations. Nonetheless, empirical comparisons (not tests ) of competing discovery operations may provide insight into the ways discovery operations may be misleading and therefore may play an important role in stimulating critical debate and eventually establishing a scientifically optimal operation. In practice, theoretical and philosophical tests are often combined to test competing discovery operations as rigorously as possible.     

Validation and optimization of viral clearance in a downstream continuous chromatography setting

Continuous bioprocessing holds the potential to improve product consistency, accelerate productivity, and lower cost of production. However, switching a bioprocess from traditional batch to continuous mode requires surmounting business and regulatory challenges. A key regulatory requirement for all biopharmaceuticals is virus safety, which is assured through a combination of testing and virus clearance through purification unit operations. For continuous processing, unit operations such as capture chromatography have aspects that could be impacted by a change to continuous multicolumn operation, for example, do they clear viruses as well as a traditional batch single column. In this study we evaluate how modifying chromatographic parameters including the linear velocity and resin capacity utilization could impact virus clearance in the context of moving from a single column to multicolumn operation. A Design of Experiment (DoE) approach was taken with two model monoclonal antibodies (mAbs) and two bacteriophages used as mammalian virus surrogates. The DoE enabled the identification of best and worst-case scenario for virus clearance overall. Using these best and worst-case conditions, virus clearance was tested in single column and multicolumn modes and found to be similar as measured by Log Reduction Values (LRV). The parameters identified as impactful for viral clearance in single column mode were predictive of multicolumn modes. Thus, these results support the hypothesis that the viral clearance capabilities of a multicolumn continuous Protein A system may be evaluated using an appropriately scaled-down single mode column and equipment.     

Start-up of chemostat: Application of fed-batch culture

A time-optimal strategy for start-up of a chemostat has been established by applying Miele's extremization method based on Green's theorem. The start-up time is always minimized by traveling along the uppermost boundary of the set of admissible domains on the (vx, μ) phase plane. The solutions are batch culture for monotone increasing growth kinetics and exponentially fed-batch culture followed by batch culture for substrate-inhibition kinetics. A time-suboptimal start-up strategy, exponentially fed-batch culture operated the same μ as that of chemostate, is proposed as more feasible than and physiologically preferable to the time-optimal strategies. The complete set of equations of operations necessary for performing each of the optimal and suboptimal start-ups is formulated. Numerical comparisons on the basis of continuous operation giving maximum productivities of cell-mass outputs show that differences in times required for start-up between optimal and suboptimal strategies are small.     

潮霉素在大麦遗传转化中的应用研究

以‘B1202’、‘U202’和‘云引1号’3个大麦品种(系)为试材,研究了潮霉素对大麦愈伤组织生长、分化及成熟种子的影响。结果表明:潮霉素对抗性愈伤的适宜筛选浓度为60 mg/L,分化阶段的适宜筛选浓度为40 mg/L,对转基因后代种子筛选以80 mg/L较为适宜,而不同材料对潮霉素的敏感性存在一定程度的差异。通过PCR检测采用农杆菌介导法转化获得的T0和T1代潮霉素抗性苗,结果证实在大麦遗传转化中采用潮霉素进行筛选是可行的。     

Optimal in-vitro expansion of chondroprogenitor cells in monolayer culture

A continuous production of large quantities of chondroprogenitor cells for the manufacture of engineered cartilage tissue products is required. Expansion of the cell population in vitro has become an essential step in the process of tissue engineering of articular cartilage and the optimization of the culture conditions is a fundamental problem that needs to be addressed. The analysis of both seeding density and passage length was considered crucial in the optimization of expansion processes, and their correct selection should be taken as a requisite to establish culture conditions for monolayer systems. The determination of the optimal seeding density and the corresponding passage length for cell expansion in a serial passaging operation was found to be a compromise between growth kinetics and process time. This optimal determination was carried out using a mathematical approach that led to values of 10(4) cell/cm(2) for seeding density and 73 h for passage length. Additional considerations concerning the running cost of the process were introduced. Although the optimal passage length gave the desired expansion factor in a minimum process time, the selection of an alternative value of 120 h was shown to reduce the cost of the expansion process in more than 60%. The optimization approach presented will contribute to the development of feasible large scale expansion operations of chondroprogenitor cells required by the cartilage tissue engineering industry.     

药用植物栀子的组织培养

栀子(Gardenia jasminoides)为药用木本植物。以栀子果皮、种子团和种子为外植体,研究不同激素配比及不同培养方式对愈伤组织诱导和芽分化的影响。研究结果表明,培养基成分为MS+0.5 mg·L–12,4-D+0.25 mg·L–16-BA较适宜果皮和种子愈伤组织的诱导,诱导率分别为83.3%和88.5%;培养基成分为MS+1.0 mg·L–12,4-D+1.0 mg·L–16-BA较适宜种子团愈伤组织的诱导,诱导率为78.1%。3种外植体诱导的愈伤组织中,只有种子愈伤组织能通过液体培养分化出芽;TDZ对芽分化有明显的促进作用;最佳的芽分化培养基为MS+0.05 mg·L–1NAA+0.10 mg·L–1TDZ,其愈伤组织分化率为8.75%。该研究以栀子种子为外植体,并获得了再生植株,为药用植物栀子转基因体系的建立奠定了基础。     

Light Amplification with Low-Gain Material: Harvesting Harmonic Resonance Modes of Surface Plasmon Polaritons on a Magnetic Meta-Surface

We theoretically show that it is feasible to utilize harmonic resonance modes of surface plasmon polaritons on a magnetic meta-surface for light amplification with low-gain material. The required threshold and optimal gain strength for active layer can be substantially reduced to about one-tenth as compared with those utilizing fundamental resonance modes. The findings provide a simple and practical metamaterial approach for light amplification with most of the existing active media.     

Change in Diet of Stoats Following Poisoning of Rats in a New- Zealand Forest

The abundance and diet of stoats (Mustela erminea) were compared before and after an aerial 1080-poison operation for possums (Trichosurus vulpecula) in a New Zealand podocarp- hardwood forest. Poisoning dramatically reduced ship rat (Rattus rattus) abundance. Although rats were the main prey item of stoats before the poisoning, stoat abundance was unaffected by the operation and there was a change in stoats' diet from rats to birds. The conservation benefits and risks of undertaking such operations are not clear. It is not known whether the predation risk for any particular species of bird (or other animal) will be higher or lower with fewer rats but the same density of stoats. As large-scale poison operations are now common in New Zealand forests, a better understanding of predator-prey relationships in these areas is required as soon as possible.     

Optimal substrate feeding policy for a fed batch fermentation with substrate and product inhibition kinetics

The optimal substrate feeding policy for the fed batch fermentation which is governed by product and substrate inhibited kinetics is presented. The conjunction point between nonsingular and singular arcs and the feeding policy along the singular arc are derived analytically in terms of the concentrations of substrate and product and the liquid volume. Thus, it is possible to determine the feeding rate by monitoring the state variables (i.e., closed loop control). As a specific example, an optimization study of the fed batch fermentation for ethanol production by Saccharomyces cerevisiae is presented. It is shown that the optimal feeding patterns are heavily dependent upon the initial conditions. The point selectivity provides the guideline for predicting the optimal feeding patterns and explaining the results of rigorous mathematical analysis.     

Plasmid transformation of Mycoplasma mycoides subspecies mycoides is promoted by high concentrations of polyethylene glycol.

The recent isolation and characterization of two plasmids from Mycoplasma mycoides subspecies mycoides has opened up new possibilities for studying mycoplasmal genetics. In order to facilitate the development of a genetic system in M. mycoides subsp. mycoides, parameters of polyethylene glycol (PEG)-mediated transformation were examined, as existing protocols prove very inefficient in this organism. The effects of PEG concentration, DNA concentration, presence of Ca2+ ions, and choice of buffers on the transformation of the Tn916-containing plasmid pAM120 into M. mycoides subsp. mycoides were examined. The stability of Tn916 in the M. mycoides subsp. mycoides chromosome was also evaluated. The optimal PEG concentration (53-62% (w/v)) in the transformation mixture was substantially higher than the PEG concentration reported to be optimal for transformation of other mycoplasmas (36% (w/v)). The PEG concentrations used here were also higher than the concentration used to promote transformation or fusion of gram-positive bacterial protoplasts. A necessity for the presence of Ca2+ ions for optimal transformation was shown, as was the possible involvement of cell culture growth stage. Our results demonstrate the need for expanding current transformation techniques for mycoplasmas. Studies also indicate that once Tn916 inserts into the M. mycoides subsp. mycoides chromosome, it can transpose to other sites at a relatively high frequency.     

药用植物栀子的组织培养

栀子(Gardenia jasminoides)为药用木本植物。以栀子果皮、种子团和种子为外植体, 研究不同激素配比及不同培养方式对愈伤组织诱导和芽分化的影响。研究结果表明, 培养基成分为MS+0.5 mg·L^–12,4-D+0.25 mg·L^–16-BA较适宜果皮和种子愈伤组织的诱导, 诱导率分别为83.3%和88.5%; 培养基成分为MS+1.0 mg·L^–12,4-D+1.0 mg·L^–16-BA较适宜种子团愈伤组织的诱导, 诱导率为78.1%。3种外植体诱导的愈伤组织中, 只有种子愈伤组织能通过液体培养分化出芽; TDZ对芽分化有明显的促进作用; 最佳的芽分化培养基为MS+0.05 mg·L^–1NAA+0.10 mg·L^–1TDZ, 其愈伤组织分化率为8.75%。该研究以栀子种子为外植体, 并获得了再生植株, 为药用植物栀子转基因体系的建立奠定了基础。     

Features of the mechanical treatment of rice husk for the performance of the solid-phase reaction of silicon dioxide with polyphenols

The distinguishing features of rice husk are high strength properties, chemical stability, high ash content, and low nutritional value, which are determined by the composition and structure of this type of raw material. The goal of the study was to determine optimal methods of mechanical treatment for the performance of a solid-phase reaction of silicon dioxide from rice husk with polyphenol compounds. Different regimes of the treatment of plant raw material have been compared. It has been shown that, for the solid-phase reaction of silicon dioxide from plant raw material with polyphenol compounds to occur, both the destruction of the supramolecular complex of silicon dioxide with the lignocellulose matrix, which is accomplished by fine grinding, and the plastic deformation of the silicon dioxide phase with the formation of reaction centers are necessary. These operations can be brought about using grinders with the attrition and shear modes of operation, including roller mills. It is preferable that silicon dioxide is in the composition of plant raw material since the reaction between silicon dioxide and polyphenols with the formation of surface complexes requires the presence of silanol groups. If silicon dioxide is derived from rice husk by conventional methods, most hydroxyl groups are eliminated, which significantly decreases the reactivity.     

The mechanism of action and interaction of regulators of cell replication

The oscillator concept of the cell cycle suggests that regulation of replication is achieved through a switch-like process. This is triggered when the values of parameters governing the behaviour of an intracellular control system exceed thresholds (bifurcations) which separate oscillatory and non-oscillatory (or damped oscillatory) modes of operation. On this basis it becomes possible to explain (a) how a given regulator can have diverse effects, (b) how distinct agents can have similar responses and (c) how various agents interact when controlling replication. The relevance of the malignant transformation is also briefly discussed.     

酵母菌半连续转化人参皂苷Rb1的条件优化

以单因素实验为基础,通过多因素方差分析实验对人参皂苷半连续转化的条件进行优化,选出最佳条件组合,得到最佳的补料方式,补料浓度为6%,补料体积为24mL,补料周期为12h,在此条件下人参皂苷Rb1生物转化达33.5%左右。在最佳补料条件下进行人参皂苷酵母菌转化,其稳定性好,转化率高,对工业生产有积极的推动作用。