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Statistics aided optimization for high-performance liquid chromatographic analysis of organic acids in tobacco.

HPLC analysis for organic acids in tobacco was optimized with the aid of statistical experimental design, a central composite face-centered design. In the design, only thirteen HPLC analyses were needed for identifying two optimal separation parameters. A Bio-Rad Aminex HPX-87H column was used for the analyses. An optimal separation for seven acids in tobacco was found at a temperature of 57 degrees C and a mobile phase of 0.032 N sulfuric acid solution, or at a temperature of 70 degrees C and a mobile phase of 0.024 N sulfuric acid solution, with a flow rate of 0.6 ml min(-1).

Chromatography, High Pressure Liquid↗

Modeling daily gas exchange of a Douglas-fir forest: comparison of three stomatal conductance models with and without a soil water stress function.

Modeling stomatal conductance is a key element in predicting tree growth and water use at the stand scale. We compared three commonly used models of stomatal conductance, the Jarvis-Loustau, Ball-Berry and Leuning models, for their suitability for incorporating soil water stress into their formulation, and for their performance in modeling forest ecosystem fluxes. We optimized the parameters of each of the three models with sap flow and soil water content data. The optimized Ball-Berry model showed clear relationships with air temperature and soil water content, whereas the optimized Leuning and Jarvis-Loustau models only showed a relationship with soil water content. We conclude that use of relative humidity instead of vapor pressure deficit, as in the Ball-Berry model, is not suitable for modeling daily gas exchange in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) in the Speulderbos forest near the village of Garderen, The Netherlands. Based on the calculated responses to soil water content, we linked a model of forest growth, FORGRO, with a model of soil water, SWIF, to obtain a forest water-balance model that satisfactorily simulated carbon and water (transpiration) fluxes and soil water contents in the Douglas-fir forest for 1995.

Journal Article↗

Discriminative learning quadratic discriminant function for handwriting recognition.

In character string recognition integrating segmentation and classification, high classification accuracy and resistance to noncharacters are desired to the underlying classifier. In a previous evaluation study, the modified quadratic discriminant function (MQDF) proposed by Kimura et al. was shown to be superior in noncharacter resistance but inferior in classification accuracy to neural networks. This paper proposes a discriminative learning algorithm to optimize the parameters of MQDF with aim to improve the classification accuracy while preserving the superior noncharacter resistance. We refer to the resulting classifier as discriminative learning QDF (DLQDF). The parameters of DLQDF adhere to the structure of MQDF under the Gaussian density assumption and are optimized under the minimum classification error (MCE) criterion. The promise of DLQDF is justified in handwritten digit recognition and numeral string recognition, where the performance of DLQDF is comparable to or superior to that of neural classifiers. The results are also competitive to the best ones reported in the literature.

Discrimination Learning↗

Multifacet structure of observed reconstructed integral images.

Three-dimensional images generated by an integral imaging system suffer from degradations in the form of grid of multiple facets. This multifacet structure breaks the continuity of the observed image and therefore reduces its visual quality. We perform an analysis of this effect and present the guidelines in the design of lenslet imaging parameters for optimization of viewing conditions with respect to the multifacet degradation. We consider the optimization of the system in terms of field of view, observer position and pupil function, lenslet parameters, and type of reconstruction. Numerical tests are presented to verify the theoretical analysis.

Algorithms↗

Optimized analytical method for cyclosporin A by high-performance liquid chromatography-electrospray ionization mass spectrometry.

The Micromass Platform LCZ mass detector parameters were optimized for simultaneous recording of the protonated (CsA-H+), sodium adduct (CsA-Na+) and potassium adduct (CsA-K+) of cyclosporin A eluted from a Symmetry Shield RP8 column. The optimized procedure allows a precise analysis of CsA in whole blood or serum without removal of salts prior to analysis. The ratio of the three forms of CsA varied depending on the assay condition and the types of specimens being analyzed. The summation of three ionic forms of CsA detected by LC-ESI-MS is a reliable and simple method to assess CsA concentration in the blood.

Cations, Monovalent↗

Optimized differential display and reamplification parameters for silver staining.

Differential display (DD) is a powerful instrument to detect differences in gene expression of malignant and normal cells. An optimized silver staining protocol was used to compare mRNA expression of keratinocytes and squamous cell carcinoma cells of the head and neck. Optimal concentration for upstream and downstream primer was 0.2 microM. Best primer concentrations for reamplification were between 40 and 60 pM. With this optimized protocol nearly 50 differentially expressed transcripts have been identified and differential display can be applied safer, easier, and faster, than by radioactive labeling.

Carcinoma, Squamous Cell↗

Isolation, characterization and optimization of antifungal activity of an actinomycete of soil origin.

About 312 actinomycetes were isolated from soil samples on chitin agar. All these isolates were purified and screened for their antifungal activity against pathogenic fungi. Out of these, 22% of the isolates exhibited activity against fungi. One promising isolate with strong antifungal activity against pathogenic fungi was selected for further studies. This isolate was from Pune, and was active against both yeasts and molds. Various fermentation parameters were optimized. Based on morphological and biochemical parameters, the isolate was identified as Streptomyces. The correlation of antifungal activity with growth indicated growth dependent production of antimetabolite. Maximum antifungal metabolite production (600 units/ml) was achieved in the late log phase, which remained constant during stationery phase, and it was extracellular in nature.

Actinobacteria↗

Inducible defense against pathogens and parasites: optimal choice among multiple options.

Defense against pathogen, parasites and herbivores is often enhanced after their invasion into the host's body. Sometimes different options are adopted depending on the identity and the quantity of the pathogen, exemplified by the switch between Th1 and Th2 systems in mammalian immunity. In this paper, we study the optimal defense of the host when two alternative responses are available, which differ in the effectiveness of suppressing the growth of pathogen (parasite, or herbivore), the damage to the host caused by the defense response, and the magnitude of time delay before the defense response becomes fully effective. The optimal defense is the one that minimizes the sum of the damages caused by the pathogen and the cost due to defense activities. The damage by pathogens increases in proportion to the time integral of the pathogen abundance, and the cost is proportional to the defense activity. We can prove that a single globally optimal combination of defense options always exists and there is no other local optimum. Depending on the parameters, the optimal is to adopt only the early response, only the late response, or both responses. The defense response with a shorter time delay is more heavily used when the pathogen grows fast, the initial pathogen abundance is large, and the difference in time delay is long. We also study the host's optimal choice between constitutive and inducible defenses. In the constitutive defense, the response to pathogen attack works without delay, but it causes the cost even when the pathogen attack does not occur. We discuss mammalian immunity and the plant chemical defense from the model's viewpoint.

Animals↗

Single-molecule fluorescence detection: autocorrelation criterion and experimental realization with phycoerythrin.

A theory for single-molecule fluorescence detection is developed and then used to analyze data from subpicomolar solutions of B-phycoerythrin (PE). The distribution of detected counts is the convolution of a Poissonian continuous background with bursts arising from the passage of individual fluorophores through the focused laser beam. The autocorrelation function reveals single-molecule events and provides a criterion for optimizing experimental parameters. The transit time of fluorescent molecules through the 120-fl imaged volume was 800 microseconds. The optimal laser power (32 mW at 514.5 nm) gave an incident intensity of 1.8 x 10(23) photons.cm-2.s-1, corresponding to a mean time of 1.1 ns between absorptions. The mean incremental count rate was 1.5 per 100 microseconds for PE monomers and 3.0 for PE dimers above a background count rate of 1.0. The distribution of counts and the autocorrelation function for 200 fM monomer and 100 fM dimer demonstrate that single-molecule detection was achieved. At this concentration, the mean occupancy was 0.014 monomer molecules in the probed volume. A hard-wired version of this detection system was used to measure the concentration of PE down to 1 fM. This single-molecule counter is 3 orders of magnitude more sensitive than conventional fluorescence detection systems.

Lasers↗

Simultaneous liquid chromatography-fluorescence analysis of type A and type B trichothecenes as fluorescent derivatives via reaction with coumarin-3-carbonyl chloride.

A method for the simultaneous LC-fluorescence detection (FLD) determination of eight trichothecenes A and B by pre-column derivatization with coumarin-3-carbonyl chloride, a highly fluorescent fluorophore, has been developed. The reaction conditions (temperature, reaction time, reactant ratios) were optimized to give a reproducible quantitative conversion. All derivatives were characterized by LC-MS. The chromatographic parameters were optimized (column, eluent) to give a very good separation of three type A (diacetoxyscirpenol, T-2 toxin, HT-2 toxin) and five type B trichothecenes [deoxynivalenol (DON), nivalenol, fusarenon-X, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol]. The best conditions were obtained on a narrow-bore C18 column with a water-methanol gradient. The detection limits (S/N = 3:1) in grain samples, with an injected volume of 5 microl, were 0.2-1 ng/g for all trichothecenes. These values are more than one order of magnitude lower than those of other LC-FLD and LC-MS methods and are similar to those obtained by GC-MS. The calibration curves were linear between 100 and 2500 ng/g. The method was successfully applied to the analysis of a certified wheat reference material, after solvent extraction and clean-up on a Mycosep column, obtaining a good recovery (89% for DON) and a high accuracy (z-score value: 0.67).

Calibration↗

[Optimizing process of four-effect countercurrent extraction of danshensu from Salvia miltiorrhiza].

OBJECTIVE: To investigate the process factors and characteristic of four-effect countercurrent extracting process of Danshensu from Salvia miltiorrhiza. Optimizing the parameters by using regression analysis and principal components analysis (PCA). METHOD: Applying the orthogonal design method to optimize the extracting process. The factors of extraction time, solvent volume, extraction temperature and the degree of grinding were considered. And also, the new process was compared with the refluence process. RESULT: The optimal condition of four-effect countercurrent extraction of Danshensu from S. miltiorrhiza was 1 h extraction time, solvent to sample ratio of 12 mL x g(-1), 80 degrees C , 20 - 40 mesh of particle size. Compared with the refluence process, the countercurrent extracting process has the advantages of higher recovery, less solvent consumption and shorter extraction time. CONCLUSION: The four-effect countercurrent extraction process has great advantages and is good for Chinese medicinal materials.

Lactates↗

Optimal conservation strategy in fluctuating environments with species interactions: resource-enhancement of the native species versus extermination of the alien species.

Alien species are often a major threat to native species. We consider optimal conservation strategies for a population whose viability is affected both by an alien species (such as a competitor, a predator, or a pathogen) and by random fluctuations of the environment (e.g. precipitation, temperature). We assume that the survivorship of the native population can be improved by providing resources such as food and shelter, and also by an extermination effort that decreases the abundance of the alien species. These efforts decrease the extinction probability of the native population, but they are accompanied by economic costs. We search for the optimal strategy that minimizes the weighted sum of the extinction probability and the economic costs over a single year. We derive conditions under which investment should be made in both resource-enhancement and extermination, and examine how the optimal effort levels change with parameters. When the optimal strategy includes both types of efforts, the optimal extermination effort level turns out to be independent of the density and economic value of the native species, or the variance of the environmental fluctuation. Furthermore, the optimal resource-enhancement effort is then independent of the density of the alien species. However, the parameter dependencies greatly change if one of the efforts becomes zero. We also examine the situation in which the impact of the alien species is uncertain. The optimal extermination effort increases with the uncertainty of this impact except when the cost of extermination is very high.

Animals↗

Development of an Efficient Regeneration and Agrobacterium-Mediated Transformation Protocol for Hosta 'Light Star' Using the RUBY Reporter Gene.

Hosta plantaginea is a perennial shade-tolerant herb of the Liliaceae family, with high ornamental and urban greening value. Hosta 'Light Star' is a newly developed ornamental cultivar with yellow-margined leaves and lilac flowers, but no efficient in vitro regeneration or genetic transformation system has been established for this cultivar to date. In this study, we established a highly efficient in vitro regeneration system for Hosta 'Light Star,' and developed an Agrobacterium-mediated genetic transformation protocol using the RUBY visual reporter gene for non-invasive screening of positive transformants. The optimal callus induction medium was MS&#x2009;+&#x2009;2&#xa0;mg/L 6-BA&#x2009;+&#x2009;0.3&#xa0;mg/L NAA&#x2009;+&#x2009;0.05&#xa0;mg/L 2, 4-D, with a callus induction rate of 53.33% for leaf explants (the optimal explant for sterile seedlings). The optimal adventitious bud proliferation medium was MS&#x2009;+&#x2009;2&#xa0;mg/L 6-BA&#x2009;+&#x2009;0.1&#xa0;mg/L NAA, with a proliferation coefficient of 5.87. The optimal rooting medium was 1/2 MS&#x2009;+&#x2009;0.5&#xa0;mg/L NAA&#x2009;+&#x2009;0.5&#xa0;mg/L IBA, with a 100% rooting rate. The optimal transplant substrate was perlite:vermiculite&#x2009;=&#x2009;2:1, with a 100% transplant survival rate after acclimatization. For Agrobacterium-mediated transformation, the optimal infection parameters were as follows: Agrobacterium suspension OD600&#x2009;=&#x2009;0.6, infection time of 10&#xa0;min, and 200&#xa0;&#x3bc;M acetosyringone; the optimal selection conditions were 300&#xa0;mg/L cefotaxime for bacteriostasis and 30&#xa0;mg/L hygromycin for transformant screening. The final stable transformation efficiency was 2.50% (95% CI 1.23-3.77%), with an escape rate of 16.13%. Transgenic plants showed distinct purplish-red coloration in roots, stems, and leaves, with significantly higher betacyanin accumulation than wild-type plants (p&#x2009;<&#x2009;0.05). Stable integration and expression of the RUBY gene were confirmed by PCR, RT-PCR, and RT-qPCR. This study establishes the first efficient regeneration and Agrobacterium-mediated transformation system for Hosta 'Light Star,' and validates the feasibility of the RUBY reporter gene as a visual marker for Hosta transformation. This system provides a solid technical platform for functional genomic studies, CRISPR/Cas9-mediated gene editing, and molecular breeding of ornamental traits in Hosta.

Transformation, Genetic↗

The effects of window delay, delinearization, and frequency on tone-burst otoacoustic emission input/output measurements.

Tone-burst otoacoustic emissions (TBOAEs) are a potential tool for objectively examining cochlear activity in humans. However, their use requires knowledge of how the TBOAE input/output depends on measurement and analysis paradigms. The present experiment examined the effect of variations in response-window timing, response delinearization, and local changes in stimulus frequency on TBOAE input/output measurement. None of these experimental manipulations had a profound effect on TBOAE measurements as long as reasonable parameter choices were made. Nonetheless, judicial choice of the experimental parameters can optimize the assessment of BM I/O functions. It is concluded that the consistency of TBOAE I/O across the parameters tested makes it a viable tool to consider for examining human cochlear activity.

Acoustic Stimulation↗

Dobutamine in severe scorpion envenomation: effects on standard hemodynamics, right ventricular performance, and tissue oxygenation.

OBJECTIVES: To document the effects of dobutamine on standard hemodynamics and right ventricular (RV) performance in patients exhibiting pulmonary edema following severe scorpion envenomation, and to characterize the tissue oxygenation profile in patients sustaining scorpion envenomation-related shock. DESIGN: Prospective cohort study. SETTING: An ICU in a university hospital. PATIENTS: Nineteen consecutive patients were admitted to the ICU for severe scorpion envenomation; all 19 patients exhibited hemodynamic pulmonary edema, and 10 patients had peripheral shock. INTERVENTIONS: All patients underwent a hemodynamic study with a Swan-Ganz catheter. In 8 of 19 patients, the thermodilution catheter was equipped with a fast-response thermistor. MEASUREMENTS AND RESULTS: Standard hemodynamic parameters were recorded on admission and following the infusion of dobutamine in all patients at a dosage, from 7 to 20 microg/kg/min, intended to achieve the best hemodynamic and tissue oxygenation compromise. RV ejection fraction (RVEF) and RV volumes were simultaneously recorded in 8 patients, and tissue oxygenation parameters were assessed in the 10 patients with peripheral shock. The clinical signs of tissue hypoperfusion improved, and optimal hemodynamic parameters were achieved at a mean +/- SD dobutamine dosage of 17 +/- 7 microg/kg/min. Dobutamine infusion evoked statistically significant increases in cardiac index, from 2.3 +/- 0.6 to 3.6 +/- 0.7 L/min/m2; stroke volume index, from 18 +/- 5 to 31 +/- 10 mL/m2; and systemic arterial pressure, from 64 +/- 12 to 78 +/- 14 mm Hg. Pulmonary artery occlusion pressure (PAOP) and venous admixture decreased significantly: from 23 +/- 4 to 15 +/- 6 mm Hg and from 29 +/- 7% to 20 +/- 5%, respectively. With respect to RV function, dobutamine infusion significantly increased the RVEF, from 24 +/- 7% to 42 +/- 9%, without significantly changing the RV end-diastolic volume index, reflecting an enhanced RV contractility. In patients with peripheral circulatory failure, the baseline tissue oxygenation profile was consistent with cardiogenic shock, showing increased oxygen extraction as a consequence of a striking depression in oxygen delivery (DO2). After dobutamine infusion, DO2 improved significantly, from 386 +/- 104 to 676 +/- 156 mL/min/m2, with a significant decrease in oxygen extraction, from 34 +/- 8% to 24 +/- 6%. CONCLUSIONS: In severe scorpion envenomation, dobutamine infusion improves impaired heart function. The effects involve both left ventricular and RV dysfunction. Impaired tissue oxygenation is also improved.

Acute Disease↗

[Utilization of the pharmacokinetic parameters of acetylsalicylic acid for optimizing its use with people of different ages].

Nomograms for application of acetylsalicylic acid in persons of different age are presented. The nomograms are based on pharmacokinetic parameters of acetylsalicylic acid with regard to the age. Proceeding from the known constants of absorption and elimination, the nomograms are made use of to screen the loading and maintenance doses, to determine the time of administering the first maintenance dose and the maximal amount of the drug in the body, which was built up as a result of the use of the accepted treatment schedule. While using the nomograms one should be guided by the known therapeutic dose and the dosage intervals or by the therapeutic or maximal allowable dose. The nomograms are unsophisticated, fairly convenient, and help determine the optimal regimen of the use of acetylsalicylic acid in persons of different age.

Adult↗

Optimization of operator and physical parameters for laser welding of dental materials.

OBJECTIVE: Interactions between lasers and materials are very complex phenomena. The success of laser welding procedures in dental metals depends on the operator's control of many parameters. The aims of this study were to evaluate factors relating to the operator's dexterity and the choice of the welding parameters (power, pulse duration and therefore energy), which are recognized determinants of weld quality. DESIGN: In vitro laboratory study. MATERIALS AND METHODS: FeNiCr dental drawn wires were chosen for these experiments because their properties are well known. Different diameters of wires were laser welded, then tested in tension and compared to the control material as extruded, in order to evaluate the quality of the welding. Scanning electron microscopy of the fractured zone and micrograph observations perpendicular and parallel to the wire axis were also conducted in order to analyse the depth penetration and the quality of the microstructure. Additionally, the micro-hardness (Vickers type) was measured both in the welded and the heat-affected zones and then compared to the non-welded alloy. RESULTS: Adequate combination of energy and pulse duration with the power set in the range between 0.8 to 1 kW appears to improve penetration depth of the laser beam and success of the welding procedure. Operator skill is also an important variable. CONCLUSION: The variation in laser weld quality in dental FeNiCr wires attributable to operator skill can be minimized by optimization of the physical welding parameters.

Chromium Alloys↗

Optimization of extracellular keratinase production by poultry farm isolate Scopulariopsis brevicaulis.

A Scopulariopsis brevicaulis poultry farm isolate was chosen to study factors influencing keratinase production. The parameters were optimized by factorial design. The highest enzyme production by this fungus was obtained at pH 7.5, a temperature of 30 degrees C and a growth period of 5 weeks. The production of the enzyme was enhanced when the culture medium was supplemented with glucose (1%), sodium nitrate (2%), feather (1.5%) and CaCl(2) (1 mM). According to the responses from the experimental design, the effects of each variable were calculated, and the interactions between them were determined. The experimental values were found to be in accordance with the predicted values, the correlation coefficient is 0.9978.

Animals↗