Comparison of various methods of KLa estimation in cultures of filamentous microorganisms

Summary Three methods, the dynamic one, the balance one, and the biological one, were used for K_La estimation in the culture ofAspergillus niger. Big differences between the values obtained by the dynamic and the other two methods were observed. The mathematical model including the diffusion of oxygen into the pellets can offer an explanation.     

A steam-sterilizable electrode of the galvanic type for the measurement of dissolved oxygen

An oxygen probe is developed, using the principle of the silver-lead galvanic cell, covered with a plastic membrane. It is steam sterilizable while filled with water; after sterilization, water is easily exchanged for the electrolyte, potassium bicarbonate. Flat plastic membranes up to a thickness of about 0.1 mm can be applied. The membrane is secured and stretched in a special way, which guarantees a leakproof seal. The outer shape of the probe has a tulip-form and consists of polished stainless steel, except for the electrode face. The electrode is suitable for laboratory and industrial use and its components are completely standardized. Almost linear response for O₂-partial pressures up to atmospheric is obtained with 0.001 in. and 0.002 in. FEP-Teflon membranes. 90% response time is of the order of 10–15 sec for the 0.001 in. membrane, Residual current is almost negligible (< 1 μA).     

Optimization of the bare platinum electrode as an oxygen measurement system in photosynthesis

This paper is concerned with the definition of the standard conditions required for optimum operation of the bare platinum electrode with photosynthetic samples. Experimental evidence shows the following: 1) Polarization circuits should have zero resistance; 2) The electrolyte layer between the electrodes should have a conductance higher than 54×10^{–6 –1} per mm² of platinum electrode area; 3) The electrodes should be polarized just before taking the measurements. All these facts can be interpreted in terms of phenomena occurring on the electrode: The adsorption of hydrogen on the electrode imposes the need for low resistances in the system, and oxygen consumption by the electrode is minimized by polarizing the electrodes as late as possible. This investigation increases the reliability of the bare platinum electrode and gives a basis for a comparison of the results from different experiments. Demonstrations of the pertinence of these conditions are made in our lab with the algae Dunaliella Tertiolecta.     

On-line measurement of oxygen uptake in cell culture using the dynamic method

Measurement of oxygen uptake rate is useful in assessing growth, viability, and metabolic activity. In cell culture, however, the oxygen demand is extremely small (typically 0.1-0.3 mM O(2)L-h) and is very difficult to measure accurately using conventional offgas analysis. In many industrial submerged cell culture systems, dissolved oxygen levels are controlled between preset limits by intermittent sparging of air or oxygen. This article describes a computational method for the automatic online determination of oxygen uptake from the dynamic dissolved oxygen probe response. Experimental measurements show that for a typical hybridoma culture, specific oxygen demand is 0.15 mM O(2)/10(9) cells/h. (c) 1996 John Wiley & Sons, Inc.     

Comparison of three different methods for measurement of tissue platinum level

We attempted to make a comparison of three methods for tissue platinum; atomic absorption spectrometry (AAS), inductively coupled plasma atomic emission spectrometry (ICP-AES), and inductively coupled plasma mass spectrometry (ICP-MS). The determination lim its were 0.05 ng/mL on ICP-MS, 50 ng/mL on ICP-AES, and 200 ngJmL on AAS, and the recovery rates were 97.7 ± 6.9% on ICP-MS, 69.0 ± 3.0% on ICP-AES, and 102.4 ± 4.0% on AAS, respectively. Plat inum was detected by ICP-AES and ICP-MS in human vertebrae, but the level was higher by ICP-AES than by ICP-MS. In the mouse kid ney treated with cisplatin, platinum was detected by ICP-MS, but not by ICP-AES. As cadmium gives the absorption peak close to plat inum, cadmium was measured together with platinum by ICP-AES in the vertebrae. From these, ICP-MS is the most sensitive for measure ment at tissue platinum. The sensitivity of ICP-AES looks worse for measuring the tissue platinum, and it is necessary to take care of the contaminant of metals, especially cadmium. AAS is not suitable for measurement of tissue platinum as in the vertebrae and kidneys, because platinum was not detectable by AAS.     

Comparison of methods of temperature measurement in swine

The purpose of these experiments was to test the equivalence of pulmonary artery, urinary bladder, tympanic, rectal and femoral artery methods of temperature measurement in healthy and critically ill swine under clinical intensive care unit (ICU) conditions using a prospective, time series design. First, sensors were tested for error and sensitivity to change in temperature with a precision-controlled water bath and a laboratory-certified digital thermometer for temperatures 34-42 degrees C. There was virtually no systematic (bias) or random (precision) error (<0.2 degrees C). The bladder sensor had the slowest response time to change in temperature (105-120 s). Next, testing was done in an experimental porcine ICU in a non-profit research institution with four male, sedated, and mechanically ventilated domestic farm pigs. The in vivo experiments were conducted over periods of 41-168 h with temperatures measured every 1-5 s. The bladder, tympanic and rectal methods had unacceptable bias (>or=0.5 degrees C) and/or precision (>or=0.2 degrees C). Response time varied from 7 s with the femoral artery method to 280 s (4.7 min) with the tympanic method. We concluded that equivalence of the methods was insufficient for them to be used interchangeably in the porcine ICU. Intravascular monitoring of core body temperature produces optimal measurement of porcine temperature under varying conditions of physiological stability.     

Comparison of colloidal gold electrode fabrication methods: the preparation of a horseradish peroxidase enzyme electrode.

In order to prepare biosensing electrodes which respond to hydrogen peroxide, horseradish peroxidase has been adsorbed to colloidal gold sols and electrodes prepared by deposition of these enzyme-gold sols onto glassy carbon using three methods: evaporation, electrodeposition and electrolyte deposition. In the latter method the enzyme-gold sol is applied to the surface of a glassy carbon disk electrode followed by an equal volume of 2 mM CaCl2. The electrolyte causes the sol to precipitate on the electrode surface, producing an immobilized enzyme electrode. Satisfactory electrodes which gave an electrochemical response to hydrogen peroxide in the presence of the electron transfer mediator ferrocenecarboxylic acid were produced by all three methods. Evaporation of horseradish peroxidase-gold sols produced electrodes with the best reproducibility and the widest linear amperometric response range. These electrodes can also easily be stored in a dry state. Although not as good as evaporation, electrodeposition also produced satisfactory electrodes. Electro-deposition provides the added advantage that it lends itself to the preparation of multi-enzyme/multi-analyte electrodes by the adsorption of different enzymes to separate gold sols, followed by sequential electrodeposition onto discrete areas of a multichannel electrode.     

Comparison of three common DNA concentration measurement methods

Accurate measurement of DNA concentration is important for DNA-based biological applications. DNA concentration is usually determined by the ultraviolet (UV) absorption, fluorescence staining, and diphenylamine reaction methods. However, the best method for quality assurance of measurements is unknown. Here, we comprehensively compared these methods using different types of samples. We found that all three methods accurately determined the concentrations of high-purity DNA solutions. After digestion of DNA samples, concentration measurements revealed that the PicoGreen dye method was very sensitive to the degradation of DNA. The three methods displayed different anti-jamming ability when contaminants such as transfer RNA (tRNA), protein, and organic chemicals were included in DNA solutions. The diphenylamine reaction method gave the highest accuracy, with an average error of approximately 10% between measured and true values. The PicoGreen dye method was influenced by tRNA and protein, and the UV absorption method was susceptible to all kinds of impurities. Overall, the diphenylamine reaction method gave the most accurate results when DNA was mixed with contaminants, the PicoGreen dye method was most suitable for degraded DNA samples or DNA extracted from processed products, and the UV absorbance method was best for evaluating the impurities in DNA solutions.     

Comparison of methods for measuring oxygen consumption in tumor cells in vitro

The oxygen consumption rate of tumor cells affects tumor oxygenation and response to therapies. Highly sensitive methods for determining cellular oxygen consumption are, therefore, needed to identify treatments that can modulate this parameter. We compared the performances of three different methods for measuring cellular oxygen consumption: electron paramagnetic resonance (EPR) oximetry, the Clark electrode, and the MitoXpress fluorescent assay. To compare the assays, we used K562 cells in the presence of rotenone and hydrocortisone, compounds that are known to inhibit the mitochondrial electron transport chain to different extents. The EPR method was the only one that could identify both rotenone and hydrocortisone as inhibitors of tumor cell oxygen consumption. The Clark electrode and the fluorescence assay demonstrated a significant decrease in cellular oxygen consumption after administration of the most potent inhibitor (rotenone) but failed to show any significant effect of hydrocortisone. EPR oximetry is, therefore, the most sensitive method for identifying inhibitors of oxygen consumption on cell assays, whereas the Clark electrode offers the unique opportunity to add external compounds during experiments and still shows great sensitivity in studying enzyme and chemical reactions that consume oxygen (non-cell assays). Finally, the MitoXpress fluorescent assay has the advantage of a high-sample throughput and low bulk requirements but at the cost of a lower sensitivity.     

三种方法测定高寒草甸生态系统蒸散比较

利用涡度相关技术(Eddy covariance technique)、小型蒸渗仪(Mini-lysimeter)和波文比-能量平衡法(BREB)对2005年和2006年夏季(7～8月份)青藏高原海北高寒草甸生态系统的昼间蒸散(E)变化进行了对比观测研究.在观测期间,存在能量不闭合现象,涡度相关系统测定的湍流通量相当于有效能量的73%.3种不同方法测定的蒸散量之间具有较好的相关性,涡度相关系统与小型蒸渗仪测定的蒸散量相关系数达0.96,与波文比法的结果相关系数为0.95.然而,波文比法计算的蒸散量最大,比涡度相关系统的观测值高43%;小型蒸渗仪法的测定值次之,比涡度相关法的观测值高19%;涡度相关法测算的蒸散值最小.研究结果表明,利用涡度相关技术测定该高寒草甸生态系统的潜热通量,可能会过小评价该生态系统的蒸散量.     

Comparison of different pointing methods for sound localizability measurement in the vision impaired subjects

In order to find out the most suitable and accurate pointing methods to study the sound localizability of persons with visual impairment, we compared the accuracy of three different pointing methods for indicating the direction of sound sources in a semi-anechoic dark room. Six subjects with visual impairment (two totally blind and four with low vision) participated in this experiment. The three pointing methods employed were (1) directing the face, (2) directing the body trunk on a revolving chair and (3) indicating a tactile cue placed horizontally in front of the subject. Seven sound emitters were arranged in a semicircle 2.0 m from the subject, 0 degrees to +/-80 degrees of the subject's midline, at a height of 1.2 m. The accuracy of the pointing methods was evaluated by measuring the deviation between the angle of the target sound source and that of the subject's response. The result was that all methods indicated that as the angle of the sound source increased from midline, the accuracy decreased. The deviations recorded toward the left and the right of midline were symmetrical. In the whole frontal area (-80 degrees to +80 degrees from midline), both the tactile cue and the body trunk methods were more accurate than the face-pointing method. There was no significant difference in the center (-40 degrees to +40 degrees from midline). In the periphery (-80 degrees and +80 degrees ), the tactile cue pointing method was the most accurate of all and the body trunk method was the next best. These results suggest that the most suitable pointing methods to study the sound localizability of the frontal azimuth for subjects who are visually impaired are the tactile cue and the body trunk methods because of their higher accuracy in the periphery.