Microcomputer tools for steady-state enzyme kinetics

Three programs useful for the investigation of steady–statekinetics have been developed. Two provide the solution to thesteady–state rate equation; the first of these is a straightforwardimplementation of the rules developed by Chou. The second isa very efficient procedure for evaluating King–Altmandiagrams and can be used for quite large mechanisms. The thirdprogram provides the numeric solution for a specific mechanismand set of initial conditions; it is well suited to extremelylarge models. Received on April 1, 1985; accepted on April 19, 1985     

海南岛南湾半岛猕猴(Macaca mulatta)种群数量动态及分布

自1981年以来,对海南岛南湾半岛猕猴进行了连续5年的调查研究。归纳1965年到1984年底的材料,该种群从100只发展到930只,其间种群的年均增长率为13％,种群增长率受着种群密度的制约,密度与增长率间存在着关系式y=-0.0003x~2+0.0857x+0.3534。该种群生长曲线可表示为指数函数式y=0.1253e~(0.1046x)或为Logistic曲线N=1850/1+e~(11.28-0.13x)。该半岛猕猴最大容纳量为1850只。各猴群的弹性核域受猴群密度和植被质量的影响,绘出了它们之间的关系图。     

A method for testing the stability of a steady-state system during the calculation of a response to large changes in regulator concentrations

A modification of the ‘finite decomposition’ method (Crabtree and Newsholme (1985) Curr. Top. Cell. Regul. 25, 21–76) for calculating physiological responses from sensitivities is described, to enable the system to be tested for stability at each step of the procedure. Instability is indicated by a change of sign of the determinant of the square matrix (N) in the governing equation for the system. The method cannot be used to predict responses beyond any step at which instability occurs.     

Daphnia revisited: local stability and bifurcation theory for physiologically structured population models explained by way of an example

We consider the interaction between a general size-structured consumer population and an unstructured resource. We show that stability properties and bifurcation phenomena can be understood in terms of solutions of a system of two delay equations (a renewal equation for the consumer population birth rate coupled to a delay differential equation for the resource concentration). As many results for such systems are available (Diekmann et al. in SIAM J Math Anal 39:1023–1069, 2007), we can draw rigorous conclusions concerning dynamical behaviour from an analysis of a characteristic equation. We derive the characteristic equation for a fairly general class of population models, including those based on the Kooijman–Metz Daphnia model (Kooijman and Metz in Ecotox Env Saf 8:254–274, 1984; de Roos et al. in J Math Biol 28:609–643, 1990) and a model introduced by Gurney–Nisbet (Theor Popul Biol 28:150–180, 1985) and Jones et al. (J Math Anal Appl 135:354–368, 1988), and next obtain various ecological insights by analytical or numerical studies of special cases.     

In situ calibration and [H+] sensitivity of the fluorescent Na+ indicator SBFI

Despite the popularity of Na+-binding benzofuran isophthalate (SBFI) to measure intracellular free Na+ concentrations ([Na+](i)), the in situ calibration techniques described to date do not favor the straightforward determination of all of the constants required by the standard equation (Grynkiewicz G, Poenie M, and Tsien RY. J Biol Chem 260: 3440-3450, 1985) to convert the ratiometric signal into [Na+]. We describe a simple method in which SBFI ratio values obtained during a "full" in situ calibration are fit by a three-parameter hyperbolic equation; the apparent dissociation constant (K(d)) of SBFI for Na+ can then be resolved by means of a three-parameter hyperbolic decay equation. We also developed and tested a "one-point" technique for calibrating SBFI ratios in which the ratio value obtained in a neuron at the end of an experiment during exposure to gramicidin D and 10 mM Na+ is used as a normalization factor for ratios obtained during the experiment; each normalized ratio is converted to [Na+](i) using a modification of the standard equation and parameters obtained from a full calibration. Finally, we extended the characterization of the pH dependence of SBFI in situ. Although the K(d) of SBFI for Na+ was relatively insensitive to changes in pH in the range 6.8-7.8, acidification resulted in an apparent decrease, and alkalinization in an apparent increase, in [Na+](i) values. The magnitudes of the apparent changes in [Na+](i) varied with absolute [Na+](i), and a method was developed for correcting [Na+](i) values measured with SBFI for changes in intracellular pH.     

A comment on "A method to measure the duration of DNA synthesis and the potential doubling time from a single sample"

Recently a method was introduced by Begg et al. (Cytometry 6:620-626, 1985) for the calculation of the DNA synthesis time, TS using flow cytometry to monitor the progression of cells that have been pulse-labeled by bromodeoxyuridine (BrdUrd). Ideally, the method uses only a single two-parameter histogram of the progressed cells to obtain TS. The essence of the method is to be able to obtain an initial point from general principles, draw a line through it and the measured point, and extrapolate to a level of progression that would be attained when the time is equal to TS. In this comment, explicit values are derived for the initial point and the progression of cells based on kinetic arguments, and a new method is presented for estimating TS. This new method is compared to earlier methods, and the effects of a finite labeling time and variation in the transit times are considered. Finally, an equation is developed for estimating TS following multiple measurements.     

Yangtzedonta是双壳类吗?

对Yangtzedonta pri mitivaYu模式标本(标本登记号:84430)的化石保存类型进行讨论,确认这应当是一枚单壳瓣实体标本,而非内模标本,由此判断,余汶(1985;Yu,2005)所描述的肌痕实际上是不存在的。所谓的"片状齿突"(余汶,1985)或"片状脊"(Yu,2005)很可能是外来的附加物,并非壳体的组成部分。无论如何,84430标本现有的形态证据尚不足于将它与双壳类联系在一起。     

Mitochondrial membrane potential estimated with the correction of probe binding

Lipophilic ions are widely used as the probe for estimation of the membrane potential. It is suggested that the correction of the probe binding to the membrane and/or intracellular constituents is a problem to be solved in order to evaluate the membrane potential accurately. Previously, we proposed a method for the correction of the probe binding (Demura, M., Kamo, N. and Kobatake, Y. (1985) Biochim. Biophys. Acta 820, 207-215). In this paper, the method was applied to the determination of the membrane potential of intact mitochondria. The probes used constitute a homologous series of (Phe)3-P+-(CH2)n-CH3 (n = 0-4) and tetraphenylphosphonium (TPP+). Binding of these probes to de-energized mitochondria followed the Langmuir isotherm. However, values of parameters determined at high (50-800 microM) and low (under 20 microM) probe concentrations were different, suggesting the existence at least two, high- and low-affinity, binding sites. With extrapolation to the 'state of no binding', the membrane potential of intact mitochondria was estimated to be -147 mV (interior-negative) when they were energized by 5 mM succinate in medium consisting of 125 mM KCl, 10 mM MgCl2, 5 mM phosphate, 0.4 mM EDTA and 50 mM Tris-HCl (pH 7.5) at 25 degrees C. Parameters appearing in the equation for the correction of probe binding were determined with the use of this value of the membrane potential. The validity of the equation and the value of the parameters were revealed by the fact that after the correction, all probes used gave approximately the same value under the same conditions. We expanded the method so as to include the langmuir adsorption isotherm. When the modified equation is used, the estimated membrane potentials were less dependent on a probe concentration less than 10 microM.     

Aflatoxin contamination of Virginia peanuts for the crop-years 1982–1986

To establish those environmental conditions which promote the growth of aflatoxin (AFT)-producing Aspergillus spp. on peanuts, a four-year (1982–1986) investigation was undertaken to examine possible relationships between air temperature (AT), precipitation (P), and AFT contamination of stored nuts. The mean percentages of nuts that possessed various AFT levels for the years 1982–1986 (June–July) ranged from 74·2 to 88·0 for 0–4 ppb, 6·3 to 14·9 for 5–15 ppb, 2·4 to 5·9 for 16–25 ppb, 2·3 to 6·4 for 26–100 ppb, and 0 to 4·7 for > 100 ppb. The mean percentages for the years which exceeded USDA/FDA regulations were 7·1 (1982–1983), 7·6 (1983–1984), 11·6 (1984–1985), and 17·0 (1985–1986). Examination of the mean percentage > 15 ppb for each month during these four years revealed that the following months fell within that range; Septtember, November, December, January, February, and May (1982–1983); July, October, April, and June (1983–1984); August, and June, (1984–1985); and July, April, and May (1985–1986). Comparisons of pooled-AFT levels, rainfall, and temperature over four years suggested a ‘better fit’ between mean monthly P and mean percentage AFTs > 15 ppb, than between the latter and mean monthly AT. However, application of a predictor equation suggested a correlation between AFT levels and monthly AT.     

Measurement of the Critical Nyctoperiod by Lemna paucicostata 6746 Grown in Continuous Light

The short-day duckweed Lemna paucicostata 6746 could be inducedto flower in two days at 26C when continuous illumination forentrainment was followed by continuous darkness. This 48-h darkperiod or the minimum darkness requirement for floral inductionwas called the induction period. The length of the inductionperiod (IP) was routinely computed as the number of 24-h cyclesusing the equation of regression of flower number in logarithmon culture time. A light pulse given about 7 h after the startof the induction period increased the apparent IP value fromtwo to three, suggesting that the interrupted first day hadfunctioned as a noninductive day. A pulse given at any otherpart of the induction period did not modify the IP value. Thelight-sensitive part is probably the inducible phase, and thefirst 7-h period of darkness terminated by it seems to be thecritical nyctoperiod. These and relevant facts suggest thatthe light-off oscillator measures the critical night length,7 h. Either red or far-red irradiation at the inducible phase extendedthe IP value by one. No red/far-red photoreversibility was detected.As expected, however, red or far-red irradiation of any otherpart of the critical nyctoperiod could not modify the IP value. (Received February 8, 1985; Accepted May 14, 1985)     

Effect of nuptiality, contraception and breast-feeding on fertility in Bangladesh

The effects of marriage, contraception, and post-partum lactational infecundability on fertility in Bangladesh are assessed by applying Bongaarts' formula to survey data for the period 1975-1985. Marriage is universal and age at marriage is low. Breastfeeding is prolonged and has a pronounced effect on fertility. The fertility-reducing effect of contraception increased over the period through increased use of modern methods. The total fertility rate (TFR) declined by 24% from 1975 to 1985. This study shows that the 3 major proximate determinants cannot account completely for variation in national fertility levels.     

On an Efficient Regression Type Estimator

The regression type estimator proposed by KAUR (1985) is considered. Another expression for the approximated mean square error (AMSE), to a first degree of approximation, is obtained. This AMSE is also minimized with respect to a parameter α. Three numerical examples are included. These numerical examples show that this estimator is not significantly more efficient than regression estimator and with respect to ratio and sample mean estimators, it does not always exhibit a high efficiency, as was contended by KAUR (1985). Moreover, an upper bound for the relative precision of the proposed estimator with respect to linear regression estimator is derived.     

Possible cooperativity in crossbridge detachment in muscle fibers having magnesium pyrophosphate at the active site.

When rabbit psoas muscle fibers bathed in solutions containing the ATP analogue magnesium pyrophosphate (MgPPi) are first stretched rapidly and then held isometric, a force is generated during the stretch which decays during the subsequent isometric period (Schoenberg, M., and E. Eisenberg. 1985. Biophys. J. 48:863-871). Previously we showed that the force decay is due to crossbridge heads detaching and reattaching in positions of lesser strain, the rate of decay of force reflecting the crossbridge detachment rate constants. Since the crossbridge detachment rate constants with MgPPi bound to the active site are so much faster than without analogue bound, at subsaturating concentrations of analogue, if the heads act independently and nucleotide association and dissociation is rapid, the rate of force decay should simply be proportional to the number of heads with bound analogue. That is, the analogue concentration dependence of the rate of force decay should have the same form as the Michaelis-Menten equation. Here we report that the concentration dependence of the rate of force decay is not described by the Michaelis equation, but is instead sigmoidal. This suggests possible cooperativity in the detachment of the crossbridge heads, the amount of cooperativity being described by an interaction coefficient of approximately 2. One idea put forward to explain the data is that both of the heads of a crossbridge may need to bind analogue before the crossbridge can relax a substantial fraction of the tension it supports.     

Slow upward drift of VO2 during constant-load cycling in untrained subjects

The oxygen uptake kinetics during constant-load exercise when sitting on a bicycle ergometer were determined in 7 untrained subjects by measuring breath-by-breath VO2 during continuous exercise to volitional exhaustion (mean endurance time = 1160 +/- 172 s) at a pedal frequency of 70 revolutions.min-1. The power output, averaging 189.5 W, was set at 82.5% of that eliciting the individual VO2max during a 5 min incremental exercise test. Throughout the exercise period, the VO2 kinetics could be appropriately described by a two-component exponential equation of the form: VO2(t) = Ya[1 - exp(-kat)] + Yb[1 - exp(-kbt)] where VO2 is net oxygen consumption and t the time from work onset. VO2 measured at the end of exercise was close to VO2max (98% VO2max) and the mean values of Ya, ka, Yb and kb amounted to 1195 ml O2.min-1, 0.034 s-1, 1562 ml O2.min-1, and 0.005 s-1 respectively. The initial rate of increase in VO2 predicted from the above equation is slower than that calculated, for the same work intensity, on the basis of the data obtained by Morton (1985) in trained subjects. For t greater than 480 s, however, the two models yield substantially equal results.     

布氏田鼠种群生态研究

1985年5-l0月用标志重捕法对布氏田鼠(Microtus brandti)的种群年龄组成、幼鼠生长率和季节迁移等进行研究,共标志2786只,重捕576只,822只次。发现布氏田鼠种群的年龄组成有较明显的季节变化。其幼鼠体重的日平均生长率随年龄增加而减慢。布氏田鼠还有季节迁移现象。     

Gymnosperm systematics and phylogeny: A reply to commentaries by C. B. Beck,C. N. Miller,and G. W. Rothwell

This paper is a reply to commentaries by Beck (1985), Miller (1985), and Rothwell (1985) on the author’s paper on basic features of gymnosperm systematics and phylogeny (Meyen, 1984). The author reasserts that his proposal to recognize three classes of gymnosperms—Ginkgoopsida, Cycadopsida, and Pinopsida—is in better agreement with the currently available paleobotanical data. The affinities of the orders Arberiales, Pentoxylales, gigantopterids, Calamopityales, and Ephedrales with the Ginkgoopsida are but tentative. The most important remaining question?etermining the understanding of relations between Ginkgoopsida and Pinopsida—concerns primary vs. secondary platyspermy in each class, and this question can be answered only by obtaining fuller information on (1) cupular or non-cupular seeds in the oldest Ginkgoopsida, and (2) the vascularization of nucellus in the oldest Pinopsida. Theoretical premises of the author’s taxonomic and phylogenetic decisions are explained.     

Structural fluctuations and current noise of ionic channels.

In the open state of the acetylcholine-receptor channel an increased current noise is observed, which may result from conformational fluctuations of the channel protein (Sigworth, F.J., 1985, Biophys. J. 47:709-720). In this study the spectrum of the current noise is analyzed assuming that low-frequency motions of structural domains of the protein give rise to conductance fluctuations. The movement of a domain is treated as the motion of an elastically bound Brownian particle, which is described by the Langevin equation. The current-noise spectrum predicted by this model is given by a sum of Lorentzians; it agrees with the observed spectrum when it is assumed that only the slowest process can be resolved in the experiment. The large value of the friction coefficient, which is obtained from the corner frequency, indicates that domain motion is restricted mainly by peptide-peptide interactions.     

Mitochondrial membrane potential estimated with the correction of probe binding

Lipophilic ions are widely used as the probe for estimation of the membrane potential. It is suggested that the correction of the probe binding to the membrane and / or intracellular constituents is a problem to be solved in order to evaluate the membrane potential accurately. Previously, we proposed a method for the correction of the probe binding (Demura, M., Kamo, N. and Kobatake, Y. (1985) Biochim. Biophys. Acta 820, 207–215). In this paper, the method was applied to the determination of the membrane potential of intact mitochondria. The probes used constitute a homologous series of (Phe)₃-P⁺-(CH₂)_n-CH₃ (n = 0−4) and tetraphenylphosphonium (TPP⁺). Binding of these probes to de-energized mitochondria followed the Langmuir isotherm. However, values of parameters determined at high (50–800 μM) and low (under 20 μM) probe concentrations were different, suggesting the existence at least two, high- and low-affinity, binding sites. With extrapolation to the ‘state of no binding’, the membrane potential of intact mitochondria was estimated to be −147 mV (interior-negative) when they were energized by 5 mM succinate in medium consisting of 125 mM KCl, 10 mM MgCl₂, 5 mM phosphate, 0.4 mM EDTA and 50 mM Tris-HCl (pH 7.5) at 25°C. Parameters appearing in the equation for the correction of probe binding were determined with the use of this value of the membrane potential. The validity of the equation and the value of the parameters were revealed by the fact that after the correction, all probes used gave approximately the same value under the same conditions. We expanded the method so as to include the Langmuir adsorption isotherm. When the modified equation is used, the estimated membrane potentials were less dependent on a probe concentration less than 10 μM.     

Estimating the Bioenergetic Cost of a Developing Kiwifruit Berry and its Growth and Maintenance Respiration Components

A response surface was developed by regression analysis to quantifythe seasonal respiratory losses by a kiwifruit [Actinidia deliciosa(A. Chev.) C. F. Liang et A. R. Ferguson var. deliciosa cv.Hayward] berry growing in Fresno, CA. The equation of the surfacewas LNRESP = 1·622 + 0·0697 x TEMP –0·0472x DAY + 0·000165 x DAYSQ, where LNRESP is the naturallogarithm of the respiration rate (nmol CO₂ g d. wt^–1s^–1), TEMP is fruit temperature (°C), DAY is the numberof days after flowering, and DAYSQ is the square of the numberof days after flowering. Respiratory losses for a fruit witha final dry mass of 18·5 g were calculated to be 5·57and 5·92 g glucose per fruit per season in 1985 and 1986,respectively. Maintenance respiration was estimated to be 2·84and 3·19 g glucose per fruit per season for 1985 and1986, respectively. The total calculated bioenergetic cost ofkiwifruit berry growth and respiration was 25·25 and25·60 g glucose per fruit per season for 1985 and 1986,respectively. Respiratory losses, expressed as a proportionof the total carbohydrate required for fruit growth, were significant(mean 22·6%). The cost of fruit growth was estimatedto be very similar for two cooler sites (Davis and Watsonville)but estimates of maintenance respiration based on Fresno fruitrespiration data were unrealistically low for the Watsonvillesite. Actinidia deliciosa (A. Chev.) C. F. Liang et A. R. Ferguson var. deliciosa cv. Hayward, kiwifruit, growth respiration, maintenance respiration, bioenergetic costs, model