Batch kinetics and oxygen consumption ofCandida lipolytica 4-1 onn-alkanes

Detailed batch kinetics ofCandida lipolytica 4-1 onn-hexadecane for varying dispersed phase volume from 0.5 to 5% v/v is presented. All batch growth curves exhibited a linear growth region, indicating a substrate uptake limit. System productivities derived from the linear region were correlated to the dispersed phase volume. The correlation coefficient was identical with that obtained on gas oil. This implies that a correlation coefficient of interfacial area to the dispersed phase volume is identical for both systems. Dissolved oxygen profile and uptake of oxygen from gas phase were also measured. The oxygen uptake rate, volumetric oxygen transfer rate and oxygen demand (requirement) were calculated by means of the balance method. Under limiting dissolved oxygen concentration the maximal oxygen transfer of the fermenter was assessed.     

Uptake of volatilen-alkanes byPseudomonas PG-I

Using a bacterial speciesPseudomonas PG-1, evidence has been obtained which indicates that uptake ofn-pentane ton-octane by microbial cells takes place primarily from the gas phase either directly orvia the aqueous phase. Specific growth rate increased along with the increase in substrate concentration but above the alkane concentration of 0.3% by volume, specific growth rate decreased indicating substrate inhibition of growth. In the case of less volatile alkanes,n-nonane andn-decane, substrate transfer is predominantly through substrate solubilization system elaborated by the cells. EDTA, a strong inhibitor of hydrocarbon solubilization by the cells, inhibited growth on these two alkanes but had negligible effect on growth onn-pentane ton-octane.     

On the theory of blood-tissue exchanges: II. Applications

The application of a previously developed theory of inert gas absorption is here outlined. Interpretation of uptake curves and the method of obtaining tissue constants from such curves is discussed, together with illustrations from actual experiment. A critique of previous analytic procedures is included. The material in this article should be construed only as the personal opinion of the writers and not as representing the opinion of the Navy Department officially.     

Time development of probability distributions for interacting species

A general solution to the dynamical equation for the probability distribution associated withn interacting species is obtained by employing the author's generic canonical expression for the rate functions. Interacting species models with limit-cycle dynamics and no stable equilibrium points feature probability distributions that are asymptotic for large values oft to Dirac δ-distributions concentrated on the limit-cycles, as illustrated here for an analytically solvable two-species model. For ann-species Volterra model, a stationary or temporally-averaged probability distribution should generally be much more complicated than the specialized Poisson form studied by Kerner and others.     

Fluid mixing and oxygen transfer in cell suspensions ofTaxus chinensis in a novel stirred bioreactor

In high-density plant cell cultures, mixing and mass transfer are two key issues, which should be emphasized for process optimization. In this work, both mixing and oxygen transfer characteristics of cell suspensions ofTaxus chinensis were studied in a new centrifugal impeller bioreactor with a working volume of 1.2 L. The mixing time (t _M) and the volumetric oxygen transfer coefficient (K _L a) under different operational conditions were determined in both tap water and cell suspensions of 100–400 g fresh weight/L (i.e., 5.65–23.1 g DW/L). At an aeration rate of 0.1 L/min,t _M decreased from 10.6s at 30 rpm to 2.89 s at 200 rpm under 100 g FW/L, and from 9.63 s (120 rpm) to 4.05 s (300 rpm) under 400 g FW/L. Compared with the effect of agitation, aeration was less significant to the suspension mixing. At a relatively high agitation speed (e.g., 200 rpm),t _M remained almost the same even though aeration rate was changed from 0.1 to 0.4 L/min. Thet _M value increased slowly from 3.98 to 5.26 s at 120 rpm when the cell density was raised from 100 to 250 g FW/L. A rapid increase of botht _M and the suspension viscosity was observed at a cell density above 300 g FW/L. As expected, theK _L a value increased with an increase of aeration rate and agitation speed, but decreased with an increase of cell density. The quantitative data obtained here are useful to investigate the effect of mixing stress on the cell physiology and metabolism ofTaxus chinensis in the bioreactor. This paper is dedicated by JJZ to his colleague Prof. Jun-Tang Yu on the occasion of his 70 birthday.     

The physiological factors which govern inert gas exchange

The decay constants (k _j ) of the equation of inert gas exchanges are the roots of an algebraic equation of degreen+1, wheren is the number of distinct absorbing tissues. The coefficients of this equation can be obtained numerically by certain independent experiments to measure the tissue parameters. Graphical solution of this equation yields theoretical values of thek _j . Combining these constants with the numerical values for the partial derivatives of thek _j then gives the per cent rate of change of thek _j as any one tissue parameter varies by a given fraction of its normal range. A numerical example of these calculations shows good conformity with experiment, and permits a quantitative estimate of variations in the speed of gas exchange from a knowledge of changes in the physiological state. The opinions expressed in this article are the private ones of the writers, and are not to be construed as reflecting the policies of the Navy Department or the Naval Service at large.     

Physiological characteristics of inorganic nitrogen uptake by spatially separate algal communities in a nitrogendeficient lake

1. The physiological characteristics of nitrogen uptake by sublittoral and eulittoral (splash zone) epilithic periphyton as well as epipelic periphyton in N-deficient Castle Lake, California were determined by evaluating the half-saturation constants (K_t) and initial slopes (V_max/K_t) of uptake kinetics curves. These results were compared to similar studies of phytoplankton nitrogen uptake in this lake. 2. The strategies of nitrogen uptake differed among the various communities and were largely determined by the proximity of each to pools of available dissolved inorganic N (DIN). 3. The sublittoral algae did not have a high biological affinity for either NH₄ or NO₃ and depended on nitrogen fixation for their N supply. The eulittoral community showed an increased capacity for DIN uptake at low substrate concentration, but not as high as measured for the phytoplankton community. Epipelic algae live immediately adjacent to a large pool of interstitial sediment NH₄ and showed no physiological adaptations for surviving in a N-deficient environment. 4. K_t values for all benthic communities were approximately two orders of magnitude greater than ambient substrate levels. In contrast, the half-saturation constants for NH₄+ NO₃ uptake by phytoplankton were very similar to in situ levels of these nutrients.     

Regeneration of resistance and ion transport in rabbit corneal epithelium after induced surface cell exfoliation

Summary Exposure of the rabbit corneal surface to a 20-m digitonin-0.9% NaCl solution leads to permeabilization of the most superficial cells of the stratified epithelium. The devitalized cells exfoliate spontaneously from the corneal surface. Detergent exposure limited to 4–8 min leads to permeabilization and rapid exfoliation of a monolayer of surface cells. Consistent with the presence of the epithelial paracellular permeability barrier in this cell layer, their permeabilization results in complete loss of transepithelial resistance (R _t ). Within minutes after detergent removal an initial recovery ofR _t can be noticed indicating generation of a new paracellular permeability barrier by the viable subsurface cells. This recovery proceeds rapidly andR _t reaches within 70 min a maximum equal to > 90% of the preexfoliation values (=2.43 k·cm²,n=22). TheR _t recovery is fully blocked in a reversible manner by 10 m dihydrocytochalasin B. The recovery is not affected by inhibition of protein synthesis with 5 m cycloheximide. When the ocular surface is treated again with digitonin the permeabilization and exfoliation of a monolayer of cells and loss ofR _t are repeated. After the second detergent exposure an initial recovery ofR _t occurs as before within minutes. However, the pace ofR _t recovery is much slower: 4–5 hr are required to reach a stable maximalR _t values amounting to about 73% of initial control. This recovery can be fully blocked by 5 m cycloheximide indicating that protein synthesis is required for generation of tight junctions by the second subcellular layer. With only a fraction ofR _t recovered, short-circuit currents amounting to, at least, 50% of control values and attributable in part to cell-to-tear movement of Cl^– through the apical surface can be measured. This suggests that apical-type Cl^– channels are either present in the apically facing membrane of subsurface cells or that they are rapidly inserted in it from preexisting intracellular pools immediately following the devitalization of the surface cells by digitonin.     

Biochemical and Pharmacological Characterization of [3H]GBR 12935 Binding In Vitro to Rat Striatal Membranes: Labeling of the Dopamine Uptake Complex

Abstract: Binding of the selective dopamine (DA) uptake inhibitor [³H]GBR 12935 to rat striatal membranes was characterized biochemically and pharmacologically. [³H]-GBR 12935 binding at 0°C was reversible and saturable and Scatchard analysis indicated a single binding site with a K_D of 5.5 nM and a B_max of 760 pmol/mg tissue. [³H]GBR 12935 labeled two binding sites. One binding site was identified as the classic DA uptake site, since methylphenidate, cocaine, diclofensine, and Lu 19–005 potently inhibited [³H]GBR 12935 binding to it. Binding to the second site was inhibited by high concentrations of the above compounds. IC₅₀ values for inhibition of [³H]GBR 12935 binding to the DA uptake site were proportional to IC50 values for inhibition of DA uptake. However, substrates of DA uptake, e.g., DA and 1-methyl-4-phenylpyridine, and DA releasers, e.g., the amphetamines, inhibited [³H]GBR 12935 binding less than DA uptake. Rate experiments excluded the possibility that these “weak” inhibitors affected the binding by alloste-ric coupled binding sites. The second binding site was not a noradrenergic, serotonergic, or GABAergic uptake site. Neither was it a dopaminergic, acetylcholinergic, histaminic, serotonergic, or adrenergic receptor. However, [³H]GBR 12935 was potently displaced from it by disubstituted piper-azine derivatives, i.e., flupentixol and piflutixol. DA uptake and the DA uptake binding site of [³H]GBR 12935 were located primarily in the striatum, but the piperazine acceptor site was distributed uniformly throughout the brain. Also only the DA uptake binding site was destroyed by 6-OH-DA. Thus, [³H]GBR 12935 labels the classic DA uptake site in rat striatum and also a piperazine acceptor site. Substrates for DA uptake and releasers of DA inhibited [³H]GBR 12935 binding with low potency, but did not alter the rate constants for [³H]GBR 12935 binding. Therefore inhibitors of DA uptake label the carrier site and prevent the carrier process.     

On the average cellular volume in synchronized cell populations

As was done by Sinclair and Ross (1969(, we consider a cellular population that consists initially (at time zero) ofN ₀ newborn cells, all with the same volumev _o. It is assumed that the occurrence of cell division is determined only by a cell’s age, and not by its volume. The frequency function of interdivision times, τ, is denoted byf(τ). If cell death is negligible, the expected number of cells,N(t), will increase according to the laws of a simple age-dependent branching process. The expression forN(t) is obtained as a sum over all generations; thevth term of this sum, in turn, is a multiple convolution integral, reflecting the life history ofvth generation cells (i.e., the lengths of thev successive interdivision periods plus the age of the cell at timet). Assuming that cell volume is a given function of cell age, e.g., linear or exponential, and that cellular volume is exactly halved at each division, it is possible to calculate the volume of a cell with a given life history, and thus the average cellular volume of the whole population as a function of time. If at time zero the volumes differ from cell to cell, the final equation must be modified by averaging over initial volumes. In the case of linear volume increase with age, a very simple asymptotic expression is found for the average cellular volume ast→∞. The case of exponential volume increase with age also leads to a simple asymptotic formula, but the resulting volume distribution is unstable. The mean cellular volume at birth and the second moment of the volume distribution can be calculated in a similar manner. Work supported by the U.S. Atomic Energy Commission.     

Respiration and nitrogenase activity in nodules ofCasuarina cunninghamiana and cultures ofFrankia sp. HFP020203: Effects of temperature and partial pressure of O2

The effects of time after exposure to acetylene and of nodule excision were examined using a flow-through system. After a transient depression in the rate of acetylene reduction that began about 1.5 min after exposure to acetylene, the rate recovered to 98% of the initial maximum value after 40 min. After nodule excision the rate stabilized to 90% of the initial maximum value observed in the intact plant.Excised nodules, measured at 6-min intervals in a closed system, with frequent changes of the gas mixture, were used for the remaining experiments. Acetylene reduction by the nodules increased rapidly as temperature was increased between 6 and 26°C. Between 26 and 36°C there was relatively little effect of temperature on acetylene reduction.Nodules and cultures ofFrankia were compared with respect to the effect of temperature and pO₂ (partial pressure of oxygen) on oxygen uptake. Cultures ofFrankia were grown on a nitrogen-free medium at either 0.3 kPa O₂ (vesicles absent) or 20 kPa O₂ (vesicles present). Oxygen uptake by nodules (vesicles absent) and by vesicle-containing cultures was strongly dependent on pO₂ at values below 20 kPa. This suggests the presence of a barrier to oxygen diffusion. Oxygen uptake was dependent on temperature as well as on pO₂, but the Q₁₀ was much larger for the cultures than for the nodules. This suggests that vesicles or related structures are not the source of the diffusion barrier in Casuarina nodules. Respiration by cultures ofFrankia lacking vesicles became O₂-saturated at low pO₂ values. Thus these cultures did not have a significant diffusion barrier. From these results it is concluded that nodules ofCasuarina cunninghamiana have a barrier to oxygen diffusion supplied by the host tissue and not byFrankia.     

Intestinal absorption of amino acids by rainbow trout,Salmo gairdneri (Richardson)

Summary A technique for the in vitro maintenance of isolated portions of rainbow trout intestine is described. Uptake of¹⁴C-L-leucine occurs by an active mechanism which is stereospecific, sodium-dependent and susceptible to inhibition by other neutral amino acids.K _t for leucine uptake is 2.72 mM with aV _max of 19.61 moles/g ethanol extracted dry wt.·10 min. L-valine and L-methionine are competitive inhibitors of L-leucine uptake withK _i values of 24.30 mM and 2.56 mM, respectively. Evidence suggests that at least two uptake sites for the transport of neutral amino acids are present in the intestine of this species.     

Determination of equilibrium constants and maximum binding capacities in complexIn vitro systems: I. The mammillary system

It is shown that in a system containingn types of mutually noninteracting binding sites, the association constants are then roots of annth order polynomial while the maximum binding capacities can be evaluated by solving a set ofn simultaneous linear equations. Thenth order polynomial and the system ofn linear equations are defined in terms of 2n intermediate coefficients, the coefficients being themselves evaluated by substituting 2n sets of appropriate experimental data into an auxiliary system of 2n linear equations. The existence and uniqueness of the solutions are established.     

Theoretical evaluation of safety of dives completed by non-stop ascent to surface

Our model of decompression sickness determines the cumulative probability of developing symptoms of this illness by the exponential equation whose index is the integral cumulative risk function of all body tissue lesions by bubbles, F _cum(t) = ΣF _n(t). The underwater dives may be considered as practically safe in the context of this model when the function F _cum(t) during its growth will not exceed some small value F _cum-max = ΣF _n-max. Using hypothetical values of parameters of tissues and functions F _n(t), we calculated the curves depth-duration for practically safe non-stop dives with air and with mixtures of oxygen with helium, neon and argon. Doing so, we obtained the distributions of values F _n-max in regard to the values of inert gas washout half-times from tissues which show that the tissues experienced the largest risks of bubble lesions are different for dives with different duration. The comparison of the indicated curves shows that the short-term dives with air are less dangerous and the long-term dives are more dangerous than the dives with helium-oxygen mixture. At the same time, the least risk of bubble lesions of tissues arises at dives with neon-oxygen mixture and the greatest risk arises at dives with argon-oxygen mixture.     

Toxicity and accumulation of thallium in bacteria and yeast

Thallium sulphate inhibited microbial growth, withBacillus megaterium KM, more sensitive to the metal thanSaccharomyces cerevisiae andEscherichia coli. Inhibition ofB. megaterium KM andS. cerevisiae, but not ofE. coli, was alleviated by increasing the potassium concentration of the medium; inhibition of respiration ofS. cerevisiae, but not ofE. coli, was similarly alleviated. Thallium was rapidly bound, presumably to cell surfaces, byS. cerevisiae andE. coli, and was progressively accumulated by energy-dependent transport systems (probably concerned primarily with potassium uptake) with both organisms. Thallium uptake kinetics suggested more than one transport system operated in yeast, possibly reflecting a multiplicity of potassium transport systems. ApparentK _m andK _i values for competitive inhibition of thallium uptake by potassium indicatedS. cerevisiae to have a higher affinity for thallium uptake than for potassium, whileE. coli had a transport system with a higher affinity for potassium than for thallium. The likely systems for thallium transport are discussed. A mutant ofE. coli with tenfold decreased sensitivity to thallium was isolated and apparently effected surface binding of thallium in amounts equivalent to the wild type organism, but showed no subsequent uptake and accumulation of the metal from buffer, even though it was able to accumulate potassium to normal intracellular concentrations during growth. Abbreviations: Metal are referred to by their recognised atomic symbols (e.g. TI = Thallium; K = potassium; Co = cobalt)     

Myocardial uptake of thiopental enantiomers by the isolated perfused rat heart

Myocardial uptake of thiopental enantiomers by an isolated perfused rat heart preparation was examined after perfusion with protein-free perfusate. Outflow perfusate samples were collected at frequent intervals for 20 min during single-pass perfusion with 10 μg/ml racemic thiopental (washin phase) and for another 45 min during perfusion with drug-free perfusate (washout phase). (+)- and (−)-thiopental concentrations were assayed by chiral high-performance liquid chromatography. Heart rate, perfusion pressure, and electrocardiogram were also monitored. During the washin phase, there was no significant difference between the mean values of the equilibration rate constants of (+)- and (−)-thiopental enantiomers (0.44 ± 0.07 min⁻¹ and 0.43 ± 0.09 min⁻¹, respectively, P > 0.05). Mean volumes of distribution of (+)- and (−)-thiopental enantiomers were similar (6.34 ± 1.20 and 6.45 ± 1.29 ml/g for the washin phase and 7.22 ± 0.71 and 7.47 ± 0.81 ml/g for the washout phase, respectively, P > 0.05). This indicates that tissue accumulation of thiopental enantiomers in the isolated perfused rat heart was not stereoselective. Uptake of thiopental by the heart was perfusion flow rate-limited and independent of capillary permeability. These findings suggest that myocardial tissue concentration of racemic thiopental should be an accurate predictor of myocardial drug effect. © 1996 Wiley-Liss, Inc.     

Inhibition by nisin of K+ uptake in a sensitivePediococcus sp. strain

Summary A nisin-sensitive strain ofPediococcus sp possessed an uptake system for K⁺ which was apparently dependent on metabolic energy and ATPase activity. K⁺ uptake rate was dependent on the glucose and K⁺ concentrations and showed approximately Michaelis-Menten kinetics with respect to both of these variables with K_t values of 1.2 mM and 599 μM respectively. The presence of nisin inhibited K⁺ uptake with the percentage inhibition proportional to the nisin activity,. Total inhibition occurred at between 4.5 and 5.0 IU ml⁻¹ and the MIC was approximately 0.6 IU ml⁻¹.     

The convergence in distribution of some simple epidemics

A group of n susceptible individuals exposed to a contagious disease isconsidered. It is assumed that at each point in time one or more susceptible individuals can contract the disease. The progress of this simple batch epidemic is modeled by a stochastic process X_n(t), t[0, ∞), representing the number of infectiveindividuals at time t. In this paper our analysis is restricted to simple batch epidemics with transition rates given by [α²X_n(t){n −X_n(t) +X_n(0)}]^1/2, t[0, ∞), α(0, ∞). This class of simple batch epidemics generalizes a model used and motivated by McNeil (1972) to describe simple epidemic situations. It is shown for this class of simple batch epidemics, that X_n(t), with suitable standardization, converges in distribution as n→∞ to a normal random variable for all t(0, t₀), and t₀ is evaluated.     

Osmotic regulation ofmyo-inositol uptake in primary astrocyte cultures

Uptake ofmyo-inositol by astrocytes in hypertonic medium (440 mosm/kg H₂O) was increased near 3-fold after incubation for 24 hours, which continued for 72 hours, as compared with the uptake by cells cultured in isotonic medium (38 nmoles/mg protein).myo-Inositol uptake by astrocytes cultured in hypotonic medium (180 mosm/kg H₂O) for periods up to 72 hours was reduced by 74% to 8 to 10 nmoles/mg protein. Astrocytes incubated in either hypotonic or hypertonic medium for 24 hours and then placed in isotonic medium reversed the initial down- or up-regulation of uptake. Activation of chronic RVD and RVI correlates with regulation ofmyo-inositol uptake. A 30 to 40 mosm/kg H₂O deviation from physiological osmolality can influencemyo-inositol homeostasis. The intracellular content ofmyo-inositol in astrocytes in isotonic medium was 25.6 ± 1.3 g/mg protein (28 mM). This level ofmyo-inositol is sufficient for this compound to function as an osmoregulator in primary astrocytes and it is likely to contribute to the maintenance of brain volume.     

A function for describing nitrogen uptake,dry matter production and rooting by wheat crops

The quantities,Y, of nitrogen taken up, and dry matter produced, at various times during the growth of six winter wheat crops at Rothamsted were shown to be related to thermal time,x, based on soil temperature, by a simple equation $$Y = (A^{ - 1/n} + \exp ( - kx))^{ - n} $$ whereA is the ultimate maximum ofY, n a shape factor andk a rate constant that is related toA andn throughx′, the inflexion point of the function. The value ofn was 1.5 for both N uptake and dry matter. The value ofA for N uptake,A _N , was well described by a multiple regression on sowing data,t _s , expressed as the number of days after August 31st, and rainfall,R _Apr , in the April before harvest, but no such regression could be found for the value ofA for dry matter,A _D . The rate constants,k _N andk _D , for N-uptake and dry-matter production respectively, could be related to the date of sowing and the weather through the corresponding inflexion points,x′ _N andx′ _D . Highly significant regressions were found, forx′ _N on the time,t _sf , between sowing and the return of the soil to field capacity and forx′ _D on the reciprocal oft _s . The function was used to generate N uptake curves from values ofA _N andk _N (obtained fromx′ _N ) given by inserting the appropriate values ofR _Apr ,t _s andt _sf in the regressions. These fitted measured N uptakes satisfactorily for the six crops used to obtain the regressions, and four grown subsequently, at Rothamsted, and also for six crops at Woburn. Values ofA _D had to be set arbitrarily because no regression had been found to predict them, but using these arbitrary values in the function gave dry matter curves that fitted the measurements satisfactorily for all ten Rothamsted crops and two of the Woburn crops. Tests for seasonal and site effects showed thatA _N was influenced more by differences between the two sites than by seasonal differences other than those inR _Apr , whereasA _D was strongly influenced by these seasonal differencess and very little by those between the sites.