Role of mitochondria in ethanol tolerance of Saccharomyces cerevisiae

The presence of active mitochondria and oxidative metabolism is shown to be essential to maintain low inhibition levels by ethanol of the growth rate (), fermentation rate (v) or respiration rate () of Saccharomyces cerevisiae wild type strain S288C. Cells which have respiratory metabolism show K _i (ethanol inhibition constant) values for , v and , higher (K _i>1 M) than those of petite mutants or grande strains grown in anaerobiosis (K _i=0.7 M). In addition, the relationship between or v and ethanol concentration is linear in cells with respiratory metabolism and exponential in cells lacking respiration. When functional mitochondria are transferred to petite mutants, the resulting strain shows K _i values similar to those of the grande strain and the inhibition of and v by increasing ethanol concentrations becomes linear.     

On the appropriateness of use of a continuous formulation for the modelling of discrete multireactant systems following Michaëlis–Menten kinetics

The possibility of solving the mass balances to a multiplicity of substrates within a CSTR in the presence of a chemical reaction following Michaelis-Menten kinetics using the assumption that the discrete distribution of said substrates is well approximated by an equivalent continuous distribution on the molecular weight is explored. The applicability of such reasoning is tested with a convenient numerical example. In addition to providing the limiting behavior of the discrete formulation as the number of homologous substrates increases, the continuous formulation yields in general simpler functional forms for the final distribution of substrates than the discrete counterpart due to the recursive nature of the solution in the latter case.List of Symbols C{N. M} mol/m³ concentration of substrate containing N monomer residues each with molecular weight M - {N, M} normalized value of C{N. M} - C {M} mol/m³ da concentration of substrate of molecular weight M - _in normalized value of C {M} at the i-th iteration of a finite difference method - {M} normalized value of C {M} - C ₀{N.M} mol/m³ inlet concentration of substrate containing N monomer residues each with molecular weight M - {N ·M} normalized value of C₀{N. M} - _{0 i} normalized value of C ₀ {M} at the i-th iteration of a finite difference method - C ₀ {M} mol/m³ da initial concentration of substrate of molecular weight M - C _tot mol/m³ (constant) overall concentration of substrates (discrete model) - C _tot mol/m³ (constant) overall concentration of substrates (continuous model) - D deviation of the continuous approach relative to the discrete approach - i dummy integer variable - I arbitrary integration constant - j dummy integer variable - k dummy integer variable - K _m mol/m³ Michaëlis-Menten constant for the substrates - l dummy integer variable - M da molecular weight of substrate - M normalized value of M - M da maximum molecular weight of a reacting substrate - N number of monomer residues of a reacting substrate - N maximum number of monomer residues of a reacting substrate - N total number of increments for the finite difference method - Q m³/s volumetric flow rate of liquid through the reactor - S inert product molecule - S _i substrate containing i monomer residues - V m³ volume of the reactor - v _max mol/m³ s reaction rate under saturating conditions of the enzyme active site with substrate - v _max{N. M} mol/m³ s reaction rate under saturating conditions of the enzyme active site with substrate containing N monomer residues with molecular weight M - _max{N · M} dimensionless value of v_max{N. M} (discrete model) - _max{M} dimensionless value of v _max {M} (continuous model) - mol/m³ s molecular weight-averaged value of v_max (discrete model) - mol.da/m³s molecular weight-averaged value of v_max (continuous model) - v _max {M} mol.da/m³s reaction rate under saturating conditions of the enzyme active site with substrate with molecular weight M - _max {M} dimensionless value of v_max{M} - _{max, (i)} dimensionless value of v_max{M} at the i-th iteration of a finite difference method - v _max mol/m³ s reference constant value of v _max Greek Symbols dimensionless operating parameter (discrete distribution) - dimensionless operating parameter (continuous distribution) - M da (average) molecular weight of a monomeric subunit - M selected increment for the finite difference method - auxiliary corrective factor (discrete model)     

Anaerobic digestion of palm oil mill effluent and condensation water waste: an overall kinetic model for methane production and substrate utilization

The process of anaerobic digestion is viewed as a series of reactions which can be described kinetically both in terms of substrate utilization and methane production. It is considered that the rate limiting factor in the digestion of complex wastewaters is hydrolysis and this cannot be adequately described using a Monod equation. In contrast readily assimilable wastewaters conform well to this approach. A generalized equation has thus been derived, based on both the Monod and Contois equations, which serves extreme cases. The model was verified experimentally using continuous feed anaerobic digesters treating palm oil mill effluent (POME) and condensation water from a thermal concentration process. POME represents a complex substrate comprising of unhydrolyzed materials whereas the condensation water is predominantly short chain volatile fatty acids. Substrate removal and methane production in both cases could be predicted accurately using the generalized equation presented.List of Symbols A (=K_skY/K_h) Kinetic parameter - B Specific methane yield, 1 of CH₄/g of substrate added B₀ Maximum specific methane yield, 1 of CH₄/g of substrate added at infinity - C Empirical constant in Contois equation - F Volumetric substrate removal rate, g/l day - k Hydrolysed substrate transport rate coefficient, 1/days - K (=YC) Kinetic parameter in Chen-Hashimoto equation - K _h Substrate hydrolysis rate coefficient, 1/days - K _s Half-saturation constant for hydrolysed substrate, g/l - M _v Volumetric methane production rate, 1 of CH₄/l day - MS Mineral solids, g/l - MSS Mineral suspended soilds, g/l - POME Palm oil mill effluent - R (=S_r/S_T0) Refractory coefficient - S _h Concentration of hydrolysed substrate, g/l - S _u Intracellular concentration of hydrolysed substrate, g/l - S ₀ Input biodegradable substrate concentration, g/l - S Biodegradable substrate concentration in the effluent or in the digester, g/l - S _r Refractory feed substrate concentration, g/l - S _T0 (=S₀+S_r) Total feed substrate concentration, g/l - S _T (S+S_r) Total substrate concentration in the effluent, g/l - TS Total solids, g/l - TSS Total suspended solids, g/l - VFA Total volatile fatty acids, g/l - VS Volatile solids, g/l - VSS Volatile suspended solids, g/l - X Biomass concentration, g/l - Y Biomass yield coefficient, biomass/substrate mass - Hydraulic retention time, days. - Specific growth rate of microorganisms, l/days - _m Maximum specific growth rate of microorganisms, l/days The authors wish to express their gratitude to the Departamento de Postgrado y Especialización del CSIC and to the Consejería de Educación y Ciencia de la Junta de Andalucia for their financial support of this work.     

Effect of time delay in specific growth rate on the periodic operation of bioreactors with input multiplicities

The effect of time delay in specific growth rate () on the periodic operation of bioreactors with input multiplicities is theoretically analyzed for productivity improvement. A periodic rectangular pulse is applied either in feed substrate concentration (S_f) or in dilution rate (D). Periodic operation under feed substrate concentration cycling gives improvement in productivity at lower value of ¯S_f of the two steady-state multiplicities of S_f only when the time delay in is larger. Whereas the larger value of ¯S_f gives improvement in average productivity for all values of time delay. Dilution rate (D) cycling gives an improvement in average productivity particularly for larger time delay in . This improvement in average productivity is obtained only at smaller value of dilution rate out of the two steady-state input multiplicities of D.List of Symbols D 1/h dilution rate - F memory function - g dummy variable - K_i g/l substrate inhibition constant - K_m g/l substrate saturation constant - P g/l product concentration - P_m g/l product saturation constant - Q g/(hl) product cell produced per unit time - S g/l substrate concentration - S_f g/l feed substrate concentration - S_f,p g/l feed substrate concentration during fraction of a period - X g/l biomass concentration - Y_X/S g/g cell mass yield - w variable either S or Z - Z g/l weighted average of substrate concentration Greek Letters 1/h time delay parameter - ₁ , ₂ product yield parameters, g/g and 1/h - pulse width expressed as a fraction of a period - 1/h specific growth rate - _m 1/h maximum specific growth rate - h period of oscillation - – average value     

Der hemmende Einfluß gleichzeitig bewegter Beuteattrappen auf das Beutefangverhalten der Erdkröte (Bufo bufo L.)

Zusammenfassung Durch simultane visuelle Bewegungsreize, die von mehreren Beutetieren (z.B. Mehlkäferlarven) ausgehen, wird der Beutefang der Erdkröte (Bufo bufo L.) gehemmt. An diese Verhaltensheobaehtung anknüpfend, wurde die Abhängigkeit dieses inhibitorischen Umfeldeffektes von verschiedenen visuellen Reizparametern im Attrappen versuch quantitativ gemessen: Im frontalen Gesichtsfeld der Kröte rotierte vor dunklem Hintergrund eine weiße, rechteckige, 2,5 × 20° große Beuteattrappe (Zentralattrappe, z) mit dem Rechteckzentrum im Drehpunkt. Zusätzlich konnten mehrere Kreisscheiben von 5 bzw. 10° (Peripherattrappen, p) um die Zentralattrappe bewegt werden.Die Beutefangaktivität R _z [Beutefangreaktionen x min^–1] auf die allein gebotene Zentralattrappe war bei einer Sehwinkelgeschwindigkeit v _s der distalen Attrappenkanten 10v _s30 [grad x see^–1] maximal und sank für kleinere oder größere Winkelgeschwindigkeiten wieder ab. Eine mit v _s=25 [grad x sec^–1] allein bewegte Peripherattrappe löste maximale Beutefangaktivität R _p aus. Mit zunehmender Anzahl n _p simultan bewegter Peripherattrappen sank die Beutefangaktivität ab.Mehrere, um den gleichen Drehpunkt bewegte Peripherattrappen, deren Abstand untereinander =10° betrug, blieben von der Kröte unbeantwortet. Sie bildeten ein inhibitorisches Umfeld und hemmten dadurch die Reaktion auf die gleichzeitig bewegte Zentralattrappe z. — Die im Simultanreizungsversuch gemessene Beutefangaktivität R _zp war abhängig vom Abstand [grad] zwischen Zentralattrappe und Peripherattrappen ( : Kürzester Abstand zwischen z und p): Für =10° war R _zp0 und stieg für >10° an. — Kontrollversuche, die jeweils auf die Simultanreizung mit z allein folgten (R _z), ließen eine -abhängige Nachhemmung erkennen. — Die hemmende Wirkung auf die Beantwortung von z war auch von der Sehwinkelgeschwindigkeit v _s der Peripherattrappen abhängig; sie war bei derjenigen Sehwinkelgeschwindigkeit (v _s=25 [grad × sec^–1]) maximal, mit der die Zentralattrappe, allein geboten, maximale Beutefangaktivität auslöste; für v _sg25 [grad × sec^–1] nahm die Hemmung wieder ab.Die Versuchsergebnisse lassen auf inhibitorische Verknüpfungen innerhalb des zentralen visuellen Systems schließen. Es wird vermutet, daß die Reiz-Verhaltens-reaktionsbeziehungen in den visuellen Simultanreizungsversuchen durch eine Art zentrale laterale Inhibition bestimmt werden.

---

Inhibitory effect of simultaneously moved prey dummies on the prey catching behaviour of the common toad (Bufo bufo L.)

Summary The prey catching behaviour of the toad (Bufo bufo L.) is generally inhibited by simultaneously visual moving stimuli caused by a group of prey animals (mealworms). According to this behavioural observation the dependence of this inhibitory effect on several visual parameters were quantitatively measured in dummy experiments: in the frontal visual field of the toad a white rectangular prey dummy of 2,5×20° (central dummy, z) was rotating in a centre against dark background. In addition several disks of 5 or 10° diameter (peripheral dummies, p) could simultaneously rotate around the central dummy (Figs. 1 and 7).The prey catching activity R _z [catching reactions x min^–1] released by rotation of only the central dummy z increased with increasing angular velocity v _s of the stimulus distal edges, reaching a maximum for 10v_s30 [degrees x sec^–1] and decreasing for v _s>30 [degrees x sec^–1] (Fig. 5).A single peripheral dummy p, moved at v _s=25 [degrees x sec^–1], released maximal catching activity R _p. The activity R _p decreased with the increasing number n _p of simultaneously offered dummies (Fig. 6).The prey catching behaviour of the toad was inhibited, when several peripheral dummies p were moved around the centre with a distance =10° from each other. They caused an inhibitory field and they also inhibited the response to a simultaneously moved central dummy z. The prey catching activity, measured in experiments in which z and p rotated simultaneously, depends on the distance [degrees] between z and p ( being the shortest distance between z and p). For =10°, R _zp was zero; R _zp increased for >10° (Figs. 9 and 10). — Control experiments carried out with z allone — after having applied the simultaneous stimulation — showed a - dependent after-inhibition (Fig. 9). — The inhibitory effect on the response to z also depended on the angular velocity v _s of p; the inhibition was at a maximum for v _s25 [degrees x sec^–1], and it decreased for v _s25 [degrees x sec^–1] (Fig. 11).The experimental results suggest inhibitory interactions within the central visual system. It is supposed that the relation between stimulus and behavioural reaction in simultaneous stimulating experiments results from some kind of central nervous lateral inhibition.

---

---

Mit Unterstützung der Deutschen Forschungsgemeinschaft (Ew 7/4+5).     

Substrate Preferences of Juvenile Hatchery-reared Lake Sturgeon, Acipenser fulvescens

Substrate preferences of juvenile, hatchery-reared lake sturgeon, Acipenser fulvescens, from a Wisconsin population were examined relative to water temperature and velocity, fish size and time of day. Given the choice of a sand, gravel, rock or smooth plastic bottom, all sturgeon were strongly attracted to the sand substrate. Water temperature did not affect substrate preference, however small sturgeon (13.5cm mean FL) acclimated to 19°C were more active than those tested in 6°C water. Small sturgeon, under all conditions, were less sedentary in the early morning and late evening hours than during the daytime. Preference of small sturgeon for sand was slightly stronger when a current of 5cmsec^-1 was present. Substrate preference of larger sturgeon (24.3cm mean FL) was not affected by any of the parameters examined. In general, hatchery-reared lake sturgeon showed similar behaviors (with respect to substrate selection) as those previously described for the same population in the wild, making it possible that substrate preferences have a genetic component and likely that hatchery rearing does not alter instinctive behavior.     

Holling's “hungry mantid” model for the invertebrate functional response considered as a Markov process. Part II: Negligible handling time

In this paper we analyse a stochastic model for invertebrate predation taking account of the predator's satiation. This model approximates Holling's hungry mantid model when handling time is negligible (see Part I). For this model we derive equations from which we can calculate the functional response and the variance of the total catch. Moreover we study a number of approximations which can be used to calculate these quantities in practical cases in a relatively simple manner.List of Notation a rate constant of digestion - b maximum of rate constant of prey encounter in the mantid - c satiation threshold for search - c satiation threshold for pursuit in the mantid - c _i (w^1/2(N- N)ⁱ) - expectation operator - f rate of change of satiation during search - F functional response: mean number of prey eaten per unit of time - g rate constant of prey capture - h probability generating function of N conditional on S = s times p - H probability generating function of N - m_i ¹ - n, N number of prey caught - p probability density of S - p_n simultaneous probability (density) of N and S - q probability of strike success - r dummy variable in generating function - s, S satiation - T _s search time - T _d digestion time - v asymptotic rate of increase of var v - V asymptotic rate of increase of var N - w weight of edible part of prey - W standard Wiener process - x prey density - z (N{S = s}-N)p - rate constant of prey escape time maximum pursuit time - (v{S = + w ^1/2}-v) - present time as a fraction of the time from the start to the end of the experiment - hazard rate of T _s - mean time between (downward) passages of S through c - v w_–1/2(N-) - edible prey biomass density - probability density of , number pi - parameter of Weibull distribution of T _s = (1/2acx(-g(c)))_1/2 - w_–1/2(S -) - satiation in the guzzler approximation: solution to d/dt = f() + g(), (0)=S(0). - biomass functional response: wF - total biomass catch in the guzzler approximation: solution to d/dt = g(), (0) = 0     

Flight of the honey bee VI: energetics of wind tunnel exhaustion flights at defined fuel content,speed adaptation and aerodynamics

To gain information on extended flight energetics, quasi-natural flight conditions imitating steady horizontal flight were set by combining the tetheredflight wind-tunnel method with the exhaustion-flight method. The bees were suspended from a two-component aerodynamic balance at different, near optimum body angle of attack and were allowed to choose their own speed: their body mass and body weight was determined before and after a flight; their speed, lift, wingbeat frequency and total flight time were measured throughout a flight. These values were used to determine thrust, resultant aerodynamic force (magnitude and tilting angle), Reynolds number, total flight distance and total flight impulse. Flights in which lift was body weight were mostly obtained. Bees, flown to complete exhausion, were refed with 5, 10, 15 or 20 l of a 1.28-mol·l^-1 glucose solution (energy content w=18.5, 37.0, 55.5 or 74.0 J) and again flown to complete exhaustion at an ambient temperature of 25±1.5°C by a flight of known duration such that the calculation of absolute and relative metabolic power was possible. Mean body mass after exhaustion was 76.49±3.52 mg. During long term flights of 7.47–31.30 min similar changes in flight velocity, lift, thrust, aerodynamic force, wingbeat frequency and tilting angle took place, independent of the volume of feeding solution. After increasing rapidly within 15 s a more or less steady phase of 60–80% of total flight time, showing only a slight decrease, was followed by a steeper, more irregular decrease, finally reaching 0 within 20–30 s. In steady phases lift was nearly equal to resultant aerodynamic force; tilting angle was 79.8±4.0°, thrust to lift radio did not vary, thrust was 18.0±7.4% of lift, lift was somewhat higher/equal/lower than body mass in 61.3%, 16.1%, 22.6% of all totally analysable flights (n=31). The following parameters were varied as functions of volume of feeding solution (5–20 l in steps of 5 l) and energy content. (18.5–74.0 J in steps of 18.5 J): total flight time, velocity, total flight distance, mean lift, thrust, mean resultant aerodynamic force, tilting angle, total flight impulse, wingbeat frequency, metabolic power and metabolic power related to body mass, the latter related to empty, full and mean (=100 mg) body mass. The following positive correlations were found: L=1.069·10^-9 f ^2.538; R=1.629·10^-9 f ^2.464; P _m=7.079·10^-8 f ^2.456; P _m=0.008v+0.008; P _m=18.996L+0.022; P _m=19.782R+0.021; P _m=82.143T+0.028; P _m=1.245·bm _f ^1.424 ; P _{mrel e}=6.471·bm _f ^1.040 ; =83.248+0.385. The following negative correlations were found: V=3.939–0.032; T=1.324·10^-4–0.038·10^-4. Statistically significant correlations were not found in T(f), L(), R(), f(), P _m(bm _e), P _{m rel e}(bm _e), P _{m rel f}(bm _e), P _{m rel f}(bm _f).Abbreviations A(m²) frontal area - bl(m) body length - bm(mg) body mass - c(mol·1^-1) glucose concentration of feeding solution - c _D (dimensionless) drag coefficient, related to A - D(N) drag - F _w(N) body weight - F _wp weight of paper fragment lost at flight start - f wingbeat frequency (s^-1) - g(=9.81 m·s^-2) gravitational acceleration - I(Ns)=R(t) dt total impulse of a flight - L(N) lift vertical sustaining force component - P _m(J·s^-1=W) metabolic power - P_{m ret} (W·g^-1) metabolic power, related to body mass - R(N) resultant aerodynamic force - Re v·bl·v ^-1 (dimensionless) Reynolds number, related to body length - s(m) v(t) dt virtual flight distance of a flight - s(km) total virtual flight distance - T (N) thrust horizontal force component of horizontal flight - T _a (°C) ambient temperature - t(s) time - t _tot (s or min) total flight time - v(m·s^-1) flight velocity - v(l) volume of feeding solution - W (J) energy and energy content of V - ( °) body angle of attack between body longitudinal axis and flow direction - ( °) tilting angle ( 90°) between R and the horizont in horizontal flight v(=1.53·10^-5m²·s^-1 for air at 25°) kinematic viscosity - (=1.2 kg·m^-3 at 25°C) air density     

Theoretical studies on the necessary number of components in mixtures

Summary Theoretical studies on the optimal numbers of components in mixtures (for example multiclonal varieties or mixtures of lines) have been performed according to phenotypic yield stability (measured by the parameter variance). For each component i, i = 1, 2,..., n, a parameter u_i with 0 u_i 1 has been introduced reflecting the different survival and yielding ability of the components. For the stochastic analysis the mean of each u_i is denoted by u ₁ and its variance by _i ² For the character total yield the phenotypic variance V can be explicitly expressed dependent on 1) the number n of components in the mixture, 2) the mean of the _i ² 3) the variance of the _i ² 4) the ratio and 5) the ratio _i ² /² where denotes the mean of the u _i and _u ² is the variance of the u _j. According to the dependence of the phenotypic stability on these factors some conclusions can be easily derived from this V-formula. Furthermore, two different approaches for a calculation of necessary or optimal numbers of components using the phenotypic variance V are discussed: A. Determination of optimal numbers in the sense that a continued increase of the number of components brings about no further significant effect according to stability. B. A reduction of b % of the number of components but nevertheless an unchanged stability can be realized by an increase of the mean of the u _i by 1% (with and _u ² assumed to be unchanged). Numerical results on n (from A) and 1 (from B) are given. Computing the coefficient of variation v for the character total yield and solving for the number n of components one obtains an explicit expression for n dependent on v and the factors 2.-5. mentioned above. In the special case of equal variances, _i ² = _o ² for each i, the number n depends on v, x = (₀/)² and y = (_u/)². Detailed numerical results for n = n (v, x, y) are given. For x 1 and y 1 one obtains n = 9, 20 and 79 for v = 0.30, 0.20 and 0.10, respectively while for x 1 and arbitrary y-values the results are n = 11, 24 and 95.This publication is an extended version of a lecture given at the 1984-EUCARPIA meeting (Section Biometrics in Plant Breeding) in Stuttgart-Hohenheim (Federal Republic of Germany)     

Synthetic substrates in the histochemical demonstration of intestinal disaccharidases

Summary The splitting of 6-Br-2-naphthyl-, -naphthyl-, and 4-Cl-5-Br-3-indolyl-glycosides which proved useful for the assessment of cytological localization of intestinal enzymes in previous studies was investigated using isolated human and rat intestinal disaccharidases as a source of enzyme activities.Previous findings based on histochemical studies were confirmed and extended. 6-Br-2naphthyl-D-glucoside is cleaved by glucoamylase and sucrase-isomaltase. The participatio of trehalase in splitting of this substrate is very low and can be neglected. The mentioned -glucosidases are responsible for the brush border staining of enterocytes with this substrate when unfixed cold microtome sections are used. Even when a differential heat inactivation of sucrase-isomaltase and of glucoamylase occurs during paraffin embedding (so that the staining in paraffin sections is due mostly to glucoamylase) the use of natural substrates is desirable for a more precise assessment of sucrase-isomaltase activity (but without the possibility of a correct localization).4-Cl-5-Br-3-indolyl--D-fucoside is the substrate of choice for the demonstration of lactase. Even when this substrate is split also by hetero--galactosidase and by acid (lysosomal) -galactosidase these activities do not disturb the histochemical demonstration of lactase. If however some doubts arise, the inhibition with p-Cl-mercuribenzoate (2 · 10^–4 M) is to be emloyed (lactase activity is not inhibited). Due to a low K_m and a high V_max of indolyl-fucoside and due to its extreme stability in solution (which enables to use the substrate solution repeatidly) this substrate is suitable in routine practice even though it is expensive. -naphthyl- and 4-Cl-5-Br-3-indolyl--D-glucosides are split by lactase and -glucosidase. Due to the fact that the mutual delineation of these activities is not easy and that K_m an V_max for lactase are not so favourable as in the case of fucoside these substrates are not recommended for the assessment of lactase.6-Br-2-naphthyl--D-glucoside is the substrate of choice for the histochemical studies concerned with hetero--galactosidase and 4-Cl-5-Br-3-indolyl--D-galactoside for acid -galactosidase.     

Response of the Photosynthetic Apparatus of the Diatom Thallassiosira weisflogii to High Irradiance Light

The response of the photosynthetic apparatus to high irradiance illumination (440–2200 W/m²) was studied in the diatom Thallassiosira weisflogii by fluorescence methods. Changes in the photosynthetic apparatus were monitored by measuring characteristics of chlorophyll fluorescence F ₀, F _m, F _v/F _m, and qN for several hours after illumination of the alga with high-intensity light. Incubation of the alga with 2 mM DTT, an inhibitor of de-epoxidase of carotenoids in the diadinoxanthin cycle, led to a decrease in the nonphotochemical quenching of chlorophyll fluorescence and a drop in the F _v/F _m ratio, a characteristic that reflects the quantum efficiency of the functioning of the photosynthetic apparatus. Light-induced absorption changes associated with transformations of carotenoids of diadinoxanthin cycle were recorded in vivo in algal suspensions in the absence and in the presence of DTT. Using the microfluorometric method, we measured cell distribution over the efficiency of the primary processes of photosynthesis (F _v/F _m) after illumination. We found cells with a high tolerance of their photosynthetic apparatus to photooxidative damage. The relatively high tolerance of a portion of the cell population to high-light illumination can be related to light-induced transformation of carotenoids and to the functioning of other protective systems of the photosynthetic apparatus in diatoms.     

Simulation of glucose isomerase reactor: optimum operating temperature

A design equation for immobilized glucose isomerase (IGI) packed bed reactor is developed assuming enzyme deactivation and substrate protection. The developed equation is used to simulate the performance of the reactor at various temperatures (50–80 °C). Enzyme deactivation is significant at high temperature. Substrate protection showed to have significant effect in reducing enzyme deactivation and increasing the enzyme half-life. Factors affecting the optimum operating temperature are discussed. The optimum operating temperature is greatly influenced by the operating period and to a lesser extent with both initial glucose concentration and glucose conversion.Two modes of reactor operation are tested i.e., constant feed flow rate and constant conversion. Reactor operating at constant conversion is more productive than reactor operating at constant flow rate if the working temperature is higher than the optimum temperature. Although at lower temperatures than the optimum, the two modes of operation give the same result.List of Symbols a residual enzyme activity - E [mg/l] concentration of active enzyme - E _a [kJ/mole] activation energy - E ₀ [mg/l] initial concentration of active enzyme - k [Specific] kinetic parameter - k _d [h^–1] first order thermal deactivation rate constant - k _e equilibrium constant - k _m [mole/l] apparent Michaelis constant - k _p [mole/l] Michaelis constant for product - k _s [mole/l] Michaelis constant for substrate - k ₀ [Specific] pre-exponential factor - Q [1/h] volumetric flow rate - ¯Q [1/h] average volumetric flow rate - R [kJ/mol·k] ideal gas constant - s [mole/l] apparent substrate concentration - s [mole/l] substrate concentration - s _e [mole/l] substrate concentration at equilibrium - s ₀ [mole/l] substrate concentration at reactor inlet - p [mole/l] product concentration - p _e [mole/l] product concentration at equilibrium - P _r [mole fructose/l·h] reactor productivity - T [k] temperature - t [h] time - t _p [h] operating time - V [l] reactor volume - v [mole/l·h] reaction rate - v [mole/l] reaction rate under enzyme deactivation and substrate protection - v _m [mole/l·h] maximum apparent reaction rate - v _p [mole/l·h] maximum reaction rate for product - v _s [mole/l·h] maximum reaction rate for substrate - x substrate fractional conversion - x _e substrate fractional conversion at equilibrium Greek Symbols effectiveness factor - mean effectiveness factor - substrate protection factor - [h] residence time - [h] average residence time - ₀ [h] initial residence time     

The electrochemical proton potential generated by the sulphur respiration of Wolinella succinogenes

Wolinella succinogenes grown on formate and elemental sulphur was found to use the polysulphide derivatives 2,2-tetrathiobispropionate (R₂S₄) or pentathionate (S₅O ₆ ⁼ ) as acceptors for formate oxidation. The specific activities of formate oxidation with these acceptors were similar to those with elemental sulphur. The main reaction products of R₂S₄ reduction were 2,2-dithiobispropionate (R₂S₂) and sulphide. Pentathionate was converted to thiosulphate and some elemental sulphur. The electrochemical proton potential across the cytoplasmic membrane of the bacterium was measured in the steady state of electron transport from formate to R₂S₄. The electrical proportion () of the determined through the distribution of labeled tetraphenylphosphonium cation was obtained as 0.17 Volt. The was zero, when a protonophore was present. The pH-difference across the membrane was negligible. Thus the generated by sulphur respiration is close to that measured earlier with fumarate as the terminal acceptor of electron transport.Abbreviations DMO 5,5-dimethyloxazolidine-2,4-dione - R₂S_n (n=2–5) 2,2-polythiobispropionate - TTFB 4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazol - TPP tetraphenylphosphonium cation     

Reactor design for the enzymatic isomerization of glucose to fructose

A comprehensive methodology is presented for the design of reactors using immobilized enzymes as catalysts. The design is based on material balances and rate equations for enzyme action and decay and considers the effect of mass transfer limitations on the expression of enzyme activity. The enzymatic isomerization of glucose into fructose with a commercial immobilized glucose isomerase was selected as a case study. Results obtained are consistent with data obtained from existing high-fructose syrup plants. The methodology may be extended to other cases, provided sound expressions for enzyme action and decay are available and a simple flow pattern within the reactor might be assumed.List of Symbols C kat/kg specific activity of the catalyst - D m²/s substrate diffusivity within the catalyst particle - Dr m reactor diameter - d d operating time of each reactor - E kat initial enzyme activity - E _i kat initial enzyme activity in each reactor - F m³/s process flowrate - F _i m³/s reactor feed flowrate at a given time - F ₀ m³/s initial feed flowrate to each reactor - H number of enzyme half-lives used in the reactors - K mole/m³ equilibrium constant - K _S mole/m³ Michaelis constant for substrate - K _P mole/m³ Michaelis constant for product - K _m mole/m³ apparent Michaelis constant f(K, K _s, K_p, s₀) - k mole/s · kat reaction rate constant - k _d d^–1 first-order thermal inactivation rate constant - L m reactor height - L _r m height of catalyst bed - N _R number of reactors - P _i kg catalyst weight in each reactor - p mole/m³ product concentration - R m particle radius - R _P ratio of minimum to maximum process flowrate - r m distance to the center of the spherical particle - s mole/m³ substrate concentration - s _0i mole/m³ substrate concentration at reactor inlet - s ₀ mole/m³ bulk substrate concentration - s mole/m³ apparent substrate concentration - T K temperature - t d time - t _i d operating time for reactor i - t _s d time elapsed between two successive charges of each reactor - V m³ reactor volumen - V _m mole/m³ s maximum apparent reaction rate - V _p mole/m³ s maximum reaction rate for product - V _R m³ actual volume of catalyst bed - V _r m³ calculated volume of catalyst bed - V _S mol/m³ s maximum reaction rate for substrate - v mol/m³ s initial reaction rate - v _i m/s linear velocity - v _m mol/m³ s apparent initial reaction rate f(K_m, s,V_m) - X substrate conversion - X _eq substrate conversion at equilibrium - =s/K dimensionless substrate concentration - ₀=s₀/K bulk dimensionless substrate concentration - _eq=s_eq/K dimensionless substrate concentration at equilibrium - local effectiveness factor - mean integrated effectiveness factor - Thiéle modulus - =r/R dimensionless radius - _s kg/m³ hydrated support density - substrate protection factor - s residence time     

Verhaltensontogenese der Habitatwahl beim Teichrohrs?nger (Acrocephalus scirpaceus)

Zusammenfassung In Anpassung an seinen aus vertikalen Vegetationsstrukturen zusammengesetzten Lebensraum besitzt der Teichrohrsänger die beste morphologische Ausstattung aller sechs mitteleuropäischer Rohrsängerarten für Vertikalklettern. An jungen Teichrohrsängern wurde überprüft, ob frühkindliche Erfahrungen auf verschiedenen Sitzstangen (auf senkrechten=K_s, senkrechten und waagerechten=K_m, waagerechten=V_w und waagerechten Sitzstangen mit Futter belohnt=V_w+) die spätere Wahl dieser Strukturelemente beeinflussen. Die lokomotorische Aktivität der vier Aufzuchtsgruppen wurde im Zweifachwahlversuch (horizontale gegen vertikale Sitzstangen) getestet. Mit Ausnahme der auf senkrechten Sitzstangen aufgezogenen Vögel (K_s), deren Wahlverhalten gleichverteilt war (Abb. 2), bevorzugten alle anderen Gruppen das vertikale Testsubstrat (Abb. 2 und 3). Auch die Belohnung mit Futter als positiver Verstärker der Horizontalelemente (V_w+) machte diese nur wenig attraktiver (Tab. 1). Alle Gruppen nutzten während der Testperiode das vertikale Substrat zunehmend stärker (Tab. 1). Die Zunahme rührte bei den Vögeln der beiden Kontrollgruppen (K_s und K_m) und der Versuchsgruppe waagerecht (V_w) alleine von der Erfahrung in der Versuchssituation her (Tab. 2). Die Nutzung vertikaler bzw. horizontaler Strukturelemente durch junge Teichrohrsänger wird somit bestimmt durch: 1. eine angeborene Präferenz für das artgemäße Substrat, 2. einen Novitätseffekt, bedingt durch die Aufzuchterfahrung (Wahlverhalten der K_s-Gruppe, Abb. 2), 3. Eigenerfahrung bei der Nutzung verschiedener Substrate im Versuch. Zusätzliche Wahlversuche später im Jahr (Oktober bis Dezember) mit denselben Versuchsvögeln zeigten keine Änderungen in der Substratwahl (Abb. 4).

---

Ontogeny of habitat choice in the Reed Warbler (Acrocephalus scirpaceus)

Summary The Reed Warbler shows the most specialized morphological traits for vertical climbing among the six central EuropeanAcrocephalus species. We consider this as an adaptation to the vertical structures in its habitat. In experiments with young Reed Warblers I tested whether early experience with different perches has an influence on the choice of these structures later on in life. Locomotory activity of the following groups was tested in double choice experiments (horizontal versus vertical perches):Control group vertical (K_s): raised on vertical perches, Control group mixed habitat (K_m): raised on vertical and horizontal perches, Test group horizontal (V_w): raised on horizontal perches, Test group horizontal, plus rewards (V_w+): raised on horizontal perches and additional feeders attached to the bars. With the exception of Control group vertical (K_s) which shows no choice preference (Fig. 2) all other birds prefered the vertical test substrate (Fig. 2 and 3). Even the reward (food) as a positive reinforcement of horizontal elements had little effect on their attractiveness (Tab. 1). An increase in the choice of the vertical substrate could be observed throughout the 3-day test period in all groups (Tab. 1). This increase was only due to the experience in the test situation for the birds in both control groups (K_s and K_m) and the Test group horizontal (V_w) (Tab. 2). Therefore the use of vertical or horizontal structures in Reed Warblers is determind by: 1. An innate preference for species-specific substrate, 2. Novelty due to early experience (choice behaviour of Control group vertical, Fig. 2), 3. Experience while using different substrates (experience in the test situation) which optimizes substrate choice according to proprioceptive learning. Additional choice experiments with the same test birds later on in the year (October to December) revealed the same results. Therefore it seems unlikely that there exists a preprogrammed change in substrate choice in the course of a year (Fig. 4). Although choice experiments of this kind can only include a limited part of the habitat-scheme, they are useful experimental designs for investigating specialized species whose habitats can be simulated closely in the laboratory.

     

Comparison of [35S]GTPγS binding to plasma membranes and endomembranes prepared from soybean and rat

Summary Plasma membranes were prepared from soybean hypocotyls and roots by aqueous two-phase partitioning and subsequent free-flow electrophoresis. The highly purified plasma membranes bound [³⁵S]GTPS with a relatively high affinity (K_d10nM). The binding was saturable and specific as it was indicated by the displacement of bound [³⁵S]GTPS by unlabeled GTPS and GTP, but not by ATPS, ATP, UTP or CTP. ITP was intermediate in its ability to displace [³⁵S]GTPS. When soybean plasma membrane proteins were separated by SDS-PAGE and displayed by autoradiography, two major [³⁵S]GTPS binding proteins were revealed with apparent molecular weights of 24 and 28 kDa. Results with plasma membranes from soybean hypocotyls and roots were similar but differed from those with plasma membranes prepared from rat liver and adipocytes where only a single major [³⁵S]GTPS binding activity with a molecular weight of 28 kDa was observed.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - G protein hetero-trimeric GTP binding protein with , , subunits - G_n protein GTP binding protein detected on nitrocellulose blots - GTPS guanosine 5-[-thio]triphosphate - IAA 3-indoleacetic acid - SDS-PAGE sodium dodecylsulfate-polyacrylamide gel electrophoresis     

Pineal melatonin rhythms in the lizardAnolis carolinensis: effects of light and temperature cycles

Summary Pineal and ocular melatonin was assessed, over 24 h periods, in male lizards (Anolis carolinensis) entrained to 24 h light-dark (LD) cycles and a constant 32 C, and in lizards entrained to both 24 h LD cycles and 24 h temperature cycles (32 C/20 C). At a constant temperature, the duration of the photoperiod has a profound effect on the duration, amplitude, and phase of the pineal melatonin rhythm (Fig. 1). The pineal melatonin rhythm under cyclic temperature peaks during the cool (20 C) phase of the cycle regardless of whether or not the cool phase occurs during the light or dark phase of a LD 1212 cycle (Fig. 3). Under a temperature cycle and constant dim illumination, a pineal melatonin rhythm is observed which peaks during the cool phase of the temperature cycle, but the amplitude of the rhythm is depressed relative to that observed under LD (Fig. 2). Illumination up to 2 h in duration does not suppress the nocturnal melatonin peak in theAnolis pineal (Fig. 4). No melatonin rhythm was observed in the eyes ofAnolis under either 24 h LD cycles and a constant temperature (Fig. 1), or under simultaneous light and temperature cycles (Fig. 3). Ocular melatonin content was, in all cases, either very low or non-detectable.Abbreviations HIOMT hydroxyindole-O-methyltransferase - NAT N-acetyltransferase     

Conformational Change of the Rieske [2Fe–2S] Protein inCytochrome bc 1 Complex

Structures of mitochondrial bc ₁ complex have been reported based on four different crystalforms by three different groups. In these structures, the extrinsic domain of the Rieske [2Fe–2S]protein, surprisingly, appeared at three different positions: the c ₁ position, where the [2Fe–2S]cluster exists in close proximity to the heme c ₁; the b position, where the [2Fe–2S] clusterexist in close proximity to the cytochrome b; and the intermediate position where the[2Fe–2S] cluster exists in between c ₁ and b positions. The conformational changes betweenthese three positions can be explained by a combination of two rotations; (1) a rotation of theentire extrinsic domain and (2) a relative rotation between the cluster-binding fold and thebase fold within the extrinsic domain. The hydroquinone oxidation and the electron bifurcationmechanism at the Q_P binding pocket of the bc ₁ complex is well explained using theseconformational changes of the Rieske [2Fe–2S] protein.     

The roles of serine and threonine sidechains in ion channels: a modelling study

The ion channel of the nicotinic acetylcholine receptor (nAChR) is believed to be lined by transmembrane M2 helices. A 4-8-12 sequence motif, comprising serine (S) or threonine (T) residues at positions 4, 8 and 12 of M2, is conserved between different members, anion and cation selective, of the nAChR superfamily. Parallel bundles of 4-8-12 motif-containing helices are considered as simplified models of ion channels. The relationship between S and T sidechain conformations and channelion interactions is explored via evaluation of interaction energies of K⁺ and of Cl^– ions with channel models. Energy calculations are used to determine optimal ₂ (C-C\-O-H) values in the presence of K⁺ or Cl^– ions. 4-8-12 motif-containing bundles may form favourable interactions with either cations or anions, dependent upon the ₂ values adopted. Parallel-helix and tilted-helix bundles are considered, as are heteromeric models designed to mimic the Torpedo nAChR. The main conclusion of the study is that conformational flexibility at ₂ enables both S and T residues to form favourable interactions with anions or cations. Consequently, there is apparently no difference between S and T residues in their interactions with permeant ions, which suggests that the presence of T vs. S residues within the 4-8-12 motif is not a major mechanism whereby anion/cation selectivity may be generated. The implications of these studies with respect to more elaborate models of nAChR and related receptors are considered.Abbreviations nAChR, GluR, NMDA-R, 5HT₃-R, GABA_AR, GlyR nicotinic acetylcholine, glutamate, NMDA, 5HT₃, GABA_A and glycine receptors, respectively - PhTx philanthotoxin - M2 second membrane-spanning helix of receptor-channel subunits