Ethanol production using nuclear petite yeast mutants

Two respiratory-deficient nuclear petites, FY23Δpet191 and FY23Δcox5a, of the yeast Saccharomyces cerevisiae were generated using polymerase-chain-reaction-mediated gene disruption, and their respective ethanol tolerance and productivity assessed and compared to those of the parental grande, FY23WT, and a mitochondrial petite, FY23ρ⁰. Batch culture studies demonstrated that the parental strain was the most tolerant to exogenously added ethanol with an inhibition constant. K _i, of 2.3% (w/v) and a specific rate of ethanol production, q _p, of 0.90 g ethanol g dry cells⁻¹ h⁻¹. FY23ρ⁰ was the most sensitive to ethanol, exhibiting a K _i of 1.71% (w/v) and q _p of 0.87 g ethanol g dry cells⁻¹ h⁻¹. Analyses of the ethanol tolerance of the nuclear petites demonstrate that functional mitochondria are essential for maintaining tolerance to the toxin with the 100% respiratory-deficient nuclear petite, FY23Δpet191, having a K _i of 2.14% (w/v) and the 85% respiratory-deficient FY23Δcox5a, having a K _i of 1.94% (w/v). The retention of ethanol tolerance in the nuclear petites as compared to that of FY23ρ⁰ is mirrored by the ethanol productivities of these nuclear mutants, being respectively 43% and 30% higher than that of the respiratory-sufficient parent strain. This demonstrates that, because of their respiratory deficiency, the nuclear petites are not subject to the Pasteur effect and so exhibit higher rates of fermentation. Received: 22 September 1997 / Accepted: 7 December 1997     

Composition and distribution of internal resistance in three types of microbial fuel cells

High internal resistance is a key problem limiting the power output of the microbial fuel cell (MFC). Therefore, more knowledge about the internal resistance is essential to enhance the performance of the MFC. However, different methods are used to determine the internal resistance, which makes the comparison difficult. In this study, three different types of MFCs were constructed to study the composition and distribution of internal resistance. The internal resistance (R _i) is partitioned into anodic resistance (R _a), cathodic resistance (R _c), and ohmic resistance () according to their origin and the design of the MFCs. These three resistances were then evaluated by the “current interrupt” method and the “steady discharging” method based on the proposed equivalent circuits for MFCs. In MFC-A, MFC-B, and MFC-C, the R _i values were 3.17, 0.35, and 0.076 Ω m², the values were 2.65, 0.085, and 0.008 Ω m², the R _a values were 0.055, 0.115, and 0.034 Ω m², and the R _c values were 0.466, 0.15, and 0.033 Ω m², respectively. For MFC-B and MFC-C, the remarkable decrease in R _i compared with the two-chamber MFC was mainly ascribed to the decline in and R _c. In MFC-C, the membrane electrodes’ assembly lowered the ohmic resistance and facilitated the mass transport through the anode and cathode electrodes, resulting in the lowest R _i among the three types.     

Evolution of discontinuous solutions to transport equations in stellarators

It was shown earlier that, in the range of rare collisions, transport equations for stellarators allow steady discontinuous solutions for the ambipolar electric field and for the plasma density and temperature gradients. Moreover, such solutions are non-single-valued; that is, their explicit form depends on the initial values of the ambipolar electric field. The time-independent transport equations are derived under the conventional quasineutrality condition; i.e., it is assumed that the electron and ion densities, N _e and N _i , are related by the relationship N _e = ZN _i (where Z is the ion charge number). In other words, the plasma charge density is assumed to be much less than the product e _i N _i . Under typical conditions, the corresponding inequality is satisfied by a large margin. However, if the electric field E has discontinuities, then it can be seen from the equation ▿·E = 4πρ that, at the discontinuity points, the charge density becomes infinite and the relationship N _e = ZN _i fails to hold, so it is necessary to replace it with N _e = ZN _i + ρ/e _e . In the transport equations, this latter replacement produces additional terms, proportional to the second radial derivative of the field E. With these additional terms, the steady solutions are modified substantially. First, the ambipolar field and the derivatives of the density and temperatures all become continuous functions of the coordinates, a result that seems to be quite obvious. The second, not-so-obvious result is that the steady solutions become single-valued, i.e., independent of the initial values of the ambipolar electric field. It turns out that, in this case, two regimes are possible, depending on the values of the plasma parameters. In the first regime, the solution is unique and is independent of the initial conditions. In the second regime, two steady solutions can exist, depending on the initial conditions. One of the solution is similar to that obtained in the first regime, and the other differs from the first one both in the ambipolar field profile and in the dependence of the density and temperatures on the minor plasma radius. It cannot be excluded that different plasma confinement modes revealed in experiments are associated with the existence of such solutions.     

Evaporative and excretory water loss during free flight in pigeons

Body water conservation is important in flying birds because the very high metabolic demands and heat dissipation requirements during flight depend on plasma-volume integrity. Wind tunnel experiments and theoretical model predictions show that evaporative water loss (EWL) depends on air temperature (T _a) and water vapor density (ρ_a), but these relationships have not been examined in free-flying birds. The contribution of excretory water loss to the total water loss of a flying bird is thought to be negligible but this assumption is untested. To study the dependence of water losses on environmental conditions in free-flying birds and to quantify the contribution of excretory water loss to total water loss, we estimated evaporative and excretory water losses in 16 trained, free-flying tippler pigeons (Columba livia, 250–340 g). We collected excreta by attaching a light latex, water-impermeable receptacle around each bird's vent. By gravimetry, we measured evaporative and excretory water losses of birds for eight flights at different T _as and compared flying to resting (control) birds for two of these flights. EWL was constant with respect to T _a when less than 15 °C, and increased with increasing T _a above 19 °C, indicating that evaporative cooling was invoked when the heat load increased. EWL increased with increasing ρ_a, possibly due to the strong correlation between ρ_a and T _a. Excretory water loss was independent of ρ_a or T _a and averaged almost 10% of the total water loss. Measurements of EWL made on pigeons during wind tunnel experiments and previous free-flight studies are consistent with our free-flight measurements made at similar T _a s. Accepted: 13 April 1999     

Impact of magnetic field inhomogeneity on electron cyclotron radiative loss in tokamak reactors

The potential importance of electron cyclotron (EC) emission in the local electron power balance in the steady-state regimes of ITER operation with high temperatures, as well as in the DEMO reactor, requires accurate calculation of the one-dimensional (over magnetic surfaces) distribution of the net radiated power density, P _EC(ρ). When the central electron temperature increases to ∼30 keV, the local EC radiative loss comprises a substantial fraction of the heating power from fusion alphas and is close to the total auxiliary NBI heating power, P _EC(0) ≃ 0.3P _α(0) ≃ P _aux(0). In the present paper, the model of EC radiative transport in an axisymmetric toroidal plasma is extended to the case of an inhomogeneous magnetic field B(R, Z). The impact of such inhomogeneity on local and total power losses is analyzed in the framework of this model by using the CYNEQ code. It is shown that, for the magnetic field B, temperature T _e , density n _e , and wall reflection coefficient R _w expected in ITER and DEMO, accurate simulations of the EC radiative loss require self-consistent 1.5D transport analysis (i.e., one-dimensional simulations of plasma transport and two-dimensional simulations of plasma equilibrium). It is shown that EC radiative transport can be described with good accuracy in the 1D approximation with the surface-averaged magnetic field, B(ρ) = 〈B(R, Z)〉 _ms . This makes it possible to substantially reduce the computational time required for time-dependent self-consistent 1.5D transport analysis. Benchmarking of the CYNEQ results with available results of the RAYTEC, EXACTEC, and CYTRAN codes is performed for various approximations of the magnetic field.     

Maximum glucoamylase production by a temperature-sensitive mutant of Saccharomyces cerevisiae in batch culture

In order to maximize the glucoamylase production by recombinant Saccharomyces cerevisiae in batch culture, first a temperature-controlled expression system for a foreign gene in S. cerevisiae was constructed. A temperature-sensitive pho80 mutant of S. cerevisiae for the PHO regulatory system, YKU131, was used for this purpose. A DNA fragment bearing the promoter of the PHO84 gene, which encodes an inorganic phosphate (P_i) transporter of S. cerevisiae and is derepressed by P_i starvation, was used as promoter. The glucoamylase gene connected with the PHO84 promoter was ligated into a YEp13 vector, designated pKU122. When the temperature-sensitive pho80 ^ts mutant harboring the plasmid pKU122 is cultivated at a lower temperature, the expression of glucoamylase gene is repressed, but at a higher temperature it is expressed. Next the effect of temperature on the specific growth rate, μ, and specific production rate, ρ, was investigated. Maximum values of ρ and ρ at various temperatures were at 30°C and 34°C, respectively. The optimal cultivation temperature strategy for maximum production of glucoamylase by this recombinant strain in batch culture was then determined by the Maximum principle using the relationships of μ and ρ to the cultivation temperature. Finally, the optimal strategy was experimentally realized by changing the cultivation temperature from Tμ (30°C) to Tρ (34°C) at the switching time, t_s. Received 18 September 1997/ Accepted in revised form 07 January 1998     

Analysis of Root Growth by Impedance Spectroscopy (EIS)

Electrical impedance spectroscopy (EIS) is investigated as a non-destructive method for monitoring root growth of tomato. This paper aims to (i) review the basic principles of EIS applied to the characterisation of the different parts of the soil–root–stem-electrode continuum, (ii) experiment the validity of the relationship between root weight and root capacitance taking into account the influence of the soil and plant electrodes position, (iii) describe an EIS analysis of the root growth of tomato plants. All experiments were carried out in 50 dm³ containers either in hydroponics at 930 μS for the test of root fresh or dry weight and root capacitance relationships, or in a potting mix (oxisol) for electrode placement tests and EIS estimation of root growth. Electrical measurements of the soil–root–stem-electrode continuum were done with a two-electrode measuring system using unpolarisable Ag–AgCl electrodes. A ‘root cutting’ and a ‘progressively immersed root system’ experiments were carried out in order to validate the relationship between root capacitance and root mass at 1 kHz. The effects of soil electrode and plant electrode placement were also tested, pointing out the sensitivity of the method to the insertion height of the “plant electrode” into the stem. For the root growth experiment, Impedance Spectra (IS) measurements were made just before harvesting the roots for dry weight and length determination. Measurements were made 14, 22, 26 and 39 days after planting, until flowering. The IS of the soil–root–stem-electrode continuum was modelled by a lumped electric circuit consisting of a series resistor R ₀ for the soil and of four parallel resistance (R _i )-capacitance (C _i ) circuits for the other components of the circuit. The model had nine parameters whose values were estimated by Complex Nonlinear Least Squares curve fitting. A stepwise ascendant regression was used to identify the electrical parameters that better correlated with root dry mass or length increment: C ₃ and C ₄ were well correlated with root dry mass with a r ² of 0.975, whereas root length was explained by the combination of 1/R ₃, C ₃, 1/R ₂ and 1/R ₁ with a r ² of 0.986. This work may be considered as a new methodological contribution to the understanding of root electrical properties in the non-destructive diagnosis of root systems.     

Elevated CO2 did not affect the hydrological balance of a mature native Eucalyptus woodland

Elevated atmospheric CO₂ concentration (eC_a) might reduce forest water‐use, due to decreased transpiration, following partial stomatal closure, thus enhancing water‐use efficiency and productivity at low water availability. If evapotranspiration (E_t) is reduced, it may subsequently increase soil water storage (ΔS) or surface runoff (R) and drainage (D_g), although these could be offset or even reversed by changes in vegetation structure, mainly increased leaf area index (L). To understand the effect of eC_a in a water‐limited ecosystem, we tested whether 2 years of eC_a (~40% increase) affected the hydrological partitioning in a mature water‐limited Eucalyptus woodland exposed to Free‐Air CO₂ Enrichment (FACE). This timeframe allowed us to evaluate whether physiological effects of eC_a reduced stand water‐use irrespective of L, which was unaffected by eC_a in this timeframe. We hypothesized that eC_a would reduce tree‐canopy transpiration (E_tree), but excess water from reduced E_tree would be lost via increased soil evaporation and understory transpiration (E_floor) with no increase in ΔS, R or D_g. We computed E_t, ΔS, R and D_g from measurements of sapflow velocity, L, soil water content (θ), understory micrometeorology, throughfall and stemflow. We found that eC_a did not affect E_tree, E_floor, ΔS or θ at any depth (to 4.5 m) over the experimental period. We closed the water balance for dry seasons with no differences in the partitioning to R and D_g between C_a levels. Soil temperature and θ were the main drivers of E_floor while vapour pressure deficit‐controlled E_tree, though eC_a did not significantly affect any of these relationships. Our results suggest that in the short‐term, eC_a does not significantly affect ecosystem water‐use at this site. We conclude that water‐savings under eC_a mediated by either direct effects on plant transpiration or by indirect effects via changes in L or soil moisture availability are unlikely in water‐limited mature eucalypt woodlands.     

Implications of an anomalous intracellular electrical response in bullfrog corneal epithelium

Summary The ionic dependencies of the transepithelial and intracellular electrical parameters were measured in the isolated frog cornea. In NaCl Ringer's the intracellular potential differenceV _sc measured under short-circuit conditions depolarized by nearly the same amount after either increasing the stromal-side KCl concentration from 2.5 to 25mm or exposure to 2mm BaCl₂ (K⁺ channel blocker). With Ba²⁺ the depolarization of theV _sc by 25mm K⁺ was reduced to one-quarter of the control change. If the Cl-permselective apical membrane resistanceR _o remained unchanged, the relative basolateral membrane resistanceR _i, which includes the lateral intercellular space, increased at the most by less than twofold after Ba²⁺. These effects in conjunction with the depolarization of theV _sc by 62 mV after increasing the stromal-side K⁺ from 2.5 to 100mm in Cl-free Ringer's as well as the increase of the apparent ratio of membrane resistances (a=R _o/R_i) from 13 to 32 are all indicative of an appreciable basolateral membrane K⁺ conductance. This ratio decreased significantly after exposure to either 25mm K⁺ or Ba²⁺. The decline ofR _o/R_i with 25mm K⁺ appears to be anomalous since this decrease is not consistent with just an increase of basolateral membrane conductance by 25mm K⁺, but rather perhaps a larger decrease ofR _o thanR _iAlso an increase of lateral space resistance may offset the effect of decreasingR _i with 25mm K⁺. In contrast,R _o/R_i did transiently increase during voltage clamping of the apical membrane potential differenceV _o and exposure to 25mm K⁺ on the stromal side. This increase and subsequent decrease ofR _o/R_i supports the idea that increases in stromal K⁺ concentration may produce secondary membrane resistance changes. These effects onR _o/R_i show that the presence of asymmetric ionic conductance properties in the apical and basolateral membranes can limit the interpretative value of this parameter. The complete substitution of Na⁺ withn-methyl-glucamine in Cl-free Ringer's on the stromal side hyperpolarized theV _sc by 6 mV whereas 10^–4 m ouabain depolarized theV _sc by 7 mV. Thus the basolateral membrane contains K⁺, Na⁺ and perhaps Cl^– pathways in parallel with the Na/K pump component.     

Accuracy of Optimized Chemical-exchange Parameters Derived by Fitting CPMG R 2 Dispersion Profiles when R 2 0a ≠ R 2 0b

The transverse relaxation rate, R₂, measured as a function of the effective field (R₂ dispersion) using a Carr-Purcell-Meiboom-Gill (CPMG) pulse train, is well suited to detect conformational exchange in proteins. The dispersion data are commonly fitted by a two-site (sites a and b) exchange model with four parameters: the relative population, p_a, the difference in chemical shifts of the two sites, δω, the correlation time for exchange, τ_ex, and the intrinsic relaxation rate (i.e., transverse relaxation rate in the absence of chemical exchange), R₂⁰. Although the intrinsic relaxation rates of the two sites, R₂^0a and R₂^0b, can differ, they are normally assumed to be the same (i.e., R₂^0a = R₂^0b = R₂⁰) when fitting dispersion data. The purpose of this investigation is to determine the magnitudes of the errors in the optimized exchange parameters that are introduced by the assumption that R₂^0a = R₂^0b. In order to accomplish this goal, we first generated synthetic constant-time CPMG R₂ dispersion data assuming two-site exchange with R₂^0a ≠ R₂^0b, and then fitted the synthetic data assuming two-site exchange with R₂⁰ = R₂^0a = R₂^0b. Although all the synthetic data generated assuming R₂^0a ≠ R₂^0b were well fitted (assuming R₂^0a = R₂^0b), the optimized values of p_a and τ _ex differed from their true values, whereas the optimized values of δω values did not. A theoretical analysis using the Carver–Richards equation explains these results, and yields simple, general equations for estimating the magnitudes of the errors in the optimized parameters, as a function of ( R₂^0a − R₂^0b).     

What is the usual internal carbon dioxide concentration in C<Subscript>4</Subscript> species under midday field conditions?

The carbon dioxide concentrating system in C₄ photosynthesis allows high net photosynthetic rates (P _N) at low internal carbon dioxide concentrations (C _i), permitting higher P _N relative to stomatal conductance (g _s) than in C₃ plants. This relation would be reflected in the ratio of C _i to external ambient (C _a) carbon dioxide concentration, which is often given as 0.3 or 0.4 for C₄ plants. For a C _a of 360 μmol mol⁻¹ that would mean a C _i about 110–140 μmol mol⁻¹. Our field observations made near midday on three weedy C₄ species, Amaranthus retroflexus, Echinochloa crus-galli, and Setaria faberi, and the C₄ crop Sorghum bicolor indicated mean values of C _i of 183–212 μ mol mol⁻¹ at C _a = 360 μmol mol⁻¹. Measurements in two other C₄ crop species grown with three levels of N fertilizer indicated that while midday values of C _i at high photon flux were higher at limiting N, even at high nitrogen C _i averaged 212 and 196 μmol mol⁻¹ for Amaranthus hypochondriacus and Zea mays, respectively. In these two crops midday C _i decreased with increasing leaf to air water vapor pressure difference. Averaged over all measurement days, the mean C _i across all C₄ species was 198 μmol mol⁻¹, for a C _i/C _a ratio of 0.55. Prior measurements on four herbaceous C₃ species using the same instrument indicated an average C _i/C _a ratio of 0.69. Hence midday C _i values in C ₄ species under field conditions may often be considerably higher and more similar to those of C₃ species than expected from measurements made on plants in controlled environments. Reducing g _s in C₄ crops at low water vapor pressure differences could potentially improve their water use efficiency without decreasing P _N.     

Radial theory of the ion current onto a probe in a low-pressure plasma with allowance for volume ionization and collisions with neutrals

The ion current onto a spherical or cylindrical probe is analyzed in the cold-ion approximation with allowance for ionization and collisions with neutrals. An expression for the ion density that takes into account both ionization and collision in the finite-size perturbed region is derived. The current-voltage characteristics for the dimensionless parameters r _p/λ _D = 0.0001−10, λ _i /λ _D = 0.01 − ∞, and zλ _D/(kT _e /M)^1/2 = 0−5 are determined by numerically solving Poisson’s equation, and the corresponding approximate expressions are obtained.     

Analysis of Chameleon Sequences by Energy Decomposition on a Pairwise Per-residue Basis

The conversion from α-helix to β-strand that has been widely observed in so-called chameleon sequences has received considerable attention since such a structural change may induce many amyloidogenic proteins to self-assemble into fibrils thus causing fatal diseases. Here we report a large scale-analysis of the energetics of secondary structural conversions in a collection of chameleon sequences retrieved from the Protein Data Bank. Major energetic contributions to the secondary structural conversion were analyzed by carrying out energy decomposition on a pairwise per-residue basis, i.e., (i,i), (i,i ± 1), (i,i ± 2), (i,i ± 3), (i,i ± 4) and > (i,i ± 4) intra-/inter-residual interactions. While the overall potential energy differences were subtle, individual residue-based interacting energy differences were observed to vary significantly depending on the specific type of secondary structural conversion. The average energy difference between α-helix and β-strand, <ΔE _α→β>, in the chameleon sequences varied significantly in (i,i), (i,i ± 1) and > (i,i ± 4) interactions. The major energetic factors in secondary structure conversions were electrostatic interactions and the polar term for solvation energy. In addition, residue-based average energy differences in α-helix → β-strand conversions were well-correlated to those in α-helix → random coil → β-strand conversions (R ² = 0.92). Assuming that three secondary structural elements can transform in either direction, this strong correlation indicates that the present energy decomposition method using database structures of chameleon sequences provides a reliable tool for the characterization of secondary structure fluctuations in amino acid sequences.     

Contributions to relational biology

The principle of biotopological mapping (Rashevsky, 1954,Bull. Math. Biophysics,16, 317–48) is given a generalized formulation, as the principle of relational epimorphism in biology. The connection between this principle and Robert Rosen’s representation of organisms by means of categories (1958,Bull. Math. Biophysics,20, 317–41) is studied. Rosen’s theory of (M,R)-systems, (1958,Bull. Math. Biophysics,20, 245–60) is generalized by dropping the assumption that only terminalM _i components are sending inputs into theR _i components. It is shown that, if the primordial organism is an (M,R)-system, then the higher organisms, obtained by a construction well discussed previously (1958,Bull. Math. Biophysics,20, 71–93), are also (M,R)-systems. Several theorems about such derived (M,R)-systems are demonstrated. It is shown that Rosen’s concept of an organism as a set of mappings throws light on phenomena of synesthesia and also leads to the conclusion that Gestalt phenomena must occur not only in the fields of visual and auditory perception but in perceptions of any modality.     

Diurnal changes in chlorophylla fluorescence and carotenoid composition inOpuntia ficus-indica,a CAM plant,and in three C3 species in Portugal during summer

Summary Diurnal changes in chlorophylla fluorescence were determined in four species, differing in life form, in Portugal during the summer of 1989. These includedOpuntia ficus-indica, a CAM plant, andHelianthus annuus, Ficus carica andArbutus unedo, three C₃ species. Steady state fluorescence yield,F _S, and maximum fluorescence yield,F _M′, were determined at different times of the day. Using the model of Genty et al. (1989), the photon use efficiency of photosystem II electron transport,φ _e, was calculated from (F _M′−F _S)/F _M′. Diurnal changes in relative rate of non-cyclic electron transport through photosystem II,J _e, were derived by multiplyingφ _e by the incident photon flux density (PFD). WhenJ _e, determined for each species for various points in time throughout the day, was plotted against corresponding values of PFD, the light response curves obtained showed thatJ _e was linearly dependent on PFD in low light and approached saturation in high light. The highest values ofJ _e were observed inHelianthus annuus, followed byOpuntia ficus-indica, Ficus carica andArbutus unedo. The proportion of the xanthophyll zeaxanthin to total carotenoids, determined around noon, was inversely related to maximum rates ofJ _e.     

Alpha and beta subunits of F1-ATPase are required for survival of petite mutants in Saccharomyces cerevisiae

Although Saccharomyces cerevisiae can form petite mutants with deletions in mitochondrial DNA (mtDNA) (ρ⁻) and can survive complete loss of the organellar genome (ρ^o), the genetic factor(s) that permit(s) survival of ρ⁻ and ρ^o mutants remain(s) unknown. In this report we show that a function associated with the F₁-ATPase, which is distinct from its role in energy transduction, is required for the petite-positive phenotype of S. cerevisiae. Inactivation of either the α or β subunit, but not the γ, δ, or ɛ subunit of F₁, renders cells petite-negative. The F₁ complex, or a subcomplex composed of the α and β subunits only, is essential for survival of ρ^o cells and those impaired in electron transport. The activity of F₁ that suppresses ρ^o lethality is independent of the membrane F_o complex, but is associated with an intrinsic ATPase activity. A further demonstration of the ability of F₁ subunits to suppress ρ^o lethality has been achieved by simultaneous expression of S. cerevisiae F₁α and γ subunit genes in Kluyveromyces lactis– which allows this petite-negative yeast to survive the loss of its mtDNA. Consequently, ATP1 and ATP2, in addition to the previously identified AAC2, YME1 and PEL1/PGS1 genes, are required for establishment of ρ⁻ or ρ^o mutations in S. cerevisiae. Received: 20 March 1999 / Accepted: 18 July 1999     

4-Functionalized 1,3-diarylpyrazoles bearing 6-aminosulfonylbenzothiazole moiety as potent inhibitors of carbonic anhydrase isoforms hCA I,II, IX and XII

A series of 24 novel heterocyclic compounds—functionalized at position 4 with aldehyde (5a–5f), carboxylic acid (6a–6f), nitrile (7a–7f) and oxime (8a–8f) functional groups—bearing 6-aminosulfonybenzothiazole moiety at position 1 of pyrazole has been synthesized and investigated for the inhibition of four isoforms of the α-class carbonic anhydrases (CAs, EC 4.2.1.1), comprising hCAs I and II (cytosolic, ubiquitous isozymes) and hCAs IX and XII (transmembrane, tumor associated isozymes). Against the human isozyme hCA I, compounds 6a–6f showed medium-weak inhibitory potential with K_i values in the range of 157–690 nM with 6a showing better potential than the standard drug acetazolamide (AZA). Against hCA II, all the compounds showed excellent to moderate inhibition with K_i values of compounds 5a, 5d, 5f, 6a–6f, 8d and 8f lower than 12 nM (K_i of AZA). Against hCA IX, all the compounds showed moderate inhibition with the exception of 6e which showed nearly 9 fold a better profile compared to AZA, whereas against hCA XII, four compounds 6e, 7a, 7b and 7d showed K_i in the same order as that of AZA. Carboxylic acid 6e was found to be an excellent inhibitor of both hCA IX and XII, with K_i values of 2.8 nM and 5.5 nM, respectively.     

A mechanobiological investigation of platelets

Understanding mechanotransduction pathways leading to thrombosis will require progressive steps, including determination of the mechanical behavior of the platelet membrane in response to applied loads. The platelet membrane deformation capacity, as quantified by membrane progression into a borosilicate glass micropipette of defined internal diameter, was probed in murine platelets using a controlled range of negative pressure (0–7 cm H₂O). Based on our observations that the platelet portion outside the micropipette was mostly spherical and that the platelet volume did not change upon aspiration, a novel continuum mechanics-based model of the platelet micropipette aspiration experiment was created, and a new hyperelastic isotropic material model including membrane residual tension was proposed for the platelet membrane. Murine platelet membranes maintained an average linear deformation behavior: L _p/R _p = 146,100p _i × R _p + 19.923, where L _p is the platelet length aspirated in the micropipette (m), R _p is micropipette radius (m) and p _i is the aspiration pressure (Pa). The theoretical model was used to generate material constants for the murine platelet membrane that allowed for an accurate simulation of the micropipette aspiration experiments. From published results, another set of material constants was established for the human platelet membrane. Limited cases of platelet lysis upon aspiration were analyzed using the theoretical model to determine preliminary membrane tension strength values.     

pH Homeostasis and Citric Acid Utilization: Differences Between Leuconostoc mesenteroides and Lactococcus lactis

This study presents the effects of citric acid and extracellular pH (pH_e) on the intracellular pH (pH_i) of wild-type and citrate negative variants (cit⁻) Leuconostoc mesenteroides subsp. mesenteroides (Ln. mesenteroides M) and Lactococcus lactis subsp. lactis bv. diacetylactis (L. lactis LD). A recent method using a pH-sensitive fluorescent indicator carboxyfluorescein succinimidyl ester (cFSE) was adapted to measure the pH_i of these two lactic acid bacteria in resting cells. Energized cells with 10 mM lactose of Ln. mesenteroides M and L. lactis LD modified their pH gradient (ΔpH) in the same manner; when the pH_e was decreased from 7 to 4, the pH_i decreased from 7 to about 5. The adjunction of 10 mM citric acid had no effect on the pH_i of wild-type and cit(−) variant of L. lactis LD, nor on the pH_i of Ln. mesenteroides cit(−) variant. Nevertheless, in Ln. mesenteroides M wild-type, citric acid utilization increased the pH_i, which was maintained at about 6.5–7.0 when the pH_e was decreased from 7 to 4. It could be concluded that citric acid allows the maintenance of pH homeostasis in Leuconostoc mesenteroides. Received: 7 March 1997 / Accepted: 14 April 1997     

Carbon dioxide supersaturation in Florida lakes

We examined data on CO₂ and related limnological and geographic information from a sample of 948 Florida freshwater lakes. The objectives for this study were (1) to determine the partial pressures of carbon dioxide (ρCO₂) in the surface waters of a large sample of Florida lakes, (2) to determine if several limnological or geographic factors are related to levels of ρCO₂ in Florida lakes, and (3) to estimate the net annual rate of loss of CO₂ to the atmosphere from the freshwater lakes of Florida. The calculated ρCO₂ for the lakes in our sample range from 0 to 81,000 μatm, with a mean of 3,550 μatm, a median of 1,030 μatm, and a geometric mean of 1,270 μatm. About 87% of the Florida lakes were supersaturated with CO₂. There were statistically significant correlations between values for ρCO₂ and several water chemistry variables; however, the R ² values were small and accounted for only a small portion of the variance. In general the ρCO₂ values were higher in the lakes with low alkalinities and low contents of dissolved salts. The best predictor of ρCO₂ is pH, with an R ² of 0.82 for a polynomial relationship. The ρCO₂ values tend to decrease from northwest to southeast across the state of Florida, which corresponds to the gradients we found for pH, alkalinity, and specific conductance. The average areal rate of carbon emission from the Florida lakes was 328 g C m⁻² y⁻¹, and the total carbon loss for the lakes and ponds of Florida was 2.0 Tg y⁻¹. This amounts to about 2% of the total carbon emissions from all the lakes of the world as estimated by previous studies. Handling editors: Darren Bade