Direct electrochemical characterization of hyperthermophilic Thermococcus celer metalloenzymes involved in hydrogen production from pyruvate

The reduction potentials of the metalloproteins pyruvate ferredoxin oxidoreductase (POR), ferredoxin, and hydrogenase isolated from hyperthermophilic Thermococcus celer (Topt = 88 degrees C) were determined as a function of temperature from 10 to 85 degrees C. Square-wave voltammetry experiments were carried out on 15 microL samples directly at an unmodified "edge-polished" pyrolytic graphite electrode using MgCl2 as an electrode promoter. POR exhibited two voltammetric waves with peaks at -280 and -403 mV at room temperature, indicating multiple redox centers, and a single wave at -420 mV at 85 degrees C. These waves displayed different temperature-dependent peak positions and peak heights, indicating that these redox centers have different thermodynamic and kinetic properties. Ferredoxin displayed a single linear temperature-dependent voltammetric wave at -280 mV at room temperature and -327 mV at 85 degrees C. Hydrogenase displayed a single biphasic temperature-dependent voltammetric wave at -197 mV at room temperature and -211 mV at 85 degrees C. Thermodynamic parameters associated with electron transfer, namely standard enthalpies and entropies for the redox centers in the various proteins, are reported.     

温周期对不同地理种群棉铃虫幼虫发育及蛹滞育的影响

为了探明不同地理种群棉铃虫Helicoverpa armigera (Hiibner)对温周期的反应,本研究系统调查了棉铃虫广东广州种群(23.08°N,113.14°E)、江西水修种群(29.04°N,115.82°E)、山东泰安种群(36.15°N,116.59°E)和辽宁喀佐种群(41.34°N,120.27°E...     

Dioxygen solubility in aqueous phosphatidylcholine dispersions

The solubility of molecular oxygen, or dioxygen, in low weight percent (1.5%) sonicated dimyristoylphosphatidylcholine (DMPC) aqueous dispersions saturated with air has been measured as a function of temperature between 10 degrees C and 40 degrees C. A modified Winkler technique was used involving a dual cell coulometric titration with voltammetric endpoint detection in a mixed solvent (methanol/water). The results indicate that dioxygen is approximately four times more soluble in the liquid crystalline bilayers (above 24 degrees C) than in the gel state bilayers (below 24 degrees C). The solubility of dioxygen in the bilayer does not appear to be strongly temperature dependent on either side of the 24 degrees C phase transition. The dioxygen solubility in gel state DMPC is approximately equal to that in water at the same temperature. Our result are contrasted with recent measurements made using EPR spin labels.     

The temperature response of C(3) and C(4) photosynthesis

We review the current understanding of the temperature responses of C(3) and C(4) photosynthesis across thermal ranges that do not harm the photosynthetic apparatus. In C(3) species, photosynthesis is classically considered to be limited by the capacities of ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco), ribulose bisphosphate (RuBP) regeneration or P(i) regeneration. Using both theoretical and empirical evidence, we describe the temperature response of instantaneous net CO(2) assimilation rate (A) in terms of these limitations, and evaluate possible limitations on A at elevated temperatures arising from heat-induced lability of Rubisco activase. In C(3) plants, Rubisco capacity is the predominant limitation on A across a wide range of temperatures at low CO(2) (<300 microbar), while at elevated CO(2), the limitation shifts to P(i) regeneration capacity at suboptimal temperatures, and either electron transport capacity or Rubisco activase capacity at supraoptimal temperatures. In C(4) plants, Rubisco capacity limits A below 20 degrees C in chilling-tolerant species, but the control over A at elevated temperature remains uncertain. Acclimation of C(3) photosynthesis to suboptimal growth temperature is commonly associated with a disproportional enhancement of the P(i) regeneration capacity. Above the thermal optimum, acclimation of A to increasing growth temperature is associated with increased electron transport capacity and/or greater heat stability of Rubisco activase. In many C(4) species from warm habitats, acclimation to cooler growth conditions increases levels of Rubisco and C(4) cycle enzymes which then enhance A below the thermal optimum. By contrast, few C(4) species adapted to cooler habitats increase Rubisco content during acclimation to reduced growth temperature; as a result, A changes little at suboptimal temperatures. Global change is likely to cause a widespread shift in patterns of photosynthetic limitation in higher plants. Limitations in electron transport and Rubisco activase capacity should be more common in the warmer, high CO(2) conditions expected by the end of the century.     

Determination of nanogram quantities of osmium-labeled nucleic acids by stripping (inverse) voltammetry

Modification of nucleic acids with OSO4 in the presence of pyridine results in a formation of a covalently bound electroactive center in a polynucleotide chain detectable by polarographic (voltammetric) methods. It has been shown that DNA modified with osmium (DNA-Os) accumulates at the hanging mercury-drop electrode during a waiting time in a wide range of potentials between 0 and -1.0 V (against the saturated calomel electrode) and produce at neutral pH a well-developed reduction peak at about -1.2 V due to scanning in the cathodic direction. Using the differential-pulse stripping (inverse) voltammetry, nanogram quantities of single-stranded DNA-Os can be determined at relatively short waiting times (1-3 min). Double-stranded DNA is modified with osmium to a much lesser extent as compared to single-stranded polynucleotides. The degree of modification of double-helical DNA is influenced by the presence of single-stranded and distorted double-stranded regions in the DNA molecules and by the environmental conditions which influence the DNA conformation. Osmium can thus be used as a probe of the DNA structure, and a few micrograms of double-helical DNA sample suffice for the voltammetric analysis.     

Effect of temperature on development and survival in Delias nigrina (Fabricius) (Lepidoptera: Pieridae)

Abstract The effect of temperature on rate of development and survival of the immature stages of a subtropical population of the black jezebel, Delias nigrina , was studied under laboratory conditions at a range of constant temperatures. Mean developmental times from first-instar larva to adult varied from 29 days at 27°C to 52 days at 19°C; the development threshold temperature and thermal constant were estimated to be 9°C and 494 degree-days, respectively. Larval developmental rates reached physiological maximum at the higher temperatures tested (25−27°C). Pupal development, by contrast, was not affected in the same way as larvae by higher temperature. Survival of the immature stages varied inversely with temperature: survival was highest at 19°C and significantly reduced at 27°C. Mortality at the higher temperature was attributable mainly to final-instar larvae and pupae. These findings indicate that, compared with other tropical pierids that have been studied, D. nigrina has: (i) a comparatively low temperature threshold; (ii) a slow rate of development; and (iii) a poor tolerance to moderately high temperatures. Physiologically, these features are more characteristic of a temperate butterfly than a tropical one. This physiological response appears to be reflected by the temperate nature of the genus as a whole, which may be related to its period of origin and evolution during past climatic events.     

Shewanella putrefaciens produces an Fe(III)-solubilizing organic ligand during anaerobic respiration on insoluble Fe(III) oxides

The mechanism of Fe(III) reduction was investigated using voltammetric techniques in anaerobic incubations of Shewanella putrefaciens strain 200 supplemented with Fe(III) citrate or a suite of Fe(III) oxides as terminal electron acceptor. Results indicate that organic complexes of Fe(III) are produced during the reduction of Fe(III) at rates that correlate with the reactivity of the Fe(III) phase and bacterial cell density. Anaerobic Fe(III) solubilization activity is detected with either Fe(III) oxides or Fe(III) citrate, suggesting that the organic ligand produced is strong enough to destabilize Fe(III) from soluble or solid Fe(III) substrates. Results also demonstrate that Fe(III) oxide dissolution is not controlled by the intrinsic chemical reactivity of the Fe(III) oxides. Instead, the chemical reaction between the endogenous organic ligand is only affected by the number of reactive surface sites available to S. putrefaciens. This report describes the first application of voltammetric techniques to demonstrate production of soluble organic-Fe(III) complexes by any Fe(III)-reducing microorganism and is the first report of a Fe(III)-solubilizing ligand generated by a metal-reducing member of the genus Shewanella.     

Effect of temperature on the biology of Creontiades dilutus (Stål) (Heteroptera: Miridae)

The egg and nymphal development, fecundity and survival of the green mirid, Creontiades dilutus were examined at a range of temperatures and a modified day-degree model fitted to the data. Day degree (DD) requirements for egg and nymphal development, and threshold temperatures were calculated from the fitted lines. Female fecundity and longevity, egg and nymphal development, and survival of C. dilutus were significantly influenced by temperature. Eggs and nymphs failed to complete development at temperatures below 17 and at 38°C. Females also failed to produce any eggs at 11 and 38°C. The optimum temperature range for female fecundity was found to be 26–32°C. The optimum temperature for the development of eggs was calculated from the model as 30.5°C and for nymphs as 31.5°C. The threshold temperature for development was 15.8°C for egg and 15.1°C for nymph; 69.4 and 156.7 DD were required for completing the egg and the nymphal development, respectively. At the optimum temperature, it was estimated that development from egg to adult took 15 days. Survival was highest at 26°C for eggs and at 30–32°C for nymphs.     

Biology of Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae) on Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and Cryptolestes ferrugineus (Stephens) (Coleoptera: Cucujidae)

The aim of the present study was to assess the effect of six different temperatures on the development of Acarophenax lacunatus (Cross & Krantz) using eggs of Tribolium castaneum (Herbst) and Cryptolestes ferrugineus (Stephens) as hosts. The temperature affected the development of A. lacunatus. The largest values for the progeny (19 mites in T. castaneum and 15 mites in C. ferrugineus) were obtained at about 30 degrees C, as also observed for the net reproductive rate (Ro), which revealed that the A. lacunatus population increased 18 times in T. castaneum and 14 times in C. ferrugineus in a generation. The intrinsic rate of increase (r m) gradually increased with temperature, reaching the maximum value at 35 degrees C in T. castaneum (1,608) and C. ferrugineus (1,289). The generation time was negatively correlated with temperature, ranging from 1,60 to 4,85 days in T. castaneum and from 1,96 to 5,34 days in C. ferrugineus. These results suggest that the mite A. lacunatus may be used in programs of biological control of T. castaneum and C. ferrugineus in the tropics.     

Modelling temperature-dependent bionomics of Bemisia tabaci (Q-biotype)

Abstract.  The influence of temperature (17, 21, 25, 30 and 35 °C) on life-history traits of a Q-biotype Bemisia tabaci population on tomato is studied. Temperature-dependent relationships are characterized for immature developmental rate, immature survival, fecundity, longevity and intrinsic rate of increase. Development time vary from 20 days at 30 °C to 56 days at 17 °C and the lowest thermal threshold is estimated at 10.2 °C. The optimal temperature for immature development is 32.5 °C. Total fecundity (eggs per female) ranges from 105.3 (at 21 °C) to 41 (at 35 °C). The longevity decreases with temperature increase. The intrinsic rate of increase ranges from 0.0450 (at 17 °C) to 0.123 (at 30 °C). The functional relationships between temperature and life-history parameters are used to evaluate the effect of temperature on the population dynamics. Such mathematical relationships could provide a basis for future development of population models.     

Metabolic physiology of the numbat (Myrmecobius fasciatus)

The numbat (Myrmecobius fasciatus) is unique amongst marsupials as it is exclusively diurnal, feeds only on termites and is semi-fossorial. This study examines the thermal and metabolic physiology of the numbat to determine if its physiology reflects its phylogeny, diet and semi-fossorial habit. Numbats (mean adult body mass 552 g) were able to regulate body temperature at ambient temperatures of 15-30 degrees C, with a body temperature at thermoneutrality (30 degrees C) of 34.1 degrees C. The thermoneutral body temperature was not significantly different from that predicted for an equivalent-sized marsupial. Basal metabolic rate, measured at 30 degrees C, was 0.389 +/- 0.025 ml O(2) g(-1) h(-1), and was slightly but not significantly lower at 82.5% of that predicted for a typical marsupial of equivalent body mass. Metabolic rate increased with decreasing ambient temperatures below 30 degrees C. Patterns of metabolic cycling observed for completely inactive numbats at ambient temperatures below 30 degrees C are likely to be related to sleep phase. Wet thermal conductance of 1.94 J g(-1) h(-1) degrees C(-1) (at 30 degrees C) was 131% of that predicted for a marsupial. Evaporative water loss of the numbat remained constant below the thermoneutral zone (<30 degrees C) at approximately 0.6 ml g(-1) h(-1), only 47.4% of that predicted for a marsupial. It increased to 1.01 +/- 0.16 ml g(-1) h(-1) at an ambient temperature of 32.5 degrees C. The thermal and metabolic physiology of the numbat is generally similar to that expected for other marsupials, and is also comparable to that of termitivorous placental mammals. Thus the reduction in body temperature and basal metabolic rate of placental termitivores is a "marsupial-like" low energy turnover physiology, and the numbat being a marsupial already has an appropriate physiology to survive exclusively on a low energy diet of termites.     

Functional response of the predators Podisus maculiventris (Say) and Podisus nigrispinus (Dallas) (Het., Pentatomidae) to the beet armyworm, Spodoptera exigua (Hübner) (Lep., Noctuidae): ef fect of temperature

Predation abilities of Podisus maculiventris and Podisus nigrispinus on caterpillars of the beet armyworm, Spodoptera exigua (Hübner), were compared at three different temperatures (18, 23 and 27°C) by performing functional response tests. In both species, predation capacity was a function of temperature and prey density: more prey were captured as temperature and number of prey offered increased. Results indicated that type II and III functional responses provided the best fit to the data obtained for P. nigrispinus at 18 and 23°C, and at 27°C, respectively. However, the data for P. maculiventris showed a better fit to type II at 18°C and to type III at higher temperatures. In both pentatomids the handling time decreased with increasing temperature. At higher temperatures, P. nigrispinus demonstrated greater predation rates than P. maculiventris . The implications of these findings for the control of caterpillar pests in glasshouses are discussed.     

Thermoregulation of water collecting honey bees (Apis mellifera)

Honey bees (Apis mellifera carnica, Apidae, Hymenoptera) visited a pond in order to collect water. During their stays at the pond the body surface temperature of water foragers was measured using contactless thermography. Irrespective of the ambient temperature (T(A)) which ranged from 13.6 to 27.2 degrees C, the water carriers reached thoracic temperatures of 36-38.8 degrees C (mean values of the measuring periods). The maximum thoracic value of an individual bee was 44.5 degrees C. At higher T(A) (20.9-27.2 degrees C) head and abdomen were only about 3 degrees C and 2 degrees C on the average higher than the surroundings, respectively. In the lower range of T(A) (13.6-16.6 degrees C), however, the bees warmed their heads up to 29.2 degrees C (13 degrees C above T(A)) and the abdomen up to 23.3 degrees C (7.1 degrees C above T(A); mean values of the measuring periods).The head and abdomen were even provided independently of one another with heat from the thorax. At a higher T(A) only little heat came from the heated thorax into the abdomen, at a cooler T(A) (13.6-16.6 degrees C) more heat reached the abdomen. In all probability, at a higher T(A) only a small amount of haemolymph was pumped from the thorax into the abdomen; the most warm blood probably circulated in the head-thorax area. The average duration of stays at the pond decreased linearly from 110 to 42 s with rising T(A). Head and thorax showed great fluctuations of temperature. For example, the head was heated by 4.6 degrees C within 25 s, the thorax by 6.1 degrees C within 30 s.Foragers drinking sucrose solution are known to increase their thoracic temperature with rising concentration of the sucrose solution. The water foragers had thoracic temperatures similar to that of bees feeding on 0.5 molar sucrose solution. It is hypothesized that the foraging motivation of both groups was similar and therefore they regulated their thoraces at the same temperature level.     

Effect of temperature on life table parameters of predatory thrips Scolothrips longicornis (Thysanoptera: Thripidae) fed on twospotted spider mites (Acari: Tetranychidae)

Environmental variables such as temperature are important factors that affect the efficiency of biological control agents. This study examined the effect of temperature on the sex ratio, longevity, oviposition periods, fecundity and life table parameters of the predatory thrips Scolothrips longicornis Priesner (Thysanoptera: Thripidae) fed on twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), at six constant temperatures: 15, 20, 26, 30, 35, and 37 degrees C. Approximately 75% of the progeny were female, except at 37 degrees C, at which temperature the proportion of males increased. Adult longevity as well as the preoviposition, oviposition, and postoviposition periods decreased significantly with temperature. Thus, adults lived for approximately 5 wk at 15 degrees C and < 1 wk at 37 degrees C with preoviposition, oviposition, and postoviposition periods ranging from 6.4 to 0.4, 24.4-3.1, and 7-0.8 d between the two temperature extremes, respectively. The maximum (56.48 eggs) and minimum (11.69 eggs) value of total fecundity was recorded at 26 and 37 degrees C, respectively. The intrinsic rate of natural increase (r(m)) of S. longicornis increased linearly with increasing temperature from 0.056 d(-1) at 15 degrees C to 0.310 d(-1) at 35 degrees C. The lower temperature threshold for the population increase of S. longicornis was estimated to be around 5 degrees C. The data suggest that the upper temperature threshold for the predatory thrips is approximately 37 degrees C. The results showed that populations of S. longicornis are able to develop at a broad range of temperatures and that this predator is well adapted to the high temperatures that occur in the Mediterranean region.     

Base protonation facilitates B-Z interconversions of poly(dG-dC) X poly(dG-dC)

Comparative studies on the salt titration and the related kinetics for poly(dG-dC) X poly(dG-dC) in pH 7.0 and 3.8 solutions clearly suggest that base protonation facilitates the kinetics of B-Z interconversion although the midpoint for such a transition in acidic solution (2.0-2.1 M NaCl) is only slightly lower than that of neutral pH. The rates for the salt-induced B to Z and the reverse actinomycin D induced Z to B transitions in pH 3.8 solutions are at least 1 order of magnitude faster than the corresponding pH 7.0 counterparts. The lowering of the B-Z transition barrier is most likely the consequence of duplex destabilization due to protonation as indicated by a striking decrease (approximately 40 degrees C) in melting temperature upon H+ binding in low salt. The thermal denaturation curve for poly(dG-dC) X poly(dG-dC) in a pH 3.8, 2.6 M NaCl solution indicates an extremely cooperative melting at 60.5 degrees C for protonated Z DNA, which is immediately followed by aggregate formation and subsequent hydrolysis to nucleotides at higher temperatures. The corresponding protonated B-form poly(dG-dC) X poly(dG-dC) in 1 M NaCl solution exhibits a melting temperature about 15 degrees C higher, suggesting further duplex destabilization upon Z formation.     

Thermal physiology of the common eelpout (Zoarces viviparus)

We investigated the temperature dependence of some physiological parameters of common eelpout (Zoarces viviparus) from different locations (North Sea, Baltic Sea and Norwegian Sea) on acclimation temperature (3 degrees C and 12 degrees C) and acute temperature variation. The lethal limit of 12 degrees C-acclimated eelpout was determined as the critical thermal maximum [loss of equilibrium (LE) and onset of muscular spasms (OS)] and it was found to be 26.6 degrees C for LE and 28.8 degrees C for OS for all populations. However, these parameters do not have any relevant ecological interpretation. We therefore investigated the effect of gradually increased water temperature on standard metabolic rate (measured as resting oxygen consumption Mo2) and critical oxygen concentration ([O2]c) of eelpouts. Acclimation to low temperature (3 degrees C) resulted in partial compensation of Mo2, paralleled by a decrease of activation energy for Mo2 (from 82 kJ mol(-1) at 12 degrees C to about 50 kJ mol(-1) at 3 degrees C) in North Sea and Baltic Sea eelpouts. At the same time, Norwegian eelpout showed no acclimation of oxygen demand to warm temperature (12 degrees C) at all. The scope for eelpout aerobic metabolism shrank considerably with increased acclimation temperature, as [O2]c approached water oxygen concentrations. At 22.5+/-1 degrees C the [O2]c reached air saturation, which is equivalent to the upper critical temperature (TcII) and at this temperature the aerobic scope for the metabolism completely disappeared. In line with previous insight, the comparative analysis of the temperature dependence of Mo2 of Z. viviparus from different populations suggests that a pejus (sub-critical) temperature for this species is about 13-15 degrees C. In conclusion, the capacity to adjust aerobic metabolism relates to thermal tolerance and the bio-geographical distribution of the species. Global warming would thus be likely to cause a shift in the distribution of this species to the North.     

Diamond-like carbon (DLC) microelectrode for electrochemical ELISA

A diamond-like carbon (DLC) microelectrode was applied to commercial ELISA kits for medical diagnosis of HIV (human immunodeficiency virus), HBV (human hepatitis B virus), HCV (human hepatitis C virus). In this work, quantification of oxidized 3,3',5,5'-tetramethylbenzidine (TMB) was carried out by using a microelectrode made of boron-doped DLC and cyclic voltammetric analysis method without the conventional quenching step which uses sulfuric acid. The microelectrode provided well-known step-shaped graphs, and limit of detection could be improved by clear determination of electrochemical oxidative and reductive peaks. To demonstrate the applicability of DLC microelectrode to conventional ELISA kits, commercial ELISA kits for detection of HIV antigen, HBV antigen and HCV antigen were also tested. These results proved that the applicability of DLC microelectrode to practical detection is feasible.     

Influence of temperature on life-table parameters of Stethorus gilvifrons (Mulsant) (Coleoptera: Coccinellidae) fed on Tetranychus urticae Koch

The influence of temperature on life-table parameters, fecundity and survivorship of the predatory ladybird, Stethorus gilvifrons , fed on Tetranychus urticae was determined at seven constant temperatures of 15°C, 20°C, 25°C, 28°C, 30°C, 35°C and 40°C. No development was observed at 40°C, thus being regarded as the threshold for the development of S. gilvifrons . The results indicate a significant decrease in male and female longevity with increasing temperature from 15°C to 35°C. The longest and shortest longevity were 18.40 and 12.75 days for males and 17.40 and 8.80 days for females, respectively. The intrinsic rate of natural increase ( r _m ) and the net reproductive rate ( R ₀) of S. gilvifrons linearly increased with increasing temperatures from 15°C to 35°C, while the mean generation time ( T ) and doubling time (DT) decreased linearly within this temperature range. The highest values of r _m (0.240 females/female/day) and R ₀ (59.27 females/female) and the lowest mean generation time (17.01 days) and DT (2.88 days) were recorded at 35°C. The maximum (185.50 eggs) and minimum (25.50 eggs) measurement of total fecundity was also recorded at 35°C and 15°C, respectively. The results indicate that temperature greatly affected fecundity, survivorship and life-table parameters of S. gilvifrons , and that 35°C is a suitable temperature for population growth of this predator.     

Chromosome condensation may enhance X-ray-related cell lethality in a temperature-sensitive mutant (tsBN2) of baby hamster kidney cells (BHK21)

In the tsBN2 cell line, which has a temperature-sensitive defect in the regulatory mechanism for chromosome condensation, the lethal effect of X rays was enhanced by incubating the cells at a nonpermissive temperature (40 degrees C) following X irradiation. This enhancement was suppressed in the presence of cycloheximide, which inhibits induction of premature chromosome condensation. The findings obtained in the case of delayed incubation at 40 degrees C and in synchronized cells indicate that X-ray-related potentially lethal damage, which can be expressed by chromosome condensation, is produced in the cells at any stage of the cell cycle, but it is repairable for all cells except those at around the late G2-M phase, where chromosome condensation occurs at a permissive temperature (33.5 degrees C). These observations suggest that the high sensitivity of late G2-M cells to X rays is caused by the events associated with chromosome condensation.     

An ESR study of the Mn(II)-heavy meromyosin system.

The Mn(II)-heavy meromyosin system was studied by measuring the ESR spectrum of Mn(II). The temperature dependence of the line width parameter W(1, t) of a freshly prepared sample changes at around 7-10 degrees C, where W(1, t) is the reciprocal of the peak-to-peak height of the lowest magnetic field component of the hyperfine structure. It is shown that the change in the slope of W(1, t) at 7-10 degrees C is due to a change in the structure of Mn(II)-heavy meromyosin or a change in the interaction between Mn(II) and heavy meromyosin without ATP. This result is in accord with the recently reported observations that heavy meromysin ATPase activity showed different temperature dependence above and below 10 degrees C in the presence of Mn(II). The characteristics of the spectrum of the Mn(II)-heavy meromyosin system in the liquid state between 2 degrees C and 20 degrees C are compared with those of a frozen sample of Mn(II)-heavy meromyosin in a low temperature region (-50-0 degrees C) and with those of the lyophilized material. The forbidden transitions are observed, and hence the zero field splitting parameter can be obtained. It is 115 +/- 15 gauss at -50 degrees C, and decreases with increase of the temperature to 70 +/- 15 gauss at 20 degrees C.