Carbon isotope effect on dehydration of bicarbonate ion catalyzed by carbonic anhydrase

The carbon-13 kinetic isotope effect on the dehydration of HCO3- by bovine carbonic anhydrase has been measured. To accomplish this, bicarbonate was added to a buffer solution at pH 8 containing carbonic anhydrase under conditions where purging of the product CO2 from the solution is rapid. Measurement of the isotopic composition of the purged CO2 as a function of the concentration of carbonic anhydrase permits calculation of the isotope effect on the enzymic reaction. The isotope effect on the dehydration is k12/k13 = 1.0101 +/- 0.0004. This effect is most consistent with a ping-pong mechanism for carbonic anhydrase action, in which proton transfer to or from the enzyme occurs in a step separate from the dehydration step. Substrate and product dissociation steps are at least 2-3-fold faster than the hydration/dehydration step.     

The spatial scale of competition from recruits on an older cohort in Atlantic salmon

Competitive effects of younger cohorts on older ones are frequently assumed to be negligible in species where older, larger individuals dominate in pairwise behavioural interactions. Here, we provide field estimates of such competition by recruits on an older age class in Atlantic salmon (Salmo salar), a species where observational studies have documented strong body size advantages which should favour older individuals in direct interactions. By creating realistic levels of spatial variation in the density of underyearling (YOY) recruits over a 1-km stretch of a stream, and obtaining accurate measurements of individual growth rates of overyearlings (parr) from capture–mark–recapture data on a fine spatial scale, we demonstrate that high YOY density can substantially decrease parr growth. Models integrating multiple spatial scales indicated that parr were influenced by YOY density within 16 m. The preferred model suggested parr daily mass increase to be reduced by 39% when increasing YOY density from 0.0 to 1.0 m⁻², which is well within the range of naturally occurring densities. Reduced juvenile growth rates will in general be expected to reduce juvenile survival (via increased length of exposure to freshwater mortality) and increase generation times (via increased age at seaward migrations). Thus, increased recruitment can significantly affect the performance of older cohorts, with important implications for population dynamics. Our results highlight that, even for the wide range of organisms that rely on defendable resources, the direction of competition among age classes cannot be assumed a priori or be inferred from behavioural observations alone.     

Effects of diurnally oscillating pCO2 on the calcification and survival of coral recruits

Manipulative studies have demonstrated that ocean acidification (OA) is a threat to coral reefs, yet no experiments have employed diurnal variations in pCO(2) that are ecologically relevant to many shallow reefs. Two experiments were conducted to test the response of coral recruits (less than 6 days old) to diurnally oscillating pCO(2); one exposing recruits for 3 days to ambient (440 μatm), high (663 μatm) and diurnally oscillating pCO(2) on a natural phase (420-596 μatm), and another exposing recruits for 6 days to ambient (456 μatm), high (837 μatm) and diurnally oscillating pCO(2) on either a natural or a reverse phase (448-845 μatm). In experiment I, recruits exposed to natural-phased diurnally oscillating pCO(2) grew 6-19% larger than those in ambient or high pCO(2). In experiment II, recruits in both high and natural-phased diurnally oscillating pCO(2) grew 16 per cent larger than those at ambient pCO(2), and this was accompanied by 13-18% higher survivorship; the stimulatory effect on growth of oscillatory pCO(2) was diminished by administering high pCO(2) during the day (i.e. reverse-phased). These results demonstrate that coral recruits can benefit from ecologically relevant fluctuations in pCO(2) and we hypothesize that the mechanism underlying this response is highly pCO(2)-mediated, night-time storage of dissolved inorganic carbon that fuels daytime calcification.     

Effect of hydrochlorothiazide on urine saturation with brushite,in vitro collagen calcification by urine,and urinary inhibitors of collagen calcification

To clarify further the beneficial effect of thiazide diuretics on recurrent calcium nephrolithiasis, the effect of short-term hydrochlorothiazide therapy on urine saturation with brushite (CaHPO₄·2H₂O), in vitro collagen calcification by urine, and urinary inhibitors of calcification was studied.In 22 patients with idiopathic calcium oxalate/phosphate stones the urine calcium excretion decreased, the urine magnesium excretion increased and the urine magnesium/calcium ratio increased significantly (P < 0.001) during hydrochlorothiazide therapy. Supersaturation of the urine with brushite, which was present in 19 of the 22 patients, was reduced significantly (P < 0.001) in all during thiazide therapy, and to the undersaturated range in 16. The ability of urine to calcify collagen in vitro also decreased significantly (P < 0.001) during thiazide therapy, a change that correlated significantly (r = 0.4513, P < 0.05) with the decrease in brushite saturation. The concentration of urinary inhibitors of calcification, as determined with an in vitro collagen calcification system, was decreased significantly (P < 0.01) by thiazide therapy.It was concluded that, in addition to decreasing urine calcium excretion and increasing urine magnesium excretion, thiazide diuretics decrease the urinary brushite saturation and thus may prevent spontaneous nucleation or crystal growth, or both, of calcium phosphate. The ability of thiazides to decrease collagen calcification in vitro suggests that they may also prevent crystal growth on a nidus of organic matrix. Thiazides do not appear to act by increasing the excretion of urinary inhibitors of calcification.     

Effect of water currents on organic matter release by two scleractinian corals

Organic matter release by scleractinian corals fulfils an important ecological role as energy carrier and particle trap in reef ecosystems, but the hypothetically stimulating impact of water currents, an essential and ubiquitous environmental factor in coral reefs, on this process has not been investigated yet. This study therefore quantifies organic matter release by two species of scleractinian corals subjected to ambient water current velocities ranging from 4 to 16?cm?s^?1 using closed-system flow-through chambers. Findings revealed that particulate organic matter (POM) concentration was significantly increased in the flow-through chambers in all investigated coral species compared to still water conditions, while no effect on dissolved organic carbon (DOC) concentration could be observed. These results suggest that POM release by corals may be controlled by hydro-mechanical impacts, while DOC fluxes are rather influenced by the physiological condition of the corals. Hence, this study indicates that previous POM release quantification results are conservative estimates and may have underestimated in situ POM release through corals in reef environments. The contribution of coral-derived POM to biogeochemical cycles in reef ecosystems, therefore, may be more pronounced than already assumed.     

Microanalysis of C and O isotopes of azooxanthellate and zooxanthellate corals by ion microprobe

We have determined the ¹⁸O and ¹³C values of azooxanthellate (Lophelia pertusa) and zooxanthellate (Porites lutea) corals at a micrometer scale using an ion microprobe (SIMS—secondary ion mass spectrometry). In P. lutea, centers of calcification are small (10 to 15 m) and difficult to locate during measurements. In L. pertusa, they are large (50 m) and arranged in lines of centers of calcification. Our results show that centers of calcification in L. pertusa have a restricted range of variation in ¹⁸O [-2.8±0.3 (PDB)], and a larger range in ¹³C [14.3 to 10.9 (PDB)]. Surrounding skeletal fibers exhibit large isotopic variation both for C and O (up to 12), and ¹³C and ¹⁸O are positively correlated. The C and O isotopic compositions of the center of calcification deviate from this linear trend at the lightest ¹⁸O values of the surrounding fibers. Ion microprobe results on P. lutea demonstrate also a large range of variation for the ¹⁸O values (up to 10). No correlation is found with C isotopes that exhibit, in comparison with L. pertusa, a small range of variation (2). This variation of ¹⁸O at a micrometer scale is probably the result of two processes: (1) an isotopic equilibrium calcification with 1 pH unit variation in the calcification fluid as indicated by direct measurements of coelenteron pH in the coral Galaxea fascicularis (Al-Horani et al. 2003) and (2) a kinetic fractionation. The ¹³C apparent disequilibrium in P. lutea may be the result of mixing between metabolic CO₂ (respiration) and dissolved inorganic carbon (DIC) coming directly from seawater.     

Effects of bicarbonate and oxygen concentration on photoinhibition of thylakoid membranes

The effects of bicarbonate and oxygen on photoinhibition of thylakoid membranes were investigated by varying their concentrations independently of each other. A pretreatment of the thylakoid suspension which lowered the bicarbonate concentration of the medium without affecting its oxygen content, increased the degree of photoinhibition upon illumination. This showed that the normal bicarbonate content of a thylakoid suspension, originating from dissolved carbondioxide from the air, protects against photoinhibition. The resistance against photoinhibition was further increased by addition of extra NaHCO₃ up to about 5 mM. The normal oxygen content can be decreased profoundly without affecting the degree of photoinhibition; in contrast, even small changes from the normal bicarbonate content affected photoinhibition.At oxygen concentrations approximately below 25 M, added NaHCO₃ not only did not protect, but caused a more severe PS 2 inactivation. This was due to a blockage by added NaHCO₃ of the recovery from a reversible photoinhibited state.Furthermore, it is shown that if the bicarbonate ions bound to high-affinity sites in PS 2 were replaced by formate ions, the thylakoid membranes became less susceptible to photoinhibition under normal oxygen tension.Abbreviations chl chlorophyll - HEPES (N-[2-Hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]) - PpBQ phenyl-p-benzoquinone - PS 2 Photosystem 2 - Q_A and Q_B primary and secondary quinone acceptors of Photosystem 2     

The effect of unconscious color hue saturation on emotional state of human

The aim of the study was to investigate influence of color hue saturation on emotional state of human. We use frontal EEG asymmetry to determine subject's emotional state. Our emotional stimuli summon opposite dynamics of frontal EEG asymmetry. Negative stimuli elicits decreasing of the value of frontal EEG asymmetry and positive stimuli increases the value of frontal EEG asymmetry in fronto-polar and frontal leads. Such dynamics of frontal EEG asymmetry point the emotional experience in accordance the stimulus modality. Blue and red color modification of stimuli leads changes in dynamics of frontal EEG asymmetry during presentation of emotional stimuli and after. In fact, that no one subject gave a report about color difference between stimuli during an experiment, we conclude that influence of color modification was unconscious. Our result shows the possibility of unconscious perception color modification to emotional state of human.     

Effect of hydrogen ion concentration in the medium on the state of chromatin in lymphoid cells

Effect of medium acidity (pH 7.2 divided by 6.6) on the state of lymphoid tissue cells was studied by microspectrofluorimetry, electrophoresis and IR-spectroscopy. Two stages of the cell structural-functional changes were found at H2CO3 acidification of the medium. The first one is a reversible transition of chromatin from the state of dense sphere to the margination state when chromatin is located along the membrane. The second one is an irreversible destruction of chromatin and the cell death. The margination stage is related to the formation of non-specific DNA-membrane contacts, while the chromatin destruction stage to the "proton breakdown" of the macromolecules and their supramolecular complexes. A relationship between the degree of margination reversibility and the value of the medium acidification and the time of cell incubation in it is shown. Different functional significance of chromatin marginated state is discussed.     

The effect of unconscious color hue saturation on the emotional state of humans

The effect of color hue saturation on the emotional state (ES) of humans has been investigated. Frontal EEG asymmetry was used to determine the subject’s ES. The emotionogenic stimuli caused converse dynamics of frontal EEG asymmetry. Negative stimuli elicited a decrease in the value of frontal EEG asymmetry, and positive stimuli increased the value of frontal EEG asymmetry in the anterofrontal and posterofrontal leads. These dynamics of frontal EEG asymmetry indicates the formation of an ES corresponding to the presented stimuli. Blue and red color balance modification of stimuli leads to changes in the dynamics of frontal EEG asymmetry during the scanning of emotionally positive and negative stimuli. None of the subjects noticed the color modification of the video stimuli used. This leads us to the conclusion that the change in the frontal asymmetry in response to modified stimuli resulted from an unconscious perception of the modification of the color characteristics of the stimuli. The results show the possibility of the effect of unconscious perception of color characteristics of video stimuli on the ES of humans.     

The impact of two pesticides on olfactory-mediated endocrine function in mature male Atlantic salmon (Salmo salar L.) parr

Short-term exposure of the olfactory epithelium of mature male Atlantic salmon parr to either the pesticide simazine (concentrations 1.0 and 2.0 microg l(-1)) or the pesticide atrazine (concentration 1.0 microg l(-1)) significantly reduced the olfactory response to the female priming pheromone, prostaglandin F(2alpha). In addition, the reproductive priming effect of the pheromone on the levels of expressible milt was also reduced after exposure to the individual pesticides (simazine 0.1, 0.5, 1.0 and 2.0 microg l(-1) and atrazine 0.5 and 2.0 microg l(-1)). When the olfactory epithelium was exposed to a mixture of simazine and atrazine, (concentrations of 0.5:0.5 and 1.0:1.0 microg l(-1)), there was no significant reduction in the olfactory response when compared to the single pesticides at equivalent concentrations. In addition, exposure to a mixture of simazine and atrazine had no synergistic effect on the priming response, and plasma levels of testosterone, 11-ketotestosterone and 17,20beta-dihydroxy-4-pregnen-3-one were similar in the groups of male parr exposed to the individual pesticides. Although the levels of expressible milt were reduced in all groups, there were no significant differences between the different pesticide treatments. The results of the study suggest that the two s-triazine pesticides have an additive and not a synergistic impact on olfactory-mediated endocrine function in mature male salmon parr.     

The regulation of extramitochondrial free calcium ion concentration by rat liver mitochondria

The mechanism whereby rat liver mitochondria regulate the extramitochondrial concentration of free Ca(2+) was investigated. At 30 degrees C and pH7.0, mitochondria can maintain a steady-state pCa(2+) (0) (the negative logarithm of the free extramitochondrial Ca(2+) concentration) of 6.1 (0.8mum). This represents a true steady state, as slight displacements in pCa(2+) (0) away from 6.1 result in net Ca(2+) uptake or efflux in order to restore pCa(2+) (0) to its original value. In the absence of added permeant weak acid, the steady-state pCa(2+) (0) is virtually independent of the Ca(2+) accumulated in the matrix until 60nmol of Ca(2+)/mg of protein has been taken up. The steady-state pCa(2+) (0) is also independent of the membrane potential, as long as the latter parameter is above a critical value. When the membrane potential is below this value, pCa(2+) (0) is variable and appears to be governed by thermodynamic equilibration of Ca(2+) across a Ca(2+) uniport. Permeant weak acids increase, and N-ethylmaleimide decreases, the capacity of mitochondria to buffer pCa(2+) (0) in the region of 6 (1mum-free Ca(2+)) while accumulating Ca(2+). Permeant acids delay the build-up of the transmembrane pH gradient as Ca(2+) is accumulated, and consequently delay the fall in membrane potential to values insufficient to maintain a pCa(2+) (0) of 6. The steady-state pCa(2+) (0) is affected by temperature, incubation pH and Mg(2+). The activity of the Ca(2+) uniport, rather than that of the respiratory chain, is rate-limiting when pCa(2+) (0) is greater than 5.3 (free Ca(2+) less than 5mum). When the Ca(2+) electrochemical gradient is in excess, the activity of the uniport decreases by 2-fold for every 0.12 increase in pCa(2+) (0) (fall in free Ca(2+)). At pCa(2+) (0) 6.1, the activity of the Ca(2+) uniport is kinetically limited to 5nmol of Ca(2+)/min per mg of protein, even when the Ca(2+) electrochemical gradient is large. A steady-state cycling of Ca(2+) through independent influx and efflux pathways provides a model which is kinetically and thermodynamically consistent with the present observations, and which predicts an extremely precise regulation of pCa(2+) (0) by liver mitochondria in vivo.     

The effect of heterotrophy on photosynthesis and tissue composition of two scleractinian corals under elevated temperature

Exogenous food can increase protein levels of coral host tissue, zooxanthellae densities, chlorophyll (chl) concentrations and rates of photosynthesis and is thought to play an important role in the resilience of bleached corals. There is however no information about the effect of heterotrophy on the bleaching susceptibility of corals under elevated temperature conditions. This study investigates potential interactions between food availability, basal metabolic functions (photosynthesis and respiration), energy status (lipid concentrations), total protein concentrations and the bleaching susceptibility (loss of chl and/or zooxanthellae) of the scleractinian corals Stylophora pistillata (Esper) and Galaxea fascicularis (Linnaeus) in response to elevated temperature (daily temperature rises of 3-4 °C) over 15 days. Feeding experiments were carried out in which the corals were either fed daily with zooplankton or starved. Compared to fed corals, starvation of both species resulted in a significant decrease in daily photosynthetic oxygen evolution over time. Gross (Pg) and net (Pn) photosynthetic production of starved corals of both species between 10:00-11:00 hrs had declined by ~50% at day 15 while there were no marked changes in Pg and Pn of fed corals. After 15 days, starved S. pistillata contained significantly lower zooxanthellae densities, lipid and protein concentrations than fed corals. Starved G. fascicularis also displayed a decrease in zooxantllae densities which was accompanied by a significant decline in algal chl concentrations. Contrary to S. pistillata, feeding treatment had no effect on the lipid concentrations of G. fascicularis. Total protein concentrations however were significantly lower in straved than in fed G. fascicularis. Furthermore, starvation resulted in a significant decrease in respiration of S. pistillata during the last four days of the experiment while treatment had no effect on the respiration rates of G. fascicularis. Overall the oxygen consumption of S. pistillata of both treatments was about 39-67% higher than the respiration of G. fascicularis indicating that low metabolic rates may have allowed starved G. fascicularis to conserve energy reserves over the course of the experiment. The combined results reveal a strong positive relationship between food availability, sustained photosynthetic activity and reduced loss in pigmentation of both species under elevated temperature conditions.