Perinatal epigenetic temperature adaptation in avian species: comparison of turkey and Muscovy duck

1. Heat production (HP) and body core temperature (CT) where measured in 1- to 10-day old Muscovy ducklings and turkey chick, incubated during the last week before hatching at a lower (34.5 °C, LT-group) or at higher (38.5 °C, HT-group), than the normal temperature of 37.5 °C (control C-group).

2. In Muscovy ducklings, on the 1st day post-hatching HP was affected by exposure to low T_a of 10 °C T_a 28.2±3.9 W kg⁻¹ in the LT-group vs. 18.1±2.4 W kg⁻¹ in normal controls. On the same day, CT was higher (39.5±1.1 °C) in the HT- than in the CT-group (37.5±2.9 °C).

3. In turkeys, the relationships between T_a and HP could be described by parabola-like functions. Apart from the first day of life, the HP of the LT-group and the HT-group was higher than of the CT-group.

4. The low prenatal temperature of incubation resulted in a decrease of the preferred temperature in the LH-group and in an increase in the HT-group.

5. It is concluded that changes in incubation temperature at the end of embryonic development may induce an epigenetic temperature adaptation, which results in a long-lasting cold- and warm-adaptation in ducks but not in turkeys.

Comparative aspects of the thermal biology of the short-tailed gerbil, Desmodillus auricularis, and the bushveld gerbil, Tatera leucogaster

1. 1. The response of oxygen consumption (VO₂), thermal conductance (C_d and C_min, body temperature (T_b), and evaporative water loss (EWL) of Tatera leucogaster and Desmodillus auricularis were measured over the range of ambient temperatures (T_a) from 5–35°C.

2. 2. Basal metabolic rate (BMR) of T. leucogaster was 0.841 ± 0.049 ml O₂ g⁻¹ h⁻¹ and lower than predicted, while that of D. auricularis was similar to the expected value (1.220 ± 0.058 ml O₂ g⁻¹ h⁻¹). D. auricularis had a high, narrow thermoneutral zone (TNZ) typical of nocturnal, xerophilic, burrowing rodents.

3. 3. D. auricularis and T. leucogaster regulated T_b over the range T_a = 5–35°C and kept EWL and dry thermal conductance at a minimum below the TNZ. However, the EWL of T. leucogaster increased rapidly above T_a = 30°C.

4. 4. After comparison with data from other species, it was concluded that there is an optimum size for xeric, nocturnal, burrowing rodents.

Shifts in thermoregulatory patterns with pregnancy in the poikilothermic mammal—The naked mole-rat (Heterocephalus glaber)

1. 1. The naked mole-rat (Heterocephalus glaber) is a poikilothermic mammal. During gestation metabolic shifts that differ from both mammalian and reptilian thermoregulatory patterns occurred.

2. 2. Body temperature was directly dependent on ambient temperature. At low ambient temperatures the temperature differential (T_b − T_a) was approximately 3°C, whereas at higher ambient temperatures the temperature differential diminished.

3. 3. In early pregnancy (prior to week 3) oxygen consumption at low ambient temperatures was greater than that of non-reproductive animals. A maximal metabolic rate (3.2 ± 1.0 ml O₂ . g⁻¹ . h⁻¹) occurred at an ambient temperature of 27°C. Thereafter the endothermic pattern of metabolism with increasing ambient temperatures was evident. Oxygen consumption decreased with increasing ambient temperature to minimal rates of 1.2 ± 0.1 ml O₂ . g⁻¹ . h⁻¹ over the ambient temperature range of 31–34°C.

4. 4. Oxygen consumption in late pregnancy (1.8 ± 0.1 ml O₂ . g⁻¹ . h⁻¹) was not correlated with ambient temperature over the entire ambient temperature range measured (24–36°C).

5. 5. Differences in thermoregulation in early and late pregnancy may be attributed to different rates of heat loss as a consequence of (a) changes in surface area and body mass or (b) vascular changes. Furthermore the thermoregulatory changes in late pregnancy may indicate that maximal overall metabolic capacity had been reached, for peak resting metabolism (expressed per animal rather than per gram body mass) in early pregnancy was similar to observed metabolism in late pregnancy.

6. 6. The dissociation of metabolism from both ambient temperature and body temperature in late pregnancy could confer an energetic advantage to this arid dwelling underground inhabitant; for it may enable the breeding female to partition a greater portion of available energy into reproduction.

Intramuscular temperatures during exercise in the heat following pre-cooling and pre-heating

Pre-cooling improves heat tolerance and time to exhaustion in the heat. We tested the possibility that reduced tissue temperatures may explain this phenomenon, using three whole-body treatments: pre-cooling, thermoneutral (control) and pre-heating. Pre-cooling reduced muscle temperature (T_m) by 6.3 °C while pre-heating increased T_m 3.4 °C, relative to control. Despite this offset, T_m climbed towards a common asymptote, with pre-cooling offering no thermal protection beyond 40 min. Following pre-cooling, exercising oesophageal temperature (T_es) initially increased at 0.09 °C min⁻¹, being significantly faster than control (0.05 °C min⁻¹) and pre-heated conditions (0.03 °C min⁻¹). Pre-cooling lowered the sweat threshold and also resulted in a reduced cardiac frequency across the exercise-heat exposure. Our observations do not support the hypothesis that pre-cooling reduces T_m at the end of an exercise-heat exposure, thereby delaying the development of fatigue.     

A convenient electrochemical preparation of reduced methyl viologen and a kinetic study of the reaction with oxygen using an anaerobic stopped-flow apparatus

1. Single reduced methyl viologen (MV^.+) acts as an electron donor in a number of enzyme systems. The large changes in extinction coefficient upon oxidation (λ_max 600 nm; MV^.+, = 1.3 · 10⁴ M⁻¹ · cm⁻¹; oxidised form of methyl viologen (MV²⁺), = 0.0) make it ideally suited to kinetic studies of electron transfer reactions using stopped-flow and standard spectrophotometric techniques.

2. A convenient electrochemical preparation of large amounts of MV^.+ has been developed.

3. A commercial stopped-flow apparatus was modified in order to obtain a high degree of anaerobicity.

4. The reaction of MV^.+ with O₂ produced H₂O₂ (k > 5 · 10⁶ M⁻¹ · s⁻¹, pH 7.5, 25 °C). H₂O₂ subsequently reacted with excess MV^.+ (k = 2.3 · 10³ M⁻¹ · s⁻¹, pH 7.5, 25 °C) to produce water. The kinetics of this reaction were complex and have only been interpreted over a limited range of concentrations.

5. The results support the theory that the herbicidal action of methyl viologen (Paraquat, Gramoxone) is due to H₂O₂ (or radicals derived from H₂O₂) induced damage of plant cell membrane.     

Behavioural and autonomic thermoregulation in hamsters during microwave-induced heat exposure

1. 1.|Preferred ambient temperature (T_a) and ventilatory frequency were measured in free-moving hamsters exposed to 2450 MHz microwaves. A waveguide exposure system which permits continuous monitoring of the absorbed heat load accrued from microwave exposure was imposed with a longitudinal temperature gradient which allowed hamsters to select their preferred T_a. Ventillatory frequency was monitored remotely by analysing the rhythmic shifts in unabsorbed microwave energy passing down the waveguide.

2. 2.|Without microwave exposure hamsters selected an average T_a of 30.2°C. This preferred T_a did not change until the rate of heat absorption (SAR) from microwave exposure exceeded approx. 2 W kg⁻¹. In a separate experiment, a SAR of 2.0 W kg⁻¹ at a T_a of 30°C was shown to promote an average 0.5°C increase in colonic temperature. Hamsters maintained their ventilatory frequency at baseline levels by selecting a cooler T_a during microwave exposure. In contrast, hamsters maintained at a T_a of 30°C (without a temperature gradient) underwent a sharp increase in ventilatory frequency compared to animals allowed to select their own T_a.

3. 3.|These data support previous studies suggesting that during thermal stress behavioural thermoregulation (i.e. preferred T_a) takes prescedence over autonomic thermoregulation (i.e. ventilatory frequency). It is apparent that selecting a cooler T_a is a more efficient and/or effective than autonomic thermoregulation for dissipating a heat load accrued from microwave exposure.

Mechanism of O2 generation in reduction and oxidation cycle of ubiquinones in a model of mitochondrial electron transport systems

O⁻₂ generation in mitochondrial electron transport systems, especially the NADPH-coenzyme Q₁₀ oxidoreductase system, was examined using a model system, NADPH-coenzyme Q₁-NADPH-dependent cytochrome P-450 reductase. One electron reduction of coenzyme Q₁ produces coenzyme Q₁ and O⁻₂ during enzyme-catalyzed reduction and O₂ + coenzyme Q₁ are in equilibrium with O⁻₂ + coenzyme Q₁ in the presence of enough O₂. The coenzyme Q₁ produced can be completely eliminated by superoxide dismutase, identical to bound coenzyme Q₁₀ radical produced in a succinate/fumarate couple-KCN-submitochondrial system in the presence of O₂. Superoxide dismutase promotes electron transfer from reduced enzyme to coenzyme Q₁ by the rapid dismutation of O₂⁻ generated, thereby preventing the reduction of coenzyme Q₁ by O⁻₂. The enzymatic reduction of coenzyme Q₁ to coenzyme Q₁H₂ via coenzyme Q₁ is smoothly achieved under anaerobic conditions. The rate of coenzyme Q₁H₂ autoxidation is extremely slow, i.e., second-order constant for [O₂][coenzyme Q₁H₂] = 1.5 M⁻¹ · s⁻¹ at 258 μM O₂, pH 7.5 and 25°C.     

The respiratory chain of Azotobacter vinelandii II. The effect of cyanide on cytochrome d

1. Cyanide causes a slow disappearance of the oxidized band (648 nm) of cytochrome d in particles of Azotobacter vinelandii and inhibits the appearance of the reduced band (631 nm). No effect of cyanide is found on the reduced band of cytochrome d.

2. The kinetics of the disappearance of the 648-nm band of cytochrome d with excess cyanide deviates from first-order kinetics at lower temperatures (22 °C) indicating that at least two conformations of the enzyme are involved. At higher temperatures (32 °C) the observed kinetics of the cyanide reaction are first order with a k_on = 0.7 M⁻¹·s⁻¹ and with an estimated k_off of approximately 5·10⁻⁵ s⁻¹.

3. The value of the k_off (7·10⁻⁴−14·10⁻⁴ s⁻¹ at 32 °C) determined from the rate of reduction of cyanocytochrome d by Na₂S₂O₄ or NADH is one order of magnitude larger than the k_off value found when the enzyme is in its oxidized state.

4. No effect of cyanide is found on the spectrum of cytochrome a₁.     

Biochemical and biophysical studies on cytochrome c oxidase XV. Reaction with fluoride

1. Fluoride is a mixed-type inhibitor of the cytochrome c oxidase activity with a K_i for the free enzyme of 10 mM and a K_i for the cytochrome c-complexed enzyme of 35 mM.

2. Fluoride shifts the γ-band of the enzyme from 423 to 421 nm and the -band from 597 to 598 nm. The difference spectrum (oxidized enzyme in the presence of fluoride minus oxidized enzyme) has peaks at 400, 453, 482, 605 and 638 nm and troughs at 430, 520, 552 and 674 nm. The changes in absorbance are small (about 3% at absorbance maxima) with respect to those of other hemoproteins.

3. On addition of fluoride to isolated cytochrome c oxidase 3 reactions can be distinguished: (I) a bimolecular binding reaction (K_on = 4 M⁻¹ · s⁻¹ and k_off = 2.9 · 10⁻²s⁻¹ at 25 °C, pH 7.4) contributing at 638 nm and 430 nm; (II) a first-order reaction (k = 2.4 · 10⁻²) s⁻¹ at 22 °C, pH 7.2) visible mainly at 430 nm and (III) a very slow reaction with a half-time in the order of 10 min.

4. The spectroscopic dissociation constants for the fluoride binding, determined from Hill plots using the absorbance changes at 638 and 430 nm, are similar (7 and 10 mM, respectively, at 22 °C, pH 7.2).

5. A mechanism for the reaction is discussed in which the bimolecular binding reaction is followed by a conformational change of the enzyme-fluoride complex.     

Trypsin immobilization by direct adsorption on metal ion chelated macroporous chitosan-silica gel beads

Silica gel bead coated with macroporous chitosan layer (CTS-SiO₂) was prepared, and the metal immobilized affinity chromatographic (IMAC) adsorbents could be obtained by chelating Cu²⁺, Zn²⁺, Ni²⁺ ions, respectively on CTS-SiO₂, and trypsin could be adsorbed on the IMAC adsorbent through metal–protein interaction forces. Batch adsorption experiments show that adsorption capacity for trypsin on these IMAC adsorbent variated with change of pH. The maximal adsorption reached when the solution was in near neutral pH in all three IMAC adsorbents. Adsorption isothermal curve indicated that maximal adsorption capacity could be found in the Cu²⁺-CTS-SiO₂ with the value of 4980 ± 125 IU g⁻¹ of the adsorbent, while the maximal adsorption capacity for trypsin on Zn²⁺ and Ni²⁺ loaded adsorbent was 3762 ± 68 IU g⁻¹ and 2636 ± 53 IU g⁻¹, respectively. Trypsin immobilized on the IMAC beads could not be desorbed by water, buffer and salt solution if the pH was kept in the range of 5–10, and could be easily desorbed from the IMAC beads by acidic solution and metal chelating species such as EDTA and imidazole. The effect of chelated metal ions species on CTS-SiO₂ beads on the activity and stability of immobilized trypsin was also evaluated and discussed. Trypsin adsorbed on Zn-IMAC beads retained highest amount of activity, about 78% of total activity could be retained. Although the Cu-IMAC showed highest affinity for trypsin, only 25.4% of the calculated activity was found on the beads, while the activity recovery found on Ni-IMAC beads was about 37.1%. A remarkable difference on stability of trypsin immobilized on three kinds of metal ion chelated beads during storage period was also found. Activity of trypsin on Cu-IMAC decreased to 24% of its initial activity after 1-week storage at 4 °C, while about 80% activity was retained on both Ni-IMAC and Zn-IMAC beads. Trypsin immobilized on Zn-CTS-SiO₂ could effectively digest BSA revealed by HPLC peptide mapping.     

A study in UF-membrane reactor on activity and stability of nitrile hydratase from Microbacterium imperiale CBS 498-74 resting cells for propionamide production

The bioconversion of propionitrile to propionamide was catalysed by nitrile hydratase (NHase) using resting cells of Microbacterium imperiale CBS 498-74 (formerly, Brevibacterium imperiale). This microorganism, cultivated in a shake flask, at 28 °C, presented a specific NHase activity of 34.4 U mg_DCW⁻¹ (dry cell weight). The kinetic parameters, K_m and V_max, tested in 50 mM sodium phosphate buffer, pH 7.0, in the propionitrile bioconversion was evaluated in batch reactor at 10 °C and resulted 21.6 mM and 11.04 μmol min⁻¹ mg_DCW⁻¹, respectively. The measured apparent activation energy, 25.54 kJ mol⁻¹, indicated a partial control by mass transport, more likely through the cell wall.

UF-membrane reactors were used for kinetic characterisation of the NHase catalysed reaction. The time dependence of enzyme deactivation on reaction temperature (from 5 to 25 °C), on substrate concentrations (from 100 to 800 mM), and on resting cell loading (from 1.5 to 200 μg  ml⁻¹) indicated: lower diffusional control (E_a=37.73 kJ mol⁻¹); and NHase irreversible damage caused by high substrate concentration. Finally, it is noteworthy that in an integral reactor continuously operating for 30 h, at 10 °C, 100% conversion of propionitrile (200 mM) was attained using 200 μg  ml⁻¹ of resting cells, with a maximum volumetric productivity of 0.5 g l⁻¹ h⁻¹.     

Finger temperatures during military field training at 0 to −29 °C

1. Skin and rectal temperatures were recorded continuously in 70 measurements during typical tasks of infantry and artillery training at 0 to −29 °C. The duration of the measurements varied from 55 min to 9.5 h.

2. The distribution of finger skin temperatures was quite similar at ambient temperature ranges 0 to −10 °C and −10 to −20 °C, while at −20 to −30 °C the finger temperatures were clearly lower.

3. At different ambient temperature ranges, 20–69% of finger temperatures were low enough to cause cold thermal sensations.

4. Sensation of cold was experienced at a finger temperature of 11.6±3.7 °C (mean±SD).     

Influence of sucrose on the rheology and granule size of cross-linked waxy maize starch dispersions heated at two temperatures

Cross-linked waxy maize (CWM) starch dispersions (STDs) of concentration 50 g kg⁻¹ were heated in sucrose solutions containing 0–600 g kg⁻¹ (g sucrose/kg dispersion) at 85 °C at low shear and in intermittently agitated cans at 110 °C. The STDs heated in 0–300 g kg⁻¹ sucrose exhibited antithixotropic behavior, while those heated in 400–600 g kg⁻¹ sucrose exhibited thixotropic behavior. The mean starch granule diameter of the starch dispersions did not show strong dependence on sucrose concentration. The dispersions, especially those with high sucrose concentrations and heated at 110 °C, exhibited G′ versus frequency (ω) profiles of gels. The STDs exhibited first normal stress differences that increased in magnitude with the concentration of sucrose. Values of the first normal stress coefficient of canned dispersions calculated from dynamic rheological data plotted against ω and experimental values plotted against shear rate of some of the STDs overlapped.     

Studies on the molecular chain morphology of konjac glucomannan

The chain geometry and parameters of konjac glucomannan were studied by using laser light scatter (LLS), gel permeation chromatography (GPC) and viscosimetry. The weight-average molecular weight (M_w), root-mean-square ratio of gyration (S^21/2), second viral coefficient (A₂) and polydispersity index (M_w/M_n) were 1.036×10⁶, 105±0.9 nm, (−1.587±0.283)×10⁻³  mol ml g⁻² and 1.015±0.003 respectively. Mark-Houwink equation was established as , and the molecular chain parameters were as follows: M_L=982.82 nm⁻¹, q=27.93 nm, d=0.74 nm, h=0.26 nm, L=1054.11 nm. To confirm the above results, konjac glucomannan was observed by using atomic force microscopy (AFM) and transmission electron microscope (TEM). The physical image showed directly that the konjac glucomannan molecule was an extending semi-flexible linear chain without branches, and than the molecular dimension also conformed to the parameters above. Therefore the image of molecular chain geometry confirmed the deduction drawn by Mark-Houwink equation and molecular chain parameters magnificently.     

Effect of lactic acid fermentation of wheat and barley whole meal flour on carbohydrate composition and digestibility in mink (Mustela vison)

Wheat and barley whole meal flours (WMFs) were subjected to treatment by fermentation, autoclaving, and fermentation followed by autoclaving. The WMFs were analysed for chemical composition, formulated into wet diets (282 g kg⁻¹) and fed to adult mink (Mustela vison) for determination of total tract digestibility of total starch, total carbohydrate, crude protein and fat. Fermentation of WMF/water mixtures inoculated with a Lactobacillus sp. (strain AD₂) was performed at 30°C for 16 h. Autoclaving was carried out for 60 min at 120°C. Fermentation increased colony-forming units (CFUs) to about 10⁸ g⁻¹ and lowered pH to 3.7–3.8 in both WMFs. All carbohydrate parameters were affected by type of cereal, and were, except for total starch, affected by treatment. Levels of total dietary fibre and β-glucans decreased by fermentation in both WMFs. The decrease in total β-glucans from 33.5 to 18.4 g kg⁻¹ in barley WMF, was mainly restricted to the soluble fraction. Glucose levels in barley WMF increased simultaneously from 0.6 to 12.3 g kg⁻¹. The main effects of autoclaving were increased levels of total dietary fibre, maltose, and increased hydration capacity. With fermentation prior to autoclaving, increases in levels of the fibre fractions and maltose were prevented while hydration capacity prevailed as an effect of autoclaving. Compared with fermentation alone, the combined treatment increased damaged starch levels and hydration capacity. Digestibilities of total carbohydrate, crude protein and fat were significantly higher for wheat than for barley. Fermentation had no effect on digestibility of total starch or total carbohydrate of wheat, but increased digestibility of total starch of barley significantly from 0.742 to 0.880, and of total carbohydrate from 0.457 to 0.616. Autoclaving had no significant effect on digestibility of total starch and total carbohydrate of wheat. Digestibility of total starch and total carbohydrate in barley increased significantly after autoclaving. Total starch and total carbohydrate digestibility of both wheat and barley were significantly enhanced by combined fermentation and autoclaving compared with fermentation alone. Compared with autoclaving alone, combined fermentation and autoclaving promoted no significant improvement of total starch and total carbohydrate digestibility in wheat, whereas total carbohydrate digestibility in barley increased from 0.605 to 0.672. Fat digestibility was slightly improved by both fermentation and autoclaving. Autoclaving of cereals reduced significantly the faecal dry matter contents of mink. This effect could be counteracted by preceding fermentation. In conclusion, lactic acid fermentation of wheat and especially barley provided chemical changes of benefit for carbohydrate digestion in the mink.     

Biochemical and biophysical studies on cytochrome c oxidase XIV. The reaction with cytochrome c as studied by pulse radiolysis

1. The reduction of cytochrome c oxidase by hydrated electrons was studied in the absence and presence of cytochrome c.

2. Hydrated electrons do not readily reduce the heme of cytochrome c oxidase. This observation supports our previous conclusion that heme a is not directly exposed to the solvent.

3. In a mixture of cytochrome c and cytochrome c oxidase, cytochrome c is first reduced by hydrated electrons (k = 4 · 10¹⁰ M⁻¹ · s⁻¹ at 22 °C and pH 7.2) after which it transfers electrons to cytochrome c oxidase with a rate constant of 6 · 10⁷ M⁻¹ · s⁻¹ at 22 °C and pH 7.2.

4. It was found that two equivalents of cytochrome c are oxidized initially per equivalent of heme a reduced, showing that one electron is accepted by a second electron acceptor, probably one of the copper atoms of cytochrome c oxidase.

5. After the initial reduction, redistribution of electrons takes place until an equilibrium is reached similar to that found in redox experiments of Tiesjema, R. H., Muijsers, A. O. and Van Gelder, B. F. (1973) Biochim. Biophys. Acta 305, 19–28.     

The thermal biology of three southern African elephant-shrews

1. 1. The thermal characteristics of Petrodromus tetradactylus, Elephantulus intufi and E. brachyrhynchus were investigated and compared with other elephant-shrews that occur in the southern African subregion.

2. 2. E. intufi and E. brachyrhynchus appear to have lower than expected basal metabolic rates (1.1185 ± 0.1623 and 0.9649 ± 0.1638 ml O₂ g⁻¹ h⁻¹, respectively) and high, narrow thermoneutral zones, similar to other elephant-shrews investigated previously. In contrast P. tetradactylus has a basal metabolic rate (0.871 ± 0.027 ml O₂ g⁻¹ h⁻¹) close to expected for body mass, and a broad, low thermoneutral zone.

3. 3. The thermal biology of macroscelids is discussed in terms of their distribution, microhabitat and body size.

     

The response of an emergent sedge Bolboschoenus medianus to salinity and nutrients

The potential for nutrient load (30, 100 and 350 g N m⁻² per year) to alter plant performance under saline conditions (control, 4.5, 9 and 13 dS m⁻¹) was examined in the sedge Bolboschoenus medianus. Relative growth rates (RGR) across nutrient loadings ranged from 30.2 to 41.8 mg g⁻¹ per day in controls and were reduced to 20.9–28.5 mg g⁻¹ per day by salinities of 13 dS m⁻¹. Whilst higher nutrient loads generally increased RGR, the response was smaller at higher salinities. Responses to salinity and nutrient load were specific. Nutrient load increased the RGR via increases in the leaf area ratio (LAR). The LAR ranged from 1.9 to 2.1 m² kg⁻¹ across salinity treatments at 30 g N m⁻² per year, and increased to 2.5–2.8 m² kg⁻¹ at 350 g N m⁻² per year. Salinity reduced the RGR via a reduction in the net assimilation rate (NAR). The NAR in control plants ranged from 14.7 to 16 g m⁻² per day across nutrient loadings and decreased to 11–12 g m⁻² per day at 13 dS m⁻¹. Carbon isotope discrimination of leaves decreased by 2–3‰ in response to 13 dS m⁻¹ at the lower nutrient loadings. A prominent response of B. medianus to salinity was a change in biomass allocation from culms to tubers. In contrast, the response to nutrient load was characterised by a shift in biomass allocation from roots to leaves.     

Thermodynamics of the disproportionation of adenosine 5'-diphosphate to adenosine 5'-triphosphate and adenosine 5'-monophosphate, II. Experimental data

High-pressure liquid-chromatography and microcalorimetry have been used to determine equilibrium constants and enthalpies of reaction for the disproportionation reaction of adenosine 5′-diphosphate (ADP) to adenosine 5′-triphosphate (ATP) andadenosine 5′-monophosphate (AMP). Adenylate kinase was used to catalyze this reaction. The measurements were carried out over the temperature range 286 to 311 K, at ionic strengths varying from 0.06 to 0.33 mol kg⁻¹, over the pH range 6.04 to 8.87, and over the pMg range 2.22 to 7.16, where pMg = -log a(Mg²⁺). The equilibrium model developed by Goldberg and Tewari (see the previous paper in this issue) was used for the analysis of the measurements. Thus, for the reference reaction: 2 ADp³⁻ (ao) AMp²⁻ (ao)+ ATp⁻ (ao), K° = 0.225 ± 0.010, ΔG° = 3.70 +- 0.11 kJ mol ⁻¹, ΔH° = −1.5 ± 1. 5 kJ mol ⁻¹, °S ° = −17 ± 5 J mol⁻¹ K⁻¹, and ACP_p°≈ = −46 J mo1l⁻¹ K⁻¹ at 298.15 K and 0.1 MPa. These results and the thermodynamic parameters for the auxiliary equilibria in solution have been used to model the thermodynamics of the disproportionation reaction over a wide range of temperature, pH, ionic strength, and magnesium ion morality. Under approximately physiological conditions (311.15 K, pH 6.94, [Mg²⁺] = 1.35 × 10⁻³ mol kg⁻¹, and I = 0.23 mol kg⁻¹) the apparent equilibrium constant (K_A′ = m(ΣAMP)m(ΣATP)/[ m(ΣADP)]²) for the overall disproportionation reaction is equal to 0.93 ± 0.02. Thermodynamic data on the disproportionation reaction and literature values for this apparent equilibrium constant in human red blood cells are used to calculate a morality of 1.94 × 10⁻⁴ mol kg⁻¹ for free magnesium ion in human red blood cells. The results are also discussed in relation to thermochemical cycles and compared with data on the hydrolysis of the guanosine phosphates.     

Metal accumulation by Halodule wrightii populations

Metal concentrations and population parameters of the seagrass Halodule wrightii were determined at three locations at Rio de Janeiro State, Brazil. The possible increase of metal availability in one of these areas, Sepetiba Bay, as a result of dredging of contaminated bottom sediments which ocurred, was evaluated by analyses of Al, Cd, Cr, Cu, Fe, Ni, Pb and Zn in root, rhizome and shoots. In addition, analyses were carried out in H. wrightii populations from non-contaminated areas located at northwestern (Cabo Frio) and southeastern (Angra do Reis) regions of Rio de Janeiro State. Concurrently, abundance and density data of the seagrass populations were obtained. It was found that concentration from Sepetiba Bay samples up to 1.6 ± 0.4 μg g⁻¹ of Cd, 12 ± 1.0 μg g⁻¹ of Cr, 27 ± 2.4 μg g⁻¹ of Pb, 291 ± 47 μg g⁻¹ of Mn, 128 ± 23 μg g⁻¹ of Zn were significantly higher than that from two other collection sites. An increase in Cd and Zn concentration was observed in H. wrightii from Sepetiba Bay indicating that metal mobilization from contaminated sediments through dredging activities were, at least in part, transferred to the biotic compartment via accumulation by the seagrass. The populations of seagrass within the region demonstrated quite substantial changes in biomass data but not in shoot or rhizome density during the study. Such changes in biomass are to be expected, as these dynamics are typical of the small, isolated monospecific populations of H. wrightii along the Rio de Janeiro coast.