Effects of environmental oxygen on development and respiration of Australian lungfish (<Emphasis Type="Italic">Neoceratodus forsteri</Emphasis>) embryos

The effects of oxygen partial pressure ( P_\textO2 P_{{{\text{O}}_{2} }} ) on development and respiration were investigated in the eggs of the Australian lungfish, Neoceratodus forsteri. At 20°C, embryonic survival and development was optimal at 15 and 20.9 kPa. Development was slowed at 5 and 10 kPa and embryos did not survive 2 kPa. At lower P_\textO2 P_{{{\text{O}}_{2} }} , the rate of oxygen consumption also decreased. Embryos responded to hypoxia by hatching at an earlier age and stage of development, and hatching wet and dry gut-free masses were reduced. The role of oxygen conductance ( G_\textO2 G_{{{\text{O}}_{2} }} ) in gas exchange was also examined under selected environmental P_\textO2 P_{{{\text{O}}_{2} }} and temperatures. The breakdown of the vitelline membrane changed capsule geometry, allowed water to be absorbed into the perivitelline space and increased capsule G_\textO2 G_{{{\text{O}}_{2} }} . This occurred at embryonic stage 32 under all treatments and was largely independent of both P_\textO2 P_{{{\text{O}}_{2} }} and temperature (15, 20 and 25°C), demonstrating that capsule G_\textO2 G_{{{\text{O}}_{2} }} cannot adaptively respond to altered environmental conditions. The membrane breakdown increased capsule diffusive G_\textO2 G_{{{\text{O}}_{2} }} and stabilised perivitelline P_\textO2 P_{{{\text{O}}_{2} }} , but reduced the convective G_\textO2 G_{{{\text{O}}_{2} }} of the perivitelline fluid, as the large perivitelline volume and inadequate convective current resulted in a P_\textO2 P_{{{\text{O}}_{2} }} gradient within the egg prior to hatch.     

Environment and feeding change the ability of heart rate to predict metabolism in resting Steller sea lions (<Emphasis Type="Italic">Eumetopias jubatus</Emphasis>)

The ability to use heart rate (fh) to predict oxygen consumption rates ( [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} ) in Steller sea lions and other pinnipeds has been investigated in fasting animals. However, it is unknown whether established fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationships hold under more complex physiological situations, such as when animals are feeding or digesting. We assessed whether fh could accurately predict [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} in trained Steller sea lions while fasting and after being fed. Using linear mixed-effects models, we derived unique equations to describe the fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship for fasted sea lions resting on land and in water. Feeding did not significantly change the fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship on land. However, Steller sea lions in water displayed a different fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship after consuming a 4-kg meal compared with the fasting condition. Incorporating comparable published fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} data from Steller sea lions showed a distinct effect of feeding after a 6-kg meal. Ultimately, our study illustrated that both feeding and physical environment are statistically relevant when deriving [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} from telemetered fh, but that only environment affects the practical ability to predict metabolism from fh. Updating current bioenergetic models with data gathered using these predictive fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} equations will yield more accurate estimates of metabolic rates of free-ranging Steller sea lions under a variety of physiological, behavioral, and environmental states.     

Changes in partial pressures of respiratory gases during submerged voluntary breath hold across odontocetes: is body mass important?

Odontocetes have an exceptional range in body mass spanning 10³ kg across species. Because, size influences oxygen utilization and carbon dioxide production rates in mammals, this lineage likely displays an extraordinary variation in oxygen store management compared to other marine mammal groups. To examine this, we measured changes in the partial pressures of respiratory gases ( P_\textO2 P_{{{\text{O}}_{2} }} , P_\textCO2 P_{{{\text{CO}}_{2} }} ), pH, and lactate in the blood during voluntary, quiescent, submerged breath holds in Pacific white-sided dolphins (Lagenorhynchus obliquidens), bottlenose dolphins (Tursiops truncatus), and a killer whale (Orcinus orca) representing a mass range of 96–3,850 kg. These measurements provided an empirical determination of the effect of body size on the variability in blood biochemistry during breath hold and experimentally determined aerobic dive limits (ADL) within one taxonomic group (odontocetes). For the species in this study, maximum voluntary breath-hold duration was positively correlated with body mass, ranging from 3.5 min in white-sided dolphins to 13.3 min for the killer whale. Variation in breath-hold duration was associated with differences in the rate of change for P_\textO2 P_{{{\text{O}}_{2} }} throughout breath hold; P_\textO2 P_{{{\text{O}}_{2} }} decreased twice as fast for the two smaller species (−0.6 mmHg O₂ min⁻¹) compared to the largest species (−0.3 mmHg O₂ min⁻¹). In contrast, the rate of increase in P_\textCO2 P_{{{\text{CO}}_{2} }} during breath hold was similar across species. These results demonstrate that large body size in odontocetes facilitates increased aerobic breath-hold capacity as mediated by decreased mass-specific metabolic rates (rates of change in P_\textO2 P_{{{\text{O}}_{2} }} served as a proxy for oxygen utilization). Indeed the experimentally determined 5 min ADL for bottlenose dolphins was surpassed by the 13.3 min maximum breath hold of the killer whale, which did not end in a rise in lactate. Rather, breath hold ended voluntarily as respiratory gases and pH fell within a narrow range for both large and small species, likely providing cues for ventilation.     

Changes in redox states of respiratory pigments recorded from the eyes of live blowflies exposed to light stimuli and hypoxia

Time courses of mitochondrial responses to illumination-induced physiological loads and to hypoxia, were recorded optically from eyes of blowflies Calliphora vicina chalky. We isolated changes in redox states of haems a₃, a, c, and b. Two types of responses to light stimulation were observed. Haems b and a₃ responded with transient oxidation and haems a and c with reduction. The same two groups emerged in response to anoxic exposure. The onset of reduction of haems a and c had virtually no latency, while haems a₃ and b exhibited a transient oxidation followed by reduction only after 10–20 s. The dependence of the steady-state reduction level on P_\textO2 P_{{{\text{O}}_{2} }} produced the same groups. Haems a and c were significantly reduced at P_\textO2 P_{{{\text{O}}_{2} }} levels around 10 kPa while with haems b and a₃ load-induced oxidation was only replaced by reduction below 2 kPa. We propose haems respond to physiological loads in accordance with their steady-state reduction, which in turn depends largely on barriers for electron transport imposed by the mitochondrial membrane potential. We also propose it may be possible to assess the values of tissue P_\textO2 P_{{{\text{O}}_{2} }} and O₂ consumption by monitoring haems that are highly oxidized at rest such as haem a.     

AMP-activated protein kinase plays a role in initiating metabolic rate suppression in goldfish hepatocytes

In the present study, we test the hypothesis that AMP-activated protein kinase (AMPK) initiates metabolic rate suppression in isolated goldfish hepatocytes. To accomplish this, we attempted to pharmacologically activate AMPK in goldfish hepatocytes with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) and the thienopyridone, A769662, to examine the effects of AMPK activation on eukaryotic elongation factor-2 (eEF2), protein synthesis, and cellular oxygen consumption rate ( [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} ). Goldfish hepatocytes treated with 1 mM AICAR under normoxic conditions (>200 μM O₂) showed a modest but significant 1.1-fold increase in AMPK phosphorylation, a 7.5-fold increase in AMPK activity, a 1.4-fold increase in eEF2 phosphorylation, and a 24% decrease in [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} . At physiologically relevant [O₂] (<40 μM O₂), the addition of 1 mM AICAR resulted in only a 13% decrease in cellular [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} with no change in sensitivity to [O₂] as assessed by estimates of cellular P₅₀ and P₉₀ values. The addition of compound C, a general protein kinase inhibitor, after AICAR incubation did not reverse the effects of AICAR on [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} in normoxia. Treatment of hepatocytes with ≤200 μM A769662 did not affect AMPK activity, AMPK phosphorylation, eEF2 phosphorylation, or cellular [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} . These data suggest that A769662 is not an activator of AMPK in goldfish hepatocytes. Although our study provides support for the hypothesis that AMPK plays a role in initiating metabolic rate suppression in goldfish hepatocytes, this support must be viewed cautiously because of the known off-target effects of the pharmacological agents used.     

Cardiorespiratory responses to hypoxia in the African catfish, <Emphasis Type="Italic">Clarias gariepinus</Emphasis> (Burchell 1822), an air-breathing fish

The African catfish, Clarias gariepinus, possesses a pair of suprabranchial chambers located in the dorsal-posterior part of the branchial cavity having extensions from the upper parts of the second and fourth gill arches, forming the arborescent organs. This structure is an air-breathing organ (ABO) and allows aerial breathing (AB). We evaluated its cardiorespiratory responses to aquatic hypoxia. To determine the mode of air-breathing (obligate or accessory), fish had the respiratory frequency (f _R) monitored and were subjected to normoxic water (PwO₂ = 140 mmHg) without becoming hyperactive for 30 h. During this period, all fish survived without displaying evidences of hyperactivity and maintained unchanged f _R, confirming that this species is a facultative air-breather. Its aquatic O₂ uptake ( [(V)\dot]\textO₂ \dot{V}{\text{O}}_{2} ) was maintained constant down to a critical PO₂ (PcO₂) of 60 mmHg, below which [(V)\dot]\textO₂ \dot{V}{\text{O}}_{2} declined linearly with further reductions of inspired O₂ tension (PiO₂). Just above the PcO₂ the ventilatory tidal volume (V _T) increased significantly along with gill ventilation ( [(V)\dot]_\textG \dot{V}_{\text{G}} ), while f _R changed little. Consequently, the water convection requirement ( [(V)\dot]_\textG /[(V)\dot]\textO₂ ) \left( {\dot{V}_{\text{G}} /\dot{V}{\text{O}}_{2} } \right) increased steeply. This threshold applied to a cardiac response that included reflex bradycardia. AB was initiated at PiO₂ = 140 mmHg (normoxia) and air-breathing episodes increased linearly with more severe hypoxia, being significantly higher at PiO₂ tensions below the PcO₂. Air-breathing episodes were accompanied by bradycardia pre air-breath, to tachycardia post air-breath.     

Nocturnal lizards from a cool-temperate environment have high metabolic rates at low temperatures

Ectotherms from low-temperature environments have higher metabolic rates at low temperatures than those from warm-temperature environments. We predicted that nocturnal lizards, which are active at much lower environmental temperatures than diurnal lizards, would also have higher metabolic rates at low temperatures, and by association a lower thermal sensitivity (Q ₁₀) than diurnal and crepuscular lizards. We measured the rate of oxygen consumption ( [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} ) of eight cool-temperate species of lizard (four nocturnal, three diurnal, and one crepuscular) at 13 and 26°C and analyzed log transformations of these data using log mass as a covariate. As expected, [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} was positively correlated with temperature in all eight species, with [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} being two to four times higher at 26°C than at 13°C. As predicted, at 13°C (but not 26°C) the [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} was significantly higher in nocturnal than diurnal lizards. Species-specific differences and mass scaling factors explain the patterns of thermal sensitivity seen among these eight lizard species. Thermal sensitivity is strongly influenced by mass, with smaller species generally having higher thermal sensitivity of their metabolic rate, and this result deserves further exploration among other ectotherms. We conclude that, along with the previously reported lower cost of locomotion found in nocturnal lizards, they also partially offset the thermal handicap of activity at low body temperatures by having an elevated [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} at lower temperatures.     

Acclimation to hypercarbia protects cardiac contractility and alters tissue carbohydrate metabolism in the Amazonian armored catfish <Emphasis Type="Italic">Pterygoplichthys pardalis</Emphasis>

The armored catfish Pterygoplichthys pardalis tolerates environmental hypercarbia, high partial pressures of CO₂ (\(P_{{{\text{CO}}_{ 2} }}\)), by preferentially protecting intracellular pH (pHi) in the face of extracellular acidosis. This response is associated with ionic changes which may disrupt contractility in cardiac muscle, and it is not known whether acclimation to hypercarbia provides protection against these changes. We studied the influence of different \(P_{{{\text{CO}}_{ 2} }}\) acclimation histories on cardiac muscle function using isometrically contracting ventricular strip preparations. Fish were held for >4 months at 21 mmHg \(P_{{{\text{CO}}_{ 2} }}\) and then exposed to normocarbia (6 mmHg \(P_{{{\text{CO}}_{ 2} }}\)) for either 15 h or 5–6 days. Acclimation to chronic hypercarbia eliminated the negative inotropic effects of in vitro hypercarbia, decreased extracellular Ca²⁺ sensitivity, and reduced maximum pacing frequency in ventricular strip preparations. Fish acclimated to chronic hypercarbia also exhibited hepatic glycogen and plasma glucose accumulation, and lower plasma lactate levels compared to fish acclimated to normocarbia for 5–6 days. We suggest chronic hypercarbia may induce cardiac remodeling to protect contractility and reduce the energetic demands of pHi regulation. The activation of HCO₃ ^? synthesis pathways may decrease glucose utilization and enhance carbohydrate stores, potentially providing protection against hypoxia, a stressor frequently encountered in conjunction with hypercarbia in the Amazon.     

Phenol biodegradation by the thermoacidophilic archaeon <Emphasis Type="Italic">Sulfolobus solfataricus</Emphasis> 98/2 in a fed-batch bioreactor

Toxic at low concentrations, phenol is one of the most common organic pollutants in air and water. In this work, phenol biodegradation was studied in extreme conditions (80°C, pH = 3.2) in a 2.7 l bioreactor with the thermoacidophilic archaeon Sulfolobus solfataricus 98/2. The strain was first acclimatized to phenol on a mixture of glucose (2000 mg l⁻¹) and phenol (94 mg l⁻¹) at a constant dissolved oxygen concentration of 1.5 mg l⁻¹. After a short lag-phase, only glucose was consumed. Phenol degradation then began while glucose was still present in the reactor. When glucose was exhausted, phenol was used for respiration and then for biomass build-up. After several batch runs (phenol < 365 mg l⁻¹), specific growth rate (μ_X) was 0.034 ± 0.001 h⁻¹, specific phenol degradation rate (q_P) was 57.5 ± 2 mg g⁻¹ h⁻¹, biomass yield (Y_X/P) was 52.2 ± 1.1 g mol⁻¹, and oxygen yield factor ( \textY_{\textX/\textO 2} ) \left( {{\text{Y}}_{{{\text{X}}/{\text{O}}_{ 2} }} } \right) was 9.2 ± 0.2 g mol⁻¹. A carbon recovery close to 100% suggested that phenol was exclusively transformed into biomass (35%) and CO₂ (65%). Molar phenol oxidation constant ( \textY_{\textO 2 /\textP} ) \left( {{\text{Y}}_{{{\text{O}}_{ 2} /{\text{P}}}} } \right) was calculated from stoichiometry of phenol oxidation and introducing experimental biomass and CO₂ conversion yields on phenol, leading to values varying between 4.78 and 5.22 mol mol⁻¹. Respiratory quotient was about 0.84 mol mol⁻¹, very close to theoretical value (0.87 mol mol⁻¹). Carbon dioxide production, oxygen demand and redox potential, monitored on-line, were good indicators of growth, substrate consumption and exhaustion, and can therefore be usefully employed for industrial phenol bioremediation in extreme environments.     

HNCA-TOCSY-CANH experiments with alternate <Superscript>13</Superscript>C-<Superscript>12</Superscript>C labeling: a set of 3D experiment with unique supra-sequential information for mainchain resonance assignment

Described here is a set of three-dimensional (3D) NMR experiments that rely on CACA-TOCSY magnetization transfer via the weak ³ \textJ_{\textCa\textCa} ^{ 3} {\text{J}}_{{{\text{C}}\alpha {\text{C}}\alpha }} coupling. These pulse sequences, which resemble recently described ¹³C detected CACA-TOCSY (Takeuchi et al. 2010) experiments, are recorded in ¹H₂O, and use ¹H excitation and detection. These experiments require alternate ¹³C-¹²C labeling together with perdeuteration, which allows utilizing the small ³ \textJ_{\textCa\textCa} ^{ 3} {\text{J}}_{{{\text{C}}\alpha {\text{C}}\alpha }} scalar coupling that is otherwise masked by the stronger ¹J_CC couplings in uniformly ¹³C labeled samples. These new experiments provide a unique assignment ladder-mark that yields bidirectional supra-sequential information and can readily straddle proline residues. Unlike the conventional HNCA experiment, which contains only sequential information to the ^{1 3} \textC^a ^{ 1 3} {\text{C}}^{\alpha } of the preceding residue, the 3D hnCA-TOCSY-caNH experiment can yield sequential correlations to alpha carbons in positions i−1, i + 1 and i−2. Furthermore, the 3D hNca-TOCSY-caNH and Hnca-TOCSY-caNH experiments, which share the same magnetization pathway but use a different chemical shift encoding, directly couple the ¹⁵N-¹H spin pair of residue i to adjacent amide protons and nitrogens at positions i−2, i−1, i + 1 and i + 2, respectively. These new experimental features make protein backbone assignments more robust by reducing the degeneracy problem associated with the conventional 3D NMR experiments.     

Effects of nitrate on intracellular nitrite and growth of Microcystis aeruginosa

Although nitrate is a macronutrient and can serve as good nitrogen source for many species of phytoplankton, high nitrate concentrations do not benefit the growth of phytoplankton. We hypothesise that algae cultured under high nitrate concentrations can accumulate intracellular nitrite, which is produced by nitrate reductase (NR) and can inhibit the growth of algae. To assess the validity of this hypothesis, Microcystis aeruginosa was grown under different nitrate concentrations from 3.57 to 21.43 mM in low CO₂ and high CO₂ conditions for 15 days. We observed that, with increasing nitrate concentrations, the intracellular nitrite concentrations of the alga increased and the growth rates and photosynthesis declined. When grown under high CO₂ conditions, M. aeruginosa showed lower intracellular nitrite concentrations and higher growth rates and \textP_\textm^\textchla {\text{P}}_{\text{m}}^{{\text{chl}}a} , \textR_\textd^\textchla {\text{R}}_{\text{d}}^{{\text{chl}}a} , α^chla than under low CO₂ conditions. These results suggest that the accumulation of intracellular nitrite could be the cause of inhibition of algal growth under high nitrate concentrations.     

Genetic effects of forest fragmentation in high-density Araucaria angustifolia populations in Southern Brazil

Araucaria angustifolia is an endangered tropical/subtropical coniferous of great interest for conservation due its economical, ecological, and social value. Only 3% of original Araucaria forests remain, which are generally confined to small forest fragments. Forest fragmentation can have serious consequences on genetic process in tree population, affecting long-term fitness and adaptability. To investigate the effects of forest fragmentation on genetic diversity and the structure of A. angustifolia populations, the genetic diversity of eight microsatellite loci was compared in four small fragmented populations (<22 ha), four tree groups (five to 11 trees) occurring in pastures and in three plots in a large continuous population. The clearest effect of fragmentation was the loss of rare alleles (p ≤ 0.05) in fragmented populations (19.4% to 47.2%) and intermediate frequency (0.05 < p ≤ 0.25) and rare alleles (p ≤ 0.05) in tree groups (19% to 86.1%) in comparison to continuous populations. Fragmented populations have significant higher fixation index ( [^(F)]_\textIS = 0.121 \widehat{F}_{\text{IS}} = 0.121 , P < 0.05) than continuous populations ( [^(F)]_\textIS = 0.083 \widehat{F}_{\text{IS}} = 0.083 , P < 0.05). High genetic differentiation was detected among tree groups ( [^(G)]_\textST^￠ = 0.258 \widehat{G}_{{{\text{ST}}}}^{\prime } = 0.258 , P < 0.01) and low among fragments ( [^(G)]_\textST^￠ = 0.031 \widehat{G}_{{{\text{ST}}}}^{\prime } = 0.031 , P < 0.05) and continuous populations ( [^(G)]_\textST^￠ = 0.026 \widehat{G}_{{{\text{ST}}}}^{\prime } = 0.026 , P < 0.05), showing a significant bottleneck effect in tree groups. Evidence that forest fragments have experienced a recent bottleneck was confirmed in at least two studied fragments. The implications of the results for conservation of the fragmented A. angustifolia populations are discussed.     

Co-regulation of dark and light reactions in three biochemical subtypes of C<Subscript>4</Subscript> species

Regulation of light harvesting in response to changes in light intensity, CO₂ and O₂ concentration was studied in C₄ species representing three different metabolic subtypes: Sorghum bicolor (NADP-malic enzyme), Amaranthus edulis (NAD-malic enzyme), and Panicum texanum (PEP-carboxykinase). Several photosynthetic parameters were measured on the intact leaf level including CO₂ assimilation rates, O₂ evolution, photosystem II activities, thylakoid proton circuit and dissipation of excitation energy. Gross rates of O₂ evolution ( J_\textO2 J_{{{\text{O}}_{2} }} , measured by analysis of chlorophyll fluorescence), net rates of O₂ evolution and CO₂ assimilation responded in parallel to changes in light and CO₂ levels. The C₄ subtypes had similar energy requirements for photosynthesis since there were no significant differences in maximal quantum efficiencies for gross rates of O₂ evolution (average value = 0.072 O₂/quanta absorbed, ~14 quanta per O₂ evolved). At saturating actinic light intensities, when photosynthesis was suppressed by decreasing CO₂, ATP synthase proton conductivity (g _H ⁺) responded strongly to changes in electron flow, decreasing linearly with J_\textO2 J_{{{\text{O}}_{2} }} , which was previously observed in C₃ plants. It is proposed that g _H ⁺ is controlled at the substrate level by inorganic phosphate availability. The results suggest development of nonphotochemical quenching in C₄ plants is controlled by a decrease in g _H ⁺, which causes an increase in proton motive force by restricting proton efflux from the lumen, rather than by cyclic or pseudocyclic electron flow.     

Energy requirements of Adélie penguin (Pygoscelis adeliae) chicks

Summary The energy requirements of Adélie penguin (Pygoscelis adeliae) chicks were analysed with respect to body mass (W, 0.145–3.35 kg, n=36) and various forms of activity (lying, standing, minor activity, locomotion, walking on a treadmill). Direct respirometry was used to measure O₂ consumption ( ) and CO₂ production. Heart rate (HR, bpm) was recorded from the ECG obtained by both externally attached electrodes and implantable HR-transmitters. The parameters measured were not affected by hand-rearing of the chicks or by implanting transmitters. HR measured in the laboratory and in the field were comparable. Oxygen uptake ranged from in lying chicks to at maximal activity, RQ=0.76. Metabolic rate in small wild chicks (0.14–0.38 kg) was not affected by time of day, nor was their feeding frequency in the colony (Dec 20–21). Regressions of HR on were highly significant (p< 0.0001) in transmitter implanted chicks (n=4), and two relationships are proposed for the pooled data, one for minor activities ( ), and one for walking ( ). Oxygen consumption, mass of the chick (2–3 kg), and duration of walking (T, s) were related as , whereas mass-specific O₂ consumption was related to walking speed (S, m·s^-1) as .Abbreviations bpm beats per minute - D distance walked (m) - ECG electrocardiogram - HR heart rate (bpm) - ns number of steps - RQ respiratory quotient - S walking speed (m·s^-1) - T time walked (s) - W body mass (kg)     

New potent biphalin analogues containing p-fluoro-L-phenylalanine at the 4,4' positions and non-hydrazine linkers

We report the synthesis and the biological evaluation of two new analogues of the potent dimeric opioid peptide biphalin. The performed modification is based on the replacement of two key structural elements of the native biphalin, namely: the hydrazine bridge which joins the two palindromic moieties and the phenylalanine residues at the 4,4′ positions of the backbone. The new analogues 9 and 10 contain 1,2-phenylenediamine and piperazine, respectively, in place of the hydrazidic linker and p-fluoro-l-phenylalanine residues at 4 and 4′ positions. Binding values are: K_\texti^m = 0.51 \textnM K_{\text{i}}^{\mu } = 0.51\,{\text{nM}} and K_\texti^d = 12.8 \textnM K_{\text{i}}^{\delta } = 12.8\,{\text{nM}} for compound 9, K_\texti^m = 0.09 \textnM K_{\text{i}}^{\mu } = 0.09\,{\text{nM}} and K_\texti^d = 0.11 \textnM K_{\text{i}}^{\delta } = 0.11\,{\text{nM}} for analogue 10.     

Longitudinal distribution of nitrate δ15N and δ18O in two contrasting tropical rivers: implications for instream nitrogen cycling

The longitudinal variations in the nitrogen (δ¹⁵N) and oxygen (δ¹⁸O) isotopic compositions of nitrate (NO₃ ^?), the carbon isotopic composition (δ¹³C) of dissolved inorganic carbon (DIC) and the δ¹³C and δ¹⁵N of particulate organic matter were determined in two Southeast Asian rivers contrasting in the watershed geology and land use to understand internal nitrogen cycling processes. The $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ became higher longitudinally in the freshwater reach of both rivers. The $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ also increased longitudinally in the river with a relatively steeper longitudinal gradient and a less cultivated watershed, while the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ gradually decreased in the other river. A simple model for the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ and the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ that accounts for simultaneous input and removal of NO₃ ^? suggested that the dynamics of NO₃ ^? in the former river were controlled by the internal production by nitrification and the removal by denitrification, whereas that in the latter river was significantly affected by the anthropogenic NO₃ ^? loading in addition to the denitrification and/or assimilation. In the freshwater-brackish transition zone, heterotrophic activities in the river water were apparently elevated as indicated by minimal dissolved oxygen, minimal δ¹³C_DIC and maximal pCO₂. The δ¹⁵N of suspended particulate nitrogen (PN) varied in parallel to the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ there, suggesting that the biochemical recycling processes (remineralization of PN coupled to nitrification, and assimilation of NO₃ ^?-N back to PN) played dominant roles in the instream nitrogen transformation. In the brackish zone of both rivers, the $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ displayed a declining trend while the $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ increased sharply. The redox cycling of NO₃ ^?/NO₂ ^? and/or deposition of atmospheric nitrogen oxides may have been the major controlling factor for the estuarine $ \delta^{15} {\text{N}}_{{{\text{NO}}_{3} }} $ and $ \delta^{18} {\text{O}}_{{{\text{NO}}_{3} }} $ , however, the exact mechanism behind the observed trends is currently unresolved.     

Long-term analysis of Hubbard Brook stable oxygen isotope ratios of streamwater and precipitation sulfate

In response to decreasing atmospheric emissions of sulfur (S) since the 1970s there has been a concomitant decrease in S deposition to watersheds in the Northeastern U.S. Previous study at the Hubbard Brook Experimental Forest, NH (USA) using chemical and isotopic analyzes ( $ \delta^{34} {\text{S}}_{{{\text{SO}}_{4} }} $ ) combined with modeling has suggested that there is an internal source of S within these watersheds that results in a net loss of S via sulfate in drainage waters. The current study expands these previous investigations by the utilization of δ¹⁸O analyzes of precipitation sulfate and streamwater sulfate. Archived stream and bulk precipitation samples at the Hubbard Brook Experimental Forest from 1968–2004 were analyzed for stable oxygen isotope ratios of sulfate ( $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ ). Overall decreasing temporal trends and seasonally low winter values of $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ in bulk precipitation are most likely attributed to similar trends in precipitation $ \delta^{18} {\text{O}}_{{{\text{H}}_{2} {\text{O}}}} $ values. Regional climate trends and changes in temperature control precipitation $ \delta^{18} {\text{O}}_{{{\text{H}}_{2} {\text{O}}}} $ values that are reflected in the $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ values of precipitation. The significant relationship between ambient temperature and the $ \delta^{18} {\text{O}}_{{{\text{H}}_{2} {\text{O}}}} $ values of precipitation is shown from a nearby site in Ottawa, Ontario (Canada). Although streamwater $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ values did not reveal temporal trends, a large difference between precipitation and streamwater $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ values suggest the importance of internal cycling of S especially through the large organic S pool and the concomitant effect on the $ \delta^{18} {\text{O}}_{{{\text{SO}}_{4} }} $ values in drainage waters.     

Genetic parameters and predicted selection responses for timber production traits in a Castanea sativa progeny trial: developing a breeding program

Total height, diameter, index volume, stem straightness, apical dominance, and survival were assessed at 8 years from seed in an open-pollinated progeny test of 36 families of European chestnut (Castanea sativa Miller) established at two sites in the Atlantic area of Galicia, Spain. Iterative spatial analysis was applied to eliminate the effect of the spatial dependence in the original data and to estimate accurately genetic parameters for evaluating the potential for selection of the measured trees. Spatial analysis was very beneficial for growth traits and survival, but less so if at all for form traits. Estimated individual heritabilities ranged from moderate to high for growth traits ([^(h)]_i² = 0.29 - 0.42 \widehat{h}_i^2 = 0.29 - 0.42 ) and stem straightness ([^(h)]_i² = 0.24 - 0.42 \widehat{h}_i^2 = 0.{24} - 0.{42} ). High coefficients of additive genetic variance were obtained for volume ( [^(\textC)]\textV_\textA = 36.5 - 41.5% \widehat{\text{C}}{{\text{V}}_{\text{A}}} = {36}.{5} - {41}.{5}\% ) and straightness ( [^(\textC)]\textV_\textA = 44.26 - 53.84% \widehat{\text{C}}{{\text{V}}_{\text{A}}} = {44}.{26} - {53}.{84}\% ). Phenotypic and estimated genetic correlations between growth traits were very high, and correlations between sites indicated that there was no important family × site interaction. No adverse correlations between traits were evident. The results indicate the ample potential for selection in the current progeny trial, where responses to within-family and combined selection for growth traits may be high. Accordingly, three selection scenarios were addressed with the aim to initiate the selection of individuals for implementing the Forest Breeding Plan of Galicia for European chestnut.