<Emphasis Type="SmallCaps">l</Emphasis>-carnitine modulates blood platelet oxidative stress

The oxidative stress induced by acute exertion may interfere with blood platelet activation. The beneficial effect of l-carnitine (γ-trimethylamino-β-hydroxybutyric acid) on oxidative stress in blood platelets has not been fully investigated; however, different studies indicate that this compound modulates platelet functions. The aim of our study was to assess the effects of l-carnitine on platelet activation and oxidative/nitrative protein damage (determined by the levels of protein carbonyl groups, thiol groups, and 3-nitrotyrosine residues) in resting blood platelets or platelets treated with peroxynitrite (ONOO⁻, a strong physiological oxidant) in vitro. We also investigated the effects of l-carnitine on the level of platelet glutathione and on the formation of superoxide anion radicals ( O₂ ^{- ·} ) \left( {{\hbox{O}}_2^{ - \bullet }} \right) , lipid peroxidation measured by thiobarbituric acid reactive substances (TBARS) in blood platelets stimulated by thrombin (a strong physiological agonist), and platelet aggregation induced by adenosine diphosphate (a strong physiological stimulator). We have observed that carnitine decreases platelet activation (measured by platelet aggregation, the generation of O₂ ^{- ·} {\hbox{O}}_2^{ - \bullet } , and TBARS production). Moreover, our results in vitro demonstrate that carnitine may protect against oxidation of thiol groups induced by ONOO⁻. Thus, carnitine may have some protectory effects against oxidative changes induced in blood platelets.     

Ventilation and gas exchange during rest and exercise in adult green sea turtles

Summary Ventilation and metabolic rate were measured during exercise in adult female green turtles at Tortuguero, Costa Rica. Six turtles were studied at night on the beach while actively covering their nests. Five turtles, captured after nesting, were studied at rest, during 20 min of spontaneous activity, and during recovery from the activity. Arterial blood samples were obtained from the latter animals and analyzed for pH, , O₂ concentration and lactate concentration. Blood was obtained by heart puncture from 8 turtles immediately after nesting and analyzed for blood lactate. Active metabolism ( ) in both groups was almost 10 times the standard resting value (0.024 l/kg·h). The increase in ventilation during exercise, due exclusively to higher breathing frequency, exceeded the increase in , so that the ratio (the air convection requirement), more than doubled. The respiratory exchange ratio, , that averaged 0.56 in the resting turtles, increased to 1.08 during exercise in the captured turtles and was 0.90 in the nesting animals. Arterial and O₂ saturation remained unchanged during exercise, indicating efficient gas exchange in the lungs. Pre-exercise values of all variables were restored 1 h after the end of exercise. Blood acid-base changes associated with activity in the captive turtles were variable and not statistically significant, but suggested partially compensated metabolic acidosis. Lactate concentrations were significantly elevated in the nesting turtles.     

A study of the effects of water carbonate alkalinity on some parameters of blood acid-base status in rainbow trout (Salmo gairdneri R.)

Summary The effects of two levels of water carbonate alkalinity (CA=0.3–0.5 meq·l⁻¹ and CA=12–13.5 meq·l⁻¹) on arterial blood acid-base status ( , pHa, [HCO ₃ ⁻ ]+[CO ₃ ⁻⁻ ]), oxygen consumption ( ) and plasma ionic composition (Na⁺, K⁺, Cl⁻) were investigated in trout living in normoxic-normocapnic water at 15°C. The results show that a high level of carbonate alkalinity induced a decrease in and a situation of mixed respiratory and metabolic acidosis compared to that in low CA water. These changes are accompanied by significant changes in ionic composition but the levels of oxygen consumption are unchanged. The role of the different capacitance coefficients of water for CO₂ and the effects of the different ionic composition of water on ionic and gascous exchanges are discussed. Equipe de Recherche associée au CNRS N° 070622     

Is the Mg2+-ATP-dependent proton pumping activity of the synaptic vesicles a factor involved in the cerebral hypoxia?

The changes in the Mg²⁺-dependent V-type ATPase activity and the Mg²⁺-ATP-dependent H⁺ pumping activity of the synaptic vesicles from the cerebral cortex of rats submitted to intermittent chronic (4 weeks) mild or severe hypoxia were evaluated. The adaptation to the chronic severe hypoxia increases both the ATPase and the H⁺ pumping activities which are inhibited by NEM with an exponential relationship between the IC₅₀ values and the in vivo O₂ concentration. The Mg²⁺-dependent increase in H⁺ pumping activity of synaptic vesicles from the rats subjected to in vivo chronic hypoxia may be antagonized by nigericin (dissipating ΔpH) and by FCCP (dissipating ΔpH and Δ_ΨSV). In contrast, valinomycin (dissipating the Δ_ΨSV and facilitating an enhancement in ΔpH) increases in vitro the H⁺ pumping activity that is inhibited by the addition of high concentration of K gluconate (reducing the rate of K⁺ efflux). The preincubation of vesicles from hypoxic rats with FCCP, but not with nigericin, inhibits the valinomycin-increased H⁺ pumping activity.l-glutamate increases the H⁺ pumping activity in synaptic vesicles from the cerebral cortex of chronic hypoxic rats, whereas other amino acids (i.e.,l-aspartate andl-homocysteate) and glutamate analogs (i.e., quisqualate and ibotenate) are ineffective. The adaptation to both chronic intermittent severe hypoxia and in vivo treatment with posatireline causes a decrease in the Mg²⁺-ATPase activity consistent with the decrease in the H⁺ pumping one of the synaptic vesicles. The addition of nigericin into incubation medium magnifies the decrease in the H⁺ pumping activity, while the addition of FCCP is ineffective, suggesting that the treatment with posatireline interferes with the Δ_ΨSV component in the of the synaptic vesicles from rats submitted to chronic hypoxia. The results of the in vivo and in vitro experiments suggest that in the synaptic vesicles from hypoxic rats the Δ_ΨSV component in may be most effective in increasing the Mg²⁺-ATP-dependent H⁺ pumping activity.     

Soil temperature and biotic factors drive the seasonal variation of soil respiration in a maize (Zea mays L.) agricultural ecosystem

The diurnal and seasonal variation of soil respiration (SR) and their driving environmental factors were studied in a maize ecosystem during the growing season 2005. The diurnal variation of SR showed asymmetric patterns, with the minimum occurring around early morning and the maximum around 13:00 h. SR fluctuated greatly during the growing season. The mean SR rate was 3.16 μmol CO₂ m⁻² s⁻¹, with a maximum of 4.87 μmol CO₂ m⁻² s⁻¹ on July 28 and a minimum of 1.32 μmol CO₂ m⁻² s⁻¹ on May 4. During the diurnal variation of SR, there was a significant exponential relationship between SR and soil temperature (T) at 10 cm depth: . At a seasonal scale, the coefficient α and β fluctuated because the biomass (B) increased α, and the net primary productivity (NPP) of maize markedly increased β of the exponential equation. Based on this, we developed the equation to estimate the magnitude of SR and to simulate its temporal variation during the growth season of maize. Most of the temporal variability (93%) in SR could be explained by the variations in soil temperature, biomass and NPP of maize. This model clearly demonstrated that soil temperature, biomass and NPP of maize combined to drive the seasonal variation of SR during the growing season. However, only taking into account the influence of soil temperature on SR, an exponential equation over- or underestimated the magnitude of SR and resulted in an erroneous representation of the seasonal variation in SR. Our results highlighted the importance of biotic factors for the estimation of SR during the growing season. It is suggested that the models of SR on agricultural sites should not only take into account the influence of soil temperature, but also incorporate biotic factors as they affect SR during the growing season.     

ENaC Activity Requires CFTR Channel Function Independently of Phosphorylation in Sweat Duct

We previously showed that activation of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) Cl⁻ conductance (gCFTR) supports parallel activation of amiloride-sensitive epithelial Na⁺ channel (ENaC) in the native human sweat duct. However, it is not clear whether phosphorylated CFTR, phosphorylated ENaC, or only Cl⁻ -channel function is required for activation. We used basilaterally α-toxin-permeabilized human sweat ducts to test the hypothesis that ENaC activation depends only on Cl⁻ -channel function and not on phosphorylation of either CFTR or ENaC. CFTR is classically activated by PKA plus millimolar ATP, but cytosolic glutamate activation of gCFTR is independent of ATP and phosphorylation. We show here that both phosphorylation-dependent (PKA) and phosphorylation-independent (glutamate) activation of CFTR Cl⁻ channel function support gENaC activation. We tested whether cytosolic application of 5 mM ATP alone, phosphorylation by cAMP, cGMP, G-protein dependent kinases (all in the presence of 100 μM ATP), or glutamate could support ENaC activation in the absence of gCFTR. We found that none of these agonists activated gENaC by themselves when Cl⁻ current ( ) through CFTR was blocked by: 1) Cl⁻ removal, 2) DIDS inhibition, 3) lowering the ATP concentration to 100 μM (instead of 5 mM required to support CFTR channel function), or 4) mutant CFTR (homozygous ΔF508 CF ducts). However, Cl⁻ gradients in the direction of absorption supported, while Cl⁻ gradients in the direction of secretion prevented ENaC activation. We conclude that the interaction between CFTR and ENaC is dependent on activated through CFTR in the direction of absorption (Cl⁻ gradient from lumen to cell). But such activation of ENaC is independent of phosphorylation and ATP. However, reversing through CFTR in the direction of secretion (Cl⁻ gradient from cell to lumen) prevents ENaC activation even in the presence of through CFTR. An erratum to this article is available at .     

Are male rhesus monkeys more aggressive than females?

The justification for the “generalization that primate males are more aggressive than females” (Fedigan & Baxter, 1984) was tested in a troop of 24 captive rhesus monkeys. Males (N=9) were more dominant than females (N=15), i.e., they had more subordinate partners ( _m=17.0 vs. _f=6.6;p<0.01); hence, their aggression rate was higher ( _m=77.6 vs. _m=38.1;p<0.05). However, adjusting the rate of aggression according to the number of subordinate targets revealed no gender difference and males directed numbers of aggressive acts against individual subordinate partners that were not higher than those of females ( _m=4.4 vs. _f=5.4;p>0.1). It was concluded that aggressiveness is an individual attribute that is (1) independent of dominance status, and (2) independent of sex.     

An evolutionary analytical model of a complementary circular code simulating the protein coding genes, the 5′ and 3′ regions

The self-complementary subset ∪{AAA,TTT} with = {AAC, AAT, ACC, ATC, ATT, CAG, CTC, CTG, GAA, GAC, GAG, GAT, GCC, GGC, GGT, GTA, GTC, GTT, TAC, TTC} of 22 trinucleotides has a preferential occurrence in the frame 0 (reading frame established by the ATG start trinucleotide) of protein (coding) genes of both prokaryotes and eukaryotes. The subsets ∪{CCC} and ∪{GGG} of 21 trinucleotides have a preferential occurrence in the shifted frames 1 and 2 respectively (frame 0 shifted by one and two nucleotides respectively in the 5′-3′ direction). and are complementary to each other. The subset contains the subset which has the rarity property (6 × 10⁻⁸) to be a complementary maximal circular code with two permutated maximal circular codes and in the frames 1 and 2 respectively. is called a C³ code. A quantitative study of these three subsets in the three frames 0, 1, 2 of protein genes, and the 5′ and 3′ regions of eukaryotes, shows that their occurrence frequencies are constant functions of the trinucleotide positions in the sequences. The frequencies of in the frame 0 of protein genes are 49, 28.5 and 22.5% respectively. In contrast, the frequencies of in the 5′ and 3′ regions of eukaryotes, are independent of the frame. Indeed, the frequency of in the three frames of 5′ (respectively 3′) regions is equal to 35.5% (respectively 38%) and is greater than the frequencies and , both equal to 32.25% (respectively 31%) in the three frames. Several frequency asymmetries unexpectedly observed (e.g. the frequency difference between and in the frame 0), are related to a new property of the subset involving substitutions. An evolutionary analytical model at three parameters (p, q, t) based on an independent mixing of the 22 codons (trinucleotides in frame 0) of with equiprobability (1/22) followed by t ≈ 4 substitutions per codon according to the proportions p ≈ 0.1; q ≈ 0.1 and r = 1 − p − q ≈ 0.8 in the three codon sites respectively, retrieves the frequencies of observed in the three frames of protein genes and explains these asymmetries. Furthermore, the same model (0.1, 0.1, t) after t ≈ 22 substitutions per codon, retrieves the statistical properties observed in the three frames of the 5′ and 3′ regions. The complex behaviour of these analytical curves is totally unexpected and a priori difficult to imagine.     

Effects of temperature and altitude on ventilation and gas exchange in chukars (Alectoris chukar)

Summary The effects of ambient temperature (T _a) on ventilation and gas exchange in chukar partridges (Alectoris chukar) were determined after acclimation to low and high altitute (LA and HA; 340 and 3,800 m, respectively).At both LA and HA, oxygen consumption ( ) increased with decreasingT _a atT _a from 20 to –20°C. AtT _a of 35 to 40°C, increased above thermoneutral values at HA but remained constant and minimal at LA. Water loss rates increased rapidly atT _a>30°C at both altitudes as birds began to pant. Ventilation rates (f) during panting were 5-to 23-fold greater than the minimalf at thermoneutralT _a.Increased atT _a below thermoneutrality was supported by increased minute volume (V i) at both altitudes. The change inV i was primarily a function of changing tidal volume (V t), althoughf increased slightly asT _a declined. Oxygen extraction ( ) remained fairly constant atT _a below 20°C at both altitudes. BothV t and were considerably lower when birds were panting than at lowerT _a.Chukars showed few obvious ventilatory adaptations to HA. The 35% change in between 340 and 3,800 m was accommodated by a corresponding change inV i (btps), most of which was accomplished by increasedf at HA, along with a slight increase in .Abbreviations and symbols HA high altitude - LA low altitude - rate of evaporative water loss - oxygen extraction efficiency - f respiratory frequency - V t tidal volume - V i minute volume - BMR basal metabolic rate - MHP metabolic heat production     

Respiratory gas exchange during thermogenesis inPhilodendron selloum Koch

During peak thermogenesis of anthesis, high rates of respiration by the sterile male florets on the spadix ofPhilodendron selloum significantly reduce the oxygen tension (P_{O 2}) and raise CO₂ tension between the florets. Nevertheless, respiration is not limited by the availability of O₂ under natural conditions. At experimental P_{O 2} below about 17 kPa, however, florets begin to show decreased O₂ consumption. A respiratory exchange ratio of 0.83 indicates that the major energy source is not starch, but is probably lipid.Abbreviations and symbols capacitance of the gas phase for O₂ (ml O₂ cm^-3 kPa^-1) - D_{O 2} binary diffusion coefficient of O₂ in air (cm² min^-1) - K_{O 2} Krogh's diffusion coefficient (ml O₂ cm^-2 min^-1 kPa^-1 cm) - P_{O 2} P_{CO 2} partial pressures of O₂ and CO₂ (kPa) - rate of O₂ consumption (ml O₂ g^-1 h^-1) - F_gas fractional gas volume - P₈₀ O₂ partial pressure at which falls below 80% of maximum - RE respiratory exchange ratio      

The proportion of genetic deviates in the tails of a normal population

Summary If genetic and environmental effects upon a quantitative phenotype X=G+E are normally and independently distributed then the probability distribution of genetic value G among individuals of fixed phenotypic value X is likewise a normal distribution. The mean of this a posteriori distribution of genetic values is +h ² (X− ) and the variance is σ _g ² (1−h ²), where is the a priori mean of X, h ² is the heritability ratio, and σ _g ² is genetic variance. For any fixed values of h ² and σ _g ² the a posteriori probability that the genetic value G associated with a given phenotype X exceeds the population mean by any specified amount can therefore be read directly from the tables of the standard normal distribution. The expected proportion of these superior genetic deviates among individuals whose phenotypic value exceeds some specified constant may also be calculated (by numerical analysis) and is presented here in graphical form. If phenotypic selection is practiced by choosing the best out of N phenotypes then N should be large enough to assure high probability of obtaining a superior genetic deviate. The operating characteristics of this type of selection are displayed in tabular form, again based upon numerical integration.

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Zusammenfassung Wenn genetische und umweltbedingte Effekte auf einen quantitativen Ph?notyp X=G+E von einander unabh?ngig und normal verteilt sind, dann entspricht die Wahrscheinlichkeitsverteilung des genetischen Werts G unter Individuen mit fixiertem ph?notypischem Wert X gleichfalls einer Normalverteilung. Das Mittel dieser a posteriori-Verteilung der genetischen Werte ist +h ² (X− ) und die Varianz ist σ _g ² (1−h ²); wobei das a priori-Mittel von X, h ² der Heritabilit?tskoeffizient und σ _g ² die genetische Varianz sind. Für jeden fixierten Wert von h ² und σ _g ² kann daher die a posteriori-Wahrscheinlichkeit, da? der genetische Wert G das Populationsmittel in Verbindung mit einem gegebenen Ph?notyp X um einen bestimmten Wert übersteigt, direkt aus den Tabellen einer standardisierten Normalverteilung abgelesen werden. Der erwartete Anteil dieser überlegenen, genetisch bedingten Abweichung unter Individuen, deren ph?notypischer Wert einen vorgegebenen Konstantwert übersteigt, kann ebenfalls numerisch errechnet werden. Er wird im vorliegenden Fall graphisch dargestellt. Wenn eine ph?notypische Selektion zur Auswahl der besten Ph?notypen aus N Individuen erfolgt, sollte N gro? genug sein, um mit hoher Wahrscheinlichkeit eine überlegene, genetisch bedingte Abweichung zu erhalten. Die wirksamen Charakteristiken dieses Typs der Selektion werden in tabellierter Form wiedergegeben, die gleichfalls auf numerischer Integration beruht.

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Dedicated to Dr. George F. Sprague on the occasion of his 65^th birthday.

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Cooperative investigations of the Colorado Agricultural Experiment Station, the Crops Research Division, Agricultural Research Service, U.S. Department of Agriculture, and the Beet Sugar Development Foundation. Approved by the Colorado Agricultural Experiment Station for publication as Scientific Series Article No. 880. Paper No. BU-78, Biometrics Unit, and Paper No. 529, Plant Breeding Department, Cornell University.

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Geneticist, Crops Research Division, Agricultural Research Service, U.S. Department of Agriculture; now deceased.

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Formerly Research Assistant, Colorado State University; now Assistant Professor of Biological Statistics, Cornell University.     

Use of the force-velocity test to determine the optimal braking force for a sprint exercise on a friction-loaded cycle ergometer

A group of 15 untrained male subjects pedalled on a friction-loaded cycle ergometer as fast as possible for 5–7 s to reach the maximal velocity (V{immax}) against different braking forces (F _B). Power was averaged during a complete crank rotation by adding the power dissipated againstF _B to the power necessary to accelerate the flywheel. For each sprint, determinations were made of peak power output ( ) power output attained atV _max ( ) calculated as the product ofV _max andF _B and the work performed to reachV _max expressed in mean power output ( ). The relationships between these parameters andF _B were examined. A biopsy taken from the vastus lateralis muscle and tomodensitometric radiographs of both thighs were taken at rest to identify muscle metabolic and morphometric properties. The value was similar for allF _B. Therefore, the average of values was defined as corrected maximal power ( ). This value was 11 higher than the maximal power output uncorrected for the acceleration. Whereas the determination did not require high loads, the highest value ( ) was produced when loading was heavy, as evidenced by the -F _B parabolic relationship. For each subject, the braking force ( ) giving was defined as optimal. The , equal to 0.844 (SD 0.108) N · kg⁻¹ bodymass, was related to thigh muscle area (r = 0.78,P < 0.05). The maximal velocity ( ) reached against this force seemed to be related more to intrinsic fibre properties (% fast twitch b fibre area and adenylate kinase activity). Thus, from the determination, it is suggested that it should be possible to predict the conditions for optimal exercise on a cycle ergometer.     

Ventilation and oxygen consumption in rosy finches and house finches at sea level and high altitude

Summary Rosy finches (Leucosticte arctoa) breed at altitudes above 3500 m in eastern California. House finches (Carpodacus mexicanus) belong to the same subfamily (Carduelinae), but breed at much lower elevations. Oxygen consumption ( ) and ventilatory parameters of these two species were measured over a wide range of ambient temperatures (T _a) at low altitude (LA; 150 m) and at high altitude (HA; 3800 m).Minimal nighttime 's of rosy finches and house finches at LA (T _a=30°C) were close to allometrically predicted values for passerine birds. At both altitudes, increased linearly with decreasingT _a betweenT _a=20 and –10°C. Resting 's were slightly higher at HA than at LA on average.In both species, minute volume ( ) was inversely related toT _a.T _a-correlated increases in resulted from significant increases in both ventilatory frequency (f) and tidal volume (V T) at both altitudes. Oxygen extraction efficiency ( ) was independent ofT _a in rosy finches at LA, but declined significantly with decreasingT _a in rosy finches at HA and in house finches at both altitudes.At a givenT _a, both species had significantly greater (BTPS) at HA than at LA. Altitude-correlated increases in resulted primarly from increases inf with little change inV T. was significantly greater at HA than at LA in both species.In spite of the difference in altitudinal distributions of rosy finches and house finches, there were few conspicuous interspecific differences in metabolic or ventilatory adaptation to altitude or lowT _a over the range of conditions examined.Symbols and abbreviations BMR basal metabolic rate - BTPS at body temperature and pressure, saturated - oxygen extraction efficiency - f ventilation frequency - h mean coefficient of heat transfer - HA high altitude - instantaneous oxygen consumption - LA low altitude - RH relative humidity - SMR standard metabolic rate - STPD standard temperature and pressure, dry - T temperature - a ambient - b body - lc lower critical of thermoneutral zone - minute volume - V T tidal volume     

Evidence for four nonallelic structural genes for the γ chain of human fetal hemoglobin

An abnormal human fetal hemoglobin not only may be either a^Gγ- or an^Aγ-chain variant but also may be present in a different proportion of the total fetal hemoglobin.^Gγ-Chain variants contribute either about one-fourth or one-eighth to the total production of HbF in the heterozygote, whereas the^Aγ-chain variants approximate either one-eighth or one-sixteenth of the total HbF. These observations may indicate the presence of four nonallelic Hb_γ structural genes (termed ) which produce γ chains in an approximate ratio of 4 : 2 : 2 : 1. HbF Malta I is considered to be the product of a mutant of the locus, an undefined HbF_x that of the locus, HbF Hull and HbF Jamaica products of mutated loci, and the newly discovered HbF Malta II a mutant of the gene. This work was supported in part by grants HL-05168 and HL-02558 from the National Institutes of Health, U.S. Public Health Service.     

Results of the oxygen Fick method in a closed blood circulation model including “total arteriovenous diffusive shunt of oxygen”

It is considered that arteriovenous diffusive shunts of oxygen may cause inaccuracy of the oxygen Fick method as where is the pulmonary oxygen uptake, is the cardiac output, and CaO₂ and CvO₂ are the arterial and venous oxygen contents, respectively. A simple circulation model, including the whole circulation with nine well-mixed compartments (C1, ... C9), is constructed: the is assigned as constant as 6000 ml min⁻¹; the blood portions of 60 ml move at an interval of 600 ms. C1 and C2 compartments, each having 60 ml volume, represent the blood of pulmonary microcirculation, C3 represents the arterial blood with a volume of 1500 ml, C4, ..., C8, each also having a volume of 60 ml, represent the blood of peripheral microcirculation, whereas C9 represents the venous blood with a volume of 3000 ml. The pulmonary oxygen uptake , related to C1 and C2, the oxygen release , related to C4,...,C8, as well as a “total arteriovenous diffusive shunt of oxygen” , from the arterial blood (C3) to the venous blood (C9), are calculated simultaneously. The alveolar gas has a constant oxygen partial pressure, and the pulmonary diffusion capacity is also constant; similar to modeling the pulmonry, oxygen diffusion, constant partial oxygen pressures for all peripheral tissues as well as constant diffusion capacities for all peripheral oxygen diffusion are also assigned. The diffusion capacities for the (between C3 and C9) are arbitrarily assigned. The Fick method gives incorrect results depending on the total arteriovenous diffusive shunt of oxygen . But the mechanism determining the magnitude of remains unclear.     

Epipelic and pelagic primary production in Alaskan Arctic lakes of varying depth

We compared on eight dates during the ice-free period physicochemical properties and rates of phytoplankton and epipelic primary production in six arctic lakes dominated by soft bottom substrate. Lakes were classified as shallow ( < 2.5 m), intermediate in depth (2.5 m <  < 4.5 m), and deep ( > 4.5 m), with each depth category represented by two lakes. Although shallow lakes circulated freely and intermediate and deep lakes stratified thermally for the entire summer, dissolved oxygen concentrations were always >70% of saturation values. Soluble reactive phosphorus and dissolved inorganic nitrogen (DIN = NO₃ ⁻–N + NH₄ ⁺–N) were consistently below the detection limit (0.05 μmol l⁻¹) in five lakes. However, one lake shallow lake (GTH 99) periodically showed elevated values of DIN (17 μmol l⁻¹), total-P (0.29 μmol l⁻¹), and total-N (33 μmol l⁻¹), suggesting wind-generated sediment resuspension. Due to increased nutrient availability or entrainment of microphytobenthos, GTH 99 showed the highest average volume-based values of phytoplankton chlorophyll a (chl a) and primary production, which for the six lakes ranged from 1.0 to 2.9 μg l⁻¹ and 0.7–3.8 μmol C l⁻¹ day⁻¹. Overall, however, increased resulted in increased area-based values of phytoplankton chl a and primary production, with mean values for the three lake classes ranging from 3.6 to 6.1 mg chl a m⁻² and 3.2–5.8 mmol C m⁻² day⁻¹. Average values of epipelic chl a ranged from 131 to 549 mg m⁻² for the three depth classes, but levels were not significantly different due to high spatial variability. However, average epipelic primary production was significantly higher in shallow lakes (12.2 mmol C m⁻² day⁻¹) than in intermediate and deep lakes (3.4 and 2.4 mmol C m⁻² day⁻¹). Total primary production (6.7–15.4 mmol C m⁻² day⁻¹) and percent contribution of the epipelon (31–66%) were inversely related to mean depth, such that values for both variables were significantly higher in shallow lakes than in intermediate or deep lakes. Handling editor: L. Naselli-Flores     

Bacterial growth and substrate degradation by BTX-oxidizing culture in response to salt stress

Interactions between microbial growth and substrate degradation are important in determining the performance of trickle-bed bioreactors (TBB), especially when salt is added to reduce biomass formation in order to alleviate media clogging. This study was aimed at quantifying salinity effects on bacterial growth and substrate degradation, and at acquiring kinetic information in order to improve the design and operation of TBB. Experiment works began by cultivating a mixed culture in a chemostat reactor receiving artificial influent containing a mixture of benzene, toluene, and xylene (BTX), followed by using the enrichment culture to degrade the individual BTX substrates under a particular salinity, which ranged 0–50 g l⁻¹ in batch mode. Then, the measured concentrations of biomass and residual substrate versus time were analyzed with the microbial kinetics; moreover, the obtained microbial kinetic constants under various salinities were modeled using noncompetitive inhibition kinetics. For the three substrates the observed bacterial yields appeared to be decreased from 0.51–0.74 to 0.20–0.22 mg mg⁻¹ and the maximum specific rate of substrate utilization, declined from 0.25–0.42 to 0.07–0.11 h⁻¹, as the salinity increased from 0 to 50 NaCl g l⁻¹. The NaCl acted as noncompetitive inhibitor, where the modeling inhibitions of the coefficients, K _T(S), were 22.7–29.7 g l⁻¹ for substrate degradation and K _T(μ), 13.0–19.0 g l⁻¹, for biomass formation. The calculated ratios for the bacterial maintenance rate, m _S, to further indicated that the percentage energy spent on maintenance increased from 19–24 to 86–91% as salinity level increased from 0 to 50 g l⁻¹. These results revealed that the bacterial growth was more inhibited than substrate degradation by the BTX oxidizers under the tested salinity levels. The findings from this study demonstrate the potential of applying NaCl salt to control excessive biomass formation in biotrickling filters.     

Effect of NaCl on kinetics ofd-glucosamine uptake in yeasts differing in halotolerance

The initial rate ofd-glucosamine uptake by the non-halotolerant yeastSaccharomyces cerevisiae was approximately halved as the apparent half saturation constant (K_m) and the apparent maximum velocity (V_max) changed from 6.6mm to 16.4mm and from 22 μmol · g⁻¹ · min⁻¹ to 16 μmol · g⁻¹ · min⁻¹, respectively, when the salinity in the medium was increased from zerom to 0.68m NaCl. Corresponding changes in a high affinity transport system in the halotolerant yeastDebaryomyces hansenii were from 1.1mm to 4.6mm and from 3.1 μmol · g⁻¹ · min⁻¹ to 4.5 μmol · g⁻¹ · min⁻¹, implying a practically unchanged transport capacity. In 2.7m NaCl, K_m and V_max in this system were 24.5mm and 1.1 μmol · g⁻¹ · min⁻¹, respectively, representing a marked decrease in transport capability. Nevertheless, the degree of affinity in this extreme salinity must still be regarded as noteworthy. In addition to the high affinity transport system inD. hansenii, a low affinity system, presumably without relevance ind-glucosamine transport, was observed.     

Insulin binding in cultured Chinese hamster kidney epithelial cells: The effect of serum in the medium

Summary [¹²⁵I]Insulin (porcine) binding to an epithelial cell line established from a Chinese hamster kidney, CHK-AC_E−100, showed an optimum at pH 8.0 and reached a maximum after 2.5 h incubation at 25°C. Dissociation of bound [¹²⁵I]insulin was facilitated by the addition of unlabeled insulin in the dilution buffer. Porcine insulin effectively competed for [¹²⁵I]insulin binding to the cultured cells and was 30 and 90 times as potent as guinea pig insulin and porcine proinsulin in causing 50% inhibition of [¹²⁵I]insulin binding; glucagon was completely ineffective. Scatchard analysis of the binding data yielded a curvilinear plot and a capacity of 0.6 ng/10⁶ cells; the average affinity of the empty receptor, , was calculated to be 1.78×10⁸ M ⁻¹ and that of the filled receptor, , 0.57×10⁸ M ⁻¹. Substitution of fetal bovine serum (FBS) in the culture medium with bovine calf, bovine newborn, or bobby calf serum altered insulin binding characteristics in the cells and reduced cell growth. Insulin binding characteristics of cells grown in hormone-supplemented medium containing 0 to 0.1% FBS were similar to those of cells grown in minimum essential medium (MEM) containing 2 to 5% FBS. The data indicated that the established Chinese hamster kidney epithelial cell line CHK-AC_E−100 possessed specific insulin receptors and the characteristics of the receptors could be manipulated by changing the serum in culture medium.     

(H)NCAHA and (H)CANNH experiments for the determination of vicinal coupling constants related to the ϕ-torsion angle

Summary A set of three-dimensional triple-resonance experiments is described which provide , , and coupling constants. The pulse sequences generate E.COSY-like multiplet patterns and comprise a magnetization transfer from the amide proton to the α-proton or vice versa via the directly bound heteronuclei. For residues with the ¹H^α spin resonating close to the H₂O signal, a modified HNCA experiment can be employed to measure the vicinal ¹H^N,¹H^α couplings. Ambiguities associated with the conversion of values into ϕ-angle constraints for protein structure determination can be resolved with the knowledge of the heteronuclear ³J-couplings. In favourable cases, stereospecific assignments of glycine α-protons can be obtained by employing the experiments described here in combination with NOE data. The methods are applied to flavodoxin from Desulfovibrio vulgaris.