Aerobic metabolism of the lizardVaranus exanthematicus: Effects of activity, temperature, and size

Summary Oxygen consumption was measured at rest and during spontaneous activity at body temperatures of 25 and 35°C in 14 fasting Savanna monitor lizards,Varanus exanthematicus ranging in weight from 172 to 7500 g. The allometric relationship between metabolic rate at 25°C and body weight (W) is given by: (ml O₂ STPD·g^–1·hr^–1)=0.88W ^–0.43 (Fig. 2). Although statistical comparisons are equivocal, this intraspecific size dependence exceeds that reported for interspecific comparisons among reptiles and other vertebrate groups (Fig 3). A reproducible diurnal pattern of activity was observed in undisturbed animals with minimum values of between 2400 and 0800 h (Fig. 1). Spontaneous activity and generally reached peak values between 1200 and 2000 hrs. The average ratio of active aerobic metabolic rate (AMR) to minimum (standard) aerobic metabolic rate (SMR) was 8.2. This voluntary AMR/SMR inVaranus exceeds the AMR/SMR for most reptiles stimulated to exhaustion. The high aerobic capacity is consistent with other evidence for efficient exchange and transport of respiratory gases inV. exanthematicus; e.g., low or absent intracardiac shunt flow resulting in high arterial saturation and low ventilation and perfusion requirements.     

The nucleotide sequence ofBeneckea harveyi 5S rRNA

Summary The complete sequence of the 5S rRNA from the bioluminescent bacterium,Beneckea harveyi has been determined to be p U G C U U G G C G C C A U A G C G A U U-G G A C C C A C U G A (U) C U U C A U U C C-G A A C C A G A A G U G A A C G A A U U A-G G C C G A U G G U G U G U G G G G C U-C C C C A U G U A G A G U A G G A A U C G-C C A G G U (U)_OH.Two sites of sensitivity to ribonuclease T₂ cleavage were identified; at A₄₁ and either A₅₄ or A₅₅. Comparison with existing sequence information fromEscherichia coli andPhotobacterium phosphoreum clarifies the amount of diversity among the bioluminescent bacteria and provides further insight into their phylogenetic position. Sequence heterogeneities were encountered and the importance of these in interpreting 5S rRNA data is discussed.     

Relationship between the octanol-water partition coefficient of tertiary amines and their effect of ‘selective’ uncoupling of photophosphorylation

A series of tertiary amines was investigated for effects on the transmembrane proton potential difference ( H), on photophosphorylation and on electron-flux control related to the intrathylakoid proton potential ( H_I), using isolated chloroplasts ofSpinacia oleracea L. As indicated by 9-aminoacridine fluorescence and [14C]methylamine uptake, all amines studied inhibited a build-up of H and, in parallel, ATP synthesis. Even when H was low, strong H₁-dependent electron-flux control was observed under the influence of tertiary amines. The strength of flux control in the presence of low H and the effectiveness of inhibition of ATP synthesis linearly increased with the lipophilicity of the amines. The most effective of the amines tested caused 50% inhibition of ATP synthesis at a concentration of 6 M, which is about 1000-fold lower than the concentration required for inhibition by methylamine. The data presented indicate the existence of two proton domains in the thylakoid vesicles, one of them feeding the ATP-synthase, the other the sites of pH-dependent electron-flux control. It is concluded that tertiary amines develop their action in a lipophilic domain of the thylakoid membrane, in the vicinity of the ATP-synthase complex. A mechanism for selective uncoupling and for the maintenance of H_I-dependent electron flux control in the presence of low H is discussed.Abbreviations and symbols coefficient for pH-dependent electron flux control - 9-AA 9-aminoacridine - Chl chlorophyll - I₅₀ amine concentration producing 50% inhibition of ATP-synthesis - J_e flux of photosynthetic electron transport - k _H apparent rate constant for proton efflux - H₁ proton potential in the thylakoid lumen - H₁ transthylakoid proton potential difference - p partition coefficient - q _AA coefficient for 9-aminoacridine fluorescence quenching - PS photosystem - Q quantum flux of photosynthetically active light Dedicated to Professor Wilhelm Simonis, on the occasion of his 80th birthday     

On the accuracy of some mark-recapture estimators

Summary The behaviour of the mark-recapture estimators of Petersen, Bailey (triple catch) and Jolly and Seber are examined theoretically and empirically by means of simulation techniques. The correlation between the parameter and its associated variance is shown to be significant for all the estimators. This correlation makes the estimated variance an insensitive measure of the accuracy of the estimate except at very high sampling intensities. Such sampling intensities are rarely achieved in experimental work and so the method of mark-recapture must be considered of very limited use. At the sampling intensities necessary to give a coefficient of variation of less than 0.05 it does not seem likely that the correlation between and its variance will produce serious underestimation but the minimum confidence limits are recommended.     

Normal modes of a two-dimensional lattice of interacting dipoles

We derive the normal mode frequency spectra for a rectangular lattice of electric dipoles oscillating about configurations corresponding to relative minima in their mutual potential energy. For two such configurations —a) all dipoles parallel and b) adjacent rows antiparallel —we obtain numerical solutions for for various choices of the lattice parameters.     

Transport of H+, K+, Na+ and Ca++ in Streptococcus

Summary The streptococci differ from other bacteria in that cation translocations (with the possible exception of one of the K⁺ uptake systems) occur by primary transport systems, i.e., by cation pumps which use directly the free energy released during hydrolysis of chemical bonds to power transport. Transport systems in other bacteria, especially for Na⁺ and Ca⁺⁺, are often secondary, using the free energy of another ion gradient to drive cation transport. In streptococci H⁺ efflux occurs via the F₁F₀-ATPase. This enzyme is composed of eight distinct subunits. Three of the subunits are embedded in the membrane and form a H⁺ channel; this is called the F₀ portion of the enzyme. The other five subunits form the catalytic part of the enzyme, called F₁, which faces the cytoplasm and can easily be stripped from the membrane. Physiologically, this enzyme functions as a H⁺-ATPase, pumping protons out of the cell to form an electrochemical proton gradient, . The F₁F₀-ATPase, however, is fully reversible and if supplied with P_i, ADP and a + of sufficient magnitude (ca –200 mv) catalyzes the synthesis of ATP. Streptococcus faecalis can accumulate K⁺ and establish a gradient of 50 000:1 (in>out) under some conditions. Uptake occurs by two transport systems. The dominant, constitutive system requires both an electrochemical proton gradient and ATP to operate. The minor, inducible K⁺ transport system, which has many similarities to the K⁺-ATPase of the Kdp transport system found in Escherichia coli, requires only ATP to power K⁺ uptake.Sodium extrusion occurs by a Na⁺/H⁺-ATPase. Exchange is electroneutral and there is no requirement for a . The possibility that the Na⁺/H⁺-ATPase may consist of two parts, a catalytic subunit and a Na⁺/H⁺ antiport subunit, is suggested by the finding that damage to the Na⁺ transport system either through mutation or protease action leads to the appearance of -requiring Na⁺/H⁺ antiporter activity.Ca⁺⁺ like Na⁺ is extruded from metabolizing, intact cells. Transport requires no but does require ATP. Reconstitution of Ca⁺⁺ transport activity with accompanying Ca⁺⁺-stimulated ATPase activity into proteoliposomes suggests that Ca⁺⁺ is transported by a Ca⁺⁺-translocating ATPase.Where respiring organelles and bacteria use secondary transport systems the streptococci have developed cation pumps. The streptococci, which are predominantly glycolyzing bacteria, generate a much inferior to that of respiring organisms and organelles. The cation pumps may have developed simply in response to an inadequate .Abbreviations electrochemical potential of protons - membrane potential - pH pH gradient - p proton-motive force - DCCD N,Na¹-dicyclohexlcarbodiimide - TCS tetrachlorosalicylanilide - FCCP carbonylcyanide-p-trifluoromethylphenylhydrazone - CCCP carbonylcyanie-m-chlorophenylhydrazone - TPMP⁺ triphenylmethyl phosphonium ion - DDA⁺ dibenzyldimethylammonium ion - Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - EGTA ethyleneglycol-bis (amino-ethyl-ether)-N,N-tetraacetic acid     

Skeletal muscle [(V)\dot]O2 \dot V_{O_2 } in rat and lemming: Effect of blood flow rate

Summary The rate of oxygen consumption ( ) by skeletal muscle was investigated in isolated perfused hindlimbs of laboratory rats and lemmings (Lemmus). In both species, increased in proportion to blood flow rate, even at flow rates 4–5 times above resting level. The slope of the line relating to skeletal muscle blood flow was significantly greater in the lemming than in the rat. This may be related to the inverse relationship between body weight and metabolic rate. These data support the hypothesis that in small animals a dependent relationship exists between blood flow and skeletal muscle .     

A method for determining the maximal steady state of blood lactate concentration from two levels of submaximal exercise

The aim of this study was to estimate the characteristic exercise intensity _CL which produces the maximal steady state of blood lactate concentration (MLSS) from submaximal intensities of 20 min carried out on the same day and separated by 40 min. Ten fit male adults [maximal oxygen uptake _max 62 (SD 7) ml · min^–1 · kg^–1] exercisOed for two 30-min periods on a cycle ergometer at 67% (test 1.1) and 82% of _max (test 1.2) separated by 40 min. They exercised 4 days later for 30 min at 82% of _max without prior exercise (test 2). Blood lactate was collected for determination of lactic acid concentration every 5 min and heart rate and O₂ uptake were measured every 30 s. There were no significant differences at the 5th, 10th, 15th, 20th, 25th, or 30th min between , lactacidaemia, and heart rate during tests 1.2 and 2. Moreover, we compared the exercise intensities _CL which produced the MLSS obtained during tests 1.1 and 1.2 or during tests 1.1 and 2 calculated from differential values of lactic acid blood concentration ([1a^–]_b) between the 30th and the 5th min or between the 20th and the 5th min. There was no significant difference between the different values of _CL [68 (SD 9), 71 (SD 7), 73 (SD 6),71 (SD 11) % of _max (ANOVA test,P<0.05). Four subjects ran for 60 min at their _CL determined from periods performed on the same day (test 1.1 and 1.2) and the difference between the [la^–]_b at 5 min and at 20 min ( ([la^–]_b)) was computed. The [la^–]_b remained constant during exercise and ranged from 2.2 to 6.7 mmol · l^–1 [mean value equal to 3.9 (SD 1) mmol · l^–1]. These data suggest that the _CL protocol did not overestimate the exercise intensity corresponding to the maximal fractional utilization of _max at MLSS. For half of the subjects the _CL was very close to the higher stage (82% of _max where an accumulation of lactate in the blood with time was observed. It can be hypothesized that _CL was very close to the real MLSS considering the level of accuracy of [la^–]_b measurement. This study showed that exercise at only two intensities, performed at 65% and 80% of _max and separated by 40 min of complete rest, can be used to determine the intensity yielding a steady state of [la^–1]_b near the real MLSS workload value.     

Breathing pattern and growth: comparative aspects

Summary The resting oxygen consumption and breathing pattern of nine newborn and adult species (ranging in body size from mouse to human) have been compared on the basis of data collected from the literature. Minute ventilation is similarly linked to at both ages, the percent of extracted as O₂ about 2.2. Tidal volume/kg is an interspecies constant in newborns and adults, approximately 8 ml/kg. Breathing frequency decreases with the increase in size in a different way at the two ages: large species have newborns breathing at rates 2–3 times above the corresponding adults' values, while in the small species newborns and adults breathe at almost the same rate. Therefore the newborns of the smallest species have both and below the expected values, implying a greater inability to cope with the external demands than newborns of larger species. Several considerations indicate that in the smallest newborns the mechanical properties of the respiratory system could be a constraint to resting ventilations larger than observed. It is therefore possible that their low is the cause, and not the effect, of the relatively small .     

Sulfur isotope values in a forested catchment over four years: Evidence for oxidation and reduction processes

A small catchment on the Swedish West Coast has been studied over four years to determine S dynamics by using S isotope ratios. A Norway spruce dominated forest covers the catchment, and small peat areas occur in the lower parts of the catchment. The runoff values varied both during the year, and from year to year. Over the period from February 1990 to December 1993, the values ranged from — 1%. to +11%. Over the same period, the throughfall values ranged from +1%. to +15%. There was no correlation (r ²= 0.01; Pr(F)=0.57) between values in throughfall and runoff. Since the only input of S to the catchment is atmospheric deposition, the long-term runoff S mass flux is controlled by the deposition. Therefore, processes in the catchment are responsible for the variation in the runoff values. During periods with enriched runoff, bacterial dissimilatory SO ₄ ^2– reduction occurs in the catchment. After very dry periods, oxidation of this reduced S, which is³²S-enriched, can be traced in runoff. Previous studies of the catchment have not been able to distinguish between: 1) oxidation of reduced S and dry deposition, and 2) reduction and adsorption. From the current study, it can be concluded that adsorption and dry deposition cannot cause the observed variation in runoff .     

Physiological responses to cold (10° C) in men after six months' practice of yoga exercises

A study was conducted on 30 healthy soldiers (age: 40–46 years) to assess the effect of selected yogic exercises (asanas) on some physiological responses to cold exposure. They were randomly divided into two groups of 15 each. One group performed regular physical exercises of physical training (PT), while the other group practised yogic exercises. At the end of 6 months of training, both the groups were exposed together to cold stress at 10°C for 2 h, and the following parameters were periodically monitored during cold exposure: heart rate (fH), blood pressure (BP), cardiac output , oral temperature (T_or), skin temperature (T _sk), respiratory rate (fR), minute ventilation , oxygen consumption , and shivering response by integrated electromyogram (EMG). There were progressive increases inBP, fR, , , and and decreases infH,T _or andT _sk during cold exposure in both the groups. However, the decrease inT _or and the increases in and were relatively lower (P<0.01) in the yoga group as compared to the PT group. The shivering response appeared much earlier and was more intense in the PT group. These findings suggest that practice of yoga exercises may improve cold tolerance.     

Activity and oxygen consumption during hypoxic exposure in high altitude and lowland sceloporine lizards

Summary Resting rates of O₂ consumption against , exercise endurance times and during recovery from vigorous exercise were measured inSceloporus occidentalis captured near sea level and inS. graciosus captured above 2850 m. Oxygen consumption against was also measured inS. occidentalis captured above 2850 m. When was recorded continuously, as ambient was slowly reduced from 155 Torr, it became directly dependent upon ambient between 110 and 120 Torr. The critical for the high altitude lizards was lower than that for the lowland lizards, which enabled the former to maintain relatively higher 's when ambient was reduced below 120 Torr. The high altitude lizards also had significantly greater endurance when stimulated to exercise at 1600 m ( 130 Torr). Both the higher under hypoxia and the greater endurance roughly parallel a significantly greater maximum in the high altitude lizards. At a simulated altitude of 3600 m ( 100 Torr), maximum and rate of recovery of the O₂ debt calculated from post active were significantly reduced in the lowland but not the high altitude lizards. The effects of simulated altitude conditions on the lowland but not the mountaine animals indicate adaptations to altitude in these sceloporine lizards. We did not find any consistent relationship between organ/body weight ratios or hematocrit and our measures of endurance or the altitude at which the lizards were captured.     

Thermoregulation,gas exchange,and ventilation in Adelie penguins (Pygoscelis adeliae)

Summary Adelie penguins (Pygoscelis adeliae) experience a wide range of ambient temperatures (T _a) in their natural habitat. We examined body temperature (T _b), oxygen consumption ( ), carbon dioxide production ( ), evaporative water loss ( ), and ventilation atT _a from –20 to 30 °C. Body temperature did not change significantly between –20 and 20°C (meanT _b=39.3°C).T _b increased slightly to 40.1 °C atT _a=30°C. Both and were constant and minimal atT _a between –10 and 20°C, with only minor increases at –20 and 30°C. The minimal of adult penguins (mean mass 4.007 kg) was 0.0112 ml/[g·min], equivalent to a metabolic heat production (MHP) of 14.9 Watt. The respiratory exchange ratio was approximately 0.7 at allT _a. Values of were low at lowT _a, but increased to 0.21 g/min at 30°C, equivalent to 0.3% of body mass/h. Dry conductance increased 3.5-fold between –20 and 30°C. Evaporative heat loss (EHL) comprised about 5% of MHP at lowT _a, rising to 47% of MHP atT _a=30°C. The means of ventilation parameters (tidal volume [VT], respiration frequency [f], minute volume [I], and oxygen extraction [ ]) were fairly stable between –20 and 10°C (VT did not change significantly over the entireT _a range). However, there was considerable inter- and intra-individual variation in ventilation patterns. AtT _a=20–30°C,f increased 7-fold over the minimal value of 7.6 breaths/min, and I showed a similar change. fell from 28–35% at lowT _a to 6% atT _a=30°C.Abbreviations C thermal conductance - EHL evaporative heat loss - oxygen extraction - f respiratory frequency - MHP metabolic heat production - evaporative water loss - LCT lower critical temperature - RE respiratory exchange ratio - T _a ambient temperature - T _b body temperature - rate of oxygen consumption - rate of carbon dioxide production - I inspiratory minute volume - VT tidal volume     

Maximal oxygen uptake,sweating and tolerance to exercise in the heat

The purpose of this experiment was to determine if tolerance to exercise in the heat is related to maximal oxygen uptake (max ₀₂) and sweating. Seven men with max ₀₂ between 42 and 66 ml/(min·kg) underwent one 2-hr exposure at 24°C T_q while working on a bicycle ergometer at rel ₀₂ of 28% ( ₀₂ = 1.23 1/min). In the hot exposures the high capacity subjects had maximal sweat rates of 800 to 1,000 g/(hr·m²) while the lower capacity men sweated 300 to 400 g/(hr·m²). These differences in sweating were not related to neuromuscular stimuli, ₀₂ (metabolic rate), T_re, T_re, _s, _s or tolerance time. Tolerance to exercise in the heat was not related to maximal ₀₂ capacity when the subjects worked at the same relative load in spite of large differences in sweating. These results question the importance of the rate of sweating for predicting work performance in hot environments.

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Zusammenfassung Das Ziel dieser Untersuchung war, zu prüfen, ob die Toleranz bei Arbeit in der Hitze in einer Beziehung steht zur maximalen O₂-Aufnahme und Schwitzen. Sieben Männer mit V₀₂ zwischen 42 – 66 ml/(min·kg) wurden belastet während 2 Stunden bei T_a 24°C und 3 × 2 Stunden bei 47°C mit Arbeit auf dem Fahrrad-Ergometer bei im Mittel von 28% V₀₂ = 1.23 1/min. Während der Hitzebelastung zeigten die leistungsfähigen Personen Schweissekretionsraten von 800 – 1000 g/(hr·m²) und die wenig leistungsfähigen 300 – 400 g/(hr·m²). Diese Unterschiede waren ohne Beziehung zu neuromuskulären Stimuli, Stoffwechselrate, T_re, T_re, _s, _s oder der Toleranzzeit. Ausdauer bei Arbeit in der Hitze war ohne Beziehung zur maximalen V₀₂-Kapazität, wenn die Personen bei der gleichen relativen Belastung arbeiteten tro grosser Unterschiede im Schwitzen. Die Ergebnisse stellen den Wert der Schweissekretionsrate zur Voraussage der Arbeitsleistung in der Hitze in Frage.

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Resume Dans cette étude, on a cherché à voir si la tolérance au travail sous contrainte de chaleur était en relation avec la possibilité maximum d'absorption de O₂ ( ₀₂) d'une part, de transpirer d'autre part. 7 hommes présentant des ₀₂ compris entre 42 et 66 ml/(min · kg) ont pédalé sur un ergomètre pendant 2 heures par une T_a de 24°C et 3 × 2 heures par 47°C et cela par une ₀₂ relative de 28% ( ₀₂ = 1,25 1/min). Durant l'effort sous contrainte de chaleur, les plus actifs ont eu des sécrétions de sueur de 800 à 1.000 g h^–1 m^–2 et les moins actifs de 300 à 400 g/h · m². Ces différences étaient sans rapport avec les stimulus neuro-musculaires, le taux de métabolisme, T_re, T_re, T_s et T_s ou la durée de tolérance. L'endurance au travail sous contrainte de chaleur n'a pas été fonction de la capacité maximum de ₀₂, lorsque les personnes travaillaient dans des conditions analogues, même si l'on a noté de grandes différences dans la transpiration. Ces résultats mettent en doute la représentativité du taux de sécrétion de sueur comme indicatif des possibilités de travailler en atmosphère chaude.

     

Effects of long scotophase and cold acclimation on heat production in two diurnal rodents

Summary Heat production of two diurnal rodents,Rhabdomys pumilio andLemniscomys griselda was measured in long scotophase-LS (8L: 16D; 25°C) acclimated and long scotophase and cold — LSAC (8L: 16D; 6°C) acclimated animals and compared to a control group (12L: 12D; 25°C).LS increased in both species. Further acclimation of LSAC increased inR. pumilio and decreased inL. griselda. LS increased body temperature (T _b) inL. griselda only. LS increased overall thermal conductance in both species. LSAC caused a further increase in this parameter inR. pumilio.A singificant (P<0.001) increase in the magnitude of maximal nonshivering thermogenesis (NST) was observed in both species due to LS acclimation. LSAC did not change this maximal NST but increased its obligatory part (minimal , P<0.05, inL. griselda, andP<0.001, inR. pumilio).The results of this study show that winter acclimatization of heat production mechanisms, in both species, may be due to extension of scotophase.Abbreviations LS long scotophase - LSAC long scotophase and cold - NA noradrenaline - NST nonshivering thermogenesis - RMR resting metabolic rate     

Laser light scattering determination of size and dispersity of synaptosomes and synaptic vesicles isolated from squid (Loligo pealei) optic lobes

Quasi-elastic laser light scattering has been used to investigate the size and dispersity of synaptosomes and synaptic vesicles isolated from optic lobes of the squid Loligo pealei. Synaptosomal fractions were highly polydisperse ( ) and the mean diameter ( ) ranged from 0.5–2.0 m. Size distribution histograms yielded two major components — smaller particles ( ) and a larger group of particles ( ). The heterogeneity of the synaptosomal particles detected in solution is in agreement with published data obtained using electron microscopy. Purified synaptic vesicle fractions also yielded complex particle size distribution data. A component with a mean diameter in the range 150–250 nm was detected, though a smaller particle ( ) dominated the scattering signal. This smaller particle closely resembles in size the electron lucent vesicles seen in the majority of squid optic lobe nerve terminals when examined by electron microscopy. Osmotically-induced shirnkage and swelling of the synptosomes was detected. Depolarization by veratridine (1.0×10^–4 M) did not result in a detectable change in the size of synaptosomal particles.     

The effect of short-term fasting and a single meal on protein synthesis and oxygen consumption in cod,Gadus morhua

Summary Rates of protein synthesis and oxygen consumption ( O₂) in cod were compared in both fasted and refed animals. During a 14-day fast both protein synthesis and respiration rates fell to stable values after 6 days. When a meal of whole sandeel at 6% body weight was fed to fish fasted for 6 days, protein synthesis and ( O₂) increased to a maximum at between 12 and 18 h after feeding. Peak ( O₂) was about twice the pre-feeding values, while whole animal protein synthesis increased four-fold. There were differences between tissues in the timing of maximum protein synthesis; the liver and stomach responded faster than the remainder of the body. Maximum protein synthesis rates in the liver and stomach occurred at 6 h after feeding, at which time their calculated contribution to total ( O₂) was 11%. Similar calculations suggested that the integrated increment in whole animal protein synthesis contributed between 23% and 44% of the post-prandial increase in ( O₂). It was concluded that protein synthesis is an important contributor to increased ( O₂) after feeding in cod.Abbreviations A _s absolute rate of protein synthesis - ASDA apparent specific dynamic action - ATP adenosine triphosphate - k _s fractional rate of protein synthesis - k _s/RNA amount of protein synthesized per unit RNA - ( O₂) oxygen consumption - PCA perchloric acid - RNA ribonucleic acid     

Two-substrates packed-bed bioreactors: inhibition of enzyme and competitive binding of substrate and product to a ligand

An analytical model is developed to describe the performance of a packed-bed immobilized enzyme reactor in which parallel processes take place. In particular, two-substrate reaction, inhibition of the enzyme by one of the reaction products, and binding of one substrate and/or one product to an added ligand are taken into account. In addition, substrates and product diffusion into the porous catalyst are also considered. Using this model, numerical simulations were performed. The results point to the fact that, when all the above processes occur concomitantly, a variety of performance characteristics can be obtained, depending on the particular values of the related parameters. Moreover, under certain conditions, the reactor performance can be improved by controlled addition of ligand.List of Symbols A total concentration of ligand - C _1,i concentration of Substrate-1 in the pores of stage i - C _2,i concentration of Substrate-2 in its free form in the pores of stage i - _2,i concentration of the Substrate-2-Ligand Complex in the pores of stage i - total concentration of Substrate-2 in the pores of stage i - _i concentration of the Product-Ligand Complex in the pores of stage i - concentration of the free Product in the pores of stage i - total concentration of the Product in the pores of stage i - internal (pore) diffusion coefficient for the Substrate-Ligand Complex - D ₁ internal (pore) diffusion coefficient of Substrate-1 - D ₂ internal (pore) diffusion coefficient of Substrate-2 - effective (pore) diffusion coefficient for Substrate-2 - internal (pore) diffusion coefficient for the Product - internal (pore) diffusion coefficient for the Product-Ligand Complex - effective (pore) diffusion coefficient for the Product - K thermodynamic equilibrium constant for binding Substrate-2 to Ligand - K _m,1,K _m,2 Michaelis constants for Substrates-1 and 2, respectively - effective Michaelis constant for Substrate-2 - K _p thermodynamic equilibrium constant for binding the reaction Product to Ligand - effective equilibrium constant for binding Substrate-2 to Ligand - effective equilibrium constant for binding the reaction Product to Ligand. - K _b inhibition constant - K _q inhibition constant - effective inhibition constant - effective inhibition constant - k _a, k _d association and dissociation rate constants for Substrate-2 — Ligand complex - association and dissociation constants for Product —Ligand complex - n total number of elementary stages in the reactor - Q volumetric flow rate throughout the reactor - R _j,i reaction rate of Substrate-j in stage i, in terms of volumetric units - S _1,0 concentration of Substrate-1 in the reactor feed - total concentration of Substrate-2 in the reactor feed - S _1,i–1,S _1,i concentration of Substrate-1 in the bulk phase leaving stages i–1 and i, respectively - S _2,i concentration of Substrate-2 in its free form, in the bulk phase leaving stage i - _2,i–1, _2,i concentration of Substrate-2 in the bulk phase leaving stage i–1 and i, respectively - total concentration of Substrate-2 in the bulk phase leaving stages i–1 and i, respectively - _i concentration of the Product-Ligand Complex in the bulk phase of stage i - concentration of free Product in the bulk phase of stage i - total concentration of Product in the bulk phase of stage i - V total volume of the reactor - V _m maximal reaction rate in terms of volumetric units - y axial coordinate of the pores - y ₀ depth of the pores Greek Symbols ₁ dimensionless parameter - dimensionless parameter - dimensionless parameter - ₁ dimensionless parameter - dimensionless parameter - _1,i dimensionless concentration of Substrate-1 in pores of stage i - dimensionless total concentration of Substrate-2 (in both free and bound form) in pores of stage i - dimensionless total concentration of the reaction product in the pores of stage i - ₁ dimensionless parameter - dimensionless parameter - dimensionless parameter - dimensionless parameter - dimensionless parameter - dimensionless position along the pore - volumetric packing density of catalytic particles (dimensionless) - porosity of the catalytic particles (dimensionless) - _1,i dimensionless concentration of Substrate-1 in the bulk phase of stage i - dimensionless total concentration of Substrate-2 (in both free and bound form) in the bulk phase of stage i     

Linkage analysis using multiple DNA polymorphic markers in normal families and in families with fragile X syndrome

Summary Linkage data, using the polymorphic markers 52A (DXS51), F9, 4D-8(DXS98), and St14(DXS52), are presented from 14 fragile X pedigrees and from 7 normal pedigrees derived from the collection of the Centre d'Étude du Polymorphisme Humaine. A multipoint linkage analysis indicates that the most probable order of these four loci in normal families is DXS51-F9-DXS98-DXS52. Recombination frequencies ( ) corresponding to maximum LOD scores ( ) were obtained by two-point linkage analysis for a nuber of linkage groups, including: DXS51-F9 ( =5.94, =0.03), F9-DXS98 ( =0.51, =0.26), F9-DXS52 ( =0.84, =0.27), and DXS98-DXS52 ( =0.32, =0.20). A multipoint linkage analysis of these loci, including the fragile X locus, was also performed for the fragile X population and the data support the relative order (DSX51, F9, DXS98)-FRAXA-DXS52. Recombination frequencies and maximum LOD scores, which again were derived from two-point linkage analyses, were obtained for the linkage groups DXS51-F9 ( =9.96, =0) and F9-DXS52 ( =0.07, =0.45), as well as for the groups DXS51-FRAXA ( =2.42, =0.15), F9-FRAXA ( =1.30, =0.18), DXS98-FRAXA ( =0.05 =0.36), and DXS52-FRAXA ( =2.42 =0.15). The linkage data was further tested for the presence of genetic heterogeneity both within and between the fragile X and normal families for the intervals DXS51-F9, F9-DXS52, F9-FRAXA, and DXS52-FRAXA using a modification of the A test. Except for the interval F9-FRAXA (P<0.10) there was no evidence of genetic heterogeneity for each of the various linkage groups examined. The heterogeneity detected for the interval F9-FRAXA, however, was most likely due to one family (Fx-28) that displayed very tight linkage between these two loci.     

Evidence that the intrinsic membrance protein LHCII in thylakoids is necessary for maintaining localized\Delta \tilde \mu _{H^ + } energy coupling

This work tested the hypothesis that thylakoid localized proton-binding domains, suggested to be involved in localized -driven ATP formation, are maintained with the involvement of several membrane proteins, including the LHCII (Laszlo, J. A., Baker, G. M., and Dilley, R. A. (1984) Biochim. Biophys. Acta 764, 160–169), which comprises about 50% of the total thylakoid protein. The concept we have in mind is that several membrane proteins cooperate to shield a localized proton diffusion pathway from direct contact with the lumen, thus providing a physical barrier to H⁺ equilibration between the sequestered domains and the lumen. A barely mutant,chlorina f ₂, that lacks Chl b and does not accumulate some of the LHCII proteins, was tested for its capacity to carry out localized-proton gradient-dependent ATP formation. Two previously developed assays permit clear discrimination between localized and delocalized gradient-driven ATP formation. Those assays include the effect of a permeable buffer, pyridine, on the number of single-turnover flashes needed to reach the energetic threshold for ATP formation and the more recently developed assay for lumen pH using 8-hydroxy-1,3,6-pyrene trisulfonic acid as a lumenally loaded pH-sensitive fluorescent probe. By those two criteria, the wild-type barley thylakoids revealed either a localized or a delocalized energy coupling mode under low- or high-salt storage conditions, respectively. Addition of Ca⁺⁺ to the high-salt storage medium caused those thylakoids to maintain a localized energy-coupling response, as previously observed for pea thylakoids. In contrast, thechlorina f ₂ mutant thylakoids had an active delocalized energy coupling activity but did not show localized energy coupling under any conditions, and added Ca⁺⁺ to the thylakoid storage medium did not alter the delocalized energy coupling mode. One interpretation of the results is that the absence of the LHCII polypeptides produces a leaky pathway for protons which allows the gradient to equilibrate with the lumen under all conditions. Another interpretation is possible but seems less likely, that being that the absence of the LHCII polypeptides in some way causes the proposed Ca⁺⁺ -gated H⁺ flux site on the membrane sector (CF₀) of the energy coupling complex to lose its gating function.