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Proximate causes of adaptive growth rates: growth efficiency variation among latitudinal populations of Rana temporaria.

In ectothermic organisms, declining season length and lower temperature towards higher latitudes often select for latitudinal variation in growth and development. However, the energetic mechanisms underlying this adaptive variation are largely unknown. We investigated growth, food intake and growth efficiency of Rana temporaria tadpoles from eight populations along a 1500 km latitudinal gradient across Sweden. To gain an insight into the mechanisms of adaptation at organ level, we also examined variation in tadpole gut length. The tadpoles were raised at two temperatures (16 and 20 degrees C) in a laboratory common garden experiment. We found increased growth rate towards higher latitudes, regardless of temperature treatment. This increase in growth was not because of a higher food intake rate, but populations from higher latitudes had higher growth efficiency, i.e. they were more efficient at converting ingested food into body mass. Low temperature reduced growth efficiency most strongly in southern populations. Relative gut length increased with latitude, and tadpoles at low temperature tended to have longer guts. However, variation in gut length was not the sole adaptive explanation for increased growth efficiency as latitude and body length still explained significant amounts of variation in growth efficiency. Hence, additional energetic adaptations are probably involved in growth efficiency variation along the latitudinal gradient.

Adaptation, Physiological↗

Migration efficiency ratios and the optimal distribution of population.

The authors present a theoretical description of the migration process and criticize the conventional interpretation of the migration efficiency ratio, which is defined as the ratio of the net number of moves of individuals between areas to the gross number of moves that take place. "The conventional interpretation of the migration efficiency ratio is that the closer it lies to zero the less efficient the migration process....However, [the authors] feel that this is a somewhat misleading conception of the notion of efficiency in migration in that it emphasizes the physical efficiency of the migration process rather than focusing on the contribution of migration to a socially efficient allocation of population. Thus, to redirect attention, [they] have chosen to judge migration efficiency on the basis of its contribution to producing an equilibrium population distribution." The focus is on internal migration.

Demography↗

Cross-linked polyethylenimine as potential DNA vector for gene delivery with high efficiency and low cytotoxicity.

Polyethylenimine (PEI) has been known as an efficient gene carrier with the highest cationic charge potential. High transfection efficiency of PEI, along with its cytotoxicity, strongly depends on its molecular weight. To enhance its gene delivery efficiency and minimize cytotoxicity, we have synthesized small cross-linked PEI with biodegradable linkages and evaluated their transfection efficiencies in vitro. In this study, branched PEI with a molecular weight of 800 Da was cross-linked by small diacrylate [1,4-butanediol diacrylate or ethyleneglycol dimethacrylate (EGDMA)] for 2-6 h. The efficiencies of the cross-linked PEI in in vitro transfection of plasmid DNA containing enhanced green fluorescent protein (EGFP) reporter gene were assessed in melanoma B16F10 cell line and other cell lines. Flow cytometry was used to quantify the cellular entry efficiency of plasmid and the transgene expression level. The cytotoxicities of the cross-linked PEI in these cells were evaluated by MTT assay. EGDMA-PEI 800-4h, a typical cross-linked PEI reported here, mediated a more efficient expression of reporter gene than the commercially available 25-kDa branched PEI control, and resulted in a 9-fold increase in gene delivery in B16F10 cells and a 16-fold increase in 293T cells, while no cytotoxicity was found at the optimized condition for gene delivery. Furthermore, the transfection activity of polyplexes was preserved in the presence of serum proteins.

Animals↗

Mechanical work and efficiency in ergometer bicycling at aerobic and anaerobic thresholds.

Internal and external mechanical work, energy consumption and mechanical efficiency were studied in constant-load ergometer bicycling at five different power outputs below, equal to, and above the aerobic (AerT) and anaerobic (AnT) thresholds. The gross, net and true efficiencies of the whole body in five male subjects were calculated. The work against the external load was defined as the external mechanical work. The internal mechanical work was calculated as the sum of the increments of kinetic and potential energy in all body segments by using methods of film analysis. Total energy consumption was measured by combining aerobic and anaerobic energy production. When the power output of the bicycle ergometer was increased from 146 +/- 15 to 283 +/- 17 W, oxygen consumption increased from 2.20 +/- 0.98 to 4.22 +/- 0.20 l min-1 (P less than 0.001), while the oxygen consumption at rest was 0.30 +/- 0.03 l min-1. The concentration of blood lactate increased from 2.2 +/- 0.4 at the lowest work load to 8.6 +/- 1.2 mmol l-1 at the highest work load (P less than 0.001). The amount of external work done per revolution increased from 139 +/- 20 to 277 +/- 29 J (P less than 0.001), while the amount of internal work per revolution remained almost constant (56 +/- 12 J). The gross efficiency in the present study was 17-20%, net efficiency 18-22% and true efficiency 21-30%, respectively. The highest gross and net efficiencies were reached at the AerT. The lowest efficiencies were obtained at highest work load.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

The efficiency of clioxanide and rafoxanide against Fasciola hepatica in sheep by different routes of administration.

Groups of sheep were infected with 100 viable metacercariae of Fasciola hepatica. The efficiencies of clioxanide and rafoxanide were evaluated against infections aged 6 and 12 weeks, by using 3 dose rates of each compound by each of 3 routes of administration. Clioxanide, at 40 mg/kg, administered orally and intra-ruminally against 6-week-old liver fluke was 85 and 90% efficient respectively. At 80 mg/kg intra-abomasally its efficiency was 43%. Clioxanide at 20 mg/kg was 96% efficient against 12-week-old liver fluke when given orally or intra-ruminally and 82% efficient when given intra-abomasally. Because of its lower efficiency intra-abomasally, clioxanide may be unsatisfactory in a proportion of sheep, for use against immature liver fluke at a dose rate of 40 mg/kg. It may be expected to give a moderate to high efficiency against mature infections at 20 mg/kg.

Abomasum↗

A nine-residue synthetic propeptide enhances secretion efficiency of heterologous proteins in Lactococcus lactis.

Lactococcus lactis, a gram-positive organism widely used in the food industry, is a potential candidate for the secretion of biologically useful proteins. We examined the secretion efficiency and capacity of L. lactis by using the Staphylococcus aureus nuclease (Nuc) as a heterologous model protein. When expressed in L. lactis from an efficient lactococcal promoter and its native signal peptide, only approximately 60% of total Nuc was present in a secreted form at approximately 5 mg per liter. The remaining 40% was found in a cell-associated precursor form. The secretion efficiency was reduced further to approximately 30% by the deletion of 17 residues of the Nuc native propeptide (resulting in NucT). We identified a modification which improved secretion efficiency of both native Nuc and NucT. A 9-residue synthetic propeptide, LEISSTCDA, which adds two negative charges at the +2 and +8 positions, was fused immediately after the signal peptide cleavage site. In the case of Nuc, secretion efficiency was increased to approximately 80% by LEISSTCDA insertion without altering the signal peptide cleavage site, and the yield was increased two- to fourfold (up to approximately 20 mg per liter). The improvement of NucT secretion efficiency was even more marked and rose from 30 to 90%. Similarly, the secretion efficiency of a third protein, the alpha-amylase of Bacillus stearothermophilus, was also improved by LEISSTCDA. These data indicate that the LEISSTCDA synthetic propeptide improves secretion of different heterologous proteins in L. lactis.

Amino Acid Sequence↗

Effect of obesity and insulin resistance on myocardial substrate metabolism and efficiency in young women.

BACKGROUND: Obesity is a risk factor for impaired cardiac performance, particularly in women. Animal studies suggest that alterations in myocardial fatty acid metabolism and efficiency in obesity can cause decreased cardiac performance. In the present study, we tested the hypothesis that myocardial fatty acid metabolism and efficiency are abnormal in obese women. METHODS AND RESULTS: We studied 31 young women (body mass index [BMI] 19 to 52 kg/m2); 19 were obese (BMI >30 kg/m2). Myocardial oxygen consumption (MVO2) and fatty acid uptake (MFAUp), utilization (MFAU), and oxidation (MFAO) were quantified by positron emission tomography. Cardiac work was measured by echocardiography, and efficiency was calculated as work/MVO2. BMI correlated with MVO2 (r=0.58, P=0.0006), MFAUp (r=0.42, P<0.05), and efficiency (r=-0.40, P<0.05). Insulin resistance, quantified by the glucose area under the curve (AUC) during an oral glucose tolerance test, correlated with MFAUp (r=0.55, P<0.005), MFAU (r=0.62, P<0.001), and MFAO (r=0.58, P<0.005). A multivariate, stepwise regression analysis showed that BMI was the only independent predictor of MVO2 and efficiency (P=0.0005 and P<0.05, respectively). Glucose AUC was the only independent predictor of MFAUp, MFAU, and MFAO (P<0.05, <0.005, and <0.005, respectively). CONCLUSIONS: In young women, obesity is a significant predictor of increased MVO2 and decreased efficiency, and insulin resistance is a robust predictor of MFAUp, MFAU, and MFAO. This increase in fatty acid metabolism and decrease in efficiency is concordant with observations made in experimental models of obesity. These metabolic changes may play a role in the pathogenesis of decreased cardiac performance in obese women.

Adult↗

Decreased contractile efficiency and increased nonmechanical energy cost in hyperthyroid rabbit heart. Relation between O2 consumption and systolic pressure-volume area or force-time integral.

Both systolic pressure-volume area (PVA) and force-time integral (FTI) have been used as measures of oxygen consumption per beat (VO2) in the isolated left ventricle. The reciprocal of the slope of the VO2-PVA relation has been considered to reflect the chemomechanical energy transduction efficiency of the contractile machinery (contractile efficiency), whereas its VO2 intercept consists of energy cost of excitation-contraction coupling and basal metabolism. To examine whether the increase in myosin isoform V1/V3 ratio in hyperthyroid rabbits decreases contractile efficiency and to determine overall mechanisms of higher oxygen consumption in hyperthyroid hearts, the VO2-PVA and VO2-FTI relations as well as the end-systolic pressure-volume relation were assessed in cross-circulated, isovolumically beating hearts isolated from normal, hyperthyroid, and hypothyroid rabbits. Normalized initial slopes of the rising limb of the curvilinearly fitted end-systolic pressure-volume relation (E'max, ventricular contractility index) were similar for normal and hyperthyroid groups. However, the slopes and VO2 intercepts of the VO2-PVA and VO2-FTI relations were greater in hyperthyroid hearts than in normal hearts. Accordingly, in the hyperthyroid hearts, the contractile efficiency (27 +/- 6%) was lower and left ventricular VO2 for excitation-contraction coupling (0.028 +/- 0.004 ml O2/beat/100 g) was higher than in normal hearts (40 +/- 4% and 0.021 +/- 0.005 ml O2/beat/100 g, respectively). This decreased contractile efficiency in the hyperthyroid hearts was attributable to myosin isoform alteration rather than to increased beta-adrenoceptors because isoproterenol did not affect the slope of the VO2-PVA relation in all groups. In contrast, the slope of the VO2-FTI relation was significantly increased by isoproterenol in all groups. Neither the VO2-PVA nor the VO2-FTI relations in hypothyroid hearts were different from those in normal hearts except for significantly lower VO2 for basal metabolism. We conclude that in hyperthyroid rabbits, the left ventricle has decreased contractile efficiency and increased energy cost of excitation-contraction coupling and that the decreased contractile efficiency in hyperthyroid hearts is probably due to the increased V1/V3 ratio of the myosin isoform component. In addition, this study demonstrates that the VO2-PVA and VO2-FTI relations dissociate depending on the intervention, even in the same isovolumic contraction mode.

Animals↗

Cardiac efficiency is improved after ischemia by altering both the source and fate of protons.

Cardiac efficiency is decreased in hearts after severe ischemia. We determined whether reducing the production of H+ from glucose metabolism or inhibiting the clearance of H+ via Na(+)-H+ exchange could increase cardiac efficiency during reperfusion. This was achieved using dichloroacetate (DCA) to stimulate glucose oxidation and 5-(N,N-dimethyl)-amiloride (DMA) to inhibit Na(+)-H+ exchange, respectively. Isolated working rat hearts were subjected to 30 minutes of global ischemia and 60 minutes of reperfusion. Glycolysis and oxidation rates of glucose, lactate, and palmitate were measured. Recovery of cardiac work, O2 consumption (MVO2), and rates of acetyl-coenzyme A and ATP production during reperfusion were determined. After ischemia, cardiac work recovered to 35 +/- 5% of preischemic values in control hearts (n = 23), although MVO2, tricarboxylic acid (TCA) cycle activity, and ATP production from glycolysis and oxidative metabolism rapidly recovered to preischemic levels. This decrease in cardiac efficiency was accompanied by a substantial production of H+ from glucose metabolism DCA caused a 2.2-fold increase in glucose oxidation, a 46 +/- 17% decrease in H+ production, a 1.6-fold increase in cardiac efficiency, and a 2.0-fold increase in cardiac work during reperfusion (n = 17). Inhibition of Na(+)-H+ exchange with DMA did not alter TCA cycle activity and ATP production rates but did result in a 1.8-fold increase in cardiac efficiency and a 1.7-fold increase in cardiac work (n = 12). These data show that cardiac efficiency and the contractile function after ischemia can be improved by either reducing the rate of H+ production from glucose metabolism during reperfusion or inhibiting the clearance of H+ via Na(+)-H+ exchange. Our data suggest that an increased requirement for ATP to restore ischemia-reperfusion-induced alterations in ion homeostasis contributes to the decrease in cardiac efficiency and contractile function after ischemia.

Acetyl Coenzyme A↗

Efficiency of the respiratory muscles in healthy individuals.

We evaluated respiratory muscle performance by determining the energy cost of breathing against incremental threshold loads and calculating the efficiency of the respiratory apparatus for handling the added work. In five subjects, the energy cost of breathing against the loads, and thus the calculated efficiency of the respiratory muscles, was reproducible on repeated measurements. In all subjects, the calculated efficiency varied while breathing with low loads but was relatively constant at loads that resulted in a mouth pressure of 20 to 60% of the subjects' maximal static inspiratory pressure (PImax). The mean efficiency calculated between 20 to 60% of PImax in 30 normal subjects (15 males and 15 females) ranged between 1.54 and 7.98%. It was significantly greater in males (5.41 +/- 0.43%) (mean +/- SEM) than in females (2.41 +/- 0.17%). There was no relationship between efficiency and body size, but the efficiency correlated with inspiratory muscle strength (PImax). We suggest that measurement of the efficiency of the respiratory muscles over a spectrum of incremental threshold loads is a simple, noninvasive, and reproducible method of assessing respiratory muscle performance and may have clinical application.

Adolescent↗

Investigation on the emission factors and removal efficiencies of heavy metals from MSW incinerators in Taiwan.

Two large-scale municipal waste incinerators (MWIs) equipped with different air pollution control devices (APCDs) were selected for conducting flue gas sampling and determination of heavy metals emissions including lead, zinc, cadmium, chromium, copper and mercury. In addition, the emission characteristics and removal efficiencies of heavy metals were evaluated via simultaneous sampling of flue gas upstream and downstream of APCDs. The average removal efficiency of particulate matter was 99.47%, and the removal efficiency of heavy metals could reach 95% or higher (except for Hg) for MWI-A. Preliminary results indicated that the average removal efficiency of particulate matter was 99.94%, and the removal efficiency of heavy metals could reach 98% or higher (except for Hg) for MWI-B. The average removal efficiencies of Hg for MWI-A and MWI-B were 47% and 30% respectively. The removal efficiencies achieved with existing APCDs were close to that reported in the US EPA document (AP-42) for MWIs. The average concentrations of heavy metals and particulate matter emitted from stack could meet current emission standards in Taiwan for both MWIs, but particulate matter, Pb and Cd concentrations of MWI-A were higher than the current emission regulations in Germany and the Netherlands.

Air Pollutants↗

Highly efficient, large volume flow electroporation.

Electroporation is widely used to transfect and load cells with various molecules. Traditional electroporation using a static mode is typically restricted to volumes less than 1 mL, which limits its use in clinical and industrial bioprocessing applications. Here we report efficient, large volume transfection results by using a scalable-volume electroporation system. Suspended (Jurkat) and adherent cells (10T1/2 and Huh-7) were tested. A large macromolecule, FITC-conjugated dextran (MW=500 kD) was used to measure cell uptake, while a plasmid carrying the gene coding for enhanced green fluorescence protein (eGFP) was used to quantitate the flow electrotransfection efficiency as determined by flow cytometry. The flow electroloading efficiency of FITC-dextran was >90%, while the cell viability was highly maintained (>90%). High flow electrotransfection efficiency (up to 75%) and cell viability (up to 90%) were obtained with processing volumes ranging from 1.5 to 50 mL. No significant difference of electrotransfection efficiency was observed between flow and static electrotransfection. When 50 mL of cell volume was processed and samples collected at different time points during electroporation, the transgene expression and cell viability results were identical. We also demonstrated that DNA plasmid containing EBNA1-OriP elements from Epstein-Barr virus were more efficient in transgene expression than standard plasmid without the elements (at least 500 too 1000-fold increase in expression level). Finally, to examine the feasibility of utilizing flow electrotransfected cells as a gene delivery vehicle, 10T1/2 cells were transfected with a DNA plasmid containing the gene coding for mIL12. mIL12 transfected cells were injected subcutaneously into mice, and produced functional mIL12, as demonstrated by anti-angiogenic activity. This is the first demonstration of efficient, large volume, flow electroporation and the in vivo efficacy of flow electrotransfected cells. This technology may be useful for clinical gene therapy and large-scale bioprocesses.

Animals↗

Technical efficiency of public district hospitals and health centres in Ghana: a pilot study.

BACKGROUND: The Government of Ghana has been implementing various health sector reforms (e.g. user fees in public health facilities, decentralization, sector-wide approaches to donor coordination) in a bid to improve efficiency in health care. However, to date, except for the pilot study reported in this paper, no attempt has been made to make an estimate of the efficiency of hospitals and/or health centres in Ghana. The objectives of this study, based on data collected in 2000, were: (i) to estimate the relative technical efficiency (TE) and scale efficiency (SE) of a sample of public hospitals and health centres in Ghana; and (ii) to demonstrate policy implications for health sector policy-makers. METHODS: The Data Envelopment Analysis (DEA) approach was used to estimate the efficiency of 17 district hospitals and 17 health centres. This was an exploratory study. RESULTS: Eight (47%) hospitals were technically inefficient, with an average TE score of 61% and a standard deviation (STD) of 12%. Ten (59%) hospitals were scale inefficient, manifesting an average SE of 81% (STD = 25%). Out of the 17 health centres, 3 (18%) were technically inefficient, with a mean TE score of 49% (STD = 27%). Eight health centres (47%) were scale inefficient, with an average SE score of 84% (STD = 16%). CONCLUSION: This pilot study demonstrated to policy-makers the versatility of DEA in measuring inefficiencies among individual facilities and inputs. There is a need for the Planning and Budgeting Unit of the Ghana Health Services to continually monitor the productivity growth, allocative efficiency and technical efficiency of all its health facilities (hospitals and health centres) in the course of the implementation of health sector reforms.

Journal Article↗

Efficiency of fast- and slow-twitch muscles of the mouse performing cyclic contractions.

The mechanical efficiency of mouse fast- and slow-twitch muscle was determined during contractions involving sinusoidal length changes. Measurements were made of muscle length, force production and initial heat output from bundles of muscle fibres in vitro at 31 degrees C. Power output was calculated as the product of the net work output per sinusoidal length cycle and the cycle frequency. The initial mechanical efficiency was defined as power output/(rate of initial heat production+power output). Both power output and rate of initial heat production were averaged over a full cycle of length change. The amplitude of length changes was +/- 5% of muscle length. Stimulus phase and duration were adjusted to maximise net work output at each cycle frequency used. The maximum initial mechanical efficiency of slow-twitch soleus muscle was 0.52 +/- 0.01 (mean +/- 1 S.E.M. N = 4) and occurred at a cycle frequency of 3 Hz. Efficiency was not significantly different from this at cycle frequencies of 1.5-4 Hz, but was significantly lower at cycle frequencies of 0.5 and 1 Hz. The maximum efficiency of fast-twitch extensor digitorum longus muscle was 0.34 +/- 0.03 (N = 4) and was relatively constant (0.32-0.34) over a broad range of frequencies (4-12 Hz). A comparison of these results with those from previous studies of the mechanical efficiency of mammalian muscles indicates that efficiency depends markedly on contraction protocol.

Animals↗

Power at the expense of efficiency in contraction of white muscle fibres from dogfish Scyliorhinus canicula

Work and heat production of white myotomal muscle fibres from dogfish were measured during sinusoidal movement (0.71-5.0 Hz) at 12 C. Stimulus phase (stimulus timing relative to movement) and duty cycle (stimulus duration as a fraction of movement cycle duration) were varied to determine the parameters optimal for power output and for efficiency (work/total energy output). Movements of 0.067 and 0.120L0 were used, where L0 is the muscle fibre length giving maximum force in an isometric tetanus. At each frequency of movement and duty cycle, the stimulus phase giving the highest power was the same as that giving the highest efficiency. In contrast, at each frequency and optimal stimulus phase, the dependence of power on duty cycle was very different from the dependence of efficiency on duty cycle. Power generally increased with increasing duty cycle, whereas efficiency decreased. Thus, high power can be achieved at the expense of efficiency by adjusting stimulus duty cycle. When stimulus phase and duty cycle were optimized, efficiency was always higher for the larger distance of movement. The efficiency of energy conversion can be maintained at a high level as the frequency of movement increases from 1.25 to 5.0 Hz.

Journal Article↗

The efficiency of an asynchronous flight muscle from a beetle.

Mechanical power output and metabolic power input were measured from an asynchronous flight muscle, the basalar muscle of the beetle Cotinus mutabilis. Mechanical power output was determined using the work loop technique and metabolic power input by monitoring CO(2) production or both CO(2) production and O(2) consumption. At 35 degrees C, and with conditions that maximized power output (60 Hz sinusoidal strain, optimal muscle length and strain amplitude, 60 Hz stimulation frequency), the peak mechanical power output during a 10 s burst was approximately 140 W kg(-1), the respiratory coefficient 0.83 and the muscle efficiency 14-16 %. The stimulus intensity used was the minimal required to achieve a maximal isometric tetanus. Increasing or decreasing the stimulus intensity from this level changed mechanical power output but not efficiency, indicating that the efficiency measurements were not contaminated by excitation of muscles adjacent to that from which the mechanical recordings were made. The CO(2) produced during an isometric tetanus was approximately half that during a bout of similar stimulation but with imposed sinusoidal strain and work output, suggesting that up to 50 % of the energy input may go to muscle activation costs. Reducing the stimulus frequency to 30 Hz from its usual value of 60 Hz reduced mechanical power output but had no significant effect on efficiency. Increasing the frequency of the sinusoidal strain from 60 to 90 Hz reduced power output but not CO(2) consumption; hence, there was a decline in efficiency. The respiratory coefficient was the same for 10 s and 30 s bursts of activity, suggesting that there was no major change in the fuel used over this time range. The mass-specific mechanical power output and the efficiency of the beetle muscle were each 2-3 times greater than values measured in previous studies, using similar techniques, from locust flight muscles, which are synchronous muscles. These results support the hypothesis that asynchronous flight muscles have evolved in several major insect taxa because they can provide greater power output and are more efficient than are synchronous muscles for operation at the high frequencies of insect flight.

Animals↗

How the efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle changes with cycle frequency.

Different species of animals require different cardiac performance and, in turn, their cardiac muscle exhibits different properties. A comparative approach can reveal a great deal about the mechanisms underlying myocardial contraction. Differences in myocardial Ca(2+) handling between fish and mammals suggest a greater energy cost of activation in fish. Further, while there is considerable evidence that heart rate (or cycle frequency) should have a profound effect on the efficiency of teleost cardiac muscle, this effect has been largely overlooked. We set out to determine how cycle frequency affects the power output and efficiency of rainbow trout (Oncorhynchus mykiss) ventricular muscle and to relate this to the heart's function in life. We measured power output and the rate of oxygen consumption ((O(2))) and then calculated efficiency over a physiologically realistic range of cycle frequencies. In contrast to mammalian cardiac muscle, in which (O(2)) increases with increasing heart rate, we found no significant change in (O(2)) in the teleost. However, power output increased by 25 % as cycle frequency was increased from 0.6 to 1.0 Hz, so net and total efficiency increased. A maximum total efficiency of 20 % was achieved at 0.8 Hz, whereas maximum power output occurred at 1.0 Hz. We propose that, since the heart operates continuously, high mechanical efficiency is a major adaptive advantage, particularly at lower heart rates corresponding to the more commonly used slower, sustainable swimming speeds. Efficiency was lower at the higher heart rates required during very fast swimming, which are used during escape or prey capture. If a fixed amount of Ca(2+) is released and then resequestered each time the muscle is activated, the activation cost should increase with frequency. We had anticipated that this would have a large effect on the total energy cost of contraction. However, since (O(2)) remains constant, less oxygen is consumed per cycle at high frequencies. We suggest that a constant (O(2)) would be observed if the amount of activator Ca(2+) were to decrease with frequency. This decrease in activation energy is consistent with the decrease in the systolic intracellular Ca(2+) ([Ca(2+)](i)) transient with increasing stimulation frequency seen in earlier studies.

Anaerobiosis↗

Asialoganglioside enhances the efficiency of gene transfection mediated by cationic liposomes with a cationic cholesterol derivative.

We investigated the transfection efficiency mediated by asialoganglioside-containing cationic liposomes. Previously we reported that monosialoganglioside GM(1) (GM(1a)) enhanced transfection efficiency. In this study, we investigated the effects of sialic acid in gangliosides on transfection efficiency. Two mammalian culture cell lines HeLa and HepG2 were transfected with luciferase plasmids (pGL3) using cationic liposomes which contain monosialoganglioside GM(1) (GM(1a)) or its asialic counterpart, asialoganglioside GM(1) (GA(1)). Both GM(1a) and GA(1) enhanced the efficiency of transfection mediated by cationic liposomes, and GA(1) exhibited higher efficiency than GM(1a) in both cell lines. Transfection efficiency of ganglioside-containing liposomes was also assessed by the effects of antisense oligonucleotides (AS-ODN) for bcl-2 gene, which suppresses apoptotic cell death. Western blotting analysis revealed that the expression of Bcl-2 was decreased by AS-ODN, and the reduction of protein expression in cells treated with GA(1)-containing liposomes was more remarkable than that with GM(1a)-containing liposomes. Furthermore, the induction rate of apoptosis was higher in cells treated with AS-ODN with GA(1)-containing liposomes. Together with the results obtained by luciferase assay mentioned above, the removal of sialic acid from ganglioside causes the enhancement of efficiency of transfection mediated by cationic liposomes.

Apoptosis↗