Heart design: free ADP scales with absolute mitochondrial and myofibrillar volumes from mouse to human

Our aim was to estimate a number of bioenergetic parameters in the beating mouse, rat and guinea pig heart in situ and compare the values to those in hearts of mammals over a 2000-fold range in body mass. For the mouse, rat and guinea pig heart, we report a phosphorylation ratio of 1005+/-50 (n=16), 460+/-32 (n=10) and 330+/-22 (n=5) mM(-1) and a free cytosolic [ADP] concentration of 13, 18 and 22 microM, respectively. When each parameter was plotted against body mass, they scaled closely to the quarter power (-0.28, r=0.99 and -0.23, r=0.97). A similar regression slope was found when the inverse of free [ADP] was plotted against absolute mitochondrial (slope=-0.26, r=0.99) and myofibrillar volumes (slope=-0.24, r=0.99). The similar slopes indicate that the ratio of absolute mitochondria and myofibrillar volumes in the healthy mammalian heart is a constant, and independent of body size. In conclusion, our study supports the hypothesis that the mammalian heart has a number of highly conserved thermodynamic and kinetic parameters that obey quarter-power laws linking the phosphorylation ratio, ATP turnover rates, free [ADP] and absolute mitochondrial volumes to body size. The results are discussed in terms of possible mechanisms and potential deviations from these laws in some disease states.     

Factors controlling vegetation structure in peatland lakes—Two conceptual models of plant zonation

Two conceptual models of plant zonation in peatland lakes are given. The first represents vegetation on slightly sloping substrate (N < 0.2) in shallow and relatively large lakes. The vegetation is not diverse (H′ = 0.0 ± 0.01). The frequency and biomass of the dominant (Sphagnum denticulatum) correlate positively with lake size, and negatively with depth and substrate slope. They are also correlated with water transparency and water color (r = −0.53), concentrations of total organic carbon (r = −0.43), Ca²⁺ (r = 0.40) and humic acids (r = −0.46), and redox potential (r = 0.44). The second model represents vegetation on steep peat walls (N > 0.3) in deep, usually small lakes. Plants occur only on the upper part of the peat wall or form a multispecies curtain hanging from the lip of peat at the top. Species diversity in this scenario is higher (H′ = 0.18 ± 0.17). The curtains usually are composed of mosses such as Warnstorfia exannulata, S. cuspidatum and S. riparium, and vascular plants are rare. The frequency and biomass of bryophytes in this type of structure are related to substrate slope (r = 0.56), lake depth (r = 0.56), Ca²⁺ concentration (r = −0.69) and water color (r = −0.51). In both models, plant biomass is correlated with temperature (r = −0.78), irradiance (r = −0.64) and water oxygenation (r = −0.54).     

Relationship between total ghrelin and inflammation in hemodialysis patients

In hemodialysis (HD) patients studies have shown that plasma ghrelin is increased and it has been speculated that ghrelin levels might be related to systemic inflammation. The present study attempted to correlate the serum levels of total ghrelin with serum TNF-α and IL-6, and with nutritional status and body composition in HD patients. Forty-seven HD patients from a single dialysis unit (18 women, mean age 55.3 ± 12.2 yr; BMI 24.4 ± 4.2 kg/m²; % body fat 29.4 ± 7.4%) were studied and compared to 21 healthy subjects (12 women, 50.7 ± 15.7 yr and BMI 25.6 ± 4.0 kg/m²; % body fat 30.0 ± 5.7%). Biochemical data, serum total ghrelin, TNF-α and IL-6 levels were measured. The body composition was evaluated by dual energy X-ray absortiometry (DEXA) and energy and protein intake were evaluated. Patients showed elevated plasma ghrelin levels when compared to healthy subjects (1.14 ± 1.0 ng/mL vs 0.58 ± 0.4; p < 0.001). There was a positive correlation between ghrelin levels and TNF-α (r = 0.25; p < 0.04), IL-6 (r = 0.42; p < 0.02), and a negative correlation between TNF-α and protein intake (r = −0.28; p < 0.03), and energy intake (r = −0.34; p < 0.01). No correlation was observed with any aspect of body composition. Plasma ghrelin levels are elevated in HD patients and associated with the state of systemic inflammation. We suggest that the inflammatory state may affect ghrelin bioactivity and metabolism in hemodialysis patients.     

Distribution and residual activity of two insecticidal proteins, avidin and aprotinin, expressed in transgenic tobacco plants, in the bodies and frass of Spodoptera litura larvae following feeding

To understand how a major cosmopolitan pest responds to two very different insecticidal proteins and to determine whether herbivorous insects and their frass could be environmental sources of recombinant proteins from transgenic plants, Spodoptera litura (Fab.) (Lepidoptera, Noctuidae) larvae were fed on tobacco leaves expressing either the biotin-binding protein, avidin, or the protease inhibitor, aprotinin. Control larvae received non-transgenic tobacco. Samples of larvae were taken after 5, 6 or 7 days’ feeding and frass was collected after two 24-h periods at 6 and 7 days. Insects in all treatments grew significantly during the experiment, but the avidin-fed larvae were significantly smaller than the others on Day 7. Avidin was found in all samples of avidin-fed larvae (7.0±0.86 ng mg⁻¹, n=45), at a lower level than in their frass (31.9±5.08 ng mg⁻¹, n=30), and these frass levels were lower than those of the the leaves fed to the larvae (69.0±6.71 ng mg⁻¹, n=45). All of the avidin detected in these samples was capable of binding biotin. On average, between 10 and 28% of avidin was recovered with the methods used, whereas almost full recovery of aprotinin was effected. Aprotinin levels in larvae (8.2±0.53 ng mg⁻¹, n=45) were also lower than aprotinin levels in frass (77.4±6.9 ng mg⁻¹, n=30), which were somewhat lower than those in the leaves fed to the larvae (88.6±2.51 ng mg⁻¹, n=45). Approximately half the trypsin-binding ability of aprotinin was lost in larvae, and in frass, aprotinin had lost about 90% of its ability to bind trypsin.     

A Single-Cell Model of Phase-Driven Control of Ventricular Fibrillation Frequency

The mechanisms controlling the rotation frequency of functional reentry in ventricular fibrillation (VF) are poorly understood. It has been previously shown that Ba²⁺ at concentrations up to 50 μmol/L slows the rotation frequency in the intact guinea pig (GP) heart, suggesting a role of the inward rectifier current (I_K1) in the mechanism governing the VF response to Ba²⁺. Given that other biological (e.g., sinoatrial node) and artificial systems display phase-locking behavior, we hypothesized that the mechanism for controlling the rotation frequency of a rotor by I_K1 blockade is phase-driven, i.e., the phase shift between transmembrane current and voltage remains constant at varying levels of I_K1 blockade. We measured whole-cell admittance in isolated GP myocytes and in transfected human embryonic kidney (HEK) cells stably expressing Kir 2.1 and 2.3 channels. The admittance phase, i.e., the phase difference between current and voltage, was plotted versus the frequency in control conditions and at 10 or 50 μmol/L Ba²⁺ (in GP heart cells) or 1 mM Ba²⁺ (in HEK cells). The horizontal distance between plots was called the “frequency shift in a single cell” and analyzed. The frequency shift in a single cell was −14.14 ± 5.71 Hz (n = 14) at 10 μM Ba²⁺ and −18.51 ± 4.00 Hz (n = 10) at 50 μM Ba²⁺, p < 0.05. The values perfectly matched the Ba²⁺-induced reduction of VF frequency observed previously in GP heart. A similar relationship was found in the computer simulations. The phase of Ba²⁺-sensitive admittance in GP cells was −2.65 ± 0.32 rad at 10 Hz and −2.79 ± 0.26 rad at 30 Hz. In HEK cells, the phase of Ba²⁺-sensitive admittance was 3.09 ± 0.03 rad at 10 Hz and 3.00 ± 0.17 rad at 30 Hz. We have developed a biological single-cell model of rotation-frequency control. The results show that although rotation frequency changes as a result of I_K1 blockade, the phase difference between transmembrane current and transmembrane voltage remains constant, enabling us to quantitatively predict the change of VF frequency resulting from I_K1 blockade, based on single-cell measurement.     

Measuring the antioxidant capacity of blood plasma using potentiometry

The use of potentiometry to measure plasma antioxidant capacity to contribute to oxidative stress evaluation is presented. In this assay, plasma (n = 60) diluted (0.3 to 1 ml) in phosphate buffer, pH 7.4, NaCl 9%, was submitted to potentiometry. A platinum wire was the working electrode and saturated calomel the reference. The results are presented as the difference between sample and buffer potential (ΔE). ΔE presented a good inverse correlation with added increasing concentrations of ascorbate (2.5−75 μmol/L; R = −0.99), urate (9.0−150 μmol/L; R = −0.99), and bilirubin (0.78−13 μmol/L; R = −0.99). Increase in the antioxidant capacity decreased ΔE. Depletion of the antioxidant capacity by tert-butylhydroperoxide (6.5−50 μmol/L) presented a direct correlation (0.97) with ΔE. Furthermore, ΔE presented an inverse correlation (R = −0.99) with increased antioxidant capacity of plasma (FRAP) induced by the addition of ascorbate (2.5−75 μmol/L). The response of the potentiometric method proved be adequate for measuring the plasma antioxidant depletion induced by acute exhaustive exercise in rats (control, n = 15; exercised, n = 15). This exercise decreased the concentration of urate (p < 0.05), decreased FRAP (p < 0.5), increased TBARS (p < 0.5), and decreased the potentiometer sensor response (p = 6.5 × 10⁻³). These results demonstrate the adequacy of potentiometry for evaluating the antioxidant capacity of blood plasma samples.     

Isolation and measurement of urinary 8-iso-prostaglandin F2α by high-performance liquid chromatography and gas chromatography–mass spectrometry

8-iso-Prostaglandin F_2α (8-iso-PGF_2α) is a product of free radical-catalyzed peroxidation of arachidonic acid. Measurement of its urinary excretion has been proposed as an index of oxidative status in vivo. A stable isotope dilution method for its quantification by gas chromatography–electron capture chemical ionization mass spectrometry is described. Sample cleanup required the combined use of high-performance liquid chromatography and thin-layer chromatography. The inter-assay R.S.D. in two separate determinations was 1.6 (n=4) and 2.3% (n=4). The accuracy of the assay was evaluated through recovery experiments. The equation of the regression plot correlating the amounts added and recovered was y=0.91x−0.31, r=0.9916 (n=12). The pair of fragment ions ([M−181]⁻) at m/z 569 and m/z 573 was monitored for quantification. The mean 8-iso-PGF_2α excretion rate was 528±127 (S.D.) ng per day in five male volunteers and 730±305 ng per day in six females. Intake of 80 mg of lycopene per day by eleven volunteers for four weeks resulted in a non-significant reduction of 8-iso-PGF_2α excretion.     

Body Composition at 6 months of Life: Comparison Of Air Displacement Plethysmography and Dual‐Energy X‐Ray Absorptiometry

Body composition assessment during infancy is important because it is a critical period for obesity risk development, thus valid tools are needed to accurately, precisely, and quickly determine both fat and fat‐free mass. The purpose of this study was to compare body composition estimates using dual‐energy x‐ray absorptiometry (DXA) and air displacement plethysmography (ADP) at 6 months old. We assessed the agreement between whole body composition using DXA and ADP in 84 full‐term average‐for‐gestational‐age boys and girls using DXA (Lunar iDXA v11–30.062; Infant whole body analysis enCore 2007 software, GE, Fairfield, CT) and ADP (Infant Body Composition System v3.1.0, COSMED USA, Concord, CA). Although the correlations between DXA and ADP for %fat (r = 0.925), absolute fat mass (r = 0.969), and absolute fat‐free mass (r = 0.945) were all significant, body composition estimates by DXA were greater for both %fat (31.1 ± 3.6% vs. 26.7 ± 4.7%; P < 0.001) and absolute fat mass (2,284 ± 449 vs. 1,921 ± 492 g; P < 0.001), and lower for fat‐free mass (5,022 ± 532 vs. 5,188 ± 508 g; P < 0.001) vs. ADP. Inter‐method differences in %fat decreased with increasing adiposity and differences in fat‐free mass decreased with increasing infant age. Estimates of body composition determined by DXA and ADP at 6 months of age were highly correlated, but did differ significantly. Additional work is required to identify the technical basis for these rather large inter‐method differences in infant body composition.     

Effect of dehydroepiandrosterone sulfate on carnitine acetyl transferase activity and l-carnitine levels in oophorectomized rats

Alteration in energy metabolism of postmenopausal women might be related to the reduction of dehydroepiandrosterone sulfate (DHEAS). DHEA and DHEAS decline with age, leveling at their nadir near menopause. DHEA and DHEAS modulate fatty acid metabolism by regulating carnitine acyltransferases and CoA. The purpose of this study was to determine whether dietary supplementation with DHEAS would also increase tissue l-carnitine levels, carnitine acetyltransferase (CAT) activity and mitochondrial respiration in oophorectomized rats. Plasma l-carnitine levels rose following oophorectomy in all groups (P<0.0001). Supplementation with DHEAS was not associated with further elevation of plasma l-carnitine levels, but with increased hepatic total and free l-carnitine (P=0.021 and P<0.0001, respectively) and cardiac total l-carnitine concentrations (P=0.045). In addition, DHEAS supplementation increased both hepatic and cardiac CAT activities (P<0.0001 and P=0.05 respectively). CAT activity positively correlated with the total and free carnitine levels in both liver and heart (r=0.764, r=0.785 and r=0.700, r=0.519, respectively). Liver mitochondrial respiratory control ratio, ADP:O ratio and oxygen uptake were similar in both control and supplemented groups. These results demonstrate that in oophorectomized rats, dietary DHEAS supplementation increases the liver and heart l-carnitine levels and CAT activities. In conclusion, DHEAS may modulate l-carnitine level and CAT activity in estrogen deficient rats. The potential role of DHEAS in the regulation of fatty acid oxidation in postmenopausal women is worthy of investigation.     

Extension of dynamic range of sensitive nanoparticle-based immunoassays

Nanoparticles have successfully been employed in immunometric assays that require high sensitivity. Certain analytes, however, require dynamic ranges (DRs) around a predetermined cut-off value. Here, we have studied the effects that antibody orientation and addition of free solid-phase and detection antibodies have on assay sensitivity and DR in traditional sandwich-type immunoassays. D-dimer and cardiac troponin I (cTnI), both routinely used in critical care testing, were applied as model analytes. The assays were performed in microtitration wells with preimmobilized solid-phase antibody. Inherently fluorescent nanoparticles coated with second antibody were used to detect the analyte. The selection of antibody orientation and addition of free solid-phase or detection antibody, with nanoparticles and calibrator, desensitized the assays and extended the DR. With D-dimer the upper limit of the DR was improved from 50 to 10,000 ng/ml, and with cTnI from 25 to 1000 ng/ml. Regression analysis with the Stago STA Liatest D-dimer assay yielded a slope (95% confidence interval) of 0.09 (0.07–0.11) and a y-intercept of −7.79 (−17.87–2.29) ng/L (n = 65, r = 0.906). Thus it is concluded that Europium(III)-chelate-doped nanoparticles can also be employed in immunoassays that require wide DRs around a certain cut-off limit.     

Towards a more objective assessment of equine personality using behavioural and physiological observations from performance test training

Current definitions of horse personality traits are rather vague, lacking clear, universally accepted guidelines for evaluation in performance tests. Therefore, the aim of the present study was to screen behavioural and physiological measurements taken during riding for potential links with scores the same horses received in the official stallion performance test for rideability and personality traits. Behaviour, heart rate (HR) and HR variability from thirty-six stallions participating in a performance test were recorded repeatedly during their performance test training. Using the coefficient of determination, regression analysis revealed that about 1/3 of variation (ranging between r = 0.26 (“constitution” (i.e. fitness, health)) and r = 0.46 (rideability)) in the personality trait scores could be explained by selecting the three most influential behaviour patterns per trait. These behaviour patterns included stumbling (with all traits except character), head-tossing (temperament, rideability), tail-swishing (willingness to work), involuntary change in gait (character) and the rider's use of her/his hands (constitution, rideability), voice (temperament) or whip (constitution). Subsequent mixed model analysis revealed a significant (P < 0.05) influence of the behaviour pattern “horse-induced change in gait” on character (−0.98 ± 0.31 scores per additional occurrence of change in gaits), of head-tossing (−0.25 ± 0.08 scores) and rider's use of voice (−0.51 ± 0.25; P = 0.0594) on temperament, and of stumbling on each of the following: willingness to work (−2.5 ± 1.2), constitution (−2.5 ± 1.2 scores; P = 0.0516) and rideability scores (−3.3 ± 1.4). In addition, constitution scores tended (P = 0.0889) to increase with higher low frequency/high frequency heart rate variation ratios (LF/HF), indicating a shift towards sympathetic dominance and thus a higher stress load in horses with higher scores for constitution. Rideability scores from the training phase were also significantly influenced by head-tossing (−0.5 ± 0.1), and in addition rideability scores from the final test were influenced by the training rider, ranging between average estimated rideability scores of 6.8 ± 0.4 for one training rider and 8.36 ± 0.3 scores for another training rider. Horses ridden with their nose-line predominantly behind the vertical received higher scores for rideability (8.3 ± 0.3) than horses ridden with their nose-line at the vertical (7.7 ± 0.2). These findings indicate that either judges perceive horses to have a better rideability when they readily offer a more extreme poll flexion, or that riders make use of horses’ better rideability by imposing a more extreme poll flexion. Several of the above described associations, but also of the non-existing links (e.g. no association between shying or heart rate and temperament) between behaviour patterns and scores for personality traits are rather surprising, warranting further investigation regarding the underlying causes of these relationships. Some of these behaviour patterns should be considered when redesigning the current guidelines for evaluation of personality traits during breeding horse performance tests, ultimately leading to improved genetic selection for equine personality traits. However, ethical implication of defining aversive behaviour such as head-tossing as an indicator of, for example, poor temperament, should not be neglected when devising new guidelines: such aversive behaviour may in fact be an indication of inadequate training techniques rather than poor horse personality.     

Effect of postpartum suppression of ovulation on uterine involution in dairy cows

The objective of this study was to investigate the effect of time of first postpartum ovulation after calving on uterine involution in dairy cows with and without uterine puerperal disease. Transvaginal follicular puncture (FP) of follicles >6 mm suppressed ovulation and development of a CL until Day 42 after calving. Fifty-three lactating Holstein Friesian cows (3.4 ± 1.2 years old, parity 2.5 ± 1.0 [median ± mean absolute deviation]) were divided into groups on the basis of the presence (UD+) or absence (UD−) of uterine disease and whether FP was carried out (FP+) or not (FP−). Uterine disease was defined as the occurrence of retained fetal membranes and/or metritis. This resulted in the following groups: UD−FP− (n = 15), UD−FP+ (n = 13), UD+FP− (n = 13), and UD+FP+ (n = 12). A general examination, vaginoscopy, transrectal palpation, and transrectal B-mode sonography of the reproductive organs were conducted on Days 8, 11, 18, and 25 and then every 10 days until Day 65 after calving. After hormonal synchronization of ovulation (cloprostenol between Days 55 and 60 postpartum and GnRH 2 days later), cows were inseminated in the next spontaneous estrus. On average, the cows ovulated on Day 21.0 ± 6.0 (UD−FP−), 50.0 ± 4.0 (UD−FP+), 16.0 ± 3.0 (UD+FP−), and 48.0 ± 2.0 (UD+FP+) postpartum. Calving-to-conception interval and first-service conception rates were not affected by FP (P > 0.05). Healthy cows with FP had smaller (P < 0.05) uterine horn and cervical diameters assessed sonographically than cows without FP. FP reduced the prevalence of purulent vaginal discharge and uterine size assessed transrectally in UD+ cows (P < 0.05). The results showed that suppression of an early ovulation by transvaginal FP improved uterine involution in cows with and without uterine disease.     

Rewarming rates of two large hibernators: Comparison of a monotreme and a eutherian

We measured body temperatures in two large hibernating mammals, the eutherian alpine marmot (Marmota marmota) and the egg-laying echidna (Tachyglossus aculeatus) from unrestrained animals in their natural environment. In both species hibernation is broken every 13 days on average by rewarming to euthermic temperatures. We found that the time course of a rewarming could be closely fitted with a sigmoid curve, allowing calculation of peak rewarming rate and corresponding body temperature. Maximum rewarming rates were twice as high in marmots as in echidnas (12.1±1.3 °C h⁻¹, n=10 cf. 6.2±1.2 °C h⁻¹, n=10). Peak rewarming rates were positively correlated with body temperature in echidnas, but negatively correlated in marmots.     

Quantitative mathematical expressions for accurate in vivo assessment of cytosolic [ADP] and ΔG of ATP hydrolysis in the human brain and skeletal muscle

Magnetic Resonance Spectroscopy affords the possibility of assessing in vivo the thermodynamic status of living tissues. The main thermodynamic variables relevant for the knowledge of the health of living tissues are: ΔG of ATP hydrolysis and cytosolic [ADP], the latter as calculated from the apparent equilibrium constant of the creatine kinase reaction. In this study we assessed the stoichiometric equilibrium constant of the creatine kinase reaction by in vitro ³¹P NMR measurements and computer calculations resulting to be: logK_CK=8.00±0.07 at T=310 K and ionic strength I=0.25 M. This value refers to the equilibrium:

PCr²⁻+ADP³⁻+H⁺=Cr+ATP⁴⁻     

Does adiposity affect muscle function during walking in children?

The biomechanical mechanisms responsible for the altered gait in obese children are not well understood, particularly as they relate to increases in adipose tissue. The purpose of this study was to test the hypotheses that as body-fat percentage (BF%) increased: (1) knee flexion during stance would decrease while pelvic obliquity would increase; (2) peak muscle forces normalized to lean-weight would increase for gluteus medius, gastrocnemius, and soleus, but decrease for the vasti; and (3) the individual muscle contributions to center of mass (COM) acceleration in the direction of their primary function(s) would not change for gluteus medius, gastrocnemius, and soleus, but decrease for the vasti. We scaled a musculoskeletal model to the anthropometrics of each participant (n=14, 8–12 years old, BF%: 16–41%) and estimated individual muscle forces and their contributions to COM acceleration. BF% was correlated with average knee flexion angle during stance (r=−0.54, p=0.024) and pelvic obliquity range of motion (r=0.78, p<0.001), as well as with relative vasti (r=−0.60, p=0.023), gluteus medius (r=0.65, p=0.012) and soleus (r=0.59, p=0.026) force production. Contributions to COM acceleration from the vasti were negatively correlated to BF% (vertical— r=−0.75, p=0.002, posterior— r=−0.68, p=0.008), but there were no correlation between BF% and COM accelerations produced by the gastrocnemius, soleus and gluteus medius. Therefore, we accept our first, partially accept our second, and accept our third hypotheses. The functional demands and relative force requirements of the hip abductors during walking in pediatric obesity may contribute to altered gait kinematics.     

Two μ1,5-dicyanamide bridged 2D and 3D metal complexes formed by covalent bonding and weak interactions

One 2D and one 3D dicyanamide bridged complexes, [Cu(dca)₂(et₂-en)]_n (1) and [Mn(dca)₂(im)₂]_n (2) [dca=dicyanamide, et₂-en=N,N-diethyl-ethylenediamine, im=imidazole], have been synthesized. Both the complexes are 1D by covalent bonding but interchain H-bonding promotes dimensionality. Moreover, π-π interaction among the imidazole ligands also plays an important role to have an interlocked 3D structure for 2. Magnetic study of both the complexes shows weak antiferromagnetic interaction between the metal centers. The magnetic data have been fitted with appropriate equations yielding best fit parameters for 1: J=−0.58±0.02 cm⁻¹, g=2.11±0.01 with R=3.2×10⁻⁶ and for 2, J=−0.21 cm⁻¹, g=2.00 and R=5.6×10⁻⁴.     

Salinity induced changes in haemolymph osmolality and total metabolic rate of the mud crab Rhithropanopeus harrisii Gould, 1841 from Baltic coastal waters

The specific metabolic rate (SMR) and haemolymph osmolality (HO) of the mud crab Rhithropanopeus harrisii Gould, 1841 from Baltic brackish waters were measured at a habitat salinity of 7 psu (T = 15 °C, full air saturation) and after step-wise acclimation to a salinity range of 3-27 psu. Values of SMR at 7 psu varied between 0.40 and 3.89 J g^− 1 WW h^− 1 (n = 25, wet weight range 0.051-1.142 g) and were significantly (p < 0.05) related to the specimen's wet weight (WW) according to the power regression SMR = 0.94 WW^− 0.41 (R² = 0.68). The SMR of females did not differ significantly (p > 0.05) from those of males. When exposed to higher salinities, the SMR of R. harrisii decreased significantly (p < 0.05) and reached a minimum value at 23 psu (0.55 ± 0.05 J g^− 1 WW h^− 1, n = 6). Mean haemolymph osmolality at 7 psu amounted to 581 ± 26 mOsm kg^− 1 (n = 5) and was 2.9 times higher than that of the external medium. R. harrisii hyperosmoregulated its body fluids up to 24 psu (727 mOsm kg^− 1) at which salinity the isosmotic point was reached.     

Leptin and leptin receptor genetic variants associate with habitual physical activity and the arm body composition response to resistance training

Purpose

We investigated the influence of Leptin (LEP) and leptin receptor (LEPR) SNPs on habitual physical activity (PA) and body composition response to a unilateral, upper body resistance training (RT) program.

Methods

European-derived American volunteers (men = 111, women = 131, 23.4 ± 5.4 yr, 24.4 ± 4.6 kg·m^− 2) were genotyped for LEP 19 G>A (rs2167270), and LEPR 326 A>G (rs1137100), 668 A>G (rs1137101), 3057 G>A (rs1805096), and 1968 G>C (rs8179183). They completed the Paffenbarger PA Questionnaire. Arm muscle and subcutaneous fat volumes were measured before and after 12 wk of supervised RT with MRI. Multivariate and repeated measures ANCOVA tested differences among phenotypes by genotype and gender with age and body mass index as covariates.

Results

Adults with the LEP 19 GG genotype reported more kcal/wk in vigorous intensity PA (1273.3 ± 176.8, p = 0.017) and sports/recreation (1922.8 ± 226.0, p < 0.04) than A allele carriers (718.0 ± 147.2, 1328.6 ± 188.2, respectively). Those with the LEP 19 GG genotype spent more h/wk in light intensity PA (39.7 ± 1.6) than A allele carriers (35.0 ± 1.4, p = 0.03). In response to RT, adults with the LEPR 668 G allele gained greater arm muscle volume (67,687.05 ± 3186.7 vs. 52,321.87 ± 5125.05 mm³, p = 0.01) and subcutaneous fat volume (10,599.89 ± 3683.57 vs. − 5224.73 ± 5923.98 mm³, p = 0.02) than adults with the LEPR 668 AA genotype, respectively.

Conclusion

LEP19 G>A and LEPR 668 A>G associated with habitual PA and the body composition response to RT. These LEP and LEPR SNPs are located in coding exons likely influencing LEP and LEPR function. Further investigation is needed to confirm our findings and establish mechanisms for LEP and LEPR genotype and PA and body composition associations we observed.     

Effect of external K+, Ca2+, and Ba2+ on membrane potential and ionic conductance in rat astrocytes

Summary 1. The purpose of this study was (a) to identify if astrocytes show a similar non-Nernstian depolarization in low K⁺ or low Ca²⁺ solutions as previously found in human glial and glioma cells, and (b) to analyze the influence of the K⁺ conductance on the membrane potential of astrocytes.2. The membrane potential (E_m) and the ionic conductance were studied with whole-cell patch-clamp technique in neonatal rat astrocytes (5–9 days in culture) and in human glioma cells (U-251MG).3. In 3.0 mM K⁺, E_m was –75 ± 1.0 mV (mean ± SEM,n=39) in rat astrocytes and –79 ± 0.7 mV (n=5) in U-251MG cells. In both cell types E_m changed linearly to the logarithm of [K⁺]₀ between 3.0 and 160 mM K⁺. K⁺ free medium caused astrocytes to hyperpolarize to –93 ± 2.7 mV (n=21) and U-251MG cells to depolarize to –27 ± 2.1 mV (n=3).4. The I-E curve did not show inward rectification in astrocytes at this developmental stage. The slope conductance (g) exhibited only a small decrease (–19%) in K⁺ free solution and no significant change in 160 mM K⁺.5. Ba²⁺ (1.0 mM) depolarized astrocytes to –45 ± 2.9 mV (n=11), decreasing the slope conductance (g) by 42.4 ± 8.3% (n=11). Ca²⁺ free solution depolarized astrocytes to –53 ± 3.4 mV (n=12) and resulted in a positive shift of the I-E curve, increasing g by 15.3 ± 8.2% (n=8).6. Calculations indicated that a block of K⁺ channels explains the depolarizing effect of Ba²⁺. The effects of K⁺ free or Ca²⁺ free solutions on E_m can be explained by a transformation of K⁺ channels to non-specific leakage channels. That astrocytes show a different reaction to low K⁺ than glioma cells can be related to the lack of inwardly rectifying K⁺ channels in astrocytes at this developmental stage.