Disk galaxy scaling relations at intermediate redshifts. I. The Tully-Fisher and velocity-size relations

NASA Astrophysics Data System (ADS)

Böhm, Asmus; Ziegler, Bodo L.

2016-07-01

Aims: Galaxy scaling relations such as the Tully-Fisher relation (between the maximum rotation velocity Vmax and luminosity) and the velocity-size relation (between Vmax and the disk scale length) are powerful tools to quantify the evolution of disk galaxies with cosmic time. Methods: We took spatially resolved slit spectra of 261 field disk galaxies at redshifts up to z ≈ 1 using the FORS instruments of the ESO Very Large Telescope. The targets were selected from the FORS Deep Field and William Herschel Deep Field. Our spectroscopy was complemented with HST/ACS imaging in the F814W filter. We analyzed the ionized gas kinematics by extracting rotation curves from the two-dimensional spectra. Taking into account all geometrical, observational, and instrumental effects, these rotation curves were used to derive the intrinsic Vmax. Results: Neglecting galaxies with disturbed kinematics or insufficient spatial rotation curve extent, Vmax was reliably determined for 124 galaxies covering redshifts 0.05 < z < 0.97. This is one of the largest kinematic samples of distant disk galaxies to date. We compared this data set to the local B-band Tully-Fisher relation and the local velocity-size relation. The scatter in both scaling relations is a factor of ~2 larger at z ≈ 0.5 than at z ≈ 0. The deviations of individual distant galaxies from the local Tully-Fisher relation are systematic in the sense that the galaxies are increasingly overluminous toward higher redshifts, corresponding to an overluminosity ΔMB = -(1.2 ± 0.5) mag at z = 1. This luminosity evolution at given Vmax is probably driven by younger stellar populations of distant galaxies with respect to their local counterparts, potentially combined with modest changes in dark matter mass fractions. The analysis of the velocity-size relation reveals that disk galaxies of a given Vmax have grown in size by a factor of ~1.5 over the past ~8 Gyr, most likely through accretion of cold gas and/or small satellites. From scrutinizing the combined evolution in luminosity and size, we find that the galaxies that show the strongest evolution toward smaller sizes at z ≈ 1 are not those that feature the strongest evolution in luminosity, and vice versa. Based on observations with the European Southern Observatory Very Large Telescope (ESO-VLT), observing run IDs 65.O-0049, 66.A-0547, 68.A-0013, 69.B-0278B, 70.B-0251A and 081.B-0107A.The full Table 1 is only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (http://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/592/A64

Inhibition by enflurane and methoxyflurane of postdrive hyperpolarization in canine Purkinje fibers.

PubMed

Pratila, M; Vogel, S; Sperelakis, N

1984-05-01

When a pacemaker cell is driven with a train of stimuli at a rate faster than its own, the termination of the drive is followed by a transient hyperpolarization, due to the activity of an electrogenic Na+-K+ pump. In this study, the effect of the halogenated ethers, enflurane and methoxyflurane, on postdrive hyperpolarization (PDH) was determined in cardiac Purkinje fibers. The fibers were removed from freshly excised canine hearts and superfused with a Tyrode's solution (containing 2.7 or 3.5 mM K+). The preparation was paced at 0.2 Hz before and after drives, and at 2 Hz during drives. Under control conditions, drives of 2 min produced a PDH of 5.5 +/- 0.2 mV. Enflurane (1.5-5%) significantly reduced the PDH. At 4 to 5%, enflurane reduced the PDH to a mean value of 42% of the control. Methoxyflurane was more potent than enflurane in affecting the PDH. At 0.5 to 0.75%, methoxyflurane reduced the PDH to 5% of the control. At higher (1-1.5%) concentrations of methoxyflurane, the PDH was converted to a depolarization, which varied between 0.5 and 8.0 mV. The PDH was restored to control levels within 10 to 20 min after washout of either anesthetic agent. Methoxyflurane (0.5 or 1%) enhanced the automaticity of spontaneously firing cells (2.35 mM K+ Tyrode's solution used). This positive chronotropic action coincided with a depolarization of 2 to 8 mV. Enflurane, at concentrations of 3 to 5%, gave similar results. On the action potential, methoxyflurane, at 1%, reduced the amplitude and duration (measured at 50% repolarization) of the plateau, and also the maximal upstroke velocity (+Vmax) of the rising phase.(ABSTRACT TRUNCATED AT 250 WORDS)

Another baryon miracle? Testing solutions to the `missing dwarfs' problem

NASA Astrophysics Data System (ADS)

Trujillo-Gomez, Sebastian; Schneider, Aurel; Papastergis, Emmanouil; Reed, Darren S.; Lake, George

2018-04-01

The dearth of dwarf galaxies in the local Universe is hard to reconcile with the large number of low-mass haloes expected within the concordance Λ cold dark matter (ΛCDM) paradigm. In this paper, we perform a systematic evaluation of the uncertainties affecting the measurement of dark matter halo abundance using galaxy kinematics. Using a large sample of dwarf galaxies with spatially resolved kinematics, we derive a correction to obtain the abundance of galaxies as a function of maximum circular velocity - a direct probe of halo mass - from the line-of-sight velocity function in the Local Volume. This method provides a direct means of comparing the predictions of theoretical models and simulations (including non-standard cosmologies and novel galaxy formation physics) to the observational constraints. The new `galactic Vmax' function is steeper than the line-of-sight velocity function but still shallower than the theoretical CDM expectation, implying that unaccounted baryonic physics may be necessary to reduce the predicted abundance of galaxies. Using the galactic Vmax function, we investigate the theoretical effects of feedback-powered outflows and photoevaporation of gas due to reionization. At the 3σ confidence level, we find that feedback and reionization are not effective enough to reconcile the disagreement. In the case of maximum baryonic effects, the theoretical prediction still deviates significantly from the observations for Vmax < 60 km s-1. CDM predicts at least 1.8 times more galaxies with Vmax = 50 km s-1 and 2.5 times more than observed at 30 km s-1. Recent hydrodynamic simulations seem to resolve the discrepancy but disagree with the properties of observed galaxies with spatially resolved kinematics. This abundance problem might point to the need to modify cosmological predictions at small scales.

Maximum shortening velocity of lymphatic muscle approaches that of striated muscle.

PubMed

Zhang, Rongzhen; Taucer, Anne I; Gashev, Anatoliy A; Muthuchamy, Mariappan; Zawieja, David C; Davis, Michael J

2013-11-15

Lymphatic muscle (LM) is widely considered to be a type of vascular smooth muscle, even though LM cells uniquely express contractile proteins from both smooth muscle and cardiac muscle. We tested the hypothesis that LM exhibits an unloaded maximum shortening velocity (Vmax) intermediate between that of smooth muscle and cardiac muscle. Single lymphatic vessels were dissected from the rat mesentery, mounted in a servo-controlled wire myograph, and subjected to isotonic quick release protocols during spontaneous or agonist-evoked contractions. After maximal activation, isotonic quick releases were performed at both the peak and plateau phases of contraction. Vmax was 0.48 ± 0.04 lengths (L)/s at the peak: 2.3 times higher than that of mesenteric arteries and 11.4 times higher than mesenteric veins. In cannulated, pressurized lymphatic vessels, shortening velocity was determined from the maximal rate of constriction [rate of change in internal diameter (-dD/dt)] during spontaneous contractions at optimal preload and minimal afterload; peak -dD/dt exceeded that obtained during any of the isotonic quick release protocols (2.14 ± 0.30 L/s). Peak -dD/dt declined with pressure elevation or activation using substance P. Thus, isotonic methods yielded Vmax values for LM in the mid to high end (0.48 L/s) of those the recorded for phasic smooth muscle (0.05-0.5 L/s), whereas isobaric measurements yielded values (>2.0 L/s) that overlapped the midrange of values for cardiac muscle (0.6-3.3 L/s). Our results challenge the dogma that LM is classical vascular smooth muscle, and its unusually high Vmax is consistent with the expression of cardiac muscle contractile proteins in the lymphatic vessel wall.

Lift production through asymmetric flapping

NASA Astrophysics Data System (ADS)

Jalikop, Shreyas; Sreenivas, K. R.

2009-11-01

At present, there is a strong interest in developing Micro Air Vehicles (MAV) for applications like disaster management and aerial surveys. At these small length scales, the flight of insects and small birds suggests that unsteady aerodynamics of flapping wings can offer many advantages over fixed wing flight, such as hovering-flight, high maneuverability and high lift at large angles of attack. Various lift generating mechanims such as delayed stall, wake capture and wing rotation contribute towards our understanding of insect flight. We address the effect of asymmetric flapping of wings on lift production. By visualising the flow around a pair of rectangular wings flapping in a water tank and numerically computing the flow using a discrete vortex method, we demonstrate that net lift can be produced by introducing an asymmetry in the upstroke-to-downstroke velocity profile of the flapping wings. The competition between generation of upstroke and downstroke tip vortices appears to hold the key to understanding this lift generation mechanism.

Models of disuse - A comparison of hindlimb suspension and immobilization

NASA Technical Reports Server (NTRS)

Fitts, R. H.; Metzger, J. M.; Riley, D. A.; Unsworth, B. R.

1986-01-01

The effects of 1 and 2 weeks of hindlimb suspension (HS) on the contractile properties of fast- and slow-twitch skeletal muscles of male Sprague Dawley rats are studied and compared with hindlimb immobilization (HI) data. The optimal length and contractile properties of the slow-twitch soleus, fast-twitch extensor digitorum longus, and the vastus lateralis are measured. It is observed that HS and HI affect slow-twitch muscles; isometric twitch duration in the slow-twitch soleus is decreased. Soleus muscle mass and peak tetanic tension declines with disuse. A major difference in the influence of HS and HI on the maximal speed of soleus muscle shortening, V(max) is detected; HS produced a twofold increase in V(max) compared to control data and HI had no significant effect on V(max). The relation between V(max) and myosin concentration is analyzed. The data reveal that HS modifies slow-twitch muscle yielding hybrid fibers with elevated shortening velocities and this change may be dependent on the elimination of load-bearing contractions.

Kinetic analysis of dihydroxyacetone production from crude glycerol by immobilized cells of Gluconobacter oxydans MTCC 904.

PubMed

Dikshit, Pritam Kumar; Moholkar, Vijayanand S

2016-09-01

The present study has investigated kinetic features of bioconversion of biodiesel-derived crude glycerol to dihydroxyacetone with immobilized Gluconobacter oxydans cells using modified Haldane substrate-inhibition model. The results have been compared against free cells and pure glycerol. Relative variations in the kinetic parameters KS, KI, Vmax, n and X reveal that immobilized G. oxydans cells (on PU foam substrate) with crude glycerol as substrate give higher order of inhibition (n) and lower maximum reaction velocities (Vmax). These results are essentially implications of substrate transport restrictions across immobilization matrix, which causes retention of substrate in the matrix and reduction in fractional available substrate (X) for the cells. This causes reduction in both KS (substrate concentration at Vmax/2) and KI (inhibition constant) as compared to free cells. For immobilized cells, substrate concentration (Smax) corresponding to Vmax is practically same for both pure and crude glycerol as substrate. Copyright © 2016 Elsevier Ltd. All rights reserved.

The monophasic action potential upstroke: a means of characterizing local conduction.

PubMed

Levine, J H; Moore, E N; Kadish, A H; Guarnieri, T; Spear, J F

1986-11-01

The upstrokes of monophasic action potentials (MAPs) recorded with an extracellular pressure electrode were characterized in isolated canine tissue preparations in vitro. The characteristics of the MAP upstroke were compared with those of the local action potential foot as well as with the characteristics of approaching electrical activation during uniform and asynchronous conduction. The upstroke of the MAP was exponential during uniform conduction. The time constant of rise of the MAP upstroke (TMAP) correlated with that of the action potential foot (Tfoot): TMAP + 1.01 Tfoot + 0.50; r2 = .80. Furthermore, changes in Tfoot with alterations in cycle length were associated with similar changes in TMAP: Tfoot = 1.06 TMAP - 0.11; r2 = .78. In addition, TMAP and Tfoot both deviated from exponential during asynchronous activation; the inflections that developed in the MAP upstroke correlated in time with intracellular action potential upstrokes that were asynchronous in onset in these tissues. Finally, the field of view of the MAP was determined and was found to be dependent in part on tissue architecture and the space constant. Specifically, the field of view of the MAP was found to be greater parallel compared with transverse to fiber orientation (6.02 +/- 1.74 vs 3.03 +/- 1.10 mm; p less than .01). These data suggest that the MAP upstroke may be used to define and characterize local electrical activation. The relatively large field of view of the MAP suggests that this technique may be a sensitive means to record focal membrane phenomena in vivo.

Movement velocity as a measure of exercise intensity in three lower limb exercises.

PubMed

Conceição, Filipe; Fernandes, Juvenal; Lewis, Martin; Gonzaléz-Badillo, Juan José; Jimenéz-Reyes, Pedro

2016-01-01

The purpose of this study was to investigate the relationship between movement velocity and relative load in three lower limbs exercises commonly used to develop strength: leg press, full squat and half squat. The percentage of one repetition maximum (%1RM) has typically been used as the main parameter to control resistance training; however, more recent research has proposed movement velocity as an alternative. Fifteen participants performed a load progression with a range of loads until they reached their 1RM. Maximum instantaneous velocity (Vmax) and mean propulsive velocity (MPV) of the knee extension phase of each exercise were assessed. For all exercises, a strong relationship between Vmax and the %1RM was found: leg press (r(2)adj = 0.96; 95% CI for slope is [-0.0244, -0.0258], P < 0.0001), full squat (r(2)adj = 0.94; 95% CI for slope is [-0.0144, -0.0139], P < 0.0001) and half squat (r(2)adj = 0.97; 95% CI for slope is [-0.0135, -0.00143], P < 0.0001); for MPV, leg press (r(2)adj = 0.96; 95% CI for slope is [-0.0169, -0.0175], P < 0.0001, full squat (r(2)adj = 0.95; 95% CI for slope is [-0.0136, -0.0128], P < 0.0001) and half squat (r(2)adj = 0.96; 95% CI for slope is [-0.0116, 0.0124], P < 0.0001). The 1RM was attained with a MPV and Vmax of 0.21 ± 0.06 m s(-1) and 0.63 ± 0.15 m s(-1), 0.29 ± 0.05 m s(-1) and 0.89 ± 0.17 m s(-1), 0.33 ± 0.05 m s(-1) and 0.95 ± 0.13 m s(-1) for leg press, full squat and half squat, respectively. Results indicate that it is possible to determine an exercise-specific %1RM by measuring movement velocity for that exercise.

Phenytoin pharmacokinetics in critically ill trauma patients.

PubMed

Boucher, B A; Rodman, J H; Jaresko, G S; Rasmussen, S N; Watridge, C B; Fabian, T C

1988-12-01

Preliminary data have suggested that phenytoin systemic clearance may increase during initial therapy in critically ill patients. The objectives for this study were to model the time-variant phenytoin clearance and evaluate concomitant changes in protein binding and urinary metabolite elimination. Phenytoin was given as an intravenous loading dose of 15 mg/kg followed by an initial maintenance dose of 6 mg/kg/day in 10 adult critically ill trauma patients. Phenytoin bound and unbound plasma concentrations were determined in 10 patients and urinary excretion of the metabolite p-hydroxyphenyl phenylhydantoin (p-HPPH) was measured in seven patients for 7 to 14 days. A Michaelis-Menten one-compartment model incorporating a time-variant maximal velocity (Vmax) was sufficient to describe the data and superior to a conventional time-invariant Michaelis-Menten model. Vmax for the time-variant model was defined as V'max + Vmax delta (1 - e(-kindt)). Vmax infinity is the value for Vmax when t is large. The median values (ranges) for the parameters were Km = 4.8 (2.6 to 20) mg/L, Vmax infinity = 1348 (372 to 4741) mg/day, and kind = 0.0115 (0.0045 to 0.132) hr-1. Phenytoin free fraction increased in a majority of patients during the study period, with a binding ratio inversely related to albumin. Measured urinary p-HPPH data were consistent with the proposed model. A loading and constant maintenance dose of phenytoin frequently yielded a substantial, clinically significant fall in plasma concentrations with a pattern of apparently increasing clearance that may be a consequence of changes in protein binding, induction of metabolism, or the influence of stress on hepatic metabolic capacity.

Relationships between maximal anaerobic power of the arms and legs and javelin performance.

PubMed

Bouhlel, E; Chelly, M S; Tabka, Z; Shephard, R

2007-06-01

The aim of this study was to examine relationships between maximal anaerobic power, as measured by leg and arm force-velocity tests, estimates of local muscle volume and javelin performance. Ten trained national level male javelin throwers (mean age 19.6+/- 2 years) participated in this study. Maximal anaerobic power, maximal force and maximal velocity were measured during leg (Wmax-L) and arm (Wmax-A) force-velocity tests, performed on appropriately modified forms of Monark cycle ergometer. Estimates of leg and arm muscle volume were made using a standard anthropometric kit. Maximal force of the leg (Fmax-L) was significantly correlated with estimated leg muscle volume (r=0.71, P<0.05). Wmax-L and Wmax-A were both significantly correlated with javelin performance (r=0.76, P<0.01; r=0.71, P <0.05, respectively). Maximal velocity of the leg (Vmax-L) was also significantly correlated with throwing performance (r=0.83; P<0.001). Wmax of both legs and arms were significantly correlated with javelin performance, the closest correlation being for Wmax-L; this emphasizes the importance of the leg muscles in this sport. Fmax-L and Vmax-L were related to muscle volume and to javelin performance, respectively. Force-velocity testing may have value in regulating conditioning and rehabilitation in sports involving throwing.

Kinetic measurement of 2-aminopurine X cytosine and 2-aminopurine X thymine base pairs as a test of DNA polymerase fidelity mechanisms.

PubMed Central

Watanabe, S M; Goodman, M F

1982-01-01

Enzyme kinetic measurements are presented showing that Km rather than maximum velocity (Vmax) discrimination governs the frequency of forming 2-aminopurine X cytosine base mispairs by DNA polymerase alpha. An in vitro system is used in which incorporation of dTMP or dCMP occurs opposite a template 2-aminopurine, and values for Km and Vmax are obtained. Results from a previous study in which dTTP and dCTP were competing simultaneously for insertion opposite 2-aminopurine indicated that dTMP is inserted 22 times more frequently than dCMP. We now report that the ratio of Km values KCm/KTm = 25 +/- 6, which agrees quantitatively with the dTMP/dCMP incorporation ratio obtained previously. We also report that VCmax is indistinguishable from VTmax. These Km and Vmax data are consistent with predictions from a model, the Km discrimination model, in which replication fidelity is determined by free energy differences between matched and mismatched base pairs. Central to this model is the prediction that the ratio of Km values for insertion of correct and incorrect nucleotides specifies the insertion fidelity, and the maximum velocities of insertion are the same for both nucleotides. PMID:6959128

Breakdown and Limit of Continuum Diffusion Velocity for Binary Gas Mixtures from Direct Simulation

NASA Astrophysics Data System (ADS)

Martin, Robert Scott; Najmabadi, Farrokh

2011-05-01

This work investigates the breakdown of the continuum relations for diffusion velocity in inert binary gas mixtures. Values of the relative diffusion velocities for components of a gas mixture may be calculated using of Chapman-Enskog theory and occur not only due to concentration gradients, but also pressure and temperature gradients in the flow as described by Hirschfelder. Because Chapman-Enskog theory employs a linear perturbation around equilibrium, it is expected to break down when the velocity distribution deviates significantly from equilibrium. This breakdown of the overall flow has long been an area of interest in rarefied gas dynamics. By comparing the continuum values to results from Bird's DS2V Monte Carlo code, we propose a new limit on the continuum approach specific to binary gases. To remove the confounding influence of an inconsistent molecular model, we also present the application of the variable hard sphere (VSS) model used in DS2V to the continuum diffusion velocity calculation. Fitting sample asymptotic curves to the breakdown, a limit, Vmax, that is a fraction of an analytically derived limit resulting from the kinetic temperature of the mixture is proposed. With an expected deviation of only 2% between the physical values and continuum calculations within ±Vmax/4, we suggest this as a conservative estimate on the range of applicability for the continuum theory.

Assessment of Portal Venous and Hepatic Artery Haemodynamic Variation in Non-Alcoholic Fatty Liver Disease (NAFLD) Patients.

PubMed

Balasubramanian, Padhmini; Boopathy, Vinoth; Govindasamy, Ezhumalai; Venkatesh, Basavaiya Prabhu

2016-08-01

Non-Alcoholic Fatty Liver Disease (NAFLD) has various spectrums of liver diseases like isolated fatty liver, steatohepatitis and cirrhosis usually progressing in a linear fashion. In this process they are known to cause certain haemodynamic changes in the portal flow and hepatic artery flow. The aim of the study was to study these haemodynamic changes in patients with NAFLD and to correlate it with the disease severity. Ninety patients diagnosed to have NAFLD based on ultrasound abdomen (30 each in grade1, grade2 and grade3 NAFLD) and 30 controls (Normal liver on ultrasound abdomen) were subjected to portal vein and hepatic artery Doppler study. Peak maximum velocity (Vmax), Peak minimum velocity (Vmin), Mean flow velocity (MFV), and Vein pulsality index (VPI) of the portal vein and hepatic artery resistivity index (HARI) of the hepatic artery were the doppler parameters which were assessed. Liver span was also assessed both for the fatty liver and controls. The mean Vmax, Vmin, MFV and VPI of the portal vein in patients with NAFLD was 12.23±1.74cm/sec, 9.31±1.45cm/sec, 10.76±1.48cm/sec, and 0.24±0.04 as compared to 14.05±2.43cm/sec, 10.01±2.27cm/sec, 12.23±2.47cm/sec, 0.3±0.08 in controls respectively. All these differences were statistically significant except for Vmin. The Mean HARI in patients with fatty liver was 0.65±0.06 when compared to controls of 0.75±0.06 (p=0.001). HARI (r-value of -0.517) had a better negative correlation followed by VPI (r-value of -0.44) and Vmax (r-value of -0.293) with the severity of NAFLD. MFV had a very weak negative correlation (r-value of -0.182) with the severity of NAFLD. The Vmax, MFV, VPI and HARI were significantly less when compared to controls suggesting a reduced portal flow and an increased hepatic arterial flow in patients with NAFLD. Among the parameters, HARI correlated better with the severity of NAFLD followed by VPI.

Universality and robustness of revivals in the transverse field XY model

NASA Astrophysics Data System (ADS)

Häppölä, Juho; Halász, Gábor B.; Hamma, Alioscia

2012-03-01

We study the structure of the revivals in an integrable quantum many-body system, the transverse field XY spin chain, after a quantum quench. The time evolutions of the Loschmidt echo, the magnetization, and the single-spin entanglement entropy are calculated. We find that the revival times for all of these observables are given by integer multiples of Trev≃L/vmax, where L is the linear size of the system and vmax is the maximal group velocity of quasiparticles. This revival structure is universal in the sense that it does not depend on the initial state and the size of the quench. Applying nonintegrable perturbations to the XY model, we observe that the revivals are robust against such perturbations: they are still visible at time scales much larger than the quasiparticle lifetime. We therefore propose a generic connection between the revival structure and the locality of the dynamics, where the quasiparticle speed vmax generalizes into the Lieb-Robinson speed vLR.

Thermodynamics of gas and steam-blast eruptions

USGS Publications Warehouse

Mastin, L.G.

1995-01-01

Eruptions of gas or steam and non-juvenile debris are common in volcanic and hydrothermal areas. From reports of non-juvenile eruptions or eruptive sequences world-wide, at least three types (or end-members) can be identified: (1) those involving rock and liquid water initially at boiling-point temperatures ('boiling-point eruptions'); (2) those powered by gas (primarily water vapor) at initial temperatures approaching magmatic ('gas eruptions'); and (3) those caused by rapid mixing of hot rock and ground- or surface water ('mixing eruptions'). For these eruption types, the mechanical energy released, final temperatures, liquid water contents and maximum theoretical velocities are compared by assuming that the erupting mixtures of rock and fluid thermally equilibrate, then decompress isentropically from initial, near-surface pressure (???10 MPa) to atmospheric pressure. Maximum mechanical energy release is by far greatest for gas eruptions (??????1.3 MJ/kg of fluid-rock mixture)-about one-half that of an equivalent mass of gunpowder and one-fourth that of TNT. It is somewhat less for mixing eruptions (??????0.4 MJ/kg), and least for boiling-point eruptions (??????0.25 MJ/kg). The final water contents of crupted boiling-point mixtures are usually high, producing wet, sloppy deposits. Final erupted mixtures from gas eruptions are nearly always dry, whereas those from mixing eruptions vary from wet to dry. If all the enthalpy released in the eruptions were converted to kinetic energy, the final velocity (vmax) of these mixtures could range up to 670 m/s for boiling-point eruptions and 1820 m/s for gas eruptions (highest for high initial pressure and mass fractions of rock (mr) near zero). For mixing eruptions, vmax ranges up to 1150 m/s. All observed eruption velocities are less than 400 m/s, largely because (1) most solid material is expelled when mr is high, hence vmax is low; (2) observations are made of large blocks the velocities of which may be less than the average for the mixture; (3) heat from solid particles is not efficiently transferred to the fluid during the eruptions; and (4) maximum velocities are reduced by choked flow or friction in the conduit. ?? 1995 Springer-Verlag.

Endurance performance and nocturnal HRV indices.

PubMed

Nummela, A; Hynynen, E; Kaikkonen, P; Rusko, H

2010-03-01

The effects of endurance training on endurance performance characteristics and cardiac autonomic modulation during night sleep were investigated. Twenty-four sedentary subjects trained over four weeks two hours per week at an average running intensity of 76+/-4% of their heart rate reserve. The R to R ECG-intervals were recorded and heart rate variability indices including high frequency power (HFP) were calculated for the nights following the training days every week. The subjects were divided into responders and non-responders according to the improvements in the maximal velocity of the incremental treadmill test (v(max)). The responders improved their v(max) by 10.9+/-46 % (p < 0.001) while no changes were observed in the non-responders (1.6+/-3.0%), although there were no differences in any training load variables between the groups. In the responders nocturnal HFP was significantly higher during the fourth training week compared to the first training week (p=0.036). Furthermore, a significant correlation was observed between the change in v(max) and the change in nocturnal HFP (r=0.482, p=0.042). It was concluded that after similar training, an increase in cardiac vagal modulation was related to improved v(max) in the sedentary subjects. Georg Thieme Verlag KG Stuttgart.New York.

Universal Dark Halo Scaling Relation for the Dwarf Spheroidal Satellites

NASA Astrophysics Data System (ADS)

Hayashi, Kohei; Ishiyama, Tomoaki; Ogiya, Go; Chiba, Masashi; Inoue, Shigeki; Mori, Masao

2017-07-01

Motivated by a recently found interesting property of the dark halo surface density within a radius, {r}\\max , giving the maximum circular velocity, {V}\\max , we investigate it for dark halos of the Milky Way’s and Andromeda’s dwarf satellites based on cosmological simulations. We select and analyze the simulated subhalos associated with Milky-Way-sized dark halos and find that the values of their surface densities, {{{Σ }}}{V\\max }, are in good agreement with those for the observed dwarf spheroidal satellites even without employing any fitting procedures. Moreover, all subhalos on the small scales of dwarf satellites are expected to obey the universal relation, irrespective of differences in their orbital evolutions, host halo properties, and observed redshifts. Therefore, we find that the universal scaling relation for dark halos on dwarf galaxy mass scales surely exists and provides us with important clues for understanding fundamental properties of dark halos. We also investigate orbital and dynamical evolutions of subhalos to understand the origin of this universal dark halo relation and find that most subhalos evolve generally along the {r}\\max \\propto {V}\\max sequence, even though these subhalos have undergone different histories of mass assembly and tidal stripping. This sequence, therefore, should be the key feature for understanding the nature of the universality of {{{Σ }}}{V\\max }.

Upstroke wing flexion and the inertial cost of bat flight

PubMed Central

Riskin, Daniel K.; Bergou, Attila; Breuer, Kenneth S.; Swartz, Sharon M.

2012-01-01

Flying vertebrates change the shapes of their wings during the upstroke, thereby decreasing wing surface area and bringing the wings closer to the body than during downstroke. These, and other wing deformations, might reduce the inertial cost of the upstroke compared with what it would be if the wings remained fully extended. However, wing deformations themselves entail energetic costs that could exceed any inertial energy savings. Using a model that incorporates detailed three-dimensional wing kinematics, we estimated the inertial cost of flapping flight for six bat species spanning a 40-fold range of body masses. We estimate that folding and unfolding comprises roughly 44 per cent of the inertial cost, but that the total inertial cost is only approximately 65 per cent of what it would be if the wing remained extended and rigid throughout the wingbeat cycle. Folding and unfolding occurred mostly during the upstroke; hence, our model suggests inertial cost of the upstroke is not less than that of downstroke. The cost of accelerating the metacarpals and phalanges accounted for around 44 per cent of inertial costs, although those elements constitute only 12 per cent of wing weight. This highlights the energetic benefit afforded to bats by the decreased mineralization of the distal wing bones. PMID:22496186

CT-1-CP-induced ventricular electrical remodeling in mice.

PubMed

Chen, Shu-fen; Wei, Tao-zhi; Rao, Li-ya; Xu, Ming-guang; Dong, Zhan-ling

2015-02-01

The chronic effects of carboxyl-terminal polypeptide of Cardiotrophin-1 (CT-1-CP) on ventricular electrical remodeling were investigated. CT-1-CP, which contains 16 amino acids in sequence of the C-terminal of Cardiotrophin-1, was selected and synthesized, and then administered to Kunming mice (aged 5 weeks) by intraperitoneal injection (500 ng·g⁻¹·day⁻¹) (4 groups, n=10 and female: male=1:1 in each group) for 1, 2, 3 and 4 weeks, respectively. The control group (n=10, female: male=1:1) was injected by physiological saline for 4 weeks. The epicardial monophasic action potential (MAP) was recorded by using a contact-type MAP electrode placed vertically on the left ventricular (LV) epicardium surface, and the electrocardiogram (ECG) signal in lead II was monitored synchronously. ECG intervals (RR, PR, QRS and QT) and the amplitude of MAP (Am), the maximum upstroke velocity (Vmax), as well as action potential durations (APDs) at different repolarization levels (APD30, APD50, APD70, and APD90) of MAP were determined and analyzed in detail. There were no significant differences in RR and P intervals between CT-1-CP-treated groups and control group, but the PR segment and the QRS complex were greater in the former than in the latter (F=2.681 and 5.462 respectively, P<0.05). Though QT interval and the corrected QT interval (QTc) were shorter in CT-1-CP-treated groups than in control group, the QT dispersion (QTd) of them was greater in the latter than in the former (F=3.090, P<0.05) and increased with the time. The ECG monitoring synchronously with the MAP showed that the compression of MAP electrode on the left ventricular epicardium induced performance similar to myocardium ischemia. As compared with those before chest-opening, the PR segment and QT intervals remained basically unchanged in control group, but prolonged significantly in all CT-1-CP-treated groups and the prolongation of QT intervals increased gradually along with the time of exposure to CT-1-CP. The QRS complex had no significant change in control group, one-week and three-week CT-1-CP-treated groups, but prolonged significantly in two-week and four-week CT-1-CP-treated groups. Interestingly, the QTd after chest-opening was significantly greater than that before chest-opening in control group (t=5.242, P<0.01), but decreased along with the time in CT-1-CP-treated groups. The mean MAP amplitude, Vmax and APD were greater in CT-1-CP-treated groups than those in control group, and became more obvious along with the time. The APD in four CT-1-CP-treat groups was prolonged mainly in middle to final repolarization phase. The difference among these groups became significant in middle phase (APD50) (F=6.076, P<0.01) and increased furthermore in late and final phases (APD70: F=10.054; APD90: F=18.691, P<0.01) along with the time of injection of CT-1-CP. The chronic action of CT-1-CP might induce the adapting alteration in cardiac conductivity and ventricular repolarization. The amplitude and the Vmax of the anterior LV epicardial MAP increased obviously, and the APD prolonged mainly in late and final phase of repolarization.

A simple theory of motor protein kinetics and energetics. II.

PubMed

Qian, H

2000-01-10

A three-state stochastic model of motor protein [Qian, Biophys. Chem. 67 (1997) pp. 263-267] is further developed to illustrate the relationship between the external load on an individual motor protein in aqueous solution with various ATP concentrations and its steady-state velocity. A wide variety of dynamic motor behavior are obtained from this simple model. For the particular case of free-load translocation being the most unfavorable step within the hydrolysis cycle, the load-velocity curve is quasi-linear, V/Vmax = (cF/Fmax-c)/(1-c), in contrast to the hyperbolic relationship proposed by A.V. Hill for macroscopic muscle. Significant deviation from the linearity is expected when the velocity is less than 10% of its maximal (free-load) value--a situation under which the processivity of motor diminishes and experimental observations are less certain. We then investigate the dependence of load-velocity curve on ATP (ADP) concentration. It is shown that the free load Vmax exhibits a Michaelis-Menten like behavior, and the isometric Fmax increases linearly with ln([ATP]/[ADP]). However, the quasi-linear region is independent of the ATP concentration, yielding an apparently ATP-independent maximal force below the true isometric force. Finally, the heat production as a function of ATP concentration and external load are calculated. In simple terms and solved with elementary algebra, the present model provides an integrated picture of biochemical kinetics and mechanical energetics of motor proteins.

New Tool to Control and Monitor Weighted Vest Training Load for Sprinting and Jumping in Soccer.

PubMed

Carlos-Vivas, Jorge; Freitas, Tomás T; Cuesta, Miguel; Perez-Gomez, Jorge; De Hoyo, Moisés; Alcaraz, Pedro E

2018-04-26

Carlos-Vivas, J, Freitas, TT, Cuesta, M, Perez-Gomez, J, De Hoyo, M, and Alcaraz, PE. New tool to control and monitor weighted vest training load for sprinting and jumping in soccer. J Strength Cond Res XX(X): 000-000, 2018-The purpose of this study was to develop 2 regression equations that accurately describe the relationship between weighted vest loads and performance indicators in sprinting (i.e., maximum velocity, Vmax) and jumping (i.e., maximum height, Hmax). Also, this study aimed to investigate the effects of increasing the load on spatio-temporal variables and power development in soccer players and to determine the "optimal load" for sprinting and jumping. Twenty-five semiprofessional soccer players performed the sprint test, whereas a total of 46 completed the vertical jump test. Two different regression equations were developed for calculating the load for each exercise. The following equations were obtained: % body mass (BM) = -2.0762·%Vmax + 207.99 for the sprint and % BM = -0.7156·%Hmax + 71.588 for the vertical jump. For both sprinting and jumping, when the load increased, Vmax and Hmax decreased. The "optimal load" for resisted training using weighted vest was unclear for sprinting and close to BM for vertical jump. This study presents a new tool to individualize the training load for resisted sprinting and jumping using weighted vest in soccer players and to develop the whole force-velocity spectrum according to the objectives of the different periods of the season.

Tidal stripping and the structure of dwarf galaxies in the Local Group

NASA Astrophysics Data System (ADS)

Fattahi, Azadeh; Navarro, Julio F.; Frenk, Carlos S.; Oman, Kyle A.; Sawala, Till; Schaller, Matthieu

2018-05-01

The shallow faint-end slope of the galaxy mass function is usually reproduced in Λ cold dark matter (ΛCDM) galaxy formation models by assuming that the fraction of baryons that turn into stars drops steeply with decreasing halo mass and essentially vanishes in haloes with maximum circular velocities Vmax < 20-30 km s-1. Dark-matter-dominated dwarfs should therefore have characteristic velocities of about that value, unless they are small enough to probe only the rising part of the halo circular velocity curve (i.e. half-mass radii, r1/2 ≪ 1 kpc). Many dwarfs have properties in disagreement with this prediction: they are large enough to probe their halo Vmax but their characteristic velocities are well below 20 km s-1. These `cold faint giants' (an extreme example is the recently discovered Crater 2 Milky Way satellite) can only be reconciled with our ΛCDM models if they are the remnants of once massive objects heavily affected by tidal stripping. We examine this possibility using the APOSTLE cosmological hydrodynamical simulations of the Local Group. Assuming that low-velocity-dispersion satellites have been affected by stripping, we infer their progenitor masses, radii, and velocity dispersions, and find them in remarkable agreement with those of isolated dwarfs. Tidal stripping also explains the large scatter in the mass discrepancy-acceleration relation in the dwarf galaxy regime: tides remove preferentially dark matter from satellite galaxies, lowering their accelerations below the amin ˜ 10-11 m s-2 minimum expected for isolated dwarfs. In many cases, the resulting velocity dispersions are inconsistent with the predictions from Modified Newtonian Dynamics, a result that poses a possibly insurmountable challenge to that scenario.

Invertase immobilization onto radiation-induced graft copolymerized polyethylene pellets

NASA Astrophysics Data System (ADS)

de Queiroz, Alvaro Antonio Alencar; Vitolo, Michele; de Oliveira, Rômulo Cesar; Higa, Olga Zazuco

1996-06-01

The graft copolymer poly(ethylene-g-acrylic acid) (LDPE-g-AA) was prepared by radiation-induced graft copolymerization of acrylic acid onto low density polyethylene (LDPE) pellets, and characterized by infrared photoacoustic spectroscopy and scanning electron microscopy (SEM). The presence of the grafted poly(acrylic acid) (PAA) was established. Invertase was immobilized onto the graft polymer and the thermodynamic parameters of the soluble and immobilized enzyme were determined. The Michaelis constant, Km, and the maximum reaction velocity, Vmax, were determined for the free and the immobilized invertase. The Michaelis constant, Km was larger for the immobilized invertase than for the free enzyme, whereas Vmax was smaller for the immobilized invertase. The thermal stability of the immobilized invertase was higher than that of the free enzyme.

N-Acetyl-2-Aminofluorene (AAF) Processing in Adult Rat Hepatocytes in Primary Culture Occurs by High-Affinity Low-Velocity and Low-Affinity High-Velocity AAF Metabolite-Forming Systems.

PubMed

Koch, Katherine S; Moran, Tom; Shier, W Thomas; Leffert, Hyam L

2018-05-01

N-acetyl-2-aminofluorene (AAF) is a procarcinogen used widely in physiological investigations of chemical hepatocarcinogenesis. Its metabolic pathways have been described extensively, yet little is known about its biochemical processing, growth cycle expression, and pharmacological properties inside living hepatocytes-the principal cellular targets of this hepatocarcinogen. In this report, primary monolayer adult rat hepatocyte cultures and high specific-activity [ring G-3 H]-N-acetyl-2-aminofluorene were used to extend previous observations of metabolic activation of AAF by highly differentiated, proliferation-competent hepatocytes in long-term cultures. AAF metabolism proceeded by zero-order kinetics. Hepatocytes processed significant amounts of procarcinogen (≈12 μg AAF/106 cells/day). Five ring-hydroxylated and one deacetylated species of AAF were secreted into the culture media. Extracellular metabolite levels varied during the growth cycle (days 0-13), but their rank quantitative order was time invariant: 5-OH-AAF > 7-OH-AAF > 3-OH-AAF > N-OH-AAF > aminofluorene (AF) > 1-OH-AAF. Lineweaver-Burk analyses revealed two principal classes of metabolism: System I (high-affinity and low-velocity), Km[APPARENT] = 1.64 × 10-7  M and VMAX[APPARENT] = 0.1 nmol/106 cells/day and System II (low-affinity and high-velocity), Km[APPARENT] = 3.25 × 10-5  M and VMAX[APPARENT] = 1000 nmol/106 cells/day. A third system of metabolism of AAF to AF, with Km[APPARENT] and VMAX[APPARENT] constants of 9.6 × 10-5  M and 4.7 nmol/106 cells/day, was also observed. Evidence provided in this report and its companion paper suggests selective roles and intracellular locations for System I- and System II-mediated AAF metabolite formation during hepatocarcinogenesis, although some of the molecules and mechanisms responsible for multi-system processing remain to be fully defined.

Effects of 8-week in-season plyometric training on upper and lower limb performance of elite adolescent handball players.

PubMed

Chelly, Mohamed Souhaiel; Hermassi, Souhail; Aouadi, Ridha; Shephard, Roy J

2014-05-01

We hypothesized that replacement of a part of the normal in-season regimen of top-level adolescent handball players by an 8-week biweekly course of lower and upper limb plyometric training would enhance characteristics important to competition, including peak power output (Wpeak), jump performance, muscle volume, and ball throwing velocity. Study participants (23 men, age: 17.4 ± 0.5 years, body mass: 79.9 ± 11.5 kg, height: 1.79 ± 6.19 m, body fat: 13.8 ± 2.1%) were randomly assigned between controls (C; n = 11) and an experimental group (E, n = 12). Measures preintervention and postintervention included force-velocity ergometer tests for upper (Wupper peak) and lower limbs (Wlower peak), force platform determinations of squat jump (SJ) and countermovement jump (CMJ) characteristics (jump height, maximal force, initial velocity, and average power), video filming of sprint velocities (first step [V1S], first 5 m [V5m], and 25-30 m [Vmax]), and anthropometric estimates of leg muscle volume. E showed gains relative to C in Wupper peak and Wlower peak (p < 0.01 and p < 0.001), SJ (height p < 0.01; force p ≤ 0.05), CMJ (height p < 0.01; force p < 0.01 and relative power p ≤ 0.05), and sprint velocities (p < 0.001 for V1S, V5m, and Vmax). E also showed increases in leg and thigh muscle volumes (p < 0.001), but arm muscle volumes did not differ from control. We conclude that introduction of biweekly plyometric training into the standard regimen improved components important to handball performance, particularly explosive actions, such as sprinting, jumping, and ball throwing velocity.

Examining the effect of galaxy evolution on the stellar-halo mass relation in the EAGLE simulation

NASA Astrophysics Data System (ADS)

Kulier, Andrea; Padilla, Nelson; Schaye, Joop; Crain, Robert; Schaller, Matthieu; Bower, Richard; Theuns, Tom; Paillas, Enrique

2018-01-01

The EAGLE hydrodynamical simulation was used in Matthee et al. 2016 to examine the scatter in the stellar mass-halo mass relation of central galaxies, finding that the stellar mass (M*) correlates well with the maximum circular velocity (Vmax) of the host halo, but with a substantial scatter that does not correlate significantly with other host halo properties. Here we further examine the scatter in the stellar mass-halo mass relation of central galaxies in EAGLE, its correlation with other properties, and its origin. We find that at fixed Vmax, galaxies with lower concentration have younger stellar populations, as expected from the relationship between concentration and halo assembly time. However, at fixed Vmax and halo concentration, galaxies with larger M* have younger stellar ages, so that combining the two effects, galaxies with younger stellar ages at fixed halo mass have higher stellar masses. The host halos of galaxies with larger M* at fixed Vmax and concentration also contain more gas than those with smaller stellar masses at z = 0.1, i.e. the baryon fraction of the halos is larger. There is an even stronger correlation between the scatter in M* at z = 0.1 and the scatter in the baryon fraction of the galaxy's progenitors at z ~ 1, such that the latter sets ~50% of the scatter in M* at z = 0.1. We conclude that most of the scatter between Vmax and M* at z = 0.1 is set at earlier redshifts by the scatter in the baryon fraction of halos, which in turn is primarily the result of differences in feedback strength within halos.

[Biomechanics and bio-energetics of smooth muscle contraction. Relation to bronchial hyperreactivity].

PubMed

Coirault, C; Blanc, F X; Chemla, D; Salmeron, S; Lecarpentier, Y

2000-06-01

Mechanical studies of isolated muscle and analysis of molecular actomyosin interactions have improved our understanding of the pathophysiology of airway smooth muscle. Mechanical properties of airway smooth muscle are similar to those of other smooth muscles. Airway smooth muscle exhibits spontaneous intrinsic tone and its maximum shortening velocity (Vmax) is 10-30 fold lower than in striated muscle. Smooth muscle myosin generates step size and elementary force per crossbridge interaction approximately similar to those of skeletal muscle myosin. Special slow cycling crossbridges, termed latch-bridges, have been attributed to myosin light chain dephosphorylation. From a mechanical point of view, it has been shown that airway hyperresponsiveness is characterized by an increased Vmax and an increased shortening capacity, with no significant change in the force-generating capacity.

Studies on long chain cis- and trans-acyl-CoA esters and Acyl-CoA dehydrogenase from rat heart mitochondria.

PubMed

Korsrud, G O; Conacher, H B; Jarvis, G A; Beare-Rogers, J L

1977-02-01

The beta-oxidation of long chain fatty acids was investigated in a preparation of rat heart mitochondria. The acyl-CoA esters of the cis and trans isomers of delta9-hexadecenoic, delta9-octadecenoic, delta11-eicosenoic, and delta13-docosenoic acids were prepared. Rates of the acyl-CoA reaction were determined with an extract from rat heart mitochondria. The apparent Michaelis constant (Km) and maximum velocity (Vmax) were calculated for each substrate. In general, apparent Vmax values decreased with increasing chain length of the monoenoic substrates. Reduced activity of acyl-CoA dehydrogenase with long chain acyl-CoA esters could have contributed to accumulation of lipids in hearts of rats fed diets containing long chain fatty acids.

How neotropical hummingbird versus bat species generate lift to hover

NASA Astrophysics Data System (ADS)

Ingersoll, Rivers; Lentink, David

2017-11-01

Both hummingbirds and nectar bats evolved the ability to hover in front of flowers providing them access to energy rich nectar. Hummingbirds have been found to generate more than a quarter of their weight support during the upstroke by inverting their wings-much more than generalist birds during slow hovering flight. In contrast to hummingbirds, bats have membrane wings which they partially fold during the upstroke. It has been hypothesized that bats generate some vertical lift force during the upstroke although the complex wake structures make it hard to quantify upstroke function through flow measurement. To compare the kinematics and aerodynamic forces generated by both groups, we caught and trained over 100 individuals spanning 18 hummingbird and 3 bat species in Coto Brus, Costa Rica. We used 3D calibrated high-speed cameras to measure wingbeat kinematics and a novel aerodynamic force platform to measure the instantaneous vertical lift force in vivo. This data gives us new insight into how ecology shapes the evolution of hovering flight across taxa in the same ecosystem. This research is supported by NSF CAREER Award 1552419 and the KACST Center of Excellence for Aeronautics and Astronautics at Stanford.

Power output and force-velocity relationship of red and white muscle fibres from the Pacific blue marlin (Makaira nigricans).

PubMed

Johnston, I A; Salamonski, J

1984-07-01

Single white fibres and small bundles (two to three) of red fibres were isolated from the trunk muscle of Pacific Blue Marlin (50-121 kg body weight). Fibres were chemically skinned with 1% Brij. Maximum Ca2+-activated force production (Po) was 57 kN m-2 for red fibres and 176 kN m-2 for white fibres at 25 degrees C. The force-velocity (P-V) characteristics of these fibres were determined at 15 and 25 degrees C. Points below 0.6 Po on the P-V curve could be fitted to a linear form of Hill's equation. The degree of curvature of the P-V curve was similar at 15 and 25 degrees C (Hill's constant a/Po = 0.24 and 0.12 for red and white fibres respectively). Extrapolated maximum contraction velocities (Vmax) were 2.5 muscle lengths s-1 (Lo S-1) (red fibres) and 5.3 Lo S-1 (white fibres) at 25 degrees C. Q10(15-25 degrees C) values for Vmax were 1.4 and 1.3 for red and white fibres respectively. Maximum power output had a similar low temperature dependence and amounted to 13 W kg-1 for red and 57 W kg-1 for white muscle at 25 degrees C. The results are briefly discussed in relation to the locomotion and ecology of marlin.

Accurate mass and velocity functions of dark matter haloes

NASA Astrophysics Data System (ADS)

Comparat, Johan; Prada, Francisco; Yepes, Gustavo; Klypin, Anatoly

2017-08-01

N-body cosmological simulations are an essential tool to understand the observed distribution of galaxies. We use the MultiDark simulation suite, run with the Planck cosmological parameters, to revisit the mass and velocity functions. At redshift z = 0, the simulations cover four orders of magnitude in halo mass from ˜1011M⊙ with 8783 874 distinct haloes and 532 533 subhaloes. The total volume used is ˜515 Gpc3, more than eight times larger than in previous studies. We measure and model the halo mass function, its covariance matrix w.r.t halo mass and the large-scale halo bias. With the formalism of the excursion-set mass function, we explicit the tight interconnection between the covariance matrix, bias and halo mass function. We obtain a very accurate (<2 per cent level) model of the distinct halo mass function. We also model the subhalo mass function and its relation to the distinct halo mass function. The set of models obtained provides a complete and precise framework for the description of haloes in the concordance Planck cosmology. Finally, we provide precise analytical fits of the Vmax maximum velocity function up to redshift z < 2.3 to push for the development of halo occupation distribution using Vmax. The data and the analysis code are made publicly available in the Skies and Universes data base.

Drug oxygenation activities mediated by liver microsomal flavin-containing monooxygenases 1 and 3 in humans, monkeys, rats, and minipigs.

PubMed

Yamazaki, Miho; Shimizu, Makiko; Uno, Yasuhiro; Yamazaki, Hiroshi

2014-07-15

Liver microsomal flavin-containing monooxygenases (FMO, EC 1.14.13.8) 1 and 3 were functionally characterized in terms of expression levels and molecular catalytic capacities in human, cynomolgus monkey, rat, and minipig livers. Liver microsomal FMO3 in humans and monkeys and FMO1 and FMO3 in rats and minipigs could be determined immunochemically with commercially available anti-human FMO3 peptide antibodies or rat FMO1 peptide antibodies. With respect to FMO-dependent N-oxygenation of benzydamine and tozasertib and S-oxygenation of methimazole and sulindac sulfide activities, rat and minipig liver microsomes had high maximum velocity values (Vmax) and high catalytic efficiency (Vmax/Km, Michaelis constant) compared with those for human or monkey liver microsomes. Apparent Km values for recombinantly expressed rat FMO3-mediated N- and S-oxygenations were approximately 10-100-fold those of rat FMO1, although these enzymes had similar Vmax values. The mean catalytic efficiencies (Vmax/Km, 1.4 and 0.4 min(-1)μM(-1), respectively) of recombinant human and monkey FMO3 were higher than those of FMO1, whereas Vmax/Km values for rat and minipig FMO3 were low compared with those of FMO1. Minipig liver microsomal FMO1 efficiently catalyzed N- and S-oxygenation reactions; in addition, the minipig liver microsomal FMO1 concentration was higher than the levels in rats, humans, and monkeys. These results suggest that liver microsomal FMO1 could contribute to the relatively high FMO-mediated drug N- and S-oxygenation activities in rat and minipig liver microsomes and that lower expression of FMO1 in human and monkey livers could be a determinant factor for species differences in liver drug N- and S-oxygenation activities between experimental animals and humans. Copyright © 2014 Elsevier Inc. All rights reserved.

High levels of leptin modulate esophageal motor characteristics in type 2 diabetic patients.

PubMed

Jorge, J X; Borges, Cláudia Iracema Cardoso; Delgado, F J; Oliveira, M H; Pereira, Maria Henriqueta; Simões, M A; Panão, E A; Coelho, A C; Silva, A L; Almeida, C C

2011-01-01

Leptin regulates gastric and intestinal motility, but its effect on oesophageal motility is unknown. We analyzed oesophageal manometric characteristics in diabetics with elevated leptin. Fasting blood leptin levels were measured in 32 type 2 individuals aged from 39-81 years. An oesophageal stationary manometry was then performed. Each manometric door (P) registered one third of the oesophageal activity. Results are presented as mean +/- SD. Twenty-one subjects had elevated leptin (HLL) while 11 displayed normal levels (NLL). Peristaltic wave distributions (%) in NLL vs. HLL were 79.4 +/- 26.3 vs. 88.6 +/- 8.3 (p = 0.2). Simultaneous and retrograde waves showed similar trends. Non-transmitted waves were 16.1 +/- 26.5 vs. 4.6 +/- 4.5% (p < 0.05). Amplitudes in NLL vs. HLL (in mm Hg) were P1: 30.2 +/- 10.8 vs. 33.2 +/- 11.7 (p = 0.4), P2: 38.4 +/- 14.4 vs. 58.0 +/- 21.2 (p = 0.01), P3: 42.4 +/- 14.4 vs. 64.7 +/-2 8.3 (p < 0.006), and average amplitudes: 37.1 +/- 12.1 vs. 52.1 +/- 17.6 (p = 0.01). Wave average upstroke (in mm Hgs) was P1: 25.6 +/- 19.1 vs. 23.3 +/- 10.1 (p = 0.6), P2: 26.8 +/- 10.7 vs. 36.2 +/- 11.6 (p < 0.03), and P3: 25.5 +/- 9.1 vs. 34.1 +/- 16.3, (p < 0.06). Wave maximum upstroke was P1: 39.0 +/- 18.6 vs. 40.5 +/- 13.8, (p = 0.8), P2: 45.5 +/- 15.5 vs. 63.8 +/- 19.2 (p = 0.01), P3: 46.6 +/- 17.8 vs. 65.0 +/- 29.1 (p <0.03). Wave duration in distal oesophagus was 4.5 +/- 0.7 vs. 5.5 +/- 1.1 s (p = 0.01), and velocity 3.3 +/- 3.3 vs. 2.96 +/- 3.7 cm/s (p = 0.6). 1--Non-transmitted waves were slightly higher in NLL. 2--In medium and distal oesophagus, the wave amplitude, medium and maximum upstroke, and duration in distal oesophagus were increased in HLL.

Galaxy kinematics in the XMMU J2235-2557 cluster field at z 1.4

NASA Astrophysics Data System (ADS)

Pérez-Martínez, J. M.; Ziegler, B.; Verdugo, M.; Böhm, A.; Tanaka, M.

2017-09-01

Aims: The relationship between baryonic and dark components in galaxies varies with the environment and cosmic time. Galaxy scaling relations describe strong trends between important physical properties. A very important quantitative tool in case of spiral galaxies is the Tully-Fisher relation (TFR), which combines the luminosity of the stellar population with the characteristic rotational velocity (Vmax) taken as proxy for the total mass. In order to constrain galaxy evolution in clusters, we need measurements of the kinematic status of cluster galaxies at the starting point of the hierarchical assembly of clusters and the epoch when cosmic star formation peaks. Methods: We took spatially resolved slit FORS2 spectra of 19 cluster galaxies at z 1.4, and 8 additional field galaxies at 1 < z < 1.2 using the ESO Very Large Telescope. The targets were selected from previous spectroscopic and photometric campaigns as [OII] and Hα emitters. Our spectroscopy was complemented with HST/ACS imaging in the F775W and F850LP filters, which is mandatory to derive the galaxy structural parameters accurately. We analyzed the ionized gas kinematics by extracting rotation curves from the two-dimensional spectra. Taking into account all geometrical, observational, and instrumental effects, we used these rotation curves to derive the intrinsic maximum rotation velocity. Results: Vmax was robustly determined for six cluster galaxies and three field galaxies. Galaxies with sky contamination or insufficient spatial rotation curve extent were not included in our analysis. We compared our sample to the local B-band TFR and the local velocity-size relation (VSR), finding that cluster galaxies are on average 1.6 mag brighter and a factor 2-3 smaller. We tentatively divided our cluster galaxies by total mass (I.e., Vmax) to investigate a possible mass dependency in the environmental evolution of galaxies. The averaged deviation from the local TFR is ⟨ ΔMB ⟩ = -0.7 for the high-mass subsample (Vmax > 200 km s-1). This mild evolution may be driven by younger stellar populations (SP) of distant galaxies with respect to their local counterparts, and thus, an increasing luminosity is expected toward higher redshifts. However, the low-mass subsample (Vmax < 200 km s-1) is made of highly overluminous galaxies that show ⟨ ΔMB ⟩ = -2.4 mag. When we repeated a similar analysis with the stellar mass TFR, we did not find significant offsets in our subsamples with respect to recent results at similar redshift. While the B-band TFR is sensitive to recent episodes of star formation, the stellar mass TFR tracks the overall evolution of the underlying stellar population. In order to understand the discrepancies between these two incarnations of the TFR, the reported B-band offsets can no longer be explained only by the gradual evolution of stellar populations with lookback time. We suspect that we instead see compact galaxies whose star formation was enhanced during their infall toward the dense regions of the cluster through interactions with the intracluster medium. Based on observations with the European Southern Observatory Very Large Telescope (ESO-VLT), observing run ID 091.B-0778(B).

Effects of acetylcholine and noradrenalin on action potentials of isolated rabbit sinoatrial and atrial myocytes.

PubMed

Verkerk, Arie O; Geuzebroek, Guillaume S C; Veldkamp, Marieke W; Wilders, Ronald

2012-01-01

The autonomic nervous system controls heart rate and contractility through sympathetic and parasympathetic inputs to the cardiac tissue, with acetylcholine (ACh) and noradrenalin (NA) as the chemical transmitters. In recent years, it has become clear that specific Regulators of G protein Signaling proteins (RGS proteins) suppress muscarinic sensitivity and parasympathetic tone, identifying RGS proteins as intriguing potential therapeutic targets. In the present study, we have identified the effects of 1 μM ACh and 1 μM NA on the intrinsic action potentials of sinoatrial (SA) nodal and atrial myocytes. Single cells were enzymatically isolated from the SA node or from the left atrium of rabbit hearts. Action potentials were recorded using the amphotericin-perforated patch-clamp technique in the absence and presence of ACh, NA, or a combination of both. In SA nodal myocytes, ACh increased cycle length and decreased diastolic depolarization rate, whereas NA decreased cycle length and increased diastolic depolarization rate. Both ACh and NA increased maximum upstroke velocity. Furthermore, ACh hyperpolarized the maximum diastolic potential. In atrial myocytes stimulated at 2 Hz, both ACh and NA hyperpolarized the maximum diastolic potential, increased the action potential amplitude, and increased the maximum upstroke velocity. Action potential duration at 50 and 90% repolarization was decreased by ACh, but increased by NA. The effects of both ACh and NA on action potential duration showed a dose dependence in the range of 1-1000 nM, while a clear-cut frequency dependence in the range of 1-4 Hz was absent. Intermediate results were obtained in the combined presence of ACh and NA in both SA nodal and atrial myocytes. Our data uncover the extent to which SA nodal and atrial action potentials are intrinsically dependent on ACh, NA, or a combination of both and may thus guide further experiments with RGS proteins.

Effects of Acetylcholine and Noradrenalin on Action Potentials of Isolated Rabbit Sinoatrial and Atrial Myocytes

PubMed Central

Verkerk, Arie O.; Geuzebroek, Guillaume S. C.; Veldkamp, Marieke W.; Wilders, Ronald

2012-01-01

The autonomic nervous system controls heart rate and contractility through sympathetic and parasympathetic inputs to the cardiac tissue, with acetylcholine (ACh) and noradrenalin (NA) as the chemical transmitters. In recent years, it has become clear that specific Regulators of G protein Signaling proteins (RGS proteins) suppress muscarinic sensitivity and parasympathetic tone, identifying RGS proteins as intriguing potential therapeutic targets. In the present study, we have identified the effects of 1 μM ACh and 1 μM NA on the intrinsic action potentials of sinoatrial (SA) nodal and atrial myocytes. Single cells were enzymatically isolated from the SA node or from the left atrium of rabbit hearts. Action potentials were recorded using the amphotericin-perforated patch-clamp technique in the absence and presence of ACh, NA, or a combination of both. In SA nodal myocytes, ACh increased cycle length and decreased diastolic depolarization rate, whereas NA decreased cycle length and increased diastolic depolarization rate. Both ACh and NA increased maximum upstroke velocity. Furthermore, ACh hyperpolarized the maximum diastolic potential. In atrial myocytes stimulated at 2 Hz, both ACh and NA hyperpolarized the maximum diastolic potential, increased the action potential amplitude, and increased the maximum upstroke velocity. Action potential duration at 50 and 90% repolarization was decreased by ACh, but increased by NA. The effects of both ACh and NA on action potential duration showed a dose dependence in the range of 1–1000 nM, while a clear-cut frequency dependence in the range of 1–4 Hz was absent. Intermediate results were obtained in the combined presence of ACh and NA in both SA nodal and atrial myocytes. Our data uncover the extent to which SA nodal and atrial action potentials are intrinsically dependent on ACh, NA, or a combination of both and may thus guide further experiments with RGS proteins. PMID:22754533

Contractile properties of rat, rhesus monkey, and human type I muscle fibers

NASA Technical Reports Server (NTRS)

Widrick, J. J.; Romatowski, J. G.; Karhanek, M.; Fitts, R. H.

1997-01-01

It is well known that skeletal muscle intrinsic maximal shortening velocity is inversely related to species body mass. However, there is uncertainty regarding the relationship between the contractile properties of muscle fibers obtained from commonly studied laboratory animals and those obtained from humans. In this study we determined the contractile properties of single chemically skinned fibers prepared from rat, rhesus monkey, and human soleus and gastrocnemius muscle samples under identical experimental conditions. All fibers used for analysis expressed type I myosin heavy chain as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Allometric coefficients for type I fibers from each muscle indicated that there was little change in peak tension (force/fiber cross-sectional area) across species. In contrast, both soleus and gastrocnemius type I fiber maximal unloaded shortening velocity (Vo), the y-intercept of the force-velocity relationship (Vmax), peak power per unit fiber length, and peak power normalized for fiber length and cross-sectional area were all inversely related to species body mass. The present allometric coefficients for soleus fiber Vo (-0.18) and Vmax (-0.11) are in good agreement with published values for soleus fibers obtained from common laboratory and domesticated mammals. Taken together, these observations suggest that the Vo of slow fibers from quadrupeds and humans scale similarly and can be described by the same quantitative relationships. These findings have implications in the design and interpretation of experiments, especially those that use small laboratory mammals as a model of human muscle function.

Screening for stroke in sickle cell anemia: comparison of transcranial Doppler imaging and nonimaging US techniques.

PubMed

Neish, Ariane S; Blews, David E; Simms, Catherine A; Merritt, Robert K; Spinks, Alice J

2002-03-01

To determine whether criteria for screening patients with sickle cell anemia for stroke established with a nonimaging transcranial Doppler ultrasonographic (US) technique are applicable to studies performed with a transcranial Doppler US imaging technique. One hundred sixty-eight examinations in 66 children were performed for sickle cell stroke screening. Children were examined with nonimaging and imaging transcranial Doppler US techniques on the same day, for a total of 84 paired examinations. The time-averaged maximum mean velocity (V(mean)) and resistive index (RI) were calculated in the middle cerebral arteries, bifurcations of the distal internal carotid arteries, distal internal carotid arteries, anterior cerebral arteries, posterior cerebral arteries, and basilar arteries. The maximum systolic velocity (V(max)) was evaluated in the distal internal carotid arteries and middle cerebral arteries. V(mean), V(max), and RI measurements were subjected to repeated-measures multivariate analysis of covariance, and the Pearson product moment correlation was used for middle cerebral artery velocity, age, and hemoglobin. V(mean) measurements obtained with nonimaging and imaging techniques varied substantially for the bifurcation of the distal internal carotid artery, the posterior cerebral artery, and the basilar artery. Substantial differences were found in RIs for every vessel. Examination time was shorter with the nonimaging technique. V(mean) measurements in the middle cerebral artery, distal internal carotid artery, and anterior cerebral artery did not vary substantially between nonimaging and imaging transcranial Doppler US. RI data did not yield comparable measurements.

Impact of afterload on the assessment of severity of aortic stenosis.

PubMed

Chang, Sung-A; Kim, Hyung-Kwan; Sohn, Dae-Won

2012-06-01

Aortic stenosis (AS) is increasingly diagnosed in current aging society. Echocardiography is the most important tool in the assessment of AS and its severity. However, load-dependency of Doppler measurement could affect the accuracy of AS severity assessment. We tried to evaluate the impact of afterload on the assessment of AS severity by modification of afterload using pneumatic compression (Pcom). Forty patients diagnosed as moderate or severe AS [effective orifice area of aortic valve (EOA(AV)) by continuity equation of < 1.5 cm(2)] were consecutively enrolled. Patients with severely uncontrolled hypertension, severe left ventricular (LV) dysfunction, and other significant valve disease were excluded. Comprehensive echocardiography was performed at baseline to assess AS severity. Then, pneumatic compression of the lower extremities by 100 mmHg was applied to increase LV afterload. After 3 minutes, echocardiography was repeated to assess AS severity. Mean blood pressure was significantly increased under Pcom (p < 0.001), while heart rate remained unchanged. Peak aortic valve velocity (V(max)) was slightly, but significantly decreased under Pcom (p = 0.03). However, Doppler velocity index and EOA(AV) by continuity equation were not affected by Pcom. AS severity assessment by echocardiography was not dependent on the change of LV afterload imposed by Pcom. AV V(max) was slightly decreased with LV afterload increment, but these changes were too small to alter treatment plan of AS patients. EOA(AV) and Doppler velocity index are more stable parameters for AS severity assessment.

Simple predictions of maximum transport rate in unsaturated soil and rock

USGS Publications Warehouse

Nimmo, John R.

2007-01-01

In contrast with the extreme variability expected for water and contaminant fluxes in the unsaturated zone, evidence from 64 field tests of preferential flow indicates that the maximum transport speed Vmax, adjusted for episodicity of infiltration, deviates little from a geometric mean of 13 m/d. A model based on constant‐speed travel during infiltration pulses of actual or estimated duration can predict Vmax with approximate order‐of‐magnitude accuracy, irrespective of medium or travel distance, thereby facilitating such problems as the prediction of worst‐case contaminant traveltimes. The lesser variability suggests that preferential flow is subject to rate‐limiting mechanisms analogous to those that impose a terminal velocity on objects in free fall and to rate‐compensating mechanisms analogous to Le Chatlier's principle. A critical feature allowing such mechanisms to dominate may be the presence of interfacial boundaries confined by neither solid material nor capillary forces.

The Star Formation Demographics of Galaxies in the Local Volume

NASA Astrophysics Data System (ADS)

Lee, Janice C.; Kennicutt, Robert C.; Funes, S. J., José G.; Sakai, Shoko; Akiyama, Sanae

2007-12-01

We examine the connections between the current global star formation activity, luminosity, dynamical mass, and morphology of galaxies in the Local Volume, using Hα data from the 11 Mpc Hα and Ultraviolet Galaxy Survey (11HUGS). Taking the equivalent width (EW) of the Hα emission line as a tracer of the specific star formation rate, we analyze the distribution of galaxies in the MB-EW and rotational velocity (Vmax)-EW planes. Star-forming galaxies show two characteristic transitions in these planes. A narrowing of the galaxy locus occurs at MB~-15 and Vmax~50 km s-1, where the scatter in the logarithmic EWs drops by a factor of 2 as the luminosities/masses increase, and galaxy morphologies shift from predominately irregular to late-type spiral. Another transition occurs at MB~-19 and Vmax~120 km s-1, above which the sequence turns off toward lower EWs and becomes mostly populated by intermediate- and early-type bulge-prominent spirals. Between these two transitions, the mean logarithmic EW appears to remain constant at 30 Å. We comment on how these features reflect established empirical relationships, and provide clues for identifying the large-scale physical processes that both drive and regulate star formation, with emphasis on the low-mass galaxies which dominate our approximately volume-limited sample.

Effects of In-Season Short-term Plyometric Training Program on Sprint and Jump Performance of Young Male Track Athletes.

PubMed

Chelly, Mohamed Souhaiel; Hermassi, Souhail; Shephard, Roy J

2015-08-01

We studied the effect of supplementing normal in-season training by a 10-week lower limb plyometric training program (hurdle and depth jumping), examining measures of competitive potential (peak power output [PP], sprint running velocity, squat jump [SJ], countermovement jump [CMJ], drop jump [DJ], and lower limb muscle volume). The subjects (27 male track athletes, aged 11.9 ± 1.0 years; body mass: 39.1 ± 6.1 kg; height: 1.56 ± 0.02 m; body fat: 12.8 ± 4.4%) were randomly assigned between a control (normal training) group (C; n = 13) and an experimental group (E; n = 14) who also performed plyometric training 3 times per week. A force-velocity ergometer test determined PP and SJ, and an Optojump apparatus evaluated CMJ height and DJ (height and power). A multiple-5-bound test assessed horizontal jumping, and video-camera analyses over a 40-m sprint yielded velocities for the first step (VS), the first 5 m (V5m), and between 35 and 40 m (Vmax). Leg muscle volume was estimated anthropometrically. Experimental group showed gains relative to C in SJ height (p < 0.001); CMJ height (p < 0.01); DJ height and power relative to body mass (p < 0.01 for both); and all sprint velocities (p < 0.01 for VS and V(5m, p) ≤ 0.05 for Vmax). There was also a significant increase (p < 0.01) in thigh muscle volume, but leg muscle volume, thigh cross-sectional area, and PP remained unchanged. We conclude that adding plyometric training improved important components of athletic performance relative to standard in-season training in young runners.

Halo Substructure and the Power Spectrum

NASA Astrophysics Data System (ADS)

Zentner, Andrew R.; Bullock, James S.

2003-11-01

We present a semianalytic model to investigate the merger history, destruction rate, and survival probability of substructure in hierarchically formed dark matter halos and use it to study the substructure content of halos as a function of input primordial power spectrum. For a standard cold dark matter ``concordance'' cosmology (ΛCDM n=1, σ8=0.95) we successfully reproduce the subhalo velocity function and radial distribution profile seen in N-body simulations and determine that the rate of merging and disruption peaks ~10-12 Gyr in the past for Milky Way-like halos, while surviving substructures are typically accreted within the last ~0-8 Gyr. We explore power spectra with normalizations and spectral ``tilts'' spanning the ranges σ8~=1-0.65 and n~=1-0.8, and include a ``running-index'' model with dn/dlnk=-0.03 similar to the best-fit model discussed in the first-year Wilkinson Microwave Anisotropy Probe (WMAP) report. We investigate spectra with truncated small-scale power, including a broken-scale inflation model and three warm dark matter cases with mW=0.75-3.0 keV. We find that the mass fraction in substructure is relatively insensitive to the tilt and overall normalization of the primordial power spectrum. All of the CDM-type models yield projected substructure mass fractions that are consistent with, but on the low side, of published estimates from strong lens systems: f9=0.4%-1.5% (64th percentile) for subhalos smaller than 109 Msolar within projected cylinders of radius r<10 kpc. Truncated models produce significantly smaller fractions, f9=0.02%-0.2% for mW~=1 keV, and are disfavored by lensing estimates. This suggests that lensing and similar probes can provide a robust test of the CDM paradigm and a powerful constraint on broken-scale inflation/warm particle masses, including masses larger than the ~1 keV upper limits of previous studies. We compare our predicted subhalo velocity functions with the dwarf satellite population of the Milky Way. Assuming that dwarfs have isotropic velocity dispersions, we find that the standard n=1 model overpredicts the number of Milky Way satellites at Vmax<~35 km s-1, as expected. Models with less small-scale power do better because subhalos are less concentrated and the mapping between observed velocity dispersion and halo Vmax is significantly altered. The running-index model, or a fixed tilt with σ8~0.75, can account for the local dwarfs without the need for differential feedback (for Vmax>~20 km s-1) however, these comparisons depend sensitively on the assumption of isotropic velocities in satellite galaxies.

Evolutionary adaptation of muscle power output to environmental temperature: force-velocity characteristics of skinned fibres isolated from antarctic, temperate and tropical marine fish.

PubMed

Johnston, I A; Altringham, J D

1985-09-01

Single fast fibres were isolated from the myotomal muscles of icefish (Chaenocephalus aceratus Lönnberg, Antarctica), North Sea Cod (Gadus morhua L.) and Pacific Blue Marlin (Makaira nigricans Wakiya, Hawaii). Fibres were chemically skinned with the non-ionic detergent Brij-58. Maximum tensions (Po, kN m-2) developed at the characteristic body temperature of each species are 231 for icefish (-1 degree C), 187 for cod (8 degrees C) and 156 for marlin (20 degrees C). At 0 degree C Po is 7 times higher for fibres from the icefish than from the marlin. Fibres from icefish and cod failed to relax completely following activations at temperatures above approximately 12 degrees C. The resultant post-contraction force is associated with a proportional increase in stiffness, suggesting the formation of a population of Ca-insensitive cross bridges. At 10 degrees C there is little interspecific variation in unloaded contraction velocity (Vmax) among the three species. Vmax (muscle lengths s-1) at normal body temperatures are 0.9 for icefish (-1 degree C), 1.0 for cod (8 degrees C) and 3.4 for marlin (20 degrees C). The force-velocity (P-V) relationship becomes progressively more curved with increasing temperature for all three species. Maximum power output for the fast muscle fibres from the Antarctic species at -1 degree C is around 60% of that of the tropical fish at 20 degrees C. Evolutionary temperature compensation of muscle power output appears largely to involve differences in the ability of cross bridges to generate force.

Computer program for the reservoir model of metabolic crossroads.

PubMed

Ribeiro, J M; Juzgado, D; Crespo, E; Sillero, A

1990-01-01

A program containing 344 sentences, written in BASIC and adapted to run in personal computers (PC) has been developed to simulate the reservoir model of metabolic crossroads. The program draws the holes of the reservoir with shapes reflecting the Vmax, Km (S0.5) and cooperativity coefficients (n) of the enzymes and calculates both the actual velocities and the percentage of contribution of every enzyme to the overall removal of their common substrate.

Optimal Protective Hypothermia in Arrested Mammalian Hearts

PubMed Central

Villet, Outi M.; Ge, Ming; Sekhar, Laigam N.; Corson, Marshall A.; Tylee, Tracy S.; Fan, Lu-Ping; Yao, Lin; Zhu, Chun; Olson, Aaron K.; Buroker, Norman E.; Xu, Cheng-Su; Anderson, David L.; Soh, Yong-Kian; Wang, Elise; Chen, Shi-Han; Portman, Michael A.

2015-01-01

Many therapeutic hypothermia recommendations have been reported, but the information supporting them is sparse, and reveals a need for the data of target therapeutic hypothermia (TTH) from well-controlled experiments. The core temperature ≤35°C is considered as hypothermia, and 29°C is a cooling injury threshold in pig heart in vivo. Thus, an optimal protective hypothermia (OPH) should be in the range 29–35°C. This study was conducted with a pig cardiopulmonary bypass preparation to decrease the core temperature to 29–35°C range at 20 minutes before and 60 minutes during heart arrest. The left ventricular (LV) developed pressure, maximum of the first derivative of LV (dP/dtmax), cardiac power, heart rate, cardiac output, and myocardial velocity (Vmax) were recorded continuously via an LV pressure catheter and an aortic flow probe. At 20 minutes of off-pump during reperfusion after 60 minutes arrest, 17 hypothermic hearts showed that the recovery of Vmax and dP/dtmax established sigmoid curves that consisted of two plateaus: a good recovery plateau at 29–30.5°C, the function recovered to baseline level (BL) (Vmax=118.4%±3.9% of BL, LV dP/dtmax=120.7%±3.1% of BL, n=6); another poor recovery plateau at 34–35°C (Vmax=60.2%±2.8% of BL, LV dP/dtmax=28.0%±5.9% of BL, p<0.05, n=6; ), which are similar to the four normothermia arrest (37°C) hearts (Vmax=55.9%±4.8% of BL, LV dP/dtmax=24.5%±2.1% of BL, n=4). The 32–32.5°C arrest hearts showed moderate recovery (n=5). A point of inflection (around 30.5–31°C) existed at the edge of a good recovery plateau followed by a steep slope. The point presented an OPH that should be the TTH. The results are concordant with data in the mammalian hearts, suggesting that the TTH should be initiated to cool core temperature at 31°C. PMID:25514569

[Effect of endogenous H2S on platelet L-Arg transport].

PubMed

Duan, Wen-zhuo; Wang, Yi-peng; Gong, Hai-min

2010-05-01

To observe the effect of novel air neuromodulator H2S on platelet function of L-Arg transport for discussing H2S of effect on platelet function. Saturate H2S solution as donate made rat rich platelet plasma and pre-incubation rat platelet with different density of H2S. To measure the velocity of L-Arg transport in platelet by radioactivity technique. At different concentrations of H2S (6.25, 12.5, 25, 50, 100 micromol/L), the velocity of L-Arg transport was lower than that in control. H2S reduced rapidly the Vmax and velocity of L-Arg transport in platelet (P < 0.05) and this effect had no effect to Km. H2S can affect platelet function by changing rapidly platelet L-Arg transport system function.

Evaluation of nefazodone-induced cardiotoxicity in human induced pluripotent stem cell-derived cardiomyocytes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Sujeong, E-mail: crystalee@gmail.com; Lee, Hyang-Ae, E-mail: hyangaelee@gmail.com; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 110-799

2016-04-01

The recent establishment of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), which express the major cardiac ion channels and recapitulate spontaneous mechanical and electrical activities, may provide a possible solution for the lack of in vitro human-based cardiotoxicity testing models. Cardiotoxicity induced by the antidepressant nefazodone was previously revealed to cause an acquired QT prolongation by hERG channel blockade. To elucidate the cellular mechanisms underlying the cardiotoxicity of nefazodone beyond hERG, its effects on cardiac action potentials (APs) and ion channels were investigated using hiPSC-CMs with whole-cell patch clamp techniques. In a proof of principle study, we examined the effectsmore » of cardioactive channel blockers on the electrophysiological profile of hiPSC-CMs in advance of the evaluation of nefazodone. Nefazodone dose-dependently prolonged the AP duration at 90% (APD{sub 90}) and 50% (APD{sub 50}) repolarization, reduced the maximum upstroke velocity (dV/dt{sub max}) and induced early after depolarizations. Voltage-clamp studies of hiPSC-CMs revealed that nefazodone inhibited various voltage-gated ion channel currents including I{sub Kr}, I{sub Ks}, I{sub Na}, and I{sub Ca}. Among them, I{sub Kr} and I{sub Na} showed relatively higher sensitivity to nefazodone, consistent with the changes in the AP parameters. In summary, hiPSC-CMs enabled an integrated approach to evaluate the complex interactions of nefazodone with cardiac ion channels. These results suggest that hiPSC-CMs can be an effective model for detecting drug-induced arrhythmogenicity beyond the current standard assay of heterologously expressed hERG K{sup +} channels. - Highlights: • Nefazodone prolonged APD and decreased upstroke velocity of APs in hiPSC-CMs. • Nefazodone inhibited cardiac ion channels, especially I{sub Kr} and I{sub Na}, in hiPSC-CMs. • Nefazodone-induced AP changes are mainly the result of I{sub Kr} and I{sub Na} inhibition. • hiPSC-CMs are sensitive model to validate nefazodone-induced cardiotoxicity. • hiPSC-CMs provide an integrated approach for evaluating mechanism of drug actions.« less

Modes of thrust generation in flying animals

NASA Astrophysics Data System (ADS)

Luo, Haoxiang; Song, Jialei; Tobalske, Bret; Luo Team; Tobalske Team

2016-11-01

For flying animals in forward flight, thrust is usually much smaller as compared with weight support and has not been given the same amount of attention. Several modes of thrust generation are discussed in this presentation. For insects performing slow flight that is characterized by low advance ratios (i.e., the ratio between flight speed and wing speed), thrust is usually generated by a "backward flick" mode, in which the wings moves upward and backward at a faster speed than the flight speed. Paddling mode is another mode used by some insects like fruit flies who row their wings backward during upstroke like paddles (Ristroph et al., PRL, 2011). Birds wings have high advance ratios and produce thrust during downstroke by directing aerodynamic lift forward. At intermediate advance ratios around one (e.g., hummingbirds and bats), the animal wings generate thrust during both downstroke and upstroke, and thrust generation during upstroke may come at cost of negative weight support. These conclusions are supported by previous experiment studies of insects, birds, and bats, as well as our recent computational modeling of hummingbirds. Supported by the NSF.

Temperature Sensitivities of Extracellular Enzyme Vmax and Km Across Thermal Environments

NASA Astrophysics Data System (ADS)

Allison, S. D.; Romero-Olivares, A.; Lu, Y.; Taylor, J.; Treseder, K. K.

2017-12-01

The magnitude and direction of carbon cycle feedbacks under climate warming remain uncertain due to insufficient knowledge about the temperature sensitivity of microbial processes in soil. Enzymatic rates could increase at higher temperatures, but this response is determined by multiple parameters that may change over time if soil microbes adapt to warming. We used the Michaelis-Menten relationship, the Arrhenius relationship, and biochemical transition state theory to construct hypotheses about the responses of extracellular enzyme Vmax and Km to temperature. Based on the Arrhenius relationship, we hypothesized that Vmax and Km would show positive temperature sensitivities. For enzymes from warmer environments, we expected to find lower Vmax, Km, and Km temperature sensitivity but higher Vmax temperature sensitivity. We tested these hypotheses with enzymes from isolates of the filamentous fungus Neurospora discreta collected around the globe and from decomposing leaf litter in a warming experiment in Alaskan boreal forest. Vmax and Km of most Neurospora extracellular enzymes were temperature sensitive with average Vmax Q10 ranging from 1.48 to 2.25 and Km Q10 ranging from 0.71 to 2.80. For both Vmax and Km, there was a tendency for the parameter to correlate negatively with its temperature sensitivity, a pattern predicted by biochemical theory. Also in agreement with theory, Vmax and Km were positively correlated for some enzymes. In contrast, there was little support for biochemical theory when comparing Vmax and Km across thermal environments. There was no relationship between temperature sensitivity of Vmax or Km and mean annual temperature of the isolation site for Neurospora strains. There was some evidence for greater Vmax under experimental warming in Alaskan litter, but the temperature sensitivities of Vmax and Km did not vary with warming as expected. We conclude that relationships among Vmax, Km, and temperature are largely consistent with biochemical theory, and our enzyme data should be useful for parameterizing trait-based models of microbial processes. However, theoretical predictions about adaptation to thermal environment were not supported by our data, suggesting that covarying edaphic and ecological factors may play a dominant role in soil enzyme responses to climate warming.

A determination and comparison of urease activity in feces and fresh manure from pig and cattle in relation to ammonia production and pH changes.

PubMed

Dai, Xiaorong; Karring, Henrik

2014-01-01

Ammonia emission from animal production is a major environmental problem and has impacts on the animal health and working environment inside production houses. Ammonia is formed in manure by the enzymatic degradation of urinary urea and catalyzed by urease that is present in feces. We have determined and compared the urease activity in feces and manure (a urine and feces mixture) from pigs and cattle at 25°C by using Michaelis-Menten kinetics. To obtain accurate estimates of kinetic parameters Vmax and K'm, we used a 5 min reaction time to determine the initial reaction velocities based on total ammoniacal nitrogen (TAN) concentrations. The resulting Vmax value (mmol urea hydrolyzed per kg wet feces per min) was 2.06±0.08 mmol urea/kg/min and 0.80±0.04 mmol urea/kg/min for pig feces and cattle feces, respectively. The K'm values were 32.59±5.65 mmol urea/l and 15.43±2.94 mmol urea/l for pig feces and cattle feces, respectively. Thus, our results reveal that both the Vmax and K'm values of the urease activity for pig feces are more than 2-fold higher than those for cattle feces. The difference in urea hydrolysis rates between animal species is even more significant in fresh manure. The initial velocities of TAN formation are 1.53 mM/min and 0.33 mM/min for pig and cattle manure, respectively. Furthermore, our investigation shows that the maximum urease activity for pig feces occurs at approximately pH 7, and in cattle feces it is closer to pH 8, indicating that the predominant fecal ureolytic bacteria species differ between animal species. We believe that our study contributes to a better understanding of the urea hydrolysis process in manure and provides a basis for more accurate and animal-specific prediction models for urea hydrolysis rates and ammonia concentration in manures and thus can be used to predict ammonia volatilization rates from animal production.

A Determination and Comparison of Urease Activity in Feces and Fresh Manure from Pig and Cattle in Relation to Ammonia Production and pH Changes

PubMed Central

Dai, Xiaorong; Karring, Henrik

2014-01-01

Ammonia emission from animal production is a major environmental problem and has impacts on the animal health and working environment inside production houses. Ammonia is formed in manure by the enzymatic degradation of urinary urea and catalyzed by urease that is present in feces. We have determined and compared the urease activity in feces and manure (a urine and feces mixture) from pigs and cattle at 25°C by using Michaelis-Menten kinetics. To obtain accurate estimates of kinetic parameters Vmax and K'm, we used a 5 min reaction time to determine the initial reaction velocities based on total ammoniacal nitrogen (TAN) concentrations. The resulting Vmax value (mmol urea hydrolyzed per kg wet feces per min) was 2.06±0.08 mmol urea/kg/min and 0.80±0.04 mmol urea/kg/min for pig feces and cattle feces, respectively. The K'm values were 32.59±5.65 mmol urea/l and 15.43±2.94 mmol urea/l for pig feces and cattle feces, respectively. Thus, our results reveal that both the Vmax and K'm values of the urease activity for pig feces are more than 2-fold higher than those for cattle feces. The difference in urea hydrolysis rates between animal species is even more significant in fresh manure. The initial velocities of TAN formation are 1.53 mM/min and 0.33 mM/min for pig and cattle manure, respectively. Furthermore, our investigation shows that the maximum urease activity for pig feces occurs at approximately pH 7, and in cattle feces it is closer to pH 8, indicating that the predominant fecal ureolytic bacteria species differ between animal species. We believe that our study contributes to a better understanding of the urea hydrolysis process in manure and provides a basis for more accurate and animal-specific prediction models for urea hydrolysis rates and ammonia concentration in manures and thus can be used to predict ammonia volatilization rates from animal production. PMID:25397404

Human beta-glucuronidase. Measurement of its activity in gallbladder bile devoid of intrinsic interference.

PubMed

Ho, Y C; Ho, K J

1988-04-01

Our purpose is to develop a standard method for preparing the bile for beta-glucuronidase determination by removal of bile acids and conjugated bilirubin which interfere with its activity. The bile acids and conjugated bilirubin in their purified solutions and in the diluted gallbladder biles could be extracted completely with cholestyramine in powder form or tetrahexylammonium chloride (THAC) in chloroform or ethyl acetate. The enzyme was, however, partially precipitated with cholestyramine and denatured by chloroform but not by ethyl acetate. A standard procedure, therefore, includes extraction of the diluted gallbladder bile with THAC in ethyl acetate, followed by determination of the maximal velocity (Vmax) of the enzyme by a kinetic method employing phenolphthalein glucuronide as the substrate. The average Vmax of beta-glucuronidase in the 20 normal gallbladder biles was 165 +/- 86 nmol/min/ml (mean +/- SD), a 23.5-fold increase over the activity before extraction. The measured activity represented the true activity of the enzyme in the bile for recovery of activity of the enzyme added to the bile was practically complete.

Peripheral Airway Smooth Muscle, but Not the Trachealis, Is Hypercontractile in an Equine Model of Asthma.

PubMed

Matusovsky, Oleg S; Kachmar, Linda; Ijpma, Gijs; Bates, Genevieve; Zitouni, Nedjma; Benedetti, Andrea; Lavoie, Jean-Pierre; Lauzon, Anne-Marie

2016-05-01

Heaves is a naturally occurring equine disease that shares many similarities with human asthma, including reversible antigen-induced bronchoconstriction, airway inflammation, and remodeling. The purpose of this study was to determine whether the trachealis muscle is mechanically representative of the peripheral airway smooth muscle (ASM) in an equine model of asthma. Tracheal and peripheral ASM of heaves-affected horses under exacerbation, or under clinical remission of the disease, and control horses were dissected and freed of epithelium to measure unloaded shortening velocity (Vmax), stress (force/cross-sectional area), methacholine effective concentration at which 50% of the maximum response is obtained, and stiffness. Myofibrillar Mg(2+)-ATPase activity, actomyosin in vitro motility, and contractile protein expression were also measured. Horses with heaves had significantly greater Vmax and Mg(2+)-ATPase activity in peripheral airway but not in tracheal smooth muscle. In addition, a significant correlation was found between Vmax and the time elapsed since the end of the corticosteroid treatment for the peripheral airways in horses with heaves. Maximal stress and stiffness were greater in the peripheral airways of the horses under remission compared with controls and the horses under exacerbation, potentially due to remodeling. Actomyosin in vitro motility was not different between controls and horses with heaves. These data demonstrate that peripheral ASM is mechanically and biochemically altered in heaves, whereas the trachealis behaves as in control horses. It is therefore conceivable that the trachealis muscle may not be representative of the peripheral ASM in human asthma either, but this will require further investigation.

Electrophysiological mechanisms of sophocarpine as a potential antiarrhythmic agent.

PubMed

Yang, Zhi-fang; Li, Ci-zhen; Wang, Wei; Chen, Ying-min; Zhang, Ying; Liu, Yuan-mou; Wang, Hong-wei

2011-03-01

To examine the electrophysiological effects of sophocarpine on action potentials (AP) and ionic currents of cardiac myocytes and to compare some of these effects with those of amiodarone. Langendorff perfusion set-up was used in isolated guinea pig heart, and responses to sophocarpine were monitored using electrocardiograph. Conventional microelectrode, voltage clamp technique and perforated patch were employed to record fast response AP (fAP), slow response AP (sAP) and ionic currents in guinea pig papillary muscle or rabbit sinus node cells. Tachyarrhythmia produced by isoprenaline (15 μmol/L) could be reversed by sophocarpine (300 μmol/L). Sophocarpine (10 μmol/L) decreased the amplitude by 4.0%, maximal depolarization velocity (V(max)) of the fAP by 24.4%, and Na(+) current (I(Na)) by 18.0%, while it prolonged the effective refractory period (ERP) by 21.1%. The same concentration of sophocarpine could also decrease the amplitude and V(max) of the sAP, by 26.8% and 25.7%, respectively, and attenuated the Ca(2+) current (I(CaL)) and the K(+) tail current substantially. Comparison of sophocarpine with amiodarone demonstrated that both prolonged the duration and the ERP of fAP and sAP, both decreased the amplitude and V(max) of the fAP and sAP, and both slowed the automatic heart rate. Sophocarpine could reverse isoprenaline-induced arrhythmia and inhibit I(Na), I(CaL), and I(Kr) currents. The electrophysiological effects of sophocarpine are similar to those of amiodarone, which might be regarded as a prospective antiarrhythmic agent.

In vivo measurement of aerodynamic weight support in freely flying birds

NASA Astrophysics Data System (ADS)

Lentink, David; Haselsteiner, Andreas; Ingersoll, Rivers

2014-11-01

Birds dynamically change the shape of their wing during the stroke to support their body weight aerodynamically. The wing is partially folded during the upstroke, which suggests that the upstroke of birds might not actively contribute to aerodynamic force production. This hypothesis is supported by the significant mass difference between the large pectoralis muscle that powers the down-stroke and the much smaller supracoracoideus that drives the upstroke. Previous works used indirect or incomplete techniques to measure the total force generated by bird wings ranging from muscle force, airflow, wing surface pressure, to detailed kinematics measurements coupled with bird mass-distribution models to derive net force through second derivatives. We have validated a new method that measures aerodynamic force in vivo time-resolved directly in freely flying birds which can resolve this question. The validation of the method, using independent force measurements on a quadcopter with pulsating thrust, show the aerodynamic force and impulse are measured within 2% accuracy and time-resolved. We demonstrate results for quad-copters and birds of similar weight and size. The method is scalable and can be applied to both engineered and natural flyers across taxa. The first author invented the method, the second and third authors validated the method and present results for quadcopters and birds.

An extended heterogeneous car-following model accounting for anticipation driving behavior and mixed maximum speeds

NASA Astrophysics Data System (ADS)

Sun, Fengxin; Wang, Jufeng; Cheng, Rongjun; Ge, Hongxia

2018-02-01

The optimal driving speeds of the different vehicles may be different for the same headway. In the optimal velocity function of the optimal velocity (OV) model, the maximum speed vmax is an important parameter determining the optimal driving speed. A vehicle with higher maximum speed is more willing to drive faster than that with lower maximum speed in similar situation. By incorporating the anticipation driving behavior of relative velocity and mixed maximum speeds of different percentages into optimal velocity function, an extended heterogeneous car-following model is presented in this paper. The analytical linear stable condition for this extended heterogeneous traffic model is obtained by using linear stability theory. Numerical simulations are carried out to explore the complex phenomenon resulted from the cooperation between anticipation driving behavior and heterogeneous maximum speeds in the optimal velocity function. The analytical and numerical results all demonstrate that strengthening driver's anticipation effect can improve the stability of heterogeneous traffic flow, and increasing the lowest value in the mixed maximum speeds will result in more instability, but increasing the value or proportion of the part already having higher maximum speed will cause different stabilities at high or low traffic densities.

Echocardiographic estimation of systemic systolic blood pressure in dogs with mild mitral regurgitation.

PubMed

Tou, Sandra P; Adin, Darcy B; Estrada, Amara H

2006-01-01

Systemic hypertension is likely underdiagnosed in veterinary medicine because systemic blood pressure is rarely measured. Systemic blood pressure can theoretically be estimated by echocardiography. According to the modified Bernoulli equation (PG = 4v(2)), mitral regurgitation (MR) velocity should approximate systolic left ventricular pressure (sLVP), and therefore systolic systemic blood pressure (sSBP) in the presence of a normal left atrial pressure (LAP) and the absence of aortic stenosis. The aim of this study was to evaluate the use of echocardiography to estimate sSBP by means of the Bernoulli equation. Systemic blood pressure can be estimated by echocardiography. Seventeen dogs with mild MR. No dogs had aortic or subaortic stenosis, and all had MR with a clear continuous-wave Doppler signal and a left atrial to aorta ratio of < or = 1.6. Five simultaneous, blinded continuous-wave measurements of maximum MR velocity (Vmax) and indirect sSBP measurements (by Park's Doppler) were obtained for each dog. Pressure gradient was calculated from Vmax by means of the Bernoulli equation, averaged, and added to an assumed LAP of 8 mm Hg to calculate sLVP. Calculated sLVP was significantly correlated with indirectly measured sSBP within a range of 121 to 218 mm Hg (P = .0002, r = .78). Mean +/- SD bias was 0.1 +/- 15.3 mm Hg with limits of agreement of -29.9 to 30.1 mm Hg. Despite the significant correlation, the wide limits of agreement between the methods hinder the clinical utility of echocardiographic estimation of blood pressure.

VizieR Online Data Catalog: Surface photometry of GHASP galaxies (Barbosa+, 2015)

NASA Astrophysics Data System (ADS)

Barbosa, C. E.; Mendes de Oliveira, C.; Amram, P.; Ferrari, F.; Russeil, D.; Epinat, B.; Perret, V.; Adami, C.; Marcelin, M.

2016-04-01

Our data set is constructed using new Rc-band observations taken at the Observatoire de Haute-Provence, supplemented with Sloan Digital Sky Survey archival data, obtained with the purpose of deriving homogeneous photometric profiles and parameters. Our results include Rc-band surface brightness profiles for 170 galaxies and ugriz profiles for 108 of these objects. We catalogue several parameters of general interest for further reference, such as total magnitude, effective radius and isophotal parameters (magnitude, position angle, ellipticity and inclination). We also perform a structural decomposition of the surface brightness profiles using a multi-component method to separate discs from bulges and bars, and to observe the main scaling relations involving luminosities, sizes and maximum velocities. We determine the Rc-band Tully-Fisher relation using maximum velocities derived solely from Hα rotation curves for a sample of 80 galaxies, resulting in a slope of -8.1+/-0.5, zero-point of -3.0+/-1.0 and an estimated intrinsic scatter of 0.28+/-0.07. We note that, unlike the Tully-Fisher relation in the near-infrared derived for the same sample, no change in the slope of the relation is seen at the low-mass end (for galaxies with Vmax<125km/s). We suggest that this different behaviour of the Tully-Fisher relation (with the optical relation being described by a single power law while the near-infrared has two), may be caused by differences in the stellar mass-to-light ratio for galaxies with Vmax<125km/s. (4 data files).

Cardiac tissue geometry as a determinant of unidirectional conduction block: assessment of microscopic excitation spread by optical mapping in patterned cell cultures and in a computer model.

PubMed

Fast, V G; Kléber, A G

1995-05-01

Unidirectional conduction block (UCB) and reentry may occur as a consequence of an abrupt tissue expansion and a related change in the electrical load. The aim of this study was to evaluate critical dimensions of the tissue necessary for establishing UCB in heart cell culture. Neonatal rat heart cell cultures with cell strands of variable width emerging into a large cell area were grown using a technique of patterned cell growth. Action potential upstrokes were measured using a voltage sensitive dye (RH-237) and a linear array of 10 photodiodes with a 15 microns resolution. A mathematical model was used to relate action potential wave shapes to underlying ionic currents. UCB (block of a single impulse in anterograde direction - from a strand to a large area - and conduction in the retrograde direction) occurred in narrow cell strands with a width of 15(SD 4) microns (1-2 cells in width, n = 7) and there was no conduction block in strands with a width of 31(8) microns (n = 9, P < 0.001) or larger. The analysis of action potential waveshapes indicated that conduction block was either due to geometrical expansion alone (n = 5) or to additional local depression of conduction (n = 2). In wide strands, action potential upstrokes during anterograde conduction were characterised by multiple rising phases. Mathematical modelling showed that two rising phases were caused by electronic current flow, whereas local ionic current did not coincide with the rising portions of the upstrokes. (1) High resolution optical mapping shows multiphasic action potential upstrokes at the region of abrupt expansion. At the site of the maximum decrement in conduction, these peaks were largely determined by the electrotonus and not by the local ionic current. (2) Unidirectional conduction block occurred in strands with a width of 15(4) microns (1-2 cells).

Labriform propulsion in fishes: kinematics of flapping aquatic flight in the bird wrasse Gomphosus varius (Labridae)

PubMed

Walker; Westneat

1997-01-01

Labriform, or pectoral fin, propulsion is the primary swimming mode for many fishes, even at high relative speeds. Although kinematic data are critical for evaluating hydrodynamic models of propulsion, these data are largely lacking for labriform swimmers, especially for species that employ an exclusively labriform mode across a broad range of speeds. We present data on pectoral fin locomotion in Gomphosus varius (Labridae), a tropical coral reef fish that uses a lift-based mechanism to fly under water at sustained speeds of 1­6 total body lengths s-1 (TL s-1). Lateral- and dorsal-view video images of three fish swimming in a flow tank at 1­4 TL s-1 were recorded at 60 Hz. From the two views, we reconstructed the three-dimensional motion of the center of mass, the fin tip and two fin chords for multiple fin beats of each fish at each of four speeds. In G. varius, the fin oscillates largely up and down: the stroke plane is tilted by approximately 20 ° from the vertical. Both frequency and the area swept by the pectoral fins increase with swimming speed. Interestingly, there are individual differences in how this area increases. Relative to the fish, the fin tip in lateral view moves along the path of a thin, inclined figure-of-eight. Relative to a stationary observer, the fin tip traces a sawtooth pattern, but the teeth are recumbent (indicating net backwards movement) only at the slowest speeds. Distal fin chords pitch nose downward during the downstroke and nose upward during the upstroke. Hydrodynamic angles of attack are largely positive during the downstroke and negative during the upstroke. The geometry of the fin and incident flow suggests that the fin is generating lift with large upward and small forward components during the downstroke. The negative incident angles during the upstroke suggest that the fin is generating largely thrust during the upstroke. In general, the large thrust is combined with a downward force during the upstroke, but the net backwards motion of the fin at slow speeds generates a small upward component during slow swimming. Both the alternating sign of the hydrodynamic angle of attack and the observed reduced frequencies suggest that unsteady effects are important in G. varius aquatic flight, especially at low speeds. This study provides a framework for the comparison of aquatic flight by fishes with aerial flight by birds, bats and insects.

Effect of Calcium on the Oxidative Phosphorylation Cascade in Skeletal Muscle Mitochondria

PubMed Central

Glancy, Brian; Willis, Wayne T; Chess, David J; Balaban, Robert S

2014-01-01

Calcium is believed to regulate mitochondrial oxidative phosphorylation, thereby contributing to the maintenance of cellular energy homeostasis. Skeletal muscle, with an energy conversion dynamic range of up to 100-fold, is an extreme case for evaluating the cellular balance of ATP production and consumption. This study examined the role of Ca2+ on the entire oxidative phosphorylation reaction network in isolated skeletal muscle mitochondria and attempted to extrapolate these results back to the muscle, in vivo. Kinetic analysis was conducted to evaluate the dose response effect of Ca2+ on the maximum velocity of oxidative phosphorylation (VmaxO) and the ADP affinity. Force-flow analysis evaluated the interplay between energetic driving forces and flux to determine the conductance, or effective activity, of individual steps within oxidative phosphorylation. Measured driving forces (extramitochondrial phosphorylation potential (ΔGATP), membrane potential, and redox states of NADH and cytochromes bH, bL, c1, c, and a,a3) were compared with flux (oxygen consumption) at 37°C. 840 nM Ca2+ generated a ∼2 fold increase in VmaxO with no change in ADP affinity (∼43 μM). Force-flow analysis revealed that Ca2+ activation of VmaxO was distributed throughout the oxidative phosphorylation reaction sequence. Specifically, Ca2+ increased the conductance of Complex IV (2.3-fold), Complexes I+III (2.2-fold), ATP production/transport (2.4-fold), and fuel transport/dehydrogenases (1.7-fold). These data support the notion that Ca2+ activates the entire muscle oxidative phosphorylation cascade, while extrapolation of these data to the exercising muscle predicts a significant role of Ca2+ in maintaining cellular energy homeostasis. PMID:23547908

Determinant Factors of the Squat Jump in Sprinting and Jumping Athletes

PubMed Central

González-Badillo, Juan José; Jiménez-Reyes, Pedro; Ramírez-Lechuga, Jorge

2017-01-01

Abstract The aim of this study was to assess the relationship between strength variables and maximum velocity (Vmax) in the squat jump (SJ) in sprinting and jumping athletes. Thirty-two sprinting and jumping athletes of national level (25.4 ± 4.5 years; 79.4 ± 6.9 kg and 180.4 ± 6.0 cm) participated in the study. Vmax in the SJ showed significant relationships with peak force 1 (PF1) (r = 0.82, p ≤ 0.001), peak force 2 (PF2) (r = 0.68, p ≤ 0.001), PF2 by controlling for PF1 (r = 0.30, non-significant), the maximum rate of force development at peak force 1 (RFDmax1) (r = 0.62, p ≤ 0.001), mean RFD 1 (RFDmean1) (r = 0.48, p ≤ 0.01), mean RFD 2 (RFDmean2) (r = 0.70, p ≤ 0.001), force at RFDmax1 (r = 0.36, p ≤ 0.05), force at RFDmax2 (r = 0.83, p ≤ 0.001) and force at RFDmax2 by controlling for PF1 (r = 0.40, p ≤ 0.05). However, Vmax in the SJ was associated negatively with the ratio PF2/PF1 (r = -0.54, p ≤ 0.01), time at peak force 2 (Tp2) (r = -0.64, p ≤ 0.001) and maximum rate of force development at peak force 2 (RFDmax2) (r = -0.71, p ≤ 0.001). These findings indicate that the peak force achieved at the beginning of the movement (PF1) is the main predictor of performance in jumping, although the RFDmax values and the ratio PF2/PF1 are also variables to be taken into account when analyzing the determinant factors of vertical jumping. PMID:28828074

Dynamical relationship between wind speed magnitude and meridional temperature contrast: Application to an interannual oscillation in Venusian middle atmosphere GCM

NASA Astrophysics Data System (ADS)

Yamamoto, Masaru; Takahashi, Masaaki

2018-03-01

We derive simple dynamical relationships between wind speed magnitude and meridional temperature contrast. The relationship explains scatter plot distributions of time series of three variables (maximum zonal wind speed UMAX, meridional wind speed VMAX, and equator-pole temperature contrast dTMAX), which are obtained from a Venus general circulation model with equatorial Kelvin-wave forcing. Along with VMAX and dTMAX, UMAX likely increases with the phase velocity and amplitude of a forced wave. In the scatter diagram of UMAX versus dTMAX, points are plotted along a linear equation obtained from a thermal-wind relationship in the cloud layer. In the scatter diagram of VMAX versus UMAX, the apparent slope is somewhat steep in the high UMAX regime, compared with the low UMAX regime. The scatter plot distributions are qualitatively consistent with a quadratic equation obtained from a diagnostic equation of the stream function above the cloud top. The plotted points in the scatter diagrams form a linear cluster for weak wave forcing, whereas they form a small cluster for strong wave forcing. An interannual oscillation of the general circulation forming the linear cluster in the scatter diagram is apparent in the experiment of weak 5.5-day wave forcing. Although a pair of equatorial Kelvin and high-latitude Rossby waves with a same period (Kelvin-Rossby wave) produces equatorward heat and momentum fluxes in the region below 60 km, the equatorial wave does not contribute to the long-period oscillation. The interannual fluctuation of the high-latitude jet core leading to the time variation of UMAX is produced by growth and decay of a polar mixed Rossby-gravity wave with a 14-day period.

Heterotrophic Potential for Amino Acid Uptake in a Naturally Eutrophic Lake1

PubMed Central

Burnison, B. Kent; Morita, Richard Y.

1974-01-01

The uptake of sixteen 14C-labeled amino acids by the indigenous heterotrophic microflora of Upper Klamath Lake, Oregon, was measured using the kinetic approach. The year-long study showed a seasonal variation in the maximum uptake velocity, Vmax, of all the amino acids which was proportional to temperature. The maximum total flux of amino acids by the heterotrophic microflora ranged from 1.2 to 11.9 μmol of C per liter per day (spring to summer). Glutamate, asparagine, aspartate, and serine had the highest Vmax values and were respired to the greatest extent. The percentages of the gross (net + respired) uptake of the amino acids which were respired to CO2 ranged from 2% for leucine to 63% for glutamate. Serine, lysine, and glycine were the most abundant amino acids found in Upper Klamath Lake surface water; at intermediate concentrations were alanine, aspartate, and threonine; and the remaining amino acids were always below 7.5 × 10-8 M (10 μg/liter). The amino acid concentrations determined chemically appear to be the sum of free and adsorbed amino acids, since the values obtained were usually greater than the (Kt + Sn) values obtained by the heterotrophic uptake experiments. PMID:4207581

Improving cardiac conduction with a skeletal muscle sodium channel by gene and cell therapy

PubMed Central

Lu, Jia; Wang, Hong-Zhan; Jia, Zhiheng; Zuckerman, Joan; Lu, Zhongju; Guo, Yuanjian; Boink, Gerard J.J.; Brink, Peter R.; Robinson, Richard B.; Entcheva, Emilia; Cohen, Ira S.

2012-01-01

The voltage-gated Na+ channel is a critical determinant of the action potential upstroke. Increasing Na+ conductance may speed action potential propagation. Here we propose use of the skeletal muscle Na+ channel SkM1 as a more favorable gene than the cardiac isoform SCN5A to enhance conduction velocity in depolarized cardiac tissue. We used cells which electrically coupled with cardiac myocytes as a delivery platform to introduce the Na+ channels. HEK293 cells were stably transfected with SkM1 or SCN5A. SkM1 had a more depolarized (18mV shift) inactivation curve than SCN5A. We also found that SkM1 recovered faster from inactivation than SCN5A. When coupled with SkM1 expressing cells, cultured myocytes showed an increase in the dV/dtmax of the action potential. Expression of SCN5A had no such effect. In an in vitro cardiac syncytium, coculture of neonatal cardiac myocytes with SkM1 expressing but not SCN5A expressing cells significantly increased the conduction velocity under both normal and depolarized conditions. In an in vitro re-entry model induced by high frequency stimulation, expression of SkM1 also enhanced angular velocity of the induced re-entry. These results suggest that cells carrying a Na+ channel with a more depolarized inactivation curve can improve cardiac excitability and conduction in depolarized tissues. PMID:22526298

In vivo maximal fascicle-shortening velocity during plantar flexion in humans.

PubMed

Hauraix, Hugo; Nordez, Antoine; Guilhem, Gaël; Rabita, Giuseppe; Dorel, Sylvain

2015-12-01

Interindividual variability in performance of fast movements is commonly explained by a difference in maximal muscle-shortening velocity due to differences in the proportion of fast-twitch fibers. To provide a better understanding of the capacity to generate fast motion, this study aimed to 1) measure for the first time in vivo the maximal fascicle-shortening velocity of human muscle; 2) evaluate the relationship between angular velocity and fascicle-shortening velocity from low to maximal angular velocities; and 3) investigate the influence of musculo-articular features (moment arm, tendinous tissues stiffness, and muscle architecture) on maximal angular velocity. Ultrafast ultrasound images of the gastrocnemius medialis were obtained from 31 participants during maximal isokinetic and light-loaded plantar flexions. A strong linear relationship between fascicle-shortening velocity and angular velocity was reported for all subjects (mean R(2) = 0.97). The maximal shortening velocity (V(Fmax)) obtained during the no-load condition (NLc) ranged between 18.8 and 43.3 cm/s. V(Fmax) values were very close to those of the maximal shortening velocity (V(max)), which was extrapolated from the F-V curve (the Hill model). Angular velocity reached during the NLc was significantly correlated with this V(Fmax) (r = 0.57; P < 0.001). This finding was in agreement with assumptions about the role of muscle fiber type, whereas interindividual comparisons clearly support the fact that other parameters may also contribute to performance during fast movements. Nevertheless, none of the biomechanical features considered in the present study were found to be directly related to the highest angular velocity, highlighting the complexity of the upstream mechanics that lead to maximal-velocity muscle contraction. Copyright © 2015 the American Physiological Society.

Optimization of Terrestrial Ecosystem Model Parameters Using Atmospheric CO2 Concentration Data With the Global Carbon Assimilation System (GCAS)

NASA Astrophysics Data System (ADS)

Chen, Zhuoqi; Chen, Jing M.; Zhang, Shupeng; Zheng, Xiaogu; Ju, Weiming; Mo, Gang; Lu, Xiaoliang

2017-12-01

The Global Carbon Assimilation System that assimilates ground-based atmospheric CO2 data is used to estimate several key parameters in a terrestrial ecosystem model for the purpose of improving carbon cycle simulation. The optimized parameters are the leaf maximum carboxylation rate at 25°C (Vmax25), the temperature sensitivity of ecosystem respiration (Q10), and the soil carbon pool size. The optimization is performed at the global scale at 1° resolution for the period from 2002 to 2008. The results indicate that vegetation from tropical zones has lower Vmax25 values than vegetation in temperate regions. Relatively high values of Q10 are derived over high/midlatitude regions. Both Vmax25 and Q10 exhibit pronounced seasonal variations at middle-high latitudes. The maxima in Vmax25 occur during growing seasons, while the minima appear during nongrowing seasons. Q10 values decrease with increasing temperature. The seasonal variabilities of Vmax25 and Q10 are larger at higher latitudes. Optimized Vmax25 and Q10 show little seasonal variabilities at tropical regions. The seasonal variabilities of Vmax25 are consistent with the variabilities of LAI for evergreen conifers and broadleaf evergreen forests. Variations in leaf nitrogen and leaf chlorophyll contents may partly explain the variations in Vmax25. The spatial distribution of the total soil carbon pool size after optimization is compared favorably with the gridded Global Soil Data Set for Earth System. The results also suggest that atmospheric CO2 data are a source of information that can be tapped to gain spatially and temporally meaningful information for key ecosystem parameters that are representative at the regional and global scales.

Variable-period surface-wave magnitudes: A rapid and robust estimator of seismic moments

USGS Publications Warehouse

Bonner, J.; Herrmann, R.; Benz, H.

2010-01-01

We demonstrate that surface-wave magnitudes (Ms), measured at local, regional, and teleseismic distances, can be used as a rapid and robust estimator of seismic moment magnitude (Mw). We used the Russell (2006) variable-period surface-wave magnitude formula, henceforth called Ms(VMAX), to estimate the Ms for 165 North American events with 3.2

Human Hepatic Cytochrome P450-Specific Metabolism of the Organophosphorus Pesticides Methyl Parathion and Diazinon

PubMed Central

Tian, Yuan; Knaak, James B.; Kostyniak, Paul J.; Olson, James R.

2012-01-01

Organophosphorus pesticides (OPs) are a public health concern due to their worldwide use and documented human exposures. Phosphorothioate OPs are metabolized by cytochrome P450s (P450s) through either a dearylation reaction to form an inactive metabolite, or through a desulfuration reaction to form an active oxon metabolite, which is a potent cholinesterase inhibitor. This study investigated the rate of desulfuration (activation) and dearylation (detoxification) of methyl parathion and diazinon in human liver microsomes. In addition, recombinant human P450s were used to determine the P450-specific kinetic parameters (Km and Vmax) for each compound for future use in refining human physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) models of OP exposure. The primary enzymes involved in bioactivation of methyl parathion were CYP2B6 (Km = 1.25 μM; Vmax = 9.78 nmol · min−1 · nmol P450−1), CYP2C19 (Km = 1.03 μM; Vmax = 4.67 nmol · min−1 · nmol P450−1), and CYP1A2 (Km = 1.96 μM; Vmax = 5.14 nmol · min−1 · nmol P450−1), and the bioactivation of diazinon was mediated primarily by CYP1A1 (Km = 3.05 μM; Vmax = 2.35 nmol · min−1 · nmol P450−1), CYP2C19 (Km = 7.74 μM; Vmax = 4.14 nmol · min−1 · nmol P450−1), and CYP2B6 (Km = 14.83 μM; Vmax = 5.44 nmol · min−1 · nmol P450−1). P450-mediated detoxification of methyl parathion only occurred to a limited extent with CYP1A2 (Km = 16.8 μM; Vmax = 1.38 nmol · min−1 · nmol P450−1) and 3A4 (Km = 104 μM; Vmax = 5.15 nmol · min−1 · nmol P450−1), whereas the major enzyme involved in diazinon detoxification was CYP2C19 (Km = 5.04 μM; Vmax = 5.58 nmol · min−1 · nmol P450−1). The OP- and P450-specific kinetic values will be helpful for future use in refining human PBPK/PD models of OP exposure. PMID:21969518

Vortex wake, downwash distribution, aerodynamic performance and wingbeat kinematics in slow-flying pied flycatchers.

PubMed

Muijres, Florian T; Bowlin, Melissa S; Johansson, L Christoffer; Hedenström, Anders

2012-02-07

Many small passerines regularly fly slowly when catching prey, flying in cluttered environments or landing on a perch or nest. While flying slowly, passerines generate most of the flight forces during the downstroke, and have a 'feathered upstroke' during which they make their wing inactive by retracting it close to the body and by spreading the primary wing feathers. How this flight mode relates aerodynamically to the cruising flight and so-called 'normal hovering' as used in hummingbirds is not yet known. Here, we present time-resolved fluid dynamics data in combination with wingbeat kinematics data for three pied flycatchers flying across a range of speeds from near hovering to their calculated minimum power speed. Flycatchers are adapted to low speed flight, which they habitually use when catching insects on the wing. From the wake dynamics data, we constructed average wingbeat wakes and determined the time-resolved flight forces, the time-resolved downwash distributions and the resulting lift-to-drag ratios, span efficiencies and flap efficiencies. During the downstroke, slow-flying flycatchers generate a single-vortex loop wake, which is much more similar to that generated by birds at cruising flight speeds than it is to the double loop vortex wake in hovering hummingbirds. This wake structure results in a relatively high downwash behind the body, which can be explained by the relatively active tail in flycatchers. As a result of this, slow-flying flycatchers have a span efficiency which is similar to that of the birds in cruising flight and which can be assumed to be higher than in hovering hummingbirds. During the upstroke, the wings of slowly flying flycatchers generated no significant forces, but the body-tail configuration added 23 per cent to weight support. This is strikingly similar to the 25 per cent weight support generated by the wing upstroke in hovering hummingbirds. Thus, for slow-flying passerines, the upstroke cannot be regarded as inactive, and the tail may be of importance for flight efficiency and possibly manoeuvrability.

Time-varying span efficiency through the wingbeat of desert locusts.

PubMed

Henningsson, Per; Bomphrey, Richard J

2012-06-07

The flight performance of animals depends greatly on the efficacy with which they generate aerodynamic forces. Accordingly, maximum range, load-lifting capacity and peak accelerations during manoeuvres are all constrained by the efficiency of momentum transfer to the wake. Here, we use high-speed particle image velocimetry (1 kHz) to record flow velocities in the near wake of desert locusts (Schistocerca gregaria, Forskål). We use the measured flow fields to calculate time-varying span efficiency throughout the wing stroke cycle. The locusts are found to operate at a maximum span efficiency of 79 per cent, typically at a plateau of about 60 per cent for the majority of the downstroke, but at lower values during the upstroke. Moreover, the calculated span efficiencies are highest when the largest lift forces are being generated (90% of the total lift is generated during the plateau of span efficiency) suggesting that the combination of wing kinematics and morphology in locust flight perform most efficiently when doing the most work.

Processing and analysis of cardiac optical mapping data obtained with potentiometric dyes

PubMed Central

Laughner, Jacob I.; Ng, Fu Siong; Sulkin, Matthew S.; Arthur, R. Martin

2012-01-01

Optical mapping has become an increasingly important tool to study cardiac electrophysiology in the past 20 years. Multiple methods are used to process and analyze cardiac optical mapping data, and no consensus currently exists regarding the optimum methods. The specific methods chosen to process optical mapping data are important because inappropriate data processing can affect the content of the data and thus alter the conclusions of the studies. Details of the different steps in processing optical imaging data, including image segmentation, spatial filtering, temporal filtering, and baseline drift removal, are provided in this review. We also provide descriptions of the common analyses performed on data obtained from cardiac optical imaging, including activation mapping, action potential duration mapping, repolarization mapping, conduction velocity measurements, and optical action potential upstroke analysis. Optical mapping is often used to study complex arrhythmias, and we also discuss dominant frequency analysis and phase mapping techniques used for the analysis of cardiac fibrillation. PMID:22821993

Actinobacterial Nitrate Reducers and Proteobacterial Denitrifiers Are Abundant in N2O-Metabolizing Palsa Peat

PubMed Central

Palmer, Katharina

2012-01-01

Palsa peats are characterized by elevated, circular frost heaves (peat soil on top of a permanently frozen ice lens) and are strong to moderate sources or even temporary sinks for the greenhouse gas nitrous oxide (N2O). Palsa peats are predicted to react sensitively to global warming. The acidic palsa peat Skalluvaara (approximate pH 4.4) is located in the discontinuous permafrost zone in northwestern Finnish Lapland. In situ N2O fluxes were spatially variable, ranging from 0.01 to −0.02 μmol of N2O m−2 h−1. Fertilization with nitrate stimulated in situ N2O emissions and N2O production in anoxic microcosms without apparent delay. N2O was subsequently consumed in microcosms. Maximal reaction velocities (vmax) of nitrate-dependent denitrification approximated 3 and 1 nmol of N2O per h per gram (dry weight [gDW]) in soil from 0 to 20 cm and below 20 cm of depth, respectively. vmax values of nitrite-dependent denitrification were 2- to 5-fold higher than the vmax nitrate-dependent denitrification, and vmax of N2O consumption was 1- to 6-fold higher than that of nitrite-dependent denitrification, highlighting a high N2O consumption potential. Up to 12 species-level operational taxonomic units (OTUs) of narG, nirK and nirS, and nosZ were retrieved. Detected OTUs suggested the presence of diverse uncultured soil denitrifiers and dissimilatory nitrate reducers, hitherto undetected species, as well as Actino-, Alpha-, and Betaproteobacteria. Copy numbers of nirS always outnumbered those of nirK by 2 orders of magnitude. Copy numbers of nirS tended to be higher, while copy numbers of narG and nosZ tended to be lower in 0- to 20-cm soil than in soil below 20 cm. The collective data suggest that (i) the source and sink functions of palsa peat soils for N2O are associated with denitrification, (ii) actinobacterial nitrate reducers and nirS-type and nosZ-harboring proteobacterial denitrifiers are important players, and (iii) acidic soils like palsa peats represent reservoirs of diverse acid-tolerant denitrifiers associated with N2O fluxes. PMID:22660709

Methylmercury-cholinesterase interactions in rats.

PubMed Central

Hastings, F L; Lucier, G W; Klein, R

1975-01-01

The interaction of methylmercury hydroxide (MMH) and cholinesterases was studied in male and female rats. MMH administered subcutaneously in doses of 10 mg/kg for 2 days reduced the level of plasma cholinesterase (ButChE) by 68% in females and 47% in males while brain acetylcholinesterase (AChE) was unaffected. Normal females had higher but more variable ButChE levels than normal males. In a time-course experiment, a single dose of MMH (10 mg/kg) reduced ButChE levels when mercury levels reached 22 mug/ml in the blood. A 10% reduction in brain AChE was observed at 72 hours; however, mercury reached a concentration of only 2.0 mug/g in brain tissue. The determination of the Michaelis constant Km and maximum velocity value Vmax for butyrylcholine and ButChE in control and MMH-treated (1 mg/kg) animals indicated that MMH reduced Vmax only. Since no loss in ButChE activity occurred when MMH and control plasma were incubated in vitro, MMH is not a direct inhibitor of ButChE. Because only the inactive monomeric form of ButChE contains free sulfhydryl groups, it is postulated that MMH combines covalently with the sulfur, preventing formation of active enzyme. By analogy, it is believed this is also the case with AChE. PMID:1227853

Characterization and regulation of glycine transport in Fusarium oxysporum var. lini.

PubMed

Castro, I M; Lima, A A; Nascimento, A F; Ruas, M M; Nicoli, J R; Brandão, R L

1996-08-01

Glycine was transported in Fusarium oxysporum cells, grown on glycine as the sole source of carbon and nitrogen, by a facilitated diffusion transport system with a half-saturation constant (Ks) of 11 mM and a maximum velocity (Vmax) of 1.2 mM (g dry weight)-1 h-1 at pH 5.0 and 26 degrees C. Under conditions of nitrogen starvation, the same system was present together with a high-affinity one (Ks) of about 47 microM and Vmax of about 60 microM (g dry weight)-1 h-1). The low-affinity system was more specific than the high-affinity system. Cells grown on gelatine showed the same behavior. In cells grown on glucose-gelatine medium, the low-affinity system was poorly expressed even after carbon and nitrogen starvation. Moreover, addition of glucose to cells grown on glycine and resuspended in mineral medium caused an increase of the glycine transport probably due to a boost in protein synthesis. This stimulation did not affect the Ks of the low-affinity system. These results demonstrate that, as is the case for other eukaryotic systems, F. oxysporum glycine transport is under control of nitrogen sources but its regulation by carbon sources appears to be more complex.

Fluid-Dynamics of Underwater Flight in Sea Butterflies: Analysis using Tomographic PIV

NASA Astrophysics Data System (ADS)

Adhikari, D.; Murphy, D. W.; Webster, D. R.; Yen, J.

2014-11-01

Sea butterflies, Limacina helicina, swim in sea water with a pair of gelatinous ``wings'' (or parapodia). Their unique propulsion mechanism has been hypothesized to consist of a combination of drag-based propulsion (rowing) and lift-based propulsion (flapping). Drag-based propulsion utilizes maximum drag on the wings during power stroke, followed by minimum drag during recovery stroke. Lift-based propulsion, in contrast, utilizes a pressure difference between the top and bottom of the wings. We present the 3D kinematics of a free-swimming sea butterfly and its induced volumetric velocity field using tomographic PIV. Both upstroke and downstroke motions propel the animal (1 - 3 mm) upward in a sawtooth-like trajectory with average speed of 5 - 15 mm/s (Re = 5 - 45) and roll the calcareous shell forwards-and-backwards at 4 - 5 Hz. The rolling motion effectively positions the wings such that they stroke downward during both the power and recovery strokes, hence inducing upward motion during both phases. A clap-and-fling mechanism is observed at the beginning of the flapping cycle. As the wings come into contact, the velocity of the organism is 2 mm/s. During fling motion, high (unsteady) lift causes the organism velocity to reach 35 mm/s. Separation vortices are observed during the fling motion, and vortices with an opposite sense of rotation form closer to the base of the wing due to the upward translation of the organism. The separation vortices shed into the wake, as the organism translates upward, in the form of separate vortex pairs.

Extracellular Matrix-Mediated Maturation of Human Pluripotent Stem Cell-Derived Cardiac Monolayer Structure and Electrophysiological Function.

PubMed

Herron, Todd J; Rocha, Andre Monteiro Da; Campbell, Katherine F; Ponce-Balbuena, Daniela; Willis, B Cicero; Guerrero-Serna, Guadalupe; Liu, Qinghua; Klos, Matt; Musa, Hassan; Zarzoso, Manuel; Bizy, Alexandra; Furness, Jamie; Anumonwo, Justus; Mironov, Sergey; Jalife, José

2016-04-01

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) monolayers generated to date display an immature embryonic-like functional and structural phenotype that limits their utility for research and cardiac regeneration. In particular, the electrophysiological function of hPSC-CM monolayers and bioengineered constructs used to date are characterized by slow electric impulse propagation velocity and immature action potential profiles. Here, we have identified an optimal extracellular matrix for significant electrophysiological and structural maturation of hPSC-CM monolayers. hPSC-CM plated in the optimal extracellular matrix combination have impulse propagation velocities ≈2× faster than previously reported (43.6±7.0 cm/s; n=9) and have mature cardiomyocyte action potential profiles, including hyperpolarized diastolic potential and rapid action potential upstroke velocity (146.5±17.7 V/s; n=5 monolayers). In addition, the optimal extracellular matrix promoted hypertrophic growth of cardiomyocytes and the expression of key mature sarcolemmal (SCN5A, Kir2.1, and connexin43) and myofilament markers (cardiac troponin I). The maturation process reported here relies on activation of integrin signaling pathways: neutralization of β1 integrin receptors via blocking antibodies and pharmacological blockade of focal adhesion kinase activation prevented structural maturation. Maturation of human stem cell-derived cardiomyocyte monolayers is achieved in a 1-week period by plating cardiomyocytes on PDMS (polydimethylsiloxane) coverslips rather than on conventional 2-dimensional cell culture formats, such as glass coverslips or plastic dishes. Activation of integrin signaling and focal adhesion kinase is essential for significant maturation of human cardiac monolayers. © 2016 American Heart Association, Inc.

Antioxidant and Inhibitory Effects of Saponin Extracts from Dianthus basuticus Burtt Davy on Key Enzymes Implicated in Type 2 Diabetes In vitro.

PubMed

Nafiu, Mikhail Olugbemiro; Ashafa, Anofi Omotayo Tom

2017-01-01

Dianthus basuticus is a plant of South African origin with various acclaimed pharmaceutical potentials. This study explored the antioxidant and antidiabetic activities of saponin extract from D. basuticus in vitro . Antioxidant activity of saponin was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (*NO)-free radical scavenging activity while antidiabetic potentials were measured by the α-amylase and α-glucosidase inhibitory activities of the saponin extract. The results showed that the saponin extract, compared with quercetin, displayed better DPPH (IC 50 = 6.95 mg/ml) and NO (IC 50 = 3.31 mg/ml) radical scavenging capabilities. Similarly, the saponin extracts elicited stronger α-glucosidase (IC 50 = 3.80 mg/ml) and moderate α-amylase (IC 50 = 4.18 mg/ml) inhibitory activities as compared to acarbose. Saponin exhibited a competitive mode of inhibition on α-amylase with same maximum velocity (Vmax) of 0.0093 mM/min for saponin compared with control 0.0095 mM/min and different the Michaelis constant (Km) values of 2.6 × 10 -6 mM and 2.1 × 10 -5 mM, respectively, while for α-glucosidase, the inhibition was uncompetitive, Vmax of 0.027 mM/min compared with control 0.039 mM/min and Km values of 1.02 × 10 -6 mM and 1.38 × 10 -6 mM, respectively. The gas chromatography-mass spectrometric analysis revealed the presence of bioactive like β- and α-amyrin, 3-O-methyl-D-glucose, methyl commate, and olean-12-en-3-beta-ol. Overall, the data suggested that the saponin extract from D. basuticus has potentials as natural antioxidants and antidiabetics. Saponin extract from Dianthus basuticus displayed promising antidiabetic and antioxidant activitySaponin competitively and uncompetitively inhibited a-amylase and a-glucosidase, respectivelyThe stronger inhibition of α-glucosidase and moderate inhibition of α-amylase by saponin extract from D. basuticus is promising good antidiabetes compared with existing drugs with associated side effects. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl, Km: The Michaelis constant, Vmax: Maximum velocity, ROS: Reactive oxygen species, NIDDM: Non-insulin-dependent diabetes mellitus, UFS: University of the Free State, GC-MS: Gas chromatography-mass spectrometric, MS: Mass spectrometry, NIST: National Institute of Standards and Technology, DNS: 3,5-dinitrosalicylic acid, NO: Nitric oxide, RNS: Reactive nitrogen species, PNPG: p-Nitrophenyl-α-D-glucopyranoside.

Antioxidant and Inhibitory Effects of Saponin Extracts from Dianthus basuticus Burtt Davy on Key Enzymes Implicated in Type 2 Diabetes In vitro

PubMed Central

Nafiu, Mikhail Olugbemiro; Ashafa, Anofi Omotayo Tom

2017-01-01

Context: Dianthus basuticus is a plant of South African origin with various acclaimed pharmaceutical potentials. Aims: This study explored the antioxidant and antidiabetic activities of saponin extract from D. basuticus in vitro. Materials and Methods: Antioxidant activity of saponin was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (*NO)-free radical scavenging activity while antidiabetic potentials were measured by the α-amylase and α-glucosidase inhibitory activities of the saponin extract. Results: The results showed that the saponin extract, compared with quercetin, displayed better DPPH (IC50 = 6.95 mg/ml) and NO (IC50 = 3.31 mg/ml) radical scavenging capabilities. Similarly, the saponin extracts elicited stronger α-glucosidase (IC50 = 3.80 mg/ml) and moderate α-amylase (IC50 = 4.18 mg/ml) inhibitory activities as compared to acarbose. Saponin exhibited a competitive mode of inhibition on α-amylase with same maximum velocity (Vmax) of 0.0093 mM/min for saponin compared with control 0.0095 mM/min and different the Michaelis constant (Km) values of 2.6 × 10-6 mM and 2.1 × 10-5 mM, respectively, while for α-glucosidase, the inhibition was uncompetitive, Vmax of 0.027 mM/min compared with control 0.039 mM/min and Km values of 1.02 × 10-6 mM and 1.38 × 10-6 mM, respectively. The gas chromatography-mass spectrometric analysis revealed the presence of bioactive like β- and α-amyrin, 3-O-methyl-D-glucose, methyl commate, and olean-12-en-3-beta-ol. Conclusion: Overall, the data suggested that the saponin extract from D. basuticus has potentials as natural antioxidants and antidiabetics. SUMMARY Saponin extract from Dianthus basuticus displayed promising antidiabetic and antioxidant activitySaponin competitively and uncompetitively inhibited a-amylase and a-glucosidase, respectivelyThe stronger inhibition of α-glucosidase and moderate inhibition of α-amylase by saponin extract from D. basuticus is promising good antidiabetes compared with existing drugs with associated side effects. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl, Km: The Michaelis constant, Vmax: Maximum velocity, ROS: Reactive oxygen species, NIDDM: Non-insulin-dependent diabetes mellitus, UFS: University of the Free State, GC-MS: Gas chromatography-mass spectrometric, MS: Mass spectrometry, NIST: National Institute of Standards and Technology, DNS: 3,5-dinitrosalicylic acid, NO: Nitric oxide, RNS: Reactive nitrogen species, PNPG: p-Nitrophenyl-α-D-glucopyranoside. PMID:29200716

Force interaction and 3D pole movement in double poling.

PubMed

Stöggl, T; Holmberg, H-C

2011-12-01

The aim of this study was to analyze double poling using combined kinetic and 3D kinematic analysis at high skiing speeds as regards pole force components, pole angles and pole behavior during the poling and swing phase. The hypothesis was that a horizontal pole force is more predictive for maximal skiing speed (V(max)) than the resultant pole force. Sixteen elite skiers performed a double-poling V(max) test while treadmill roller skiing. Pole forces and 3D kinematics of pole movement at a speed of 30 km/h were analyzed and related to V(max). The duration of the "preparation phase" showed the strongest relationship with V(max) (r=0.87, P<0.001). Faster skiers generated longer cycle lengths with longer swing and poling times, had less inclined pole angles at pole plant and a later peak pole force. Horizontal pole forces were not more highly related to V(max) compared with the resultant pole force. Impact force was not related to V(max). At high skiing speeds, skiers should aim to combine high pole forces with appropriate timing of pole forces and appropriate pole and body positions during the swing and poling phase. The emphasis in training should be on the development of specific strength capacities for pole force production and the utilization of these capacities in double-poling training sessions. © 2011 John Wiley & Sons A/S.

Three-dimensional simulation for fast forward flight of a calliope hummingbird

PubMed Central

Song, Jialei; Powers, Donald R.; Hedrick, Tyson L.; Luo, Haoxiang

2016-01-01

We present a computational study of flapping-wing aerodynamics of a calliope hummingbird (Selasphorus calliope) during fast forward flight. Three-dimensional wing kinematics were incorporated into the model by extracting time-dependent wing position from high-speed videos of the bird flying in a wind tunnel at 8.3 m s−1. The advance ratio, i.e. the ratio between flight speed and average wingtip speed, is around one. An immersed-boundary method was used to simulate flow around the wings and bird body. The result shows that both downstroke and upstroke in a wingbeat cycle produce significant thrust for the bird to overcome drag on the body, and such thrust production comes at price of negative lift induced during upstroke. This feature might be shared with bats, while being distinct from insects and other birds, including closely related swifts. PMID:27429779

The topology of large-scale structure. III - Analysis of observations

NASA Astrophysics Data System (ADS)

Gott, J. Richard, III; Miller, John; Thuan, Trinh X.; Schneider, Stephen E.; Weinberg, David H.; Gammie, Charles; Polk, Kevin; Vogeley, Michael; Jeffrey, Scott; Bhavsar, Suketu P.; Melott, Adrian L.; Giovanelli, Riccardo; Hayes, Martha P.; Tully, R. Brent; Hamilton, Andrew J. S.

1989-05-01

A recently developed algorithm for quantitatively measuring the topology of large-scale structures in the universe was applied to a number of important observational data sets. The data sets included an Abell (1958) cluster sample out to Vmax = 22,600 km/sec, the Giovanelli and Haynes (1985) sample out to Vmax = 11,800 km/sec, the CfA sample out to Vmax = 5000 km/sec, the Thuan and Schneider (1988) dwarf sample out to Vmax = 3000 km/sec, and the Tully (1987) sample out to Vmax = 3000 km/sec. It was found that, when the topology is studied on smoothing scales significantly larger than the correlation length (i.e., smoothing length, lambda, not below 1200 km/sec), the topology is spongelike and is consistent with the standard model in which the structure seen today has grown from small fluctuations caused by random noise in the early universe. When the topology is studied on the scale of lambda of about 600 km/sec, a small shift is observed in the genus curve in the direction of a 'meatball' topology.

The topology of large-scale structure. III - Analysis of observations. [in universe

NASA Technical Reports Server (NTRS)

Gott, J. Richard, III; Weinberg, David H.; Miller, John; Thuan, Trinh X.; Schneider, Stephen E.

1989-01-01

A recently developed algorithm for quantitatively measuring the topology of large-scale structures in the universe was applied to a number of important observational data sets. The data sets included an Abell (1958) cluster sample out to Vmax = 22,600 km/sec, the Giovanelli and Haynes (1985) sample out to Vmax = 11,800 km/sec, the CfA sample out to Vmax = 5000 km/sec, the Thuan and Schneider (1988) dwarf sample out to Vmax = 3000 km/sec, and the Tully (1987) sample out to Vmax = 3000 km/sec. It was found that, when the topology is studied on smoothing scales significantly larger than the correlation length (i.e., smoothing length, lambda, not below 1200 km/sec), the topology is spongelike and is consistent with the standard model in which the structure seen today has grown from small fluctuations caused by random noise in the early universe. When the topology is studied on the scale of lambda of about 600 km/sec, a small shift is observed in the genus curve in the direction of a 'meatball' topology.

Effect of right ventricular pacing on cardiac apex rotation assessed by a gyroscopic sensor.

PubMed

Marcelli, Emanuela; Cercenelli, Laura; Parlapiano, Mario; Fumero, Roberto; Bagnoli, Paola; Costantino, Maria Laura; Plicchi, Gianni

2007-01-01

To quantify cardiac apex rotation (CAR), the authors recently proposed the use of a Coriolis force sensor (gyroscope) as an alternative to other complex techniques. The aim of this study was to evaluate the effects of right ventricular (RV) pacing on CAR. A sheep heart was initially paced from the right atrium to induce a normal activation sequence at a fixed heart rate (AAI mode) and then an atrioventricular pacing was performed (DOO mode, AV delay = 60 ms). A small gyroscope was epicardially glued on the cardiac apex to measure the angular velocity (Ang V). From AAI to DOO pacing mode, an increase (+9.2%, p < 0.05) of the maximum systolic twisting velocity (Ang VMAX) and a marked decrease (-19.9%, p < 0.05) of the maximum diastolic untwisting velocity (Ang VMIN) resulted. RV pacing had negligible effects (-3.1%, p = 0.09) on the maximum angle of CAR, obtained by integrating Ang V. The hemodynamic parameters of systolic (LVdP/dtMAX) and diastolic (LVdP/dtMIN) cardiac function showed slight variations (-3.8%, p < 0.05 and +3.9%, p < 0.05, respectively). Results suggest that cardiac dyssynchrony induced by RV pacing can alter the normal physiological ventricular twist patterns, particularly affecting diastolic untwisting velocity.

Hummingbird wing efficacy depends on aspect ratio and compares with helicopter rotors

PubMed Central

Kruyt, Jan W.; Quicazán-Rubio, Elsa M.; van Heijst, GertJan F.; Altshuler, Douglas L.; Lentink, David

2014-01-01

Hummingbirds are the only birds that can sustain hovering. This unique flight behaviour comes, however, at high energetic cost. Based on helicopter and aeroplane design theory, we expect that hummingbird wing aspect ratio (AR), which ranges from about 3.0 to 4.5, determines aerodynamic efficacy. Previous quasi-steady experiments with a wing spinner set-up provide no support for this prediction. To test this more carefully, we compare the quasi-steady hover performance of 26 wings, from 12 hummingbird taxa. We spun the wings at angular velocities and angles of attack that are representative for every species and measured lift and torque more precisely. The power (aerodynamic torque × angular velocity) required to lift weight depends on aerodynamic efficacy, which is measured by the power factor. Our comparative analysis shows that AR has a modest influence on lift and drag forces, as reported earlier, but interspecific differences in power factor are large. During the downstroke, the power required to hover decreases for larger AR wings at the angles of attack at which hummingbirds flap their wings (p < 0.05). Quantitative flow visualization demonstrates that variation in hover power among hummingbird wings is driven by similar stable leading edge vortices that delay stall during the down- and upstroke. A side-by-side aerodynamic performance comparison of hummingbird wings and an advanced micro helicopter rotor shows that they are remarkably similar. PMID:25079868

Pigeons steer like helicopters and generate down- and upstroke lift during low speed turns.

PubMed

Ros, Ivo G; Bassman, Lori C; Badger, Marc A; Pierson, Alyssa N; Biewener, Andrew A

2011-12-13

Turning is crucial for animals, particularly during predator-prey interactions and to avoid obstacles. For flying animals, turning consists of changes in (i) flight trajectory, or path of travel, and (ii) body orientation, or 3D angular position. Changes in flight trajectory can only be achieved by modulating aerodynamic forces relative to gravity. How birds coordinate aerodynamic force production relative to changes in body orientation during turns is key to understanding the control strategies used in avian maneuvering flight. We hypothesized that pigeons produce aerodynamic forces in a uniform direction relative to their bodies, requiring changes in body orientation to redirect those forces to turn. Using detailed 3D kinematics and body mass distributions, we examined net aerodynamic forces and body orientations in slowly flying pigeons (Columba livia) executing level 90° turns. The net aerodynamic force averaged over the downstroke was maintained in a fixed direction relative to the body throughout the turn, even though the body orientation of the birds varied substantially. Early in the turn, changes in body orientation primarily redirected the downstroke aerodynamic force, affecting the bird's flight trajectory. Subsequently, the pigeon mainly reacquired the body orientation used in forward flight without affecting its flight trajectory. Surprisingly, the pigeon's upstroke generated aerodynamic forces that were approximately 50% of those generated during the downstroke, nearly matching the relative upstroke forces produced by hummingbirds. Thus, pigeons achieve low speed turns much like helicopters, by using whole-body rotations to alter the direction of aerodynamic force production to change their flight trajectory.

Pigeons steer like helicopters and generate down- and upstroke lift during low speed turns

PubMed Central

Ros, Ivo G.; Bassman, Lori C.; Badger, Marc A.; Pierson, Alyssa N.; Biewener, Andrew A.

2011-01-01

Turning is crucial for animals, particularly during predator–prey interactions and to avoid obstacles. For flying animals, turning consists of changes in (i) flight trajectory, or path of travel, and (ii) body orientation, or 3D angular position. Changes in flight trajectory can only be achieved by modulating aerodynamic forces relative to gravity. How birds coordinate aerodynamic force production relative to changes in body orientation during turns is key to understanding the control strategies used in avian maneuvering flight. We hypothesized that pigeons produce aerodynamic forces in a uniform direction relative to their bodies, requiring changes in body orientation to redirect those forces to turn. Using detailed 3D kinematics and body mass distributions, we examined net aerodynamic forces and body orientations in slowly flying pigeons (Columba livia) executing level 90° turns. The net aerodynamic force averaged over the downstroke was maintained in a fixed direction relative to the body throughout the turn, even though the body orientation of the birds varied substantially. Early in the turn, changes in body orientation primarily redirected the downstroke aerodynamic force, affecting the bird’s flight trajectory. Subsequently, the pigeon mainly reacquired the body orientation used in forward flight without affecting its flight trajectory. Surprisingly, the pigeon’s upstroke generated aerodynamic forces that were approximately 50% of those generated during the downstroke, nearly matching the relative upstroke forces produced by hummingbirds. Thus, pigeons achieve low speed turns much like helicopters, by using whole-body rotations to alter the direction of aerodynamic force production to change their flight trajectory. PMID:22123982

Sprint swimming performance of wild bull trout (Salvelinus confluentus)

USGS Publications Warehouse

Mesa, M.G.; Phelps, J.; Weiland, L.K.

2008-01-01

We conducted laboratory experiments to determine the sprint swimming performance of wild juvenile and adult bull trout Salvelinus confluentus. Sprint swimming speeds were estimated using high-speed digital video analysis. Thirty two bull trout were tested in sizes ranging from about 10 to 31 cm. Of these, 14 fish showed at least one motivated, vigorous sprint. When plotted as a function of time, velocity of fish increased rapidly with the relation linear or slightly curvilinear. Their maximum velocity, or Vmax, ranged from 1.3 to 2.3 m/s, was usually achieved within 0.8 to 1.0 s, and was independent of fish size. Distances covered during these sprints ranged from 1.4 to 2.4 m. Our estimates of the sprint swimming performance are the first reported for this species and may be useful for producing or modifying fish passage structures that allow safe and effective passage of fish without overly exhausting them. ?? 2008 by the Northwest Scientific Association. All rights reserved.

Preliminary study of the association between the elimination parameters of phenytoin and phenobarbital.

PubMed

Methaneethorn, Janthima; Panomvana, Duangchit; Vachirayonstien, Thaveechai

2017-09-26

Therapeutic drug monitoring is essential for both phenytoin and phenobarbital therapy given their narrow therapeutic indexes. Nevertheless, the measurement of either phenytoin or phenobarbital concentrations might not be available in some rural hospitals. Information assisting individualized phenytoin and phenobarbital combination therapy is important. This study's objective was to determine the relationship between the maximum rate of metabolism of phenytoin (Vmax) and phenobarbital clearance (CLPB), which can serve as a guide to individualized drug therapy. Data on phenytoin and phenobarbital concentrations of 19 epileptic patients concurrently receiving both drugs were obtained from medical records. Phenytoin and phenobarbital pharmacokinetic parameters were studied at steady-state conditions. The relationship between the elimination parameters of both drugs was determined using simple linear regression. A high correlation coefficient between Vmax and CLPB was found [r=0.744; p<0.001 for Vmax (mg/kg/day) vs. CLPB (L/kg/day)]. Such a relatively strong linear relationship between the elimination parameters of both drugs indicates that Vmax might be predicted from CLPB and vice versa. Regression equations were established for estimating Vmax from CLPB, and vice versa in patients treated with combination of phenytoin and phenobarbital. These proposed equations can be of use in aiding individualized drug therapy.

Identification of functional amino acid residues involved in polyamine and agmatine transport by human organic cation transporter 2.

PubMed

Higashi, Kyohei; Imamura, Masataka; Fudo, Satoshi; Uemura, Takeshi; Saiki, Ryotaro; Hoshino, Tyuji; Toida, Toshihiko; Kashiwagi, Keiko; Igarashi, Kazuei

2014-01-01

Polyamine (putrescine, spermidine and spermine) and agmatine uptake by the human organic cation transporter 2 (hOCT2) was studied using HEK293 cells transfected with pCMV6-XL4/hOCT2. The Km values for putrescine and spermidine were 7.50 and 6.76 mM, and the Vmax values were 4.71 and 2.34 nmol/min/mg protein, respectively. Spermine uptake by hOCT2 was not observed at pH 7.4, although it inhibited both putrescine and spermidine uptake. Agmatine was also taken up by hOCT2, with Km value: 3.27 mM and a Vmax value of 3.14 nmol/min/mg protein. Amino acid residues involved in putrescine, agmatine and spermidine uptake by hOCT2 were Asp427, Glu448, Glu456, Asp475, and Glu516. In addition, Glu524 and Glu530 were involved in putrescine and spermidine uptake activity, and Glu528 and Glu540 were weakly involved in putrescine uptake activity. Furthermore, Asp551 was also involved in the recognition of spermidine. These results indicate that the recognition sites for putrescine, agmatine and spermidine on hOCT2 strongly overlap, consistent with the observation that the three amines are transported with similar affinity and velocity. A model of spermidine binding to hOCT2 was constructed based on the functional amino acid residues.

[Evaluation of the arterial blood flow parameters in the eye of myopic patients].

PubMed

Mrugacz, Małgorzata; Bryl, Anna

2013-04-01

Myopia is a common refractive defect. Has a good vision from near and deterioration of vision with increasing distance. The main reason for its occurrence is too long axis of the eyeball. The consequence of elongation of the eyeball is the development of degenerative changes in the retina. Despite much research has failed to clearly identify the causes of degenerative changes in those short-sighted. The aim of the study was to evaluate the maximum and minimum speed in arterial blood vessels of the eye in people with myopia. The study included 70 patients (138 eyes), 53 women and 17 men, aged from 18 to 79 years, with myopia of -0.25 to -18.0 Dsph and length of the eyeball from 22.61 to 33.36 mm. Depending on the kind and the degree of the progress of degenerative changes, patients were divided in 4 groups: I - without degenerative changes on the fundus (n=32; K-23, M-9); II- with the short-sighted sickle (n=20; K-14, M-6); Ill - with the structure thinned down of the retina, accompanying the short-sighted sickle (n = 8; K-6, M-2); IV - with extensive choroidal-retina disappearances in the fundus (n = 10; K-7, M-3). In all individuals enrolled underwent Color Doppler ultrasound with apparatus SSA 770A Toshiba Aplio with linear probe frequency 12 MHz, judging maximum (Vmax) and minimum (Vmin) speed in the arteries of the eye: ophthalmic artery (OA), central retinal artery (CRA) and short posterior ciliary arteries (SPCA) located on the nasal and temporal side of the optic disc. The results were statistically analyzed. No statistically significant relationship between the nature of degenerative changes of the eye, and blood velocity in the OA. There was a increase in Vmax and Vmin blood in OA in Group IV, but these changes were not statistically significant. Statistically significant correlation was observed while in the CRA. With a decrease in Vmax and Vmin of blood flowing through a vessel exacerbation of retinal degeneration. Vmax and Vmin changes in the blood did not correlate significantly SPCA with retinal degeneration, although the results were much worse in the temporal vessels. With the deterioration of blood flow parameters of the central retinal artery and short posterior ciliary arteries comes to the severity of the retinal degeneration in myopic patients. More severe impairment of blood circulation in the temporal ciliary explain higher incidence of degenerative changes in the temporal side of the optic disc.

Acute Effects of Lateral Thigh Foam Rolling on Arterial Tissue Perfusion Determined by Spectral Doppler and Power Doppler Ultrasound.

PubMed

Hotfiel, Thilo; Swoboda, Bernd; Krinner, Sebastian; Grim, Casper; Engelhardt, Martin; Uder, Michael; Heiss, Rafael U

2017-04-01

Hotfiel, T, Swoboda, B, Krinner, S, Grim, C, Engelhardt, M, Uder, M, and Heiss, R. Acute effects of lateral thigh foam rolling on arterial tissue perfusion determined by spectral Doppler and power Doppler ultrasound. J Strength Cond Res 31(4): 893-900, 2017-Foam rolling has been developed as a popular intervention in training and rehabilitation. However, evidence on its effects on the cellular and physiological level is lacking. The aim of this study was to assess the effect of foam rolling on arterial blood flow of the lateral thigh. Twenty-one healthy participants (age, 25 ± 2 years; height, 177 ± 9 cm; body weight, 74 ± 9 kg) were recruited from the medical and sports faculty. Arterial tissue perfusion was determined by spectral Doppler and power Doppler ultrasound, represented as peak flow (Vmax), time average velocity maximum (TAMx), time average velocity mean (TAMn), and resistive index (RI), and with semiquantitative grading that was assessed by 4 blindfolded investigators. Measurement values were assessed under resting conditions and twice after foam rolling exercises of the lateral thigh (0 and 30 minutes after intervention). The trochanteric region, mid portion, and distal tibial insertion of the lateral thigh were representative for data analysis. Arterial blood flow of the lateral thigh increased significantly after foam rolling exercises compared with baseline (p ≤ 0.05). We detected a relative increase in Vmax of 73.6% (0 minutes) and 52.7% (30 minutes) (p < 0.001), in TAMx of 53.2% (p < 0.001) and 38.3% (p = 0.002), and in TAMn of 84.4% (p < 0.001) and 68.2% (p < 0.001). Semiquantitative power Doppler scores at all portions revealed increased average grading of 1.96 after intervention and 2.04 after 30 minutes compared with 0.75 at baseline. Our results may contribute to the understanding of local physiological reactions to self-myofascial release.

SURFS: Riding the waves with Synthetic UniveRses For Surveys

NASA Astrophysics Data System (ADS)

Elahi, Pascal J.; Welker, Charlotte; Power, Chris; Lagos, Claudia del P.; Robotham, Aaron S. G.; Cañas, Rodrigo; Poulton, Rhys

2018-04-01

We present the Synthetic UniveRses For Surveys (SURFS) simulations, a set of N-body/Hydro simulations of the concordance Λ Cold Dark Matter (ΛCDM) cosmology. These simulations use Planck cosmology, contain up to 10 billion particles, and sample scales and halo masses down to 1 kpc and 108 M⊙. We identify and track haloes from z = 24 to today using a state-of-the-art 6D halo finder and merger tree builder. We demonstrate that certain properties of haloes merger trees are numerically converged for haloes composed of ≳100 particles. Haloes smoothly grow in mass, Vmax, with the mass history characterized by log M(a) ∝ exp [-(a/β)α], where a is the scale factor, α(M) ≈ 0.8 & β(M) ≈ 0.024, with these parameters decreasing with decreasing halo mass. Subhaloes follow power-law cumulative mass and velocity functions, i.e. n( > f) ∝ f-α with αM = 0.83 ± 0.01 and α _{V_max}=2.13± 0.03 for mass and velocity, respectively, independent of redshift, as seen in previous studies. The halo-to-halo scatter in amplitude is 0.9 dex. The number of subhaloes in a halo weakly correlates with a halo's concentration c and spin λ:haloes of high c and low λ have 60 per cent more subhaloes than similar mass haloes of low c and high λ. High cadence tracking shows subhaloes are dynamic residents, with 25 per cent leaving their host halo momentarily, becoming a backsplash subhalo, and another 20 per cent changing hosts entirely, in agreement with previous studies. In general, subhaloes have elliptical orbits, e ≈ 0.6, with periods of 2.3^{+2.1}_{-1.7} Gyrs. Subhaloes lose most of their mass at pericentric passage with mass loss rates of ˜ 40 per cent Gyr-1. These catalogues will be made publicly available.

The flutter-by effect: a comprehensive study of the fluttering cusps of the Perceval heart valve prosthesis.

PubMed

Aljalloud, Ali; Shoaib, Mohamed; Egron, Sandrine; Arias, Jessica; Tewarie, Lachmandath; Schnoering, Heike; Lotfi, Shahram; Goetzenich, Andreas; Hatam, Nima; Pott, Desiree; Zhong, Zhaoyang; Steinseifer, Ulrich; Zayat, Rachad; Autschbach, Ruediger

2018-05-17

Sutureless aortic valve prostheses are gaining popularity due to the substantial reduction in cross-clamp time. In this study, we report our observations on the cusp-fluttering phenomenon of the Perceval bioprosthesis (LivaNova, London, UK) using a combination of technical and medical perspectives. Between August 2014 and December 2016, a total of 108 patients (69% women) with a mean age of 78 years had aortic valve replacement using the Perceval bioprosthesis (34 combined procedures). All patients underwent transoesophageal echocardiography (TOE) intraoperatively. TOE was performed postoperatively to detect paravalvular leakage and to measure gradients, acceleration time, Doppler velocity indices (Vmax and LVOT/Vmax AV) and effective orifice area indices. In addition, a TOE examination was performed in 21 patients postoperatively. Data were collected retrospectively from our hospital database. The retrospective evaluation of the intraoperative TOE examinations revealed consistent fluttering in all patients with the Perceval bioprosthesis. The echocardiographic postoperative measurements showed a mean effective orifice area index of 0.91 ± 0.12 cm2/m2. The overall mean pressure and peak pressure gradients were in a higher range (13.5 ± 5.1 mmHg and 25.5 ± 8.6 mmHg, respectively), whereas acceleration time (62.8 ± 16.4 ms) and Doppler velocity indices (0.43 ± 0.11) were within the normal range according to the American Society of Echocardiography or european association of echocardiography (EAE) guidelines. The 2-dimensional TOE in Motion Mode (M-Mode) that was performed in patients with elevated lactate dehydrogenase (LDH) levels revealed remarkable fluttering of the cusps of the Perceval bioprosthesis. In our study cohort, we observed the fluttering phenomenon in all patients who received the Perceval bioprosthesis, which was correlated with elevated LDH levels and higher pressure gradients.

Hawkmoth flight performance in tornado-like whirlwind vortices.

PubMed

Ortega-Jimenez, Victor Manuel; Mittal, Rajat; Hedrick, Tyson L

2014-06-01

Vertical vortex systems such as tornadoes dramatically affect the flight control and stability of aircraft. However, the control implications of smaller scale vertically oriented vortex systems for small fliers such as animals or micro-air vehicles are unknown. Here we examined the flapping kinematics and body dynamics of hawkmoths performing hovering flights (controls) and maintaining position in three different whirlwind intensities with transverse horizontal velocities of 0.7, 0.9 and 1.2 m s(-1), respectively, generated in a vortex chamber. The average and standard deviation of yaw and pitch were respectively increased and reduced in comparison with hovering flights. Average roll orientation was unchanged in whirlwind flights but was more variable from wingbeat to wingbeat than in hovering. Flapping frequency remained unchanged. Wingbeat amplitude was lower and the average stroke plane angle was higher. Asymmetry was found in the angle of attack between right and left wings during both downstroke and upstroke at medium and high vortex intensities. Thus, hawkmoth flight control in tornado-like vortices is achieved by a suite of asymmetric and symmetric changes to wingbeat amplitude, stroke plane angle and principally angle of attack.

The Major Role of IK1 in Mechanisms of Rotor Drift in the Atria: A Computational Study

PubMed Central

Berenfeld, Omer

2016-01-01

Maintenance of paroxysmal atrial fibrillation (AF) by fast rotors in the left atrium (LA) or at the pulmonary veins (PVs) is not fully understood. This review describes the role of the heterogeneous distribution of transmembrane currents in the PVs and LA junction (PV-LAJ) in the localization of rotors in the PVs. Experimentally observed heterogeneities in IK1, IKs, IKr, Ito, and ICaL in the PV-LAJ were incorporated into models of human atrial kinetics to simulate various conditions and investigate rotor drifting mechanisms. Spatial gradients in the currents resulted in shorter action potential duration, less negative minimum diastolic potential, slower upstroke and conduction velocity for rotors in the PV region than in the LA. Rotors under such conditions drifted toward the PV and stabilized at the less excitable region. Our simulations suggest that IK1 heterogeneity is dominant in determining the drift direction through its impact on the excitability gradient. These results provide a novel framework for understanding the complex dynamics of rotors in AF. PMID:28096699

Estimates of circulation and gait change based on a three-dimensional kinematic analysis of flight in cockatiels (Nymphicus hollandicus) and ringed turtle-doves (Streptopelia risoria).

PubMed

Hedrick, Tyson L; Tobalske, Bret W; Biewener, Andrew A

2002-05-01

Birds and bats are known to employ two different gaits in flapping flight, a vortex-ring gait in slow flight and a continuous-vortex gait in fast flight. We studied the use of these gaits over a wide range of speeds (1-17 ms(-1)) and transitions between gaits in cockatiels (Nymphicus hollandicus) and ringed turtle-doves (Streptopelia risoria) trained to fly in a recently built, variable-speed wind tunnel. Gait use was investigated via a combination of three-dimensional kinematics and quasi-steady aerodynamic modeling of bound circulation on the distal and proximal portions of the wing. Estimates of lift from our circulation model were sufficient to support body weight at all but the slowest speeds (1 and 3 ms(-1)). From comparisons of aerodynamic impulse derived from our circulation analysis with the impulse estimated from whole-body acceleration, it appeared that our quasi-steady aerodynamic analysis was most accurate at intermediate speeds (5-11 ms(-1)). Despite differences in wing shape and wing loading, both species shifted from a vortex-ring to a continuous-vortex gait at 7 ms(-1). We found that the shift from a vortex-ring to a continuous-vortex gait (i) was associated with a phase delay in the peak angle of attack of the proximal wing section from downstroke into upstroke and (ii) depended on sufficient forward velocity to provide airflow over the wing during the upstroke similar to that during the downstroke. Our kinematic estimates indicated significant variation in the magnitude of circulation over the course the wingbeat cycle when either species used a continuous-vortex gait. This variation was great enough to suggest that both species shifted to a ladder-wake gait as they approached the maximum flight speed (cockatiels 15 ms(-1), doves 17 ms(-1)) that they would sustain in the wind tunnel. This shift in flight gait appeared to reflect the need to minimize drag and produce forward thrust in order to fly at high speed. The ladder-wake gait was also employed in forward and vertical acceleration at medium and fast flight speeds.

Whole-body kinematics of a fruit bat reveal the influence of wing inertia on body accelerations.

PubMed

Iriarte-Díaz, José; Riskin, Daniel K; Willis, David J; Breuer, Kenneth S; Swartz, Sharon M

2011-05-01

The center of mass (COM) of a flying animal accelerates through space because of aerodynamic and gravitational forces. For vertebrates, changes in the position of a landmark on the body have been widely used to estimate net aerodynamic forces. The flapping of relatively massive wings, however, might induce inertial forces that cause markers on the body to move independently of the COM, thus making them unreliable indicators of aerodynamic force. We used high-speed three-dimensional kinematics from wind tunnel flights of four lesser dog-faced fruit bats, Cynopterus brachyotis, at speeds ranging from 2.4 to 7.8 m s(-1) to construct a time-varying model of the mass distribution of the bats and to estimate changes in the position of their COM through time. We compared accelerations calculated by markers on the trunk with accelerations calculated from the estimated COM and we found significant inertial effects on both horizontal and vertical accelerations. We discuss the effect of these inertial accelerations on the long-held idea that, during slow flights, bats accelerate their COM forward during 'tip-reversal upstrokes', whereby the distal portion of the wing moves upward and backward with respect to still air. This idea has been supported by the observation that markers placed on the body accelerate forward during tip-reversal upstrokes. As in previously published studies, we observed that markers on the trunk accelerated forward during the tip-reversal upstrokes. When removing inertial effects, however, we found that the COM accelerated forward primarily during the downstroke. These results highlight the crucial importance of the incorporation of inertial effects of wing motion in the analysis of flapping flight.

Computational investigation of cicada aerodynamics in forward flight.

PubMed

Wan, Hui; Dong, Haibo; Gai, Kuo

2015-01-06

Free forward flight of cicadas is investigated through high-speed photogrammetry, three-dimensional surface reconstruction and computational fluid dynamics simulations. We report two new vortices generated by the cicada's wide body. One is the thorax-generated vortex, which helps the downwash flow, indicating a new phenomenon of lift enhancement. Another is the cicada posterior body vortex, which entangles with the vortex ring composed of wing tip, trailing edge and wing root vortices. Some other vortex features include: independently developed left- and right-hand side leading edge vortex (LEV), dual-core LEV structure at the mid-wing region and near-wake two-vortex-ring structure. In the cicada forward flight, approximately 79% of the total lift is generated during the downstroke. Cicada wings experience drag in the downstroke, and generate thrust during the upstroke. Energetics study shows that the cicada in free forward flight consumes much more power in the downstroke than in the upstroke, to provide enough lift to support the weight and to overcome drag to move forward.

Computational investigation of cicada aerodynamics in forward flight

PubMed Central

Wan, Hui; Dong, Haibo; Gai, Kuo

2015-01-01

Free forward flight of cicadas is investigated through high-speed photogrammetry, three-dimensional surface reconstruction and computational fluid dynamics simulations. We report two new vortices generated by the cicada's wide body. One is the thorax-generated vortex, which helps the downwash flow, indicating a new phenomenon of lift enhancement. Another is the cicada posterior body vortex, which entangles with the vortex ring composed of wing tip, trailing edge and wing root vortices. Some other vortex features include: independently developed left- and right-hand side leading edge vortex (LEV), dual-core LEV structure at the mid-wing region and near-wake two-vortex-ring structure. In the cicada forward flight, approximately 79% of the total lift is generated during the downstroke. Cicada wings experience drag in the downstroke, and generate thrust during the upstroke. Energetics study shows that the cicada in free forward flight consumes much more power in the downstroke than in the upstroke, to provide enough lift to support the weight and to overcome drag to move forward. PMID:25551136

Noninvasive calculation of the aortic blood pressure waveform from the flow velocity waveform: a proof of concept

PubMed Central

Vennin, Samuel; Mayer, Alexia; Li, Ye; Fok, Henry; Clapp, Brian; Alastruey, Jordi

2015-01-01

Estimation of aortic and left ventricular (LV) pressure usually requires measurements that are difficult to acquire during the imaging required to obtain concurrent LV dimensions essential for determination of LV mechanical properties. We describe a novel method for deriving aortic pressure from the aortic flow velocity. The target pressure waveform is divided into an early systolic upstroke, determined by the water hammer equation, and a diastolic decay equal to that in the peripheral arterial tree, interposed by a late systolic portion described by a second-order polynomial constrained by conditions of continuity and conservation of mean arterial pressure. Pulse wave velocity (PWV, which can be obtained through imaging), mean arterial pressure, diastolic pressure, and diastolic decay are required inputs for the algorithm. The algorithm was tested using 1) pressure data derived theoretically from prespecified flow waveforms and properties of the arterial tree using a single-tube 1-D model of the arterial tree, and 2) experimental data acquired from a pressure/Doppler flow velocity transducer placed in the ascending aorta in 18 patients (mean ± SD: age 63 ± 11 yr, aortic BP 136 ± 23/73 ± 13 mmHg) at the time of cardiac catheterization. For experimental data, PWV was calculated from measured pressures/flows, and mean and diastolic pressures and diastolic decay were taken from measured pressure (i.e., were assumed to be known). Pressure reconstructed from measured flow agreed well with theoretical pressure: mean ± SD root mean square (RMS) error 0.7 ± 0.1 mmHg. Similarly, for experimental data, pressure reconstructed from measured flow agreed well with measured pressure (mean RMS error 2.4 ± 1.0 mmHg). First systolic shoulder and systolic peak pressures were also accurately rendered (mean ± SD difference 1.4 ± 2.0 mmHg for peak systolic pressure). This is the first noninvasive derivation of aortic pressure based on fluid dynamics (flow and wave speed) in the aorta itself. PMID:26163442

Cellobiose Dehydrogenase Inhibition of Polymerization of Phenolic Compounds and Enhancing Lignin Degradation by Lignina.

PubMed

Fang, Jing; Liu, Wen; Gao, Pei-Ji

1999-01-01

The kinetic behavior of cellobiose dehydrogenase (CDH) was investigated by steady-state initial velocity studies. Variation in the concentration of one substrate led to changes in K(m) and V(max) of the other substrate. The results were consistent with a ping-pong mechanism. In the presence of cellobiose, CDH could reduce many oxidized products catalyzed by soybean hull peroxidase (SHP). The oxidation product of 1-hydroxybenzotriazole (HBT) catalyzed by SHP inactivated the enzyme itself however, CDH could prevent SHP from inactivation by reducing the oxidation product of HBT. CDH could also inhibit the polymerization of phenolic compounds catalyzed by SHP. It was found that the addition of CDH could enhance kraft pulp lignin degradation by ligninases.

Effect of dietary fatty acids on jejunal and ileal oleic acid uptake by rat brush border membrane vesicles.

PubMed

Prieto, R M; Stremmel, W; Sales, C; Tur, J A

1996-04-18

To test the effect of dietary fatty acids on fatty acid uptake, the influx kinetics of a representative long-chain fatty acid, 3H-oleic acid, in both the jejunum and ileum of rats has been studied using brush border membrane vesicles (BBMV). Animals were fed with semipurified diets containing 5 g fat/100 g diet, as corn oil (control group), safflower oil (unsaturated group) and coconut oil hydrogenated (saturated group). With increasing unbound oleate concentration in the medium, the three dietary groups showed saturable kinetics in both jejunal and ileal BBMV (controls: Vmax = 0.15 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 136 +/- 29.1 nmol for jejunum, and Vmax = 0.23 +/- 0.03 nmol x mg protein-1 x 5 min-1 and Km = 196 +/- 50.3 nmol for ileum; unsaturated: Vmax = 0.28 +/- 0.05 nmol x mg protein-1 x 5 min-1 and Km = 242.7 +/- 91.8 nmol for jejunum, and Vmax = 1.29 +/- 0.06 nmol x mg protein-1 x 5 min-1 and Km = 509.8 +/- 97.5 nmol for ileum; saturated: Vmax = 0.03 +/- 0.01 nmol x mg protein-1 x 5 min-1 and Km = 124.5 +/- 72.6 nmol for jejunum, and Vmax = 0.04 +/- 0.01 nmol x mg protein -1.5 min-1 and Km = 205.6 +/- 85.3 nmol for ileum). These results support the theory that feeding an isocaloric diet containing only unsaturated fatty acids enhanced oleic acid uptake, and feeding an isocaloric diet containing only saturated fatty acids decreased oleic acid uptake. The results obtained in the present work also show the adaptative ability of jejunum and ileum to the type of dietary fat.

Abnormal sodium current properties contribute to cardiac electrical and contractile dysfunction in a mouse model of myotonic dystrophy type 1.

PubMed

Algalarrondo, Vincent; Wahbi, Karim; Sebag, Frédéric; Gourdon, Geneviève; Beldjord, Chérif; Azibi, Kamel; Balse, Elise; Coulombe, Alain; Fischmeister, Rodolphe; Eymard, Bruno; Duboc, Denis; Hatem, Stéphane N

2015-04-01

Myotonic dystrophy type 1 (DM1) is the most common neuromuscular disorder and is associated with cardiac conduction defects. However, the mechanisms of cardiac arrhythmias in DM1 are unknown. We tested the hypothesis that abnormalities in the cardiac sodium current (INa) are involved, and used a transgenic mouse model reproducing the expression of triplet expansion observed in DM1 (DMSXL mouse). The injection of the class-I antiarrhythmic agent flecainide induced prominent conduction abnormalities and significantly lowered the radial tissular velocities and strain rate in DMSXL mice compared to WT. These abnormalities were more pronounced in 8-month-old mice than in 3-month-old mice. Ventricular action potentials recorded by standard glass microelectrode technique exhibited a lower maximum upstroke velocity [dV/dt](max) in DMSXL. This decreased [dV/dt](max) was associated with a 1.7 fold faster inactivation of INa in DMSXL myocytes measured by the whole-cell patch-clamp technique. Finally in the DMSXL mouse, no mutation in the Scn5a gene was detected and neither cardiac fibrosis nor abnormalities of expression of the sodium channel protein were observed. Therefore, alterations in the sodium current markedly contributed to electrical conduction block in DM1. This result should guide pharmaceutical and clinical research toward better therapy for the cardiac arrhythmias associated with DM1. Copyright © 2014 Elsevier B.V. All rights reserved.

Cycling of Dissolved Organic Phosphorus and Alkaline Phosphatase Activity in Euphotic Zone of the Western North Pacific

NASA Astrophysics Data System (ADS)

Suzumura, M.

2010-12-01

Phosphorus is an essential nutrient for marine organisms. In oligotrophic environments, concentrations of dissolved inorganic phosphate (SRP), the most bioavailable form of phosphorus, are low and have been hypothesized to constrain the primary productivity. Evidence has been found that dissolved organic phosphorus (DOP) supports a significant fraction of primary production through hydrolytic remineralization of DOP to SRP by alkaline phosphatase (APA). In this study, DOP biogeochemistry was investigated at three locations of the open-ocean environment in the Kuroshio region and at a semi-eutrophic coastal site of the western North Pacific. Concentrations of SRP, DOP and hydrolyzable ester-P were measured in the euphotic zone. Kinetic parameters of APA were determined using a fluorogenic substrate, including potential maximum velocity (Vmax), apparent Michaelis-Menten half-saturation constant (Km), and turnover time (TA) of APA hydrolyzable DOP. SRP concentrations were quite low (≤ 10 nM) in the surface seawater and rapidly increased below the chlorophyll a maximum layer (CML). DOP concentration ranged from 29 to 223 nM. Above the CML, DOP composed a major fraction accounting for 60-100% of dissolved total P. A significant linear relationship was found between the concentrations of SRP and hydrolyzable ester-P (R2 = 0.83, P < 0.01). This suggests active utilization of ester-P under phosphate-depleted conditions. In the Kuroshio region, Vmax of APA exhibited the highest value at the surface water (0 m) and decreased rapidly with depth, while at the coastal site the peak value was found at CML. TA of hydrolyzable DOP was quite variable among the locations and increased with depth especially below CML. The estimated values of in situ hydrolysis rate were much lower (2-34%) than the potential Vmax which was determined with the addition of an excess amount of the substrate. The results suggest that marine microbes can efficiently and rapidly utilize hydrolyzable DOP under phosphate-depleted conditions and that there is still room in the in situ APA activity. Utilization of DOP, however, is likely regulated by the ambient concentrations of hydrolyzable ester-P lower than the apparent Km.

Recessive constitutive mutant Chinese hamster ovary cells (CHO-K1) with an altered A system for amino acid transport and the mechanism of gene regulation of the A system.

PubMed Central

Moffett, J; Englesberg, E

1984-01-01

Chinese hamster ovary cells (CHO-K1) starved for 24 h for amino acids show a severalfold increase in velocity of proline transport through the A system (Vmax is five times that of unstarved cells). This increase is inhibited by cycloheximide, actinomycin D, N-methyl-alpha-amino isobutyric acid (MeAIB, a non-metabolizable specific A system amino acid analog), and by other amino acids that are generally transported by the A system. However, transport by the A system is not a prerequisite for this repression, and all compounds that have affinity for the A system do not necessarily act as "co-repressors." The addition of proline, MeAIB, or other amino acids, as described above, to derepressed cells results in a rapid decrease in A system activity. As shown with proline and MeAIB, this decrease in activity is in part due to a rapid trans-inhibition and a slow, irreversible inactivation of the A system. Neither process is inhibited by cycloheximide or actinomycin D. Alanine antagonizes the growth of CHO-K1 pro cells by preventing proline transport, and alanine-resistant mutants (alar) have been isolated (Moffett et al., Somatic Cell Genet. 9:189-213, 1983). alar2 and alar4 are partial and full constitutive mutants for the A system and have two and six times the Vmax for proline uptake by the A system, respectively. The A system in alar4 is also immune to the co-repressor-induced inactivation. Both alar2 and alar4 phenotypes are recessive. Alar3 shows an increase in Vmax and Km for proline transport through the A system, and this phenotype is codominant. All three mutants have a pleiotropic effect, producing increases in activity of the ASC and P systems of amino acid transport. This increase is not due to an increase in the Na+ gradient. The ASC and P phenotypes behave similarly to the A system in hybrids. A model has been proposed incorporating these results. PMID:6538929

Functional and Structural Correlates of Motor Speed in the Cerebellar Anterior Lobe

PubMed Central

Wenzel, Uwe; Taubert, Marco; Ragert, Patrick; Krug, Jürgen; Villringer, Arno

2014-01-01

In athletics, motor performance is determined by different abilities such as technique, endurance, strength and speed. Based on animal studies, motor speed is thought to be encoded in the basal ganglia, sensorimotor cortex and the cerebellum. The question arises whether there is a unique structural feature in the human brain, which allows “power athletes” to perform a simple foot movement significantly faster than “endurance athletes”. We acquired structural and functional brain imaging data from 32 track-and-field athletes. The study comprised of 16 “power athletes” requiring high speed foot movements (sprinters, jumpers, throwers) and 16 endurance athletes (distance runners) which in contrast do not require as high speed foot movements. Functional magnetic resonance imaging (fMRI) was used to identify speed specific regions of interest in the brain during fast and slow foot movements. Anatomical MRI scans were performed to assess structural grey matter volume differences between athletes groups (voxel based morphometry). We tested maximum movement velocity of plantarflexion (PF-Vmax) and acquired electromyographical activity of the lateral and medial gastrocnemius muscle. Behaviourally, a significant difference between the two groups of athletes was noted in PF-Vmax and fMRI indicates that fast plantarflexions are accompanied by increased activity in the cerebellar anterior lobe. The same region indicates increased grey matter volume for the power athletes compared to the endurance counterparts. Our results suggest that speed-specific neuro-functional and -structural differences exist between power and endurance athletes in the peripheral and central nervous system. PMID:24800742

Identification of Functional Amino Acid Residues Involved in Polyamine and Agmatine Transport by Human Organic Cation Transporter 2

PubMed Central

Higashi, Kyohei; Imamura, Masataka; Fudo, Satoshi; Uemura, Takeshi; Saiki, Ryotaro; Hoshino, Tyuji; Toida, Toshihiko; Kashiwagi, Keiko; Igarashi, Kazuei

2014-01-01

Polyamine (putrescine, spermidine and spermine) and agmatine uptake by the human organic cation transporter 2 (hOCT2) was studied using HEK293 cells transfected with pCMV6-XL4/hOCT2. The Km values for putrescine and spermidine were 7.50 and 6.76 mM, and the Vmax values were 4.71 and 2.34 nmol/min/mg protein, respectively. Spermine uptake by hOCT2 was not observed at pH 7.4, although it inhibited both putrescine and spermidine uptake. Agmatine was also taken up by hOCT2, with Km value: 3.27 mM and a Vmax value of 3.14 nmol/min/mg protein. Amino acid residues involved in putrescine, agmatine and spermidine uptake by hOCT2 were Asp427, Glu448, Glu456, Asp475, and Glu516. In addition, Glu524 and Glu530 were involved in putrescine and spermidine uptake activity, and Glu528 and Glu540 were weakly involved in putrescine uptake activity. Furthermore, Asp551 was also involved in the recognition of spermidine. These results indicate that the recognition sites for putrescine, agmatine and spermidine on hOCT2 strongly overlap, consistent with the observation that the three amines are transported with similar affinity and velocity. A model of spermidine binding to hOCT2 was constructed based on the functional amino acid residues. PMID:25019617

Endothelins activate Ca(2+)-gated K(+) channels via endothelin B receptors in CD-1 mouse erythrocytes.

PubMed

Rivera, A; Rotter, M A; Brugnara, C

1999-10-01

Cell dehydration mediated by Ca(2+)-activated K(+) channels plays an important role in the pathogenesis of sickle cell disease. CD-1 mouse erythrocytes possess a Ca(2+)-activated K(+) channel (Gardos channel) with maximal velocity (V(max)) of 0.154 +/- 0.02 mmol. l cells(-1). min(-1) and an affinity constant (K(0.5)) for Ca(2+) of 286 +/- 83 nM in the presence of A-23187. Cells pretreated with 500 nM endothelin-1 (ET-1) increased their V(max) by 88 +/- 9% (n = 8) and decreased their K(0.5) for Ca(2+) to 139 +/- 63 nM (P < 0.05; n = 4). Activation of the Gardos channel resulted in an EC(50) of 75 +/- 20 nM for ET-1 and 374 +/- 97 nM for ET-3. Analysis of the affinity of unlabeled ET-1 for its receptor showed two classes of binding sites with apparent dissociation constants of 167 +/- 51 and 785 +/- 143 nM and with capacity of binding sites of 298 +/- 38 and 1,568 +/- 211 sites/cell, respectively. The Gardos channel was activated by the endothelin B (ET(B)) receptor agonist IRL 1620 and inhibited by BQ-788, demonstrating the involvement of ET(B) receptors. Calphostin C inhibited 73% of ET-1-induced Gardos activation and 84% of the ET-1-induced membrane protein kinase C activity. Thus endothelins regulate erythrocyte Gardos channels via ET(B) receptors and a calphostin-sensitive mechanism.

Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones.

PubMed

Yan, Feng-Ying; Xia, Wei; Zhang, Xiao-Xu; Chen, Sha; Nie, Xin-Zheng; Qian, Li-Chun

2016-06-01

An extracellular β-glucosidase produced by Aspergillus terreus was identified, purified, characterized and was tested for the hydrolysis of soybean isoflavone. Matrix-assisted laser desorption/ionization with tandem time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) revealed the protein to be a member of the glycosyl hydrolase family 3 with an apparent molecular mass of about 120 kDa. The purified β-glucosidase showed optimal activity at pH 5.0 and 65 °C and was very stable at 50 °C. Moreover, the enzyme exhibited good stability over pH 3.0-8.0 and possessed high tolerance towards pepsin and trypsin. The kinetic parameters Km (apparent Michaelis-Menten constant) and Vmax (maximal reaction velocity) for p-nitrophenyl-β-D-glucopyranoside (pNPG) were 1.73 mmol/L and 42.37 U/mg, respectively. The Km and Vmax for cellobiose were 4.11 mmol/L and 5.7 U/mg, respectively. The enzyme efficiently converted isoflavone glycosides to aglycones, with a hydrolysis rate of 95.8% for daidzin, 86.7% for genistin, and 72.1% for glycitin. Meanwhile, the productivities were 1.14 mmol/(L·h) for daidzein, 0.72 mmol/(L·h) for genistein, and 0.19 mmol/(L·h) for glycitein. This is the first report on the application of A. terreus β-glucosidase for converting isoflavone glycosides to their aglycones in soybean products.

THE IMPACT OF SCALING FACTOR VARIABILITY ON RISK-RELEVANT TOXICOKINETIC OUTCOMES IN CHILDREN: A CASE STUDY USING BROMODICHLOROMETHANE (BDCM)

EPA Science Inventory

Biotransformation rates (Vmax) extrapolated from in vitro data are used increasingly in human physiologically based pharmacokinetic (PBPK) models. Extrapolation of Vmax from in vitro data requires use of scaling factors, including mg of microsomal protein/g liver (MPPGL), nmol of...

The Impact of Immediate Verbal Feedback on the Improvement of Swimming Technique

PubMed Central

Zatoń, Krystyna; Szczepan, Stefan

2014-01-01

The present research attempts to ascertain the impact of immediate verbal feedback (IVF) on modifications of stroke length (SL). In all swimming styles, stroke length is considered an essential kinematic parameter of the swimming cycle. It is important for swimming mechanics and energetics. If SL shortens while the stroke rate (SR) remains unchanged or decreases, the temporal-spatial structure of swimming is considered erroneous. It results in a lower swimming velocity. Our research included 64 subjects, who were divided into two groups: the experimental – E (n=32) and the control – C (n=32) groups. A pretest and a post-test were conducted. The subjects swam the front crawl over the test distance of 25m at Vmax. Only the E group subjects were provided with IVF aiming to increase their SL. All tests were filmed by two cameras (50 samples•s-1). The kinematic parameters of the swimming cycle were analyzed using the SIMI Reality Motion Systems 2D software (SIMI Reality Motion Systems 2D GmbH, Germany). The movement analysis allowed to determine the average horizontal swimming velocity over 15 meters. The repeated measures analysis of variance ANOVA with a post-hoc Tukey range test demonstrated statistically significant (p<0.05) differences between the two groups in terms of SL and swimming velocity. IVF brought about a 6.93% (Simi method) and a 5.09% (Hay method) increase in SL, as well as a 2.92% increase in swimming velocity. PMID:25114741

A molecular dynamics study on the role of attractive and repulsive forces in internal energy, internal pressure and structure of dense fluids

NASA Astrophysics Data System (ADS)

Goharshadi, Elaheh K.; Morsali, Ali; Mansoori, G. Ali

2007-01-01

Isotherms of experimental data of internal pressure of dense fluids versus molar volume, Vm are shown to have each a maximum point at a Vmax below the critical molar volume. In this study, we investigated the role of attractive and repulsive intermolecular energies on this behavior using a molecular dynamics simulation technique. In the simulation, we choose the Lennard-Jones (LJ) intermolecular potential energy function. The LJ potential is known to be an effective potential representing a statistical average of the true pair and many-body interactions in simple molecular systems. The LJ potential function is divided into attractive and repulsive parts. MD calculations have produced internal energy, potential energy, transitional kinetic energy, and radial distribution function (RDF) for argon at 180 K and 450 K using LJ potential, LJ repulsive, and LJ attractive parts. It is shown that the LJ potential function is well capable of predicting the inflection point in the internal energy-molar volume curve as well as maximum point in the internal pressure-molar volume curve. It is also shown that at molar volumes higher than Vmax, the attractive forces have strong influence on determination of internal energy and internal pressure. At volumes lower than Vmax, neither repulsive nor attractive forces are dominating. Also, the coincidence between RDFs resulting from LJ potential and repulsive parts of LJ potential improves as molar volume approaches Vmax from high molar volumes. The coincidence becomes complete at Vmax ⩾ V.

THE IMPORTANCE OF OBTAINING INFORMATION ON THE SPECIFIC CONTENT OF TISSUE ENZYMES METABOLIZING ORGANOPHOSPHORUS PESTICIDES, PRIOR TO DETERMINE VMAX, KM VALUES FOR USE IN PBPK MODELS

EPA Science Inventory

Physiological pharmacokinetic/pharmacodynamic models require Vmax, Km values for the metabolism of OPs by tissue enzymes. Current literature values cannot be easily used in OP PBPK models (i.e., parathion and chlorpyrifos) because standard methodologies were not used in their ...

THE IMPORTANCE OF OBTAINING INFORMATION ON THE SPECIFIC CONTENT OF TISSUE ENZYMES METABOLIZING ORGANOPHOSPHORUS PESTICIDES, PRIOR TO DETERMINING VMAX, KM VALUES FOR USE IN PBPK MODELS

EPA Science Inventory

Physiological pharmacokinetic\\pharmacodynamic models require Vmax, Km values for the metabolism of OPs by tissue enzymes. Current literature values cannot be easily used in OP PBPK models (i.e., parathion and chlorpyrifos) because standard methodologies were not used in their ...

Monocarboxylate transporters, blood lactate removal after supramaximal exercise, and fatigue indexes in humans.

PubMed

Thomas, C; Perrey, S; Lambert, K; Hugon, G; Mornet, D; Mercier, J

2005-03-01

The present study investigated whether muscular monocarboxylate transporter (MCT) 1 and 4 contents are related to the blood lactate removal after supramaximal exercise, fatigue indexes measured during different supramaximal exercises, and muscle oxidative parameters in 15 humans with different training status. Lactate recovery curves were obtained after a 1-min all-out exercise. A biexponential time function was then used to determine the velocity constant of the slow phase (gamma(2)), which denoted the blood lactate removal ability. Fatigue indexes were calculated during 1-min all-out (FI(AO)) and repeated 10-s (FI(Sprint)) cycling sprints. Biopsies were taken from the vastus lateralis muscle. MCT1 and MCT4 contents were quantified by Western blots, and maximal muscle oxidative capacity (V(max)) was evaluated with pyruvate + malate and glutamate + malate as substrates. The results showed that the blood lactate removal ability (i.e., gamma(2)) after a 1-min all-out test was significantly related to MCT1 content (r = 0.70, P < 0.01) but not to MCT4 (r = 0.50, P > 0.05). However, greater MCT1 and MCT4 contents were negatively related with a reduction of blood lactate concentration at the end of 1-min all-out exercise (r = -0.56, and r = -0.61, P < 0.05, respectively). Among skeletal muscle oxidative indexes, we only found a relationship between MCT1 and glutamate + malate V(max) (r = 0.63, P < 0.05). Furthermore, MCT1 content, but not MCT4, was inversely related to FI(AO) (r = -0.54, P < 0.05) and FI(Sprint) (r = -0.58, P < 0.05). We concluded that skeletal muscle MCT1 expression was associated with the velocity constant of net blood lactate removal after a 1-min all-out test and with the fatigue indexes. It is proposed that MCT1 expression may be important for blood lactate removal after supramaximal exercise based on the existence of lactate shuttles and, in turn, in favor of a better tolerance to muscle fatigue.

Kinematics of flap-bounding flight in the zebra finch over a wide range of speeds

PubMed

Tobalske; Peacock; Dial

1999-07-01

It has been proposed elsewhere that flap-bounding, an intermittent flight style consisting of flapping phases interspersed with flexed-wing bounds, should offer no savings in average mechanical power relative to continuous flapping unless a bird flies 1.2 times faster than its maximum range speed (Vmr). Why do some species use intermittent bounds at speeds slower than 1.2Vmr? The 'fixed-gear hypothesis' suggests that flap-bounding is used to vary mean power output in small birds that are otherwise constrained by muscle physiology and wing anatomy to use a fixed muscle shortening velocity and pattern of wing motion at all flight speeds; the 'body-lift hypothesis' suggests that some weight support during bounds could make flap-bounding flight aerodynamically advantageous in comparison with continuous flapping over most forward flight speeds. To test these predictions, we studied high-speed film recordings (300 Hz) of wing and body motion in zebra finches (Taenopygia guttata, mean mass 13.2 g, N=4) taken as the birds flew in a variable-speed wind tunnel (0-14 m s-1). The zebra finches used flap-bounding flight at all speeds, so their flight style was unique compared with that of birds that facultatively shift from continuous flapping or flap-gliding at slow speeds to flap-bounding at fast speeds. There was a significant effect of flight speed on all measured aspects of wing motion except percentage of the wingbeat spent in downstroke. Changes in angular velocity of the wing indicated that contractile velocity in the pectoralis muscle changed with flight speed, which is not consistent with the fixed-gear hypothesis. Although variation in stroke-plane angle relative to the body, pronation angle of the wing and wing span at mid-upstroke showed that the zebra finch changed within-wingbeat geometries according to speed, a vortex-ring gait with a feathered upstroke appeared to be the only gait used during flapping. In contrast, two small species that use continuous flapping during slow flight (0-4 m s-1) either change wingbeat gait according to flight speed or exhibit more variation in stroke-plane and pronation angles relative to the body. Differences in kinematics among species appear to be related to wing design (aspect ratio, skeletal proportions) rather than to pectoralis muscle fiber composition, indicating that the fixed-gear hypothesis should perhaps be modified to exclude muscle physiology and to emphasize constraints due to wing anatomy. Body lift was produced during bounds at speeds from 4 to 14 m s-1. Maximum body lift was 0.0206 N (15.9 % of body weight) at 10 m s-1; body lift:drag ratio declined with increasing air speed. The aerodynamic function of bounds differed with increasing speed from an emphasis on lift production (4-10 m s-1) to an emphasis on drag reduction with a slight loss in lift (12 and 14 m s-1). From a mathematical model of aerodynamic costs, it appeared that flap-bounding offered the zebra finch an aerodynamic advantage relative to continuous flapping at moderate and fast flight speeds (6-14 m s-1), with body lift augmenting any savings offered solely by flap-bounding at speeds faster than 7.1 m s-1. The percentage of time spent flapping during an intermittent flight cycle decreased with increasing speed, so the mechanical cost of transport was likely to be lowest at faster flight speeds (10-14 m s-1).

An inverse approach to determining spatially varying arterial compliance using ultrasound imaging

NASA Astrophysics Data System (ADS)

Mcgarry, Matthew; Li, Ronny; Apostolakis, Iason; Nauleau, Pierre; Konofagou, Elisa E.

2016-08-01

The mechanical properties of arteries are implicated in a wide variety of cardiovascular diseases, many of which are expected to involve a strong spatial variation in properties that can be depicted by diagnostic imaging. A pulse wave inverse problem (PWIP) is presented, which can produce spatially resolved estimates of vessel compliance from ultrasound measurements of the vessel wall displacements. The 1D equations governing pulse wave propagation in a flexible tube are parameterized by the spatially varying properties, discrete cosine transform components of the inlet pressure boundary conditions, viscous loss constant and a resistance outlet boundary condition. Gradient descent optimization is used to fit displacements from the model to the measured data by updating the model parameters. Inversion of simulated data showed that the PWIP can accurately recover the correct compliance distribution and inlet pressure under realistic conditions, even under high simulated measurement noise conditions. Silicone phantoms with known compliance contrast were imaged with a clinical ultrasound system. The PWIP produced spatially and quantitatively accurate maps of the phantom compliance compared to independent static property estimates, and the known locations of stiff inclusions (which were as small as 7 mm). The PWIP is necessary for these phantom experiments as the spatiotemporal resolution, measurement noise and compliance contrast does not allow accurate tracking of the pulse wave velocity using traditional approaches (e.g. 50% upstroke markers). Results from simulations indicate reflections generated from material interfaces may negatively affect wave velocity estimates, whereas these reflections are accounted for in the PWIP and do not cause problems.

Kinematics of slow turn maneuvering in the fruit bat Cynopterus brachyotis.

PubMed

Iriarte-Díaz, José; Swartz, Sharon M

2008-11-01

Maneuvering abilities have long been considered key factors that influence habitat selection and foraging strategies in bats. To date, however, very little experimental work has been carried out to understand the mechanisms that bats use to perform maneuvers. In the present study, we examined the kinematics of slow-speed turning flight in the lesser short-nosed fruit bat, Cynopterus brachyotis, to understand the basic mechanics employed to perform maneuvers and to compare them with previous findings in bats and other flying organisms. Four individuals were trained to fly in L-shaped flight enclosure that required them to make a 90 deg. turn midway through each flight. Flights were recorded with three low-light, high-speed videocameras, allowing the three-dimensional reconstruction of the body and wing kinematics. For any flying organisms, turning requires changes of the direction of travel and the reorientation of the body around the center of mass to maintain the alignment with the flight direction. In C. brachyotis, changes in body orientation (i.e. heading) took place during upstroke and preceded the changes in flight direction, which were restricted to the downstroke portion of the wingbeat cycle. Mean change in flight direction was significantly correlated to the mean heading angular velocity at the beginning of the downstroke and to the mean bank angle during downstroke, although only heading velocity was significant when both variables were considered. Body reorientation prior to changes in direction might be a mechanism to maintain the head and body aligned with the direction of travel and, thus, maximizing spatial accuracy in three-dimensionally complex environments.

Anti-lipase and antioxidant properties of 30 medicinal plants used in Oaxaca, México.

PubMed

Villa-Ruano, Nemesio; Zurita-Vásquez, Guilibaldo G; Pacheco-Hernández, Yesenia; Betancourt-Jiménez, Martha G; Cruz-Durán, Ramiro; Duque-Bautista, Horacio

2013-01-01

We report the results of in vitro anti-lipase and antioxidant assays using crude ethanolic extracts from 30 plants grown in Oaxaca, México. Anti-lipase tests were performed by using porcine pancreatic lipase (PPL) [EC 3.1.1.3] from Affymetrix/USB. The extracts of Solanum erianthum, Salvia microphylla, Brungmansia suaveolens and Cuphea aequipetala showed up to 60% PPL inhibition. The effect of these extracts on the kinetic parameters of PPL (Km= 0.36 mM, and Vmax=0.085 mM min -1) revealed that the alcoholic preparations of S. erianthum and C. aequipetala engendered a non-competitive inhibition (Vmax=0.055 mM min -1; Vmax= 0.053 mM min -1), whereas those of S. microphylla and B. suaveolens produced a mixed inhibition (Km= 0.567 mM, Vmax=0.051 mM min _1; Km=0.643 mM, Vmax= 0.042 mM min ¹). In addition to these findings, seven extracts from different plants were able to inhibit PPL in the range of 30-50%. Antioxidant tests against 2,2-Diphenyl-1-picryl hydrazyl (DPPH) confirmed that Arctostaphylos pungens, Gnaphalium roseum, Crotalaria pumila, Cuphea aequipetala, Rhus chondroloma, and Satureja laevigata possess relevant antioxidant activity (IC(5)0=50-80 μg mL¹). The general composition of the most effective ethanolic extracts was obtained in order to confirm their known chemistry reported by previous works. Comprehensive chemical analysis of the ethanolic extracts and their poisoning effects suggests that S. microphylla, C. aequipetala and A. pungens could be considered as the best sources with both desired properties.

Addressing the too big to fail problem with baryon physics and sterile neutrino dark matter

NASA Astrophysics Data System (ADS)

Lovell, Mark R.; Gonzalez-Perez, Violeta; Bose, Sownak; Boyarsky, Alexey; Cole, Shaun; Frenk, Carlos S.; Ruchayskiy, Oleg

2017-07-01

N-body dark matter simulations of structure formation in the Λ cold dark matter (ΛCDM) model predict a population of subhaloes within Galactic haloes that have higher central densities than inferred for the Milky Way satellites, a tension known as the 'too big to fail' problem. Proposed solutions include baryonic effects, a smaller mass for the Milky Way halo and warm dark matter (WDM). We test these possibilities using a semi-analytic model of galaxy formation to generate luminosity functions for Milky Way halo-analogue satellite populations, the results of which are then coupled to the Jiang & van den Bosch model of subhalo stripping to predict the subhalo Vmax functions for the 10 brightest satellites. We find that selecting the brightest satellites (as opposed to the most massive) and modelling the expulsion of gas by supernovae at early times increases the likelihood of generating the observed Milky Way satellite Vmax function. The preferred halo mass is 6 × 1011 M⊙, which has a 14 per cent probability to host a Vmax function like that of the Milky Way satellites. We conclude that the Milky Way satellite Vmax function is compatible with a CDM cosmology, as previously found by Sawala et al. using hydrodynamic simulations. Sterile neutrino-WDM models achieve a higher degree of agreement with the observations, with a maximum 50 per cent chance of generating the observed Milky Way satellite Vmax function. However, more work is required to check that the semi-analytic stripping model is calibrated correctly for each sterile neutrino cosmology.

Glutamine, glutamate, and other possible regulators of alpha-ketoglutarate and malate uptake by synaptic terminals.

PubMed

Shank, R P; Campbell, G L

1984-04-01

The uptake of alpha-ketoglutarate and malate by rat brain synaptosomal preparations was found to be affected by a variety of substances at physiologically relevant concentrations. Glutamine altered the uptake of alpha-ketoglutarate by causing an apparent reduction in the substrate-carrier affinity and an increase in Vmax. In contrast, glutamine did not appear to affect the Vmax of malate uptake, but it did increase markedly the uptake velocity at low concentrations of malate. L-Glutamate and L-aspartate were comparatively strong inhibitors of alpha-ketoglutarate and malate uptake. N-Acetylaspartate was a weak inhibitor of alpha-ketoglutarate uptake, a finding that contrasts with our previous observation that this compound potently inhibited alpha-ketoglutarate uptake by synaptosomes obtained from the cerebellum of 8- to 14-day-old mice. Ca2+ exhibited a variable effect but usually enhanced the uptake of alpha-ketoglutarate. The addition of small amounts of postmicrosomal supernatant to the incubation medium enhanced the uptake of alpha-ketoglutarate by low-density synaptosomes. By comparison, the uptake of glutamate, glutamine, gamma-aminobutyric acid, and several other amino acids was not affected. The enhancement of alpha-ketoglutarate uptake by the supernatant was due to a heat labile substance that was retained by dialysis tubing (MW cutoff = 8,000) and Amicon filter cones (CF 25), and was precipitated by ammonium sulfate at 60% saturation. In experiments in which the metabolic conversion of [U-14C] alpha-ketoglutarate to glutamate, aspartate, glutamine, and gamma-aminobutyric acid was determined, the presence of glutamine and glutamate in the incubation medium did not affect the pattern of labelling appreciably.

Spreading out and staying sharp - creating diverse rotation curves via baryonic and self-interaction effects

NASA Astrophysics Data System (ADS)

Creasey, Peter; Sameie, Omid; Sales, Laura V.; Yu, Hai-Bo; Vogelsberger, Mark; Zavala, Jesús

2017-06-01

Galactic rotation curves are a fundamental constraint for any cosmological model. We use controlled N-body simulations of galaxies to study the gravitational effect of baryons in a scenario with collisionless cold dark matter (CDM) versus one with a self-interacting dark matter (SIDM) component. In particular, we examine the inner profiles of the rotation curves in the velocity range Vmax = [30-250] km s-1, whose diversity has been found to be greater than predicted by the ΛCDM scenario. We find that the scatter in the observed rotation curves exceeds that predicted by dark matter only mass-concentration relations in either the CDM nor SIDM models. Allowing for realistic baryonic content and spatial distributions, however, helps create a large variety of rotation curve shapes, which is in a better agreement with observations in the case of self-interactions due to the characteristic cored profiles being more accommodating to the slowly rising rotation curves than CDM. We find individual fits to model two of the most remarkable outliers of similar Vmax, UGC 5721 and IC 2574 - the former a cusp-like rotation curve and the latter a seemingly 8-kpc-cored profile. This diversity in SIDM arises as permutations of overly concentrated haloes with compact baryonic distributions versus underdense haloes with extended baryonic discs. The SIDM solution is promising and its feasibility ultimately depends on the sampling of the halo mass-concentration relation and its interplay with the baryonic profiles, emphasizing the need for a better understanding of the frequency of extreme outliers present in current observational samples.

Change of heart dimensions and function during pregnancy in goats.

PubMed

Szaluś-Jordanow, Olga; Czopowicz, Michał; Witkowski, Lucjan; Moroz, Agata; Mickiewicz, Marcin; Frymus, Tadeusz; Markowska-Daniel, Iwona; Bagnicka, Emilia; Kaba, Jarosław

2018-03-08

The study aimed to evaluate the effect of pregnancy on heart diameters and function in goats. Transthoracic echocardiography of 12 female dairy goats of two Polish regional breeds was performed. A Mindray M7 diagnostic ultrasound system with Phased Array transducer was used. Simultaneously, electrocardiography was recorded. All animals were examined four times - at mating season, at the end of the first trimester, at the end of the second trimester and just before kidding. Eleven measurements were taken each time: aortic and left atrial diameter (AoD and LAD), right and left ventricular internal diameter in diastole (RVIDd and LVIDd), left ventricular internal diameter in systole (LVIDs), inter-ventricular septum thickness in diastole and systole (IVSd and IVSd) and left ventricular posterior wall in diastole and systole (LVPWd and LVPWs), maximum left and right ventricular outflow tract velocity (RVOT Vmax and LVOT Vmax). Nine consecutive measurements were derived: the ratio of the left atrial diameter to the aortic diameter (AoD/LAD), left ventricular fractional shortening (FS%), left ventricular ejection fraction (EF%), maximum outflow tract pressure gradients (RVOT PGmax and LVOT PGmax), left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV), stroke volume (SV) and cardiac output (CO). HR, LAD, LVPWs, IVSs increased significantly in the first trimester. AoD and RVIDd were significantly higher around parturition. LVIDd, FS%, EF%, SV and CO rose both in the first and third trimester. No measurement decreased during pregnancy. The study confirms that pregnancy causes changes in the heart size and functioning. Copyright © 2018. Published by Elsevier Ltd.

Lion (Panthera leo) and caracal (Caracal caracal) type IIx single muscle fibre force and power exceed that of trained humans.

PubMed

Kohn, Tertius A; Noakes, Timothy D

2013-03-15

This study investigated for the first time maximum force production, shortening velocity (Vmax) and power output in permeabilised single muscle fibres at 12°C from lion, Panthera leo (Linnaeus 1758), and caracal, Caracal caracal (Schreber 1776), and compared the values with those from human cyclists. Additionally, the use and validation of previously frozen tissue for contractile experiments is reported. Only type IIx muscle fibres were identified in the caracal sample, whereas type IIx and only two type I fibres were found in the lion sample. Only pure type I and IIa, and hybrid type IIax fibres were identified in the human samples - there were no pure type IIx fibres. Nevertheless, compared with all the human fibre types, the lion and caracal fibres were smaller (P<0.01) in cross-sectional area (human: 6194±230 μm(2), lion: 3008±151 μm(2), caracal: 2583±221 μm(2)). On average, the felid type IIx fibres produced significantly greater force (191-211 kN m(-2)) and ~3 times more power (29.0-30.3 kN m(-2) fibre lengths s(-1)) than the human IIax fibres (100-150 kN m(-2), 4-11 kN m(-2) fibre lengths s(-1)). Vmax values of the lion type IIx fibres were also higher than those of human type IIax fibres. The findings suggest that the same fibre type may differ substantially between species and potential explanations are discussed.

Motility assays using myosin attached to surfaces through specific binding to monoclonal antibodies.

PubMed Central

Winkelmann, D. A.; Bourdieu, L.; Kinose, F.; Libchaber, A.

1995-01-01

We have analyzed the dependence of actin filament movement on the mode of myosin attachment to surfaces. Monoclonal antibodies that bind to three distinct sites were used to tether myosin to nitrocellulose-coated glass. One antibody reacts with an epitope on the regulatory light chain located at the head-rod junction. The other two react with sites in the rod domain, one in the S2 region near the S2-LMM hinge, and the other at the C terminus of the myosin rod. These monoclonal antibodies were used to provide increasing flexibility in the mode of attachment. Fast skeletal muscle myosin monomers were bound to the surfaces through the specific interaction with these monoclonal antibodies and the sliding movement of fluorescently labeled actin filaments analyzed by video microscopy. Each of these antibodies produced stable, myosin-coated surfaces that supported uniform movement of actin over the course of several hours. Attachment of myosin through the anti-S2 and anti-LMM monoclonal antibodies yielded a maximum velocity of 10 microns/s at 30 degrees C, whereas attachment through anti-LC2 produced a lower velocity of 4-5 microns/s. Each antibody showed a characteristic minimum myosin density below which sliding movement was no longer supported and an exponential dependence of actin filament velocity on myosin surface density below Vmax. Maximum sliding velocity was achieved over a range of myosin surface densities. Thus, the specific mode of attachment can influence the characteristic velocity of actin filament movement and the surface density needed to support movement. These data are being used to analyze the dynamics of sliding filament assays and evaluate estimates of the average number of motor molecules per unit length of actin required to support movement. PMID:7787107

High intensity interval training vs. high-volume running training during pre-season conditioning in high-level youth football: a cross-over trial.

PubMed

Faude, Oliver; Schnittker, Reinhard; Schulte-Zurhausen, Roman; Müller, Florian; Meyer, Tim

2013-01-01

We aimed at comparing the endurance effects of high-intensity interval training (HIIT) with high-volume running training (HVT) during pre-season conditioning in 20 high-level youth football players (15.9 (s 0.8) years). Players either conducted HVT or HIIT during the summer preparation period. During winter preparation they performed the other training programme. Before and after each training period several fitness tests were conducted: multi-stage running test (to assess the individual anaerobic threshold (IAT) and maximal running velocity (Vmax)), vertical jumping height, and straight sprinting. A significant increase from pre- to post-test was observed in IAT velocity (P < 0.001) with a greater increase after HVT (+0.8 km · h(-1) vs. +0.5 km · h(-1) after HIIT, P = 0.04). Maximal velocity during the incremental exercise test also slightly increased with time (P = 0.09). Forty per cent (HIIT) and 15% (HVT) of all players did not improve IAT beyond baseline variability. The players who did not respond to HIIT were significantly slower during 30 m sprinting than responders (P = 0.02). No further significant differences between responders and non-responders were observed. Jump heights deteriorated significantly after both training periods (P < 0.003). Both training programmes seem to be promising means to improve endurance capacity in high-level youth football players during pre-season conditioning.

Pedagogical view of model metabolic cycles.

PubMed

García-Herrero, Victor; Sillero, Antonio

2015-01-01

The main purpose of this study was to present a simplified view of model metabolic cycles. Although the models have been elaborated with the Mathematica Program, and using a system of differential equations, the main conclusions were presented in a rather intuitive way, easily understandable by students of general courses of Biochemistry, and without any need of mathematical support. A change in any kinetic constant (Km or Vmax) of only one enzyme affected the metabolic profile of all the substrates of the cycle. In addition, it is shown how an increase in the Km or a decrease in the Vmax values of any particular enzyme promoted an increase of its substrate; the contrary occurred decreasing the Km or increasing the Vmax values. © 2015 The International Union of Biochemistry and Molecular Biology.

Cyanide removal by Chinese vegetation--quantification of the Michaelis-Menten kinetics.

PubMed

Yu, Xiaozhang; Zhou, Puhua; Zhou, Xishi; Liu, Yunda

2005-07-01

Little is known about metabolism rates of environmental chemicals by vegetation. A good model compound to study the variation of rates among plant species is cyanide. Vascular plants possess an enzyme system that detoxifies cyanide by converting it to the amino acid asparagine. Knowledge of the kinetic parameters, the half-saturation constant (Km) and the maximum metabolic capacity (vmax), is very useful for enzyme characterization and biochemical purposes. The goal of this study is to find the enzyme kinetics (K(M) and vmax) during cyanide metabolism in the presence of Chinese vegetation, to provide quantitative data for engineered phytoremediation, and to investigate the variation of metabolic rates of plants. Detached leaves (1.0 g fresh weight) from 12 species out of 9 families were kept in glass vessels with 100 mL of aqueous solution spiked with potassium cyanide at 23 degrees C for 28 h. Four different treatment concentrations of cyanide were used, ranging from 0.44 to 7.69 mg CN/L. The disappearance of cyanide from the aqueous solution was analyzed spectrophotometrically. Realistic values of the half-saturation constant (KM) and the maximum metabolic capacity (vmax) were estimated by a computer program using non-linear regression treatments. As a comparison, Lineweaver-Burk plots were also used to estimate the kinetic parameters. The values obtained for K(M) and vmax varied with plant species. Using non-linear regression treatments, values of vmax and K(M) were found in a range between 6.68 and 21.91 mg CN/kg/h and 0.90 to 3.15 mg CN/L, respectively. The highest vmax was by Chinese elder (Sambucus chinensis), followed by upright hedge-parsley (Torilis japonica). The lowest Vmax was demonstrated by the hybrid willow (Salix matssudana x alba). However, the highest K(M) was found in the water lily (Nymphea teragona), followed by the poplar (Populus deltoides Marsh). The lowest K(M) was demonstrated by corn (Zea mays L.). The values of vmax were normally distributed with a mean of 13 mg CN/kg/h. Significant removal of cyanide from aqueous solution was observed in the presence of plant materials without phytotoxicity, even at high doses of cyanide. This gives rise to the conclusion that the Chinese plant species used in this study are all able to efficiently metabolize cyanide, although with different maximum metabolic capacities. A second conclusion is that the variation of metabolism rates between species is small. All these plants had a similar K(M), indicating the same enzyme is active in all plants. Detoxification of cyanide with trees seems to be a feasible option for cleaning soils and water contaminated with cyanide. For phytoremediation projects, screening appropriate plant species adapted to local conditions should be seriously considered. More chemicals should be investigated to find common principles of the metabolism of environmental chemicals by plants.

Noninvasive calculation of the aortic blood pressure waveform from the flow velocity waveform: a proof of concept.

PubMed

Vennin, Samuel; Mayer, Alexia; Li, Ye; Fok, Henry; Clapp, Brian; Alastruey, Jordi; Chowienczyk, Phil

2015-09-01

Estimation of aortic and left ventricular (LV) pressure usually requires measurements that are difficult to acquire during the imaging required to obtain concurrent LV dimensions essential for determination of LV mechanical properties. We describe a novel method for deriving aortic pressure from the aortic flow velocity. The target pressure waveform is divided into an early systolic upstroke, determined by the water hammer equation, and a diastolic decay equal to that in the peripheral arterial tree, interposed by a late systolic portion described by a second-order polynomial constrained by conditions of continuity and conservation of mean arterial pressure. Pulse wave velocity (PWV, which can be obtained through imaging), mean arterial pressure, diastolic pressure, and diastolic decay are required inputs for the algorithm. The algorithm was tested using 1) pressure data derived theoretically from prespecified flow waveforms and properties of the arterial tree using a single-tube 1-D model of the arterial tree, and 2) experimental data acquired from a pressure/Doppler flow velocity transducer placed in the ascending aorta in 18 patients (mean ± SD: age 63 ± 11 yr, aortic BP 136 ± 23/73 ± 13 mmHg) at the time of cardiac catheterization. For experimental data, PWV was calculated from measured pressures/flows, and mean and diastolic pressures and diastolic decay were taken from measured pressure (i.e., were assumed to be known). Pressure reconstructed from measured flow agreed well with theoretical pressure: mean ± SD root mean square (RMS) error 0.7 ± 0.1 mmHg. Similarly, for experimental data, pressure reconstructed from measured flow agreed well with measured pressure (mean RMS error 2.4 ± 1.0 mmHg). First systolic shoulder and systolic peak pressures were also accurately rendered (mean ± SD difference 1.4 ± 2.0 mmHg for peak systolic pressure). This is the first noninvasive derivation of aortic pressure based on fluid dynamics (flow and wave speed) in the aorta itself. Copyright © 2015 the American Physiological Society.

Leaf chlorophyll constraint on model simulated gross primary productivity in agricultural systems

NASA Astrophysics Data System (ADS)

Houborg, Rasmus; McCabe, Matthew F.; Cescatti, Alessandro; Gitelson, Anatoly A.

2015-12-01

Leaf chlorophyll content (Chll) may serve as an observational proxy for the maximum rate of carboxylation (Vmax), which describes leaf photosynthetic capacity and represents the single most important control on modeled leaf photosynthesis within most Terrestrial Biosphere Models (TBMs). The parameterization of Vmax is associated with great uncertainty as it can vary significantly between plants and in response to changes in leaf nitrogen (N) availability, plant phenology and environmental conditions. Houborg et al. (2013) outlined a semi-mechanistic relationship between Vmax25 (Vmax normalized to 25 °C) and Chll based on inter-linkages between Vmax25, Rubisco enzyme kinetics, N and Chll. Here, these relationships are parameterized for a wider range of important agricultural crops and embedded within the leaf photosynthesis-conductance scheme of the Community Land Model (CLM), bypassing the questionable use of temporally invariant and broadly defined plant functional type (PFT) specific Vmax25 values. In this study, the new Chll constrained version of CLM is refined with an updated parameterization scheme for specific application to soybean and maize. The benefit of using in-situ measured and satellite retrieved Chll for constraining model simulations of Gross Primary Productivity (GPP) is evaluated over fields in central Nebraska, U.S.A between 2001 and 2005. Landsat-based Chll time-series records derived from the Regularized Canopy Reflectance model (REGFLEC) are used as forcing to the CLM. Validation of simulated GPP against 15 site-years of flux tower observations demonstrate the utility of Chll as a model constraint, with the coefficient of efficiency increasing from 0.91 to 0.94 and from 0.87 to 0.91 for maize and soybean, respectively. Model performances particularly improve during the late reproductive and senescence stage, where the largest temporal variations in Chll (averaging 35-55 μg cm-2 for maize and 20-35 μg cm-2 for soybean) are observed. While prolonged periods of vegetation stress did not occur over the studied fields, given the usefulness of Chll as an indicator of plant health, enhanced GPP predictabilities should be expected in fields exposed to longer periods of moisture and nutrient stress. While the results support the use of Chll as an observational proxy for Vmax25, future work needs to be directed towards improving the Chll retrieval accuracy from space observations and developing consistent and physically realistic modeling schemes that can be parameterized with acceptable accuracy over spatial and temporal domains.

A Quantitative Comparison of the Behavior of Human Ventricular Cardiac Electrophysiology Models in Tissue

PubMed Central

Elshrif, Mohamed M.; Cherry, Elizabeth M.

2014-01-01

Numerical integration of mathematical models of heart cell electrophysiology provides an important computational tool for studying cardiac arrhythmias, but the abundance of available models complicates selecting an appropriate model. We study the behavior of two recently published models of human ventricular action potentials, the Grandi-Pasqualini-Bers (GPB) and the O'Hara-Virág-Varró-Rudy (OVVR) models, and compare the results with four previously published models and with available experimental and clinical data. We find the shapes and durations of action potentials and calcium transients differ between the GPB and OVVR models, as do the magnitudes and rate-dependent properties of transmembrane currents and the calcium transient. Differences also occur in the steady-state and S1–S2 action potential duration and conduction velocity restitution curves, including a maximum conduction velocity for the OVVR model roughly half that of the GPB model and well below clinical values. Between single cells and tissue, both models exhibit differences in properties, including maximum upstroke velocity, action potential amplitude, and minimum diastolic interval. Compared to experimental data, action potential durations for the GPB and OVVR models agree fairly well (although OVVR epicardial action potentials are shorter), but maximum slopes of steady-state restitution curves are smaller. Although studies show alternans in normal hearts, it occurs only in the OVVR model, and only for a narrow range of cycle lengths. We find initiated spiral waves do not progress to sustained breakup for either model. The dominant spiral wave period of the GPB model falls within clinically relevant values for ventricular tachycardia (VT), but for the OVVR model, the dominant period is longer than periods associated with VT. Our results should facilitate choosing a model to match properties of interest in human cardiac tissue and to replicate arrhythmia behavior more closely. Furthermore, by indicating areas where existing models disagree, our findings suggest avenues for further experimental work. PMID:24416228

Novel transporters from Kluyveromyces marxianus and Pichia guilliermondii expressed in Saccharomyces cerevisiae enable growth on L-arabinose and D-xylose.

PubMed

Knoshaug, Eric P; Vidgren, Virve; Magalhães, Frederico; Jarvis, Eric E; Franden, Mary Ann; Zhang, Min; Singh, Arjun

2015-10-01

Genes encoding L-arabinose transporters in Kluyveromyces marxianus and Pichia guilliermondii were identified by functional complementation of Saccharomyces cerevisiae whose growth on L-arabinose was dependent on a functioning L-arabinose transporter, or by screening a differential display library, respectively. These transporters also transport D-xylose and were designated KmAXT1 (arabinose-xylose transporter) and PgAXT1, respectively. Transport assays using L-arabinose showed that KmAxt1p has K(m) 263 mM and V(max) 57 nM/mg/min, and PgAxt1p has K(m) 0.13 mM and V(max) 18 nM/mg/min. Glucose, galactose and xylose significantly inhibit L-arabinose transport by both transporters. Transport assays using D-xylose showed that KmAxt1p has K(m) 27 mM and V(max) 3.8 nM/mg/min, and PgAxt1p has K(m) 65 mM and V(max) 8.7 nM/mg/min. Neither transporter is capable of recovering growth on glucose or galactose in a S. cerevisiae strain deleted for hexose and galactose transporters. Transport kinetics of S. cerevisiae Gal2p showed K(m) 371 mM and V(max) 341 nM/mg/min for L-arabinose, and K(m) 25 mM and V(max) 76 nM/mg/min for galactose. Due to the ability of Gal2p and these two newly characterized transporters to transport both L-arabinose and D-xylose, one scenario for the complete usage of biomass-derived pentose sugars would require only the low-affinity, high-throughput transporter Gal2p and one additional high-affinity general pentose transporter, rather than dedicated D-xylose or L-arabinose transporters. Additionally, alignment of these transporters with other characterized pentose transporters provides potential targets for substrate recognition engineering. Copyright © 2015 John Wiley & Sons, Ltd.

Microbial Group Specific Uptake Kinetics of Inorganic Phosphate and Adenosine-5′-Triphosphate (ATP) in the North Pacific Subtropical Gyre

PubMed Central

Björkman, Karin; Duhamel, Solange; Karl, David M.

2012-01-01

We investigated the concentration dependent uptake of inorganic phosphate (Pi) and adenosine-5′-triphosphate (ATP) in microbial populations in the North Pacific Subtropical Gyre (NPSG). We used radiotracers to measure substrate uptake into whole water communities, differentiated microbial size classes, and two flow sorted groups; Prochlorococcus (PRO) and non-pigmented bacteria (NPB). The Pi concentrations, uptake rates, and Pi pool turnover times (Tt) were (mean, ±SD); 54.9 ± 35.0 nmol L−1 (n = 22), 4.8 ± 1.9 nmol L−1 day−1 (n = 19), and 14.7 ± 10.2 days (n = 19), respectively. Pi uptake into >2 μm cells was on average 12 ± 7% (n = 15) of the total uptake. The kinetic response to Pi (10–500 nmol L−1) was small, indicating that the microorganisms were close to their maximum uptake velocity (Vmax). Vmax averaged 8.0 ± 3.6 nmol L−1 day−1 (n = 19) in the >0.2 μm group, with half saturation constants (Km) of 40 ± 28 nmol L−1 (n = 19). PRO had three times the cell specific Pi uptake rate of NPB, at ambient concentrations, but when adjusted to cells L−1 the rates were similar, and these two groups were equally competitive for Pi. The Tt of γ-P-ATP in the >0.2 μm group were shorter than for the Pi pool (4.4 ± 1.0 days; n = 6), but this difference diminished in the larger size classes. The kinetic response to ATP was large in the >0.2 μm class with Vmax exceeding the rates at ambient concentrations (mean 62 ± 27 times; n = 6) with a mean Vmax for γ-P-ATP of 2.8 ± 1.0 nmol L−1 day−1, and Km at 11.5 ± 5.4 nmol L−1 (n = 6). The NPB contribution to γ-P-ATP uptake was high (95 ± 3%, n = 4) at ambient concentrations but decreased to ∼50% at the highest ATP amendment. PRO had Km values 5–10 times greater than NPB. The above indicates that PRO and NPB were in close competition in terms of Pi acquisition, whereas P uptake from ATP could be attributed to NPB. This apparent resource partitioning may be a niche separating strategy and an important factor in the successful co-existence within the oligotrophic upper ocean of the NPSG. PMID:22701449

Energetic aspects of skeletal muscle contraction: implications of fiber types.

PubMed

Rall, J A

1985-01-01

In this chapter fundamental energetic properties of skeletal muscles as elucidated from isolated muscle preparations are described. Implications of these intrinsic properties for the energetic characterization of different fiber types and for the understanding of locomotion have been considered. Emphasis was placed on the myriad of physical and chemical techniques that can be employed to understand muscle energetics and on the interrelationship of results from different techniques. The anaerobic initial processes which liberate energy during contraction and relaxation are discussed in detail. The high-energy phosphate (approximately P) utilized during contraction and relaxation can be distributed between actomyosin ATPase or cross-bridge cycling (70%) and the Ca2+ ATPase of the sacroplasmic reticulum (30%). Muscle shortening increases the rate of approximately P hydrolysis, and stretching a muscle during contraction suppresses the rate of approximately P hydrolysis. The economy of an isometric contraction is defined as the ratio of isometric mechanical response to energetic cost and is shown to be a fundamental intrinsic parameter describing muscle energetics. Economy of contraction varies across the animal kingdom by over three orders of magnitude and is different in different mammalian fiber types. In mammalian skeletal muscles differences in economy of contraction can be attributed mainly to differences in the specific actomyosin and Ca2+ ATPase of muscles. Furthermore, there is an inverse relationship between economy of contraction and maximum velocity of muscle shortening (Vmax) and maximum power output. This is a fundamental relationship. Muscles cannot be economical at developing and maintaining force and also exhibit rapid shortening. Interestingly, there appears to be a subtle system of unknown nature that modulates the Vmax and economy of contraction. Efficiency of a work-producing contraction is defined and contrasted to the economy of contraction. Unlike economy, maximum efficiency of work production varies little across the animal kingdom. There are difficulties associated with the measurement of maximum efficiency of contraction, and it has yet to be determined unequivocally if the maximum efficiency of contraction varies in different fiber types. The intrinsic properties of force per cross-sectional area, economy, and Vmax determine the basic energetic properties of skeletal muscles. Nonetheless, the mechanics and energetics of skeletal muscles in the body are profoundly influenced by muscle architecture, attachment of muscles to the skeleton, and motor unit organization.(ABSTRACT TRUNCATED AT 400 WORDS)

pH regulation in barnacle muscle fibers: dependence on extracellular sodium and bicarbonate.

PubMed

Boron, W F; McCormick, W C; Roos, A

1981-01-01

Intracellular pH (pHi) regulation was studied in barnacle muscle fibers with pH-sensitive microelectrodes. The cells were acid loaded, and the subsequent recovery of pHi was monitored. The rate of recovery was reduced by one-third when external Na+ ([Na+]o) was replaced by Li+, but recovery was completely abolished when Na+ was replaced by choline or N-methyl-D-glucamine. In other experiments, varying amounts of Na+ were replaced by choline, and the acid extrusion rate, derived from the recovery rate of pHi, was calculated at a single value of pHi, 6.80. The dependence of the acid extrusion rate on [Na+]o could be described by Michaelis-Menten kinetics; at pHo (extracellular) = 8.0 and [HCO3-]o (extracellular) = 10 mM, the apparent Km and Vmax were 59 mM and 1.3 mmol x l(-1) x min-1. When [HCO3-]o was reduced to 2.5 mM at the same pHo, Km did not change significantly, but Vmax was substantially reduced. On the other hand, when pHo was reduced to 7.4 at constant [HCO3-]o, Vmax changed only slightly, but Km increased substantially. In similar experiments, we examined the dependence of the acid extrusion rate on [HCO3-]o. At pHo = 8.0 and [Na+]o = 440 mM, the apparent Km and Vmax were 4.1 mM and 2.1 mmol x 1-1 x min-1. When pHo was reduced to 7.4, Vmax was not altered, but Km substantially increased. The kinetic data are discussed in terms of the role of pHo, [Na+]o, and [HCO3-]o in the pHi-regulating system.

Predicting power-optimal kinematics of avian wings

PubMed Central

Parslew, Ben

2015-01-01

A theoretical model of avian flight is developed which simulates wing motion through a class of methods known as predictive simulation. This approach uses numerical optimization to predict power-optimal kinematics of avian wings in hover, cruise, climb and descent. The wing dynamics capture both aerodynamic and inertial loads. The model is used to simulate the flight of the pigeon, Columba livia, and the results are compared with previous experimental measurements. In cruise, the model unearths a vast range of kinematic modes that are capable of generating the required forces for flight. The most efficient mode uses a near-vertical stroke–plane and a flexed-wing upstroke, similar to kinematics recorded experimentally. In hover, the model predicts that the power-optimal mode uses an extended-wing upstroke, similar to hummingbirds. In flexing their wings, pigeons are predicted to consume 20% more power than if they kept their wings full extended, implying that the typical kinematics used by pigeons in hover are suboptimal. Predictions of climbing flight suggest that the most energy-efficient way to reach a given altitude is to climb as steeply as possible, subjected to the availability of power. PMID:25392398

Phenytoin intoxication during concurrent diazepam therapy

PubMed Central

Rogers, Howard J.; Haslam, Robert A.; Longstreth, James; Lietman, Paul S.

1977-01-01

Phenytoin elimination is a saturable process obeying Michaelis-Menten kinetics. Plasma phenytoin levels are not related linearly to dose, and small changes in enzyme activity produced by concurrent drug therapy could alter plasma levels. Two cases of phenytoin intoxication associated with simultaneous administration of diazepam are reported. Intravenous phenytoin infusions were given and the apparent Km and Vmax computed from the resulting plasma phenytoin levels. In one case `Km' and `Vmax' were 0.8 μmol/1 and 1.3 μmol/1/hour respectively during concurrent diazepam administration, and 50.3 μmol/1 and 4.4 μmol/1/hour after discontinuation of diazepam. In the second case phenytoin infusion with diazepam gave `Km' and `Vmax' values of 0.012 μmol/1 and 0.95 μmol/1/hour. Without diazepam these were 28.8 μmol/1 and 0.92 μmol/1/hour respectively. PMID:599366

Effects of deoxygenation on active and passive Ca2+ transport and cytoplasmic Ca2+ buffering in normal human red cells.

PubMed Central

Tiffert, T; Etzion, Z; Bookchin, R M; Lew, V L

1993-01-01

1. The effects of deoxygenation on cytoplasmic Ca2+ buffering, saturated Ca2+ extrusion rate through the Ca2+ pump (Vmax), passive Ca2+ influx and physiological [Ca2+]i level were investigated in human red cells to assess whether or not their Ca2+ metabolism might be altered by deoxygenation in capillaries and venous circulation. 2. The study was performed in fresh human red cells maintained in a tonometer either fully oxygenated or deoxygenated. Cytoplasmic Ca2+ buffering was estimated from the equilibrium distribution of 45Ca2+ induced by the divalent cation ionophore A23187 and the Vmax of the Ca2+ pump was measured either by the Co(2+)-exposure method or following ionophore wash-out. The passive Ca2+ influx and physiological [Ca2+]i were determined in cells preloaded with the Ca2+ chelator benz-2 and resuspended in autologous plasma. 3. Deoxygenation increased the fraction of ionized Ca2+ in cell water by 34-74% and reduced the Vmax of the Ca2+ pump by 18-32%. 4. To elucidate whether or not these effects were secondary to deoxygenation-induced pH shifts, the effects of deoxygenation on cell and medium pH, and of pH on cytoplasmic Ca2+ binding and Ca2+ pump Vmax in oxygenated cells were examined in detail. 5. Deoxygenation generated large alkaline pH shifts that could be explained if the apparent isoelectric point (pI) of haemoglobin increased by 0.2-0.4 pH units in intact cells, consistently higher than the value of 0.15 reported for pure haemoglobin solutions. 6. In oxygenated cells, the fraction of ionized cell calcium, alpha, was little affected by pH within the 7.0-7.7 range. Ca2+ pump Vmax was maximal at a medium pH of about 7.55. Comparison between pH effects elicited by HCl-NaOH additions and by replacing Cl- with gluconate suggested that Vmax was inhibited by both internal acidification and external alkalinization. Since deoxygenation alkalinized cells and medium within a range stimulatory for Vmax, the inhibition observed was not due to pH. 7. There was no significant effect of deoxygenation on passive Ca2+ uptake, or steady-state physiological [Ca2+]i level. 8. The deoxygenation-induced reduction in Ca2+ binding capacity may result from the increased protonation of haemoglobin on deoxygenation and from binding of 2,3-diphosphoglyceric acid (2,3-DPG) and ATP to deoxyhaemoglobin; inhibition of the Ca2+ pump may result from shifts in the [Mg2+]i/[ATP]i ratio away from a near optimal stimulatory value in the oxygenated state. PMID:8229816

Simplified numerical approach for estimation of effective segregation coefficient at the melt/crystal interface

NASA Astrophysics Data System (ADS)

Prostomolotov, A. I.; Verezub, N. A.; Voloshin, A. E.

2014-09-01

A thermo-gravitational convection and impurity transfer in the melt were investigated using a simplified numerical model for Bridgman GaSb(Te) crystal growth in microgravity conditions. Simplifications were as follows: flat melt/crystal interface, fixed melt sizes and only lateral ampoule heating. Calculations were carried out by Ansys®Fluent® code employing a two-dimensional Navier-Stokes-Boussinesq and heat and mass transfer equations in a coordinate system moving with the melt/crystal interface. The parametric dependence of the effective segregation coefficient Keff at the melt/crystal interface was studied for various ampoule sizes and for microgravity conditions. For the uprising one-vortex flow, the resulting dependences were presented as Keff vs. Vmax-the maximum velocity value. These dependences were compared with the formulas by Burton-Prim-Slichter's, Ostrogorsky-Muller's, as well as with the semi-analytical solutions.

Formation of a Malin 1 analogue in IllustrisTNG by stimulated accretion

NASA Astrophysics Data System (ADS)

Zhu, Qirong; Xu, Dandan; Gaspari, Massimo; Rodriguez-Gomez, Vicente; Nelson, Dylan; Vogelsberger, Mark; Torrey, Paul; Pillepich, Annalisa; Zjupa, Jolanta; Weinberger, Rainer; Marinacci, Federico; Pakmor, Rüdiger; Genel, Shy; Li, Yuexing; Springel, Volker; Hernquist, Lars

2018-06-01

The galaxy Malin 1 contains the largest stellar disk known but the formation mechanism of this structure has been elusive. In this paper, we report a Malin 1 analogue in the 100 Mpc IllustrisTNG simulation and describe its formation history. At redshift zero, this massive galaxy, having a maximum circular velocity Vmax of 430 km s-1, contains a 100 kpc gas/stellar disk with morphology similar to Malin 1. The simulated galaxy reproduces well many observed features of Malin 1's vast disk, including its stellar ages, metallicities, and gas rotation curve. We trace the extended disk back in time and find that a large fraction of the cold gas at redshift zero originated from the cooling of hot halo gas, triggered by the merger of a pair of intruding galaxies. Our finding provides a novel way to form large galaxy disks as extreme as Malin 1 within the current galaxy formation framework.

[Effects of dauricine on action potentials and slow inward currents of guinea pig ventricular papillary muscles].

PubMed

Li, S N; Zhang, K Y

1992-11-01

Effects of dauricine (Dau) on the action potentials (AP), the slow action potentials (SAP), and the slow inward currents (Isi) of guinea pig ventricular papillary muscles were observed by means of intracellular microelectrode and single sucrose gap voltage clamp technique. In the early stage, Dau shortened action potential duration 100 (APD100) and effective refractory period (ERP) (ERP/APD < 1; P < 0.01), but did not affect APD20 and other parameters. In the late stage, Dau prolonged APD100, ERP, and APD20, significantly decreased action potential amplitude (APA), maximum velocity (Vmax), and overshot (OS) (ERP/APD > 1; P < 0.01), greatly diminished APA and OS of SAP induced by isoprenaline (P < 0.01), and remarkably inhibited Isi (P < 0.01). The results suggested that Dau exerted an inhibitory effect on Na+, Ca2+, and K+ channels.

Mind the Gap: A Semicontinuum Model for Discrete Electrical Propagation in Cardiac Tissue.

PubMed

Costa, Caroline Mendonca; Silva, Pedro Andre Arroyo; dos Santos, Rodrigo Weber

2016-04-01

Electrical propagation in cardiac tissue is a discrete or discontinuous phenomenon that reflects the complexity of the anatomical structures and their organization in the heart, such as myocytes, gap junctions, microvessels, and extracellular matrix, just to name a few. Discrete models or microscopic and discontinuous models are, so far, the best options to accurately study how structural properties of cardiac tissue influence electrical propagation. These models are, however, inappropriate in the context of large scale simulations, which have been traditionally performed by the use of continuum and macroscopic models, such as the monodomain and the bidomain models. However, continuum models may fail to reproduce many important physiological and physiopathological aspects of cardiac electrophysiology, for instance, those related to slow conduction. In this study, we develop a new mathematical model that combines characteristics of both continuum and discrete models. The new model was evaluated in scenarios of low gap-junctional coupling, where slow conduction is observed, and was able to reproduce conduction block, increase of the maximum upstroke velocity and of the repolarization dispersion. None of these features can be captured by continuum models. In addition, the model overcomes a great disadvantage of discrete models, as it allows variation of the spatial resolution within a certain range.

Ankle-Brachial Index, Toe-Brachial Index, and Pulse Volume Recording in Healthy Young Adults

PubMed Central

Masaki, Hisao; Yunoki, Yasuhiro; Tabuchi, Atushi; Morita, Ichiro; Mohri, Satoshi; Tanemoto, Kazuo

2015-01-01

Objective: To clarify the characteristics of ankle-brachial index (ABI), toe-brachial index (TBI), and pulse volume recording (PVR) of the ankle with brachial-ankle pulse wave velocity (baPWV) in healthy young adults. Material and Methods: We analyzed ABI, TBI, baPWV, and PVR in the ankle of healthy adults aged 20 to 25 years (median, 20 years) using an automatic oscillometric device between 2002 and 2013. The ABI, baPWV, and PVR in 1282 legs of 641 subjects (301 men and 340 women) and the TBI in 474 toes of 237 subjects (117 men and 120 women) were evaluated. Results: The measured values showed no bilateral differences. ABI and baPWV were higher in men than in women, but TBI was similar in both sexes. ABI <1.0 was observed in 18.1% of the legs in men and in 25.6% in women. TBI <0.7 was observed in 16.2% of the toes in men and 19.1% in women. For ankle PVR, the % mean arterial pressure was higher in women than in men. The upstroke time was <180 ms in most subjects. Conclusions: For young people, ABI <1.0 or TBI <0.7 may not always indicate vascular abnormalities. When evaluating circulatory indexes, age and sex should be considered. PMID:26421072

Seasonal acclimatization of the cardiac action potential in the Arctic navaga cod (Eleginus navaga, Gadidae).

PubMed

Hassinen, Minna; Abramochkin, Denis V; Vornanen, Matti

2014-04-01

Freshwater fishes of north-temperate latitudes adjust electrical excitability of the heart to seasonal temperature changes by changing expression levels of ion channel isoforms. However, little is known about thermal responses of action potential (AP) in the hearts of marine polar fishes. To this end, we examined cardiac AP in the atrial myocardium of the Arctic navaga cod (Eleginus navaga) from the White Sea (Russia) acclimatized to winter (March) and summer (September) seasons. Acute increases in temperature from 4 to 10 °C were associated with increases in heart rate, maximum velocity of AP upstroke and negative resting membrane potential, while duration of AP was shortened in both winter-acclimatized and summer-acclimatized navaga hearts. In winter, there was a compensatory shortening (41.1%) of atrial AP duration and this was associated with a strong increase in transcript expression of Erg K(+) channels, known to produce the rapid component of the delayed rectifier K(+) current, I(Kr). Smaller increases were found in the expression of Kir2.1 channels that produce the inward rectifier K(+) current, I(K1). These findings indicate that the heart of navaga cod has a good acclimatory capacity in electrical excitation of cardiac myocytes, which enables cardiac function in the cold-eurythermal waters of the subarctic White Sea.

Na+-dependent and Na+-independent betaine transport across the apical membrane of rat renal epithelium.

PubMed

Cano, Mercedes; Calonge, María L; Ilundáin, Anunciación A

2015-10-01

The low renal excretion of betaine indicates that the kidney efficiently reabsorbs the betaine filtered by the glomeruli but the mechanisms involved in such a process have been scarcely investigated. We have detected concentrative and non-concentrative betaine transport activity in brush-border membrane vesicles (BBMV) from rat renal cortex and medulla. The concentrative system is the Sodium/Imino-acid Transporter 1 (SIT1) because it is Na+- and Cl--dependent, electrogenic and is inhibited by an anti-SIT1 antibody. Its apparent affinity constant for betaine, Kt, is 1.1±0.5 mM and its maximal transport velocity, Vmax, 0.5±0.1 nmol betaine/mg protein/s. Inhibitors of the Na+/Cl-/betaine uptake are L-proline (75%) and cold betaine, L-carnitine and choline (40-60%). Neither creatine, TEA, taurine, β-alanine, GABA nor glycine significantly inhibited Na+/Cl-/betaine uptake. The non-concentrative betaine transport system is Na+- and H+-independent, electroneutral, with a Kt for betaine of 47±7 μM and a Vmax of 7.8±1 pmol betaine/mg protein/s. Its transport activity is nearly abolished by betaine, followed by L-carnitine (70-80%) and proline (40-50%), but a difference from the Na+/Cl-/betaine transport is that it is inhibited by TEA (approx. 50%) and unaffected by choline. The underlying carrier functions as an antiporter linking betaine entry into the BBMV with the efflux of either L-carnitine or betaine, an exchange unaffected by the anti-SIT1 antibody. As far as we know this is the first work reporting that betaine crosses the apical membrane of rat renal epithelium by SIT1 and by a Na+- and H+-independent transport system. Copyright © 2015 Elsevier B.V. All rights reserved.

Critical evaluation of a badminton-specific endurance test.

PubMed

Fuchs, Michael; Faude, Oliver; Wegmann, Melissa; Meyer, Tim

2014-03-01

To overcome the limitations of traditional 1-dimensional fitness tests in analyzing physiological properties of badminton players, a badminton-specific endurance test (BST) was created. This study aimed at analyzing the influence of various fitness dimensions on BST performance. 18 internationally competing male German badminton players (22.4 ± 3.2 y, 79.2 ± 7.7 kg, 1.84 ± 0.06 m, world-ranking position [WRP] 21-501) completed a straight-sprint test, a change-of-direction speed test, various jump tests (countermovement jump, drop jump, standing long jump), a multistage running test (MST), and the BST. During this on-court field test players have to respond to a computerized sign indicating direction and speed of badminton-specific movements by moving into the corresponding corners. Significant correlations were found between performance in MST and BST (individual anaerobic threshold [IAT], r = .63, P = .005; maximum velocity [Vmax], r = .60, P = .009). A negative correlation (r = -.59, P = .014) was observed between IAT in BST and drop-jump contact time. No further associations between performance indices could be detected. Apart from a small portion explained by MST results (IAT, R2 = .40; Vmax, R2 = .36), the majority of BST performance cannot be explained by the determined physiological correlates. Moreover, it was impossible to predict the WRP of a player on the basis of BST results (r = -.15, P = .55). Neither discipline-specific performance nor basic physiological properties were appropriately reflected by a BST in elite badminton players. This does not substantiate its validity for regular use as a testing tool. However, it may be useful for monitoring on-court training sessions.

Numerical simulation of oxygen delivery to muscle tissue in the presence of hemoglobin-based oxygen carriers.

PubMed

Patton, Jaqunda N; Palmer, Andre F

2006-01-01

This work represents a culmination of research on oxygen transport to muscle tissue, which takes into account oxygen transport due to convection, diffusion, and the kinetics of simultaneous reactions between oxygen and hemoglobin and myoglobin. The effect of adding hemoglobin-based oxygen carriers (HBOCs) to the plasma layer of blood in a single capillary surrounded by muscle tissue based on the geometry of the Krogh tissue cylinder is examined for a range of HBOC oxygen affinity, HBOC concentration, capillary inlet oxygen tension (pO(2)), and hematocrit. The full capillary length of the hamster retractor muscle was modeled under resting (V(max) = 1.57 x 10(-4) mLO(2) mL(-1) s(-1), cell velocity (v(c)) = 0.015 cm/s) and working (V(max) = 1.57 x 10(-3) mLO(2) mL(-1) s(-1), v(c) = 0.075 cm/s) conditions. Two spacings between the red blood cell (RBC) and the capillary wall were examined, corresponding to a capillary with and without an endothelial surface layer. Simulations led to the following conclusions, which lend physiological insight into oxygen transport to muscle tissue in the presence of HBOCs: (1) The reaction kinetics between oxygen and myoglobin in the tissue region, oxygen and HBOCs in the plasma, and oxygen and RBCs in the capillary lumen should not be neglected. (2) Simulation results yielded new insight into possible mechanisms of oxygen transport in the presence of HBOCs. (3) HBOCs may act as a source or sink for oxygen in the capillary and may compete with RBCs for oxygen. (4) HBOCs return oxygen delivery to muscle tissue to normal for varying degrees of hypoxia (inlet capillary pO(2) < 30 mmHg) and anemia (hematocrit < 46%) for the hamster model.

[Origin of motion in the human ureter: mechanics, energetics and kinetics of the myosin molecular motors].

PubMed

Vargiu, Romina; Perinu, Anna; De Lisa, Antonello; Tintrup, Frank; Manca, Francesco; Mancinelli, Rino

2012-01-01

Ureteral peristalsis is the result of coordinated mechanical motor performance of longitudinal and circular smooth muscle layer of the ureter wall. The main aim of this study was to characterize in smooth muscle of proximal segments of human ureter, the mechanical properties at level of muscle tissue and at level of myosin molecular motors. Ureteral samples were collected from 15 patients, who underwent nephrectomy for renal cancer. Smooth muscle strips longitudinally and circularly oriented from proximal segments of human ureter were used for the in vitro experiments. Mechanical indices including the maximum unloaded shortening velocity (Vmax), and the maximum isometric tension (P0) normalized per cross-sectional area, were determined in vitro determined in electrically evoked contractions of longitudinal and circular smooth muscle strips. Myosin cross-bridge (CB) number per mm2 (Ψ) the elementary force per single CB (Ψ) and kinetic parameters were calculated in muscle strips, using Huxley's equations adapted to nonsarcomeric muscles. Longitudinal smooth muscle strips exhibited a significantly (p<0.05) faster Vmax (63%) and a higher P0 (40%), if compared to circular strips. Moreover, longitudinal muscle strips showed a significantly higher unitary force (Ψ) per CB. However, no significant differences were observed in CB number, the attachment (f1) and the detachment (g2) rate constants between longitudinal and circular muscle strips. The main result obtained in the present work documents that the mechanical, energetic and unitary forces per CB of longitudinal layer of proximal ureter are better compared to the circular one; these preliminary findings suggested, unlike intestinal smooth muscle, a major role of longitudinal smooth muscle layer in the ureter peristalsis.

Administration of progesterone after trauma and hemorrhagic shock prevents hepatocellular injury.

PubMed

Kuebler, Joachim F; Yokoyama, Yukihiro; Jarrar, Doraid; Toth, Balazs; Rue, Loring W; Bland, Kirby I; Wang, Ping; Chaudry, Irshad H

2003-07-01

Administration of a single dose of progesterone following trauma and hemorrhage in progesterone-deficient rats would ameliorate the inflammatory response and hepatocellular damage. A university laboratory. Ovariectomized female Sprague-Dawley rats (250-350 g; Charles River Laboratories, Wilmington, Mass) underwent a 5-cm midline laparotomy (ie, induction of soft tissue trauma), were bled to a mean arterial blood pressure of 35 mm Hg for about 90 minutes, and then were resuscitated using Ringer lactate solution. Progesterone (25 mg/kg of body weight) or vehicle was administered subcutaneously at the end of resuscitation. In additional animals, Kupffer cells were isolated following trauma, hemorrhage, and resuscitation and treated in vitro with progesterone, lipopolysaccharide, or both. Six hours following resuscitation, plasma tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) levels and liver myeloperoxidase activity were determined. Hepatocellular function (maximum velocity of indocyanine green clearance [Vmax] and the efficiency of the active transport or Michaelis-Menten constant [Km]) and plasma levels of transaminases were measured 20 hours after resuscitation. Kupffer cell IL-6 and TNF-alpha production were assessed. Plasma levels of TNF-alpha, IL-6, aspartate aminotransferase, and alanine aminotransferase, as well as hepatic myeloperoxidase activity were increased, whereas indocyanine green clearance was depressed in vehicle-treated rats following trauma-hemorrhage. Animals treated with progesterone showed significantly reduced levels of the TNF-alpha, IL-6, and transaminases as well as reduced myeloperoxidase activity in the liver. Progesterone-treated animals showed increased Vmax and Kmax values for indocyanine green. In vitro treatment of Kupffer cells with progesterone decreased TNF-alpha production but did not affect the production of IL-6. Progesterone administration following trauma-hemorrhage ameliorates the proinflammatory response and, subsequently, the hepatocellular injury via direct action on immunocompetent cells.

Is gut the "motor" for producing hepatocellular dysfunction after trauma and hemorrhagic shock?

PubMed

Wang, P; Ba, Z F; Cioffi, W G; Bland, K I; Chaudry, I H

1998-02-01

Although studies suggest that the gut may be the "motor" responsible for producing sepsis and multiple organ failure after injury, it is not known whether enterectomy prior to the onset of hemorrhage alters proinflammatory cytokines TNF and IL-6 and, if so, whether hepatocellular dysfunction and damage are prevented or attenuated under such conditions. Under methoxyflurane anesthesia, an enterectomy in the rat was performed by excision of the duodenum, jejunum, and ileum. The rats were then bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the maximal shed volume was returned in the form of Ringer's lactate. The animals were then resuscitated with four times the volume of shed blood with Ringer's lactate over 1 h. At 1.5 h after the completion of resuscitation, hepatocellular function [i.e., the maximal velocity (Vmax) and transport efficiency (Km) of indocyanine green (ICG) clearance] was assessed by an in vivo ICG clearance technique. Blood samples were taken for the measurement of TNF, IL-6, and liver enzymes (i.e., SGPT and SGOT). Cardiac output and microvascular blood flow were determined by ICG dilution and laser Doppler flowmetry, respectively. The increase in circulating levels of TNF but not IL-6 was prevented by enterectomy prior to hemorrhage. The reduced Vmax and K(m) and elevated SGPT and SGOT following hemorrhage and resuscitation, however, were not significantly affected by prior enterectomy. Moreover, enterectomy before hemorrhage further reduced hepatic perfusion. Since enterectomy prior to the onset of hemorrhage does not prevent or attenuate the reduced ICG clearance and elevated liver enzymes despite downregulation of TNF production, it appears that the small intestine does not play a significant role in producing hepatocellular dysfunction and injury following trauma and hemorrhagic shock.

High- and Low-Order Overtaking-Ability Affordances: Drivers Rely on the Maximum Velocity and Acceleration of Their Cars to Perform Overtaking Maneuvers.

PubMed

Basilio, Numa; Morice, Antoine H P; Marti, Geoffrey; Montagne, Gilles

2015-08-01

The aim of this study was to answer the question, Do drivers take into account the action boundaries of their car when overtaking? The Morice et al. affordance-based approach to visually guided overtaking suggests that the "overtake-ability" affordance can be formalized as the ratio of the "minimum satisfying velocity" (MSV) of the maneuver to the maximum velocity (V(max)) of the driven car. In this definition, however, the maximum acceleration (A(max)) of the vehicle is ignored. We hypothesize that drivers may be sensitive to an affordance redefined with the ratio of the "minimum satisfying acceleration" (MSA) to the A(max) of the car. Two groups of nine drivers drove cars differing in their A(max). They were instructed to attempt overtaking maneuvers in 25 situations resulting from the combination of five MSA and five MSV values. When overtaking frequency was expressed as a function of MSV and MSA, maneuvers were found to be initiated differently for the two groups. However, when expressed as a function of MSV/V(max) and MSA/A(max), overtaking frequency was quite similar for both groups. Finally, a multiple regression coefficient analysis demonstrated that overtaking decisions are fully explained by a composite variable comprising MSA/A(max) and the time required to reach MSV. Drivers reliably decide whether overtaking is safe (or not) by using low- and high-order variables taking into account their car's maximum velocity and acceleration, respectively, as predicted by "affordance-based control" theory. Potential applications include the design of overtaking assistance, which should exploit the MSA/A(max) variables in order to suggest perceptually relevant overtaking solutions. © 2015, Human Factors and Ergonomics Society.

ATP sulfurylase from higher plants: kinetic and structural characterization of the chloroplast and cytosol enzymes from spinach leaf.

PubMed

Renosto, F; Patel, H C; Martin, R L; Thomassian, C; Zimmerman, G; Segel, I H

1993-12-01

Two forms of ATP sulfurylase were purified from spinach leaf. The major (chloroplast) form accounts for 85 to 90% of the total leaf activity (0.03 +/- 0.01 adenosine-5'-phosphosulfate (APS) synthesis units x gram fresh weight-1). Both enzyme forms appear to be tetramers composed of 49- to 50-kDa subunits with the minor (cytosolic) form being slightly larger than the chloroplast form. The specific activities (units x milligram protein-1) of the chloroplast form at pH 8.0, 30 degrees C, were as follows: APS synthesis, 16; molybdolysis, 229; ATP synthesis, 267; selenolysis, 4.1; fluorophosphate activation, 11. Kinetic constants for the physiological reaction were as follows: KmA = 0.046 mM, K(ia) = 0.85 mM, KmB = 0.25 mM, KmQ = 0.37 microM, K(iq) = 64-85 nM, and KmP = 10 microM, where A = MgATP, B = SO4(2-), P = total PPi at 5 mM Mg2+, and Q = APS. The kinetic constants for molybdolysis were similar to those of the APS synthesis reaction. The kinetic constants of the minor (cytosol) form were similar to those of the major form with two exceptions: (a) The molybdolysis activity was 120 units x milligram protein-1, yielding a Vmax (ATP synthesis)/Vmax (molybdolysis) ratio close to 2 (compared to about unity for the chloroplast form) and (b) KmA was greater (0.24 and 0.15 mM for APS synthesis and molybdolysis, respectively). Initial velocity measurements (made over an extended range of MgATP and SO4(2-) concentrations), product inhibition studies (by initial velocity methods and by reaction progress curve analyses), dead end inhibition studies (with monovalent and divalent oxyanions), and kcat/Km comparisons (for SO4(2-) and MoO4(2-) support a random AB-ordered PQ kinetic mechanism in which MgATP and SO4(2-) bind in a highly synergistic manner. Equilibrium binding studies indicated the presence of one APS site per subunit. HPLC elution profiles of chymotryptic and tryptic peptides were essentially the same for both enzyme forms. The N-terminal sequence of residues 5-20 of the cytosol enzyme was identical to residues 1-16 of the chloroplast enzyme.

Kinetic properties of wild-type and altered recombinant amidases by the use of ion-selective electrode assay method.

PubMed

Martins, S; Karmali, A; Serralheiro, M L

2006-08-15

A novel assay method was investigated for wild-type and recombinant mutant amidases (EC 3.5.1.4) from Pseudomonas aeruginosa by ammonium ion-selective electrode (ISE). The initial velocity is proportional to the enzyme concentration by using the wild-type enzyme. The specific activities of the purified amidase were found to be 88.2 and 104.2 U mg protein(-1) for the linked assay and ISE methods, respectively. The kinetic constants--Vmax, Km, and Kcat--determined by Michaelis-Menten plot were 101.13 U mg protein(-1), 1.12x10(-2) M, and 64.04 s(-1), respectively, for acrylamide as the substrate. On the other hand, the lower limit of detection and range of linearity of enzyme concentration were found to be 10.8 and 10.8 to 500 ng, respectively, for the linked assay method and 15.0 and 15.0 to 15,000 ng, respectively, for the ISE method. Hydroxylamine was found to act as an uncompetitive activator of hydrolysis reaction catalyzed by amidase given that there is an increase in Vmax and Km when acetamide was used as the substrate. However, the effect of hydroxylamine on the hydrolysis reaction was dependent on the type of amidase and substrate involved in the reaction mixture. The degrees of activation (epsilon(a)) of the wild-type and mutant (T103I and C91A) enzymes were found to be 2.54, 12.63, and 4.33, respectively, for acetamide as the substrate. However, hydroxylamine did not activate the reaction catalyzed by wild-type and altered (C91A and W138G) amidases by using acrylamide and acetamide, respectively, as the substrate. The activating effect of hydroxylamine on the hydrolysis of acetamide, acrylamide, and p-nitrophenylacetamide can be explained by the fact that additional formation of ammonium ions occurred due to the transferase activity of amidases. However, the activating effect of hydroxylamine on the hydrolysis of p-nitroacetanilide may be due to a change in conformation of enzyme molecule. Therefore, the use of ISE permitted the study of the kinetic properties of wild-type and mutant amidases because it was possible to measure initial velocity of the enzyme-catalyzed reaction in real time.

Active Recovery After High-Intensity Interval-Training Does Not Attenuate Training Adaptation.

PubMed

Wiewelhove, Thimo; Schneider, Christoph; Schmidt, Alina; Döweling, Alexander; Meyer, Tim; Kellmann, Michael; Pfeiffer, Mark; Ferrauti, Alexander

2018-01-01

Objective: High-intensity interval training (HIIT) can be extremely demanding and can consequently produce high blood lactate levels. Previous studies have shown that lactate is a potent metabolic stimulus, which is important for adaptation. Active recovery (ACT) after intensive exercise, however, enhances blood lactate removal in comparison with passive recovery (PAS) and, consequently, may attenuate endurance performance improvements. Therefore, the aim of this study was to examine the influence of regular ACT on training adaptations during a HIIT mesocycle. Methods: Twenty-six well-trained male intermittent sport athletes (age: 23.5 ± 2.5 years; O 2 max: 55.36 ± 3.69 ml min kg -1 ) participated in a randomized controlled trial consisting of 4 weeks of a running-based HIIT mesocycle with a total of 12 HIIT sessions. After each training session, participants completed 15 min of either moderate jogging (ACT) or PAS. Subjects were matched to the ACT or PAS groups according to age and performance. Before the HIIT program and 1 week after the last training session, the athletes performed a progressive incremental exercise test on a motor-driven treadmill to determine O 2 max, maximum running velocity (vmax), the running velocity at which O 2 max occurs (vO 2 max), and anaerobic lactate threshold (AT). Furthermore, repeated sprint ability (RSA) were determined. Results: In the whole group the HIIT mesocycle induced significant or small to moderate changes in vmax ( p < 0.001, effect size [ES] = 0.65,), vO 2 max ( p < 0.001, ES = 0.62), and AT ( p < 0.001, ES = 0.56) compared with the values before the intervention. O 2 max and RSA remained unchanged throughout the study. In addition, no significant differences in the changes were noted in any of the parameters between ACT and PAS except for AT ( p < 0.05, ES = 0.57). Conclusion: Regular use of individualized ACT did not attenuate training adaptations during a HIIT mesocycle compared to PAS. Interestingly, we found that the ACT group obtained a significantly higher AT following the training program compared to the PAS group. This could be because ACT allows a continuation of the training at a low intensity and may activate specific adaptive mechanisms that are not triggered during PAS.

Active Recovery After High-Intensity Interval-Training Does Not Attenuate Training Adaptation

PubMed Central

Wiewelhove, Thimo; Schneider, Christoph; Schmidt, Alina; Döweling, Alexander; Meyer, Tim; Kellmann, Michael; Pfeiffer, Mark; Ferrauti, Alexander

2018-01-01

Objective: High-intensity interval training (HIIT) can be extremely demanding and can consequently produce high blood lactate levels. Previous studies have shown that lactate is a potent metabolic stimulus, which is important for adaptation. Active recovery (ACT) after intensive exercise, however, enhances blood lactate removal in comparison with passive recovery (PAS) and, consequently, may attenuate endurance performance improvements. Therefore, the aim of this study was to examine the influence of regular ACT on training adaptations during a HIIT mesocycle. Methods: Twenty-six well-trained male intermittent sport athletes (age: 23.5 ± 2.5 years; O2max: 55.36 ± 3.69 ml min kg-1) participated in a randomized controlled trial consisting of 4 weeks of a running-based HIIT mesocycle with a total of 12 HIIT sessions. After each training session, participants completed 15 min of either moderate jogging (ACT) or PAS. Subjects were matched to the ACT or PAS groups according to age and performance. Before the HIIT program and 1 week after the last training session, the athletes performed a progressive incremental exercise test on a motor-driven treadmill to determine O2max, maximum running velocity (vmax), the running velocity at which O2max occurs (vO2max), and anaerobic lactate threshold (AT). Furthermore, repeated sprint ability (RSA) were determined. Results: In the whole group the HIIT mesocycle induced significant or small to moderate changes in vmax (p < 0.001, effect size [ES] = 0.65,), vO2max (p < 0.001, ES = 0.62), and AT (p < 0.001, ES = 0.56) compared with the values before the intervention. O2max and RSA remained unchanged throughout the study. In addition, no significant differences in the changes were noted in any of the parameters between ACT and PAS except for AT (p < 0.05, ES = 0.57). Conclusion: Regular use of individualized ACT did not attenuate training adaptations during a HIIT mesocycle compared to PAS. Interestingly, we found that the ACT group obtained a significantly higher AT following the training program compared to the PAS group. This could be because ACT allows a continuation of the training at a low intensity and may activate specific adaptive mechanisms that are not triggered during PAS. PMID:29720949

Effects of fasting and semistarvation on the kinetics of active and passive sugar absorption across the small intestine in vivo.

PubMed Central

Debnam, E S; Levin, R J

1975-01-01

The effects of dietary restriction on the kinetics of absorption in vivo of glucose, galactose and alpha-methyl glucoside were assessed by electrical and chemical methods in the rat jejunum. 2. The 'apparent Km', maximum absorption or Vmax (mu-mole/10 cm. 15 min) and maximum potential difference (p.d.max) were obtained for the jejunal electrogenic active transfer mechanism from the transfer p.d.s and the chemical absorption data corrected for diffusion using various graphical kinetic plots. 3. Fasting for 3 days greatly decreased the 'apparent Kms', obtained from electrical or chemical data, for all the sugars but had no effect on those for L-valine or L-methionine. Semistarvation caused a less pronounced reduction of the 'apparent Kms' for the sugars. The dietary-induced change in 'apparent Km' for glucose was also observed in the fasted hamster. One interpretation of these changes is that the affinity of the carriers for sugars increases during dietary restriction; the greater the level of restriction the greater the increase. 4. Fasting and semistarvation caused large reductions in the Vmax. These reductions were correlated with a reduced enterocyte population estimated by changes in enterocyte column size. 5. The reduction in the Vmax for galactose was mainly accounted for by the decrease in enterocyte population. In the case of glucose, other factors such as reduced enterocyte metabolism or changes in the carriers must be involved to explain the discrepancy between the large decrease in Vmax and the enterocyte column size. 6. Fasting and semi-starvation had complex, differential actions on the p.d.max for glucose, galactose and alpha-methyl glucoside. These changes did not correlate with those observed in the Vmax measured chemically. 7. A standard diet obtained from two commercial sources was found to differ greatly in its effect on the electrogenic transfer system for alpha-methyl glucoside but had no effect on those for galactose and glucose. PMID:1206572

How Insects Initiate Flight: Computational Analysis of a Damselfly in Takeoff Flight

NASA Astrophysics Data System (ADS)

Bode-Oke, Ayodeji; Zeyghami, Samane; Dong, Haibo; Flow Simulation Research Group Team

2017-11-01

Flight initiation is essential for survival in biological fliers and can be classified into jumping and non-jumping takeoffs. During jumping takeoffs, the legs generate most of the initial impulse. Whereas the wings generate most of the forces in non-jumping takeoffs, which are usually voluntary, slow, and stable. It is of interest to understand how non-jumping takeoffs occur and what strategies insects use to generate the required forces. Using a high fidelity computational fluid dynamics simulation, we identify the flow features and compute the wing aerodynamic forces to elucidate how flight forces are generated by a damselfly performing a non-jumping takeoff. Our results show that a damselfly generates about three times its bodyweight during the first half-stroke for liftoff while flapping through a steeply inclined stroke plane and slicing the air at high angles of attack. Consequently, a Leading Edge Vortex (LEV) is formed during both the downstroke and upstroke on all the four wings. The formation of the LEV, however, is inhibited in the subsequent upstrokes following takeoff. Accordingly, we observe a drastic reduction in the magnitude of the aerodynamic force, signifying the importance of LEV in augmenting force production. This work was supported by National Science Foundation [CBET-1313217] and Air Force Research Laboratory [FA9550-12-1-007].

Characteristics of butanol metabolism in alcohol dehydrogenase-deficient deermice.

PubMed Central

Alderman, J A; Kato, S; Lieber, C S

1989-01-01

Deermice lacking the low-Km alcohol dehydrogenase eliminated butan-1-ol, a substrate for microsomal oxidation but not for catalase, at 117 mumol/min per kg body wt. Microsomal fractions and hepatocytes metabolized butan-1-ol also (Vmax. = 6.7 nmol/min per nmol of cytochrome P-450, Km = 0.85 mM; Vmax. = 5.3 nmol/min per 10(6) cells, Km = 0.71 mM respectively). These results are consistent with alcohol oxidation by the microsomal system in these deermice. PMID:2930472

WPVS 007: Dramatic Broad Absorption Line Variability in a Narrow-line Seyfert 1

NASA Astrophysics Data System (ADS)

Cooper, Erin M.; Leighly, K.; Hamann, F. W.; Grupe, D.; Dietrich, M.

2014-01-01

Blue-shifted broad absorption lines are the manifestation of gaseous outflows in astrophysical phenomena. In active galaxies, these outflowing winds may play a key role in the central engine physics by removing angular momentum and in influencing host galaxy evolution by imparting energy and chemically enriched gas to the surrounding medium. AGN wind variability affords us a valuable tool to study this still poorly understood phenomenon. The existence of a high velocity broad line outflow in WPVS007 is especially extraordinary, as Seyfert-luminosity active galaxies are unexpected to produce them. With its lower luminosity and compact size, the NLS1 galaxy WPVS007 (M_V=-19.7, z=0.02882) provides us the ability to study even colossal variability on merely human timescales. Since its 1996 FOS observation, displaying miniBALs but no true broad absorption lines, WPVS007 has experienced a short but rich history of UV BAL variability. By the 2003 FUSE observation, WPVS007 had developed a BAL with v_max ~ 6000km/s, indicating an optically thick, high velocity outflow. We present the 2010 and 2013 June and December HST COS spectra. Between 2003 and 2010, both the maximum and minimum outflow velocity had increased substantially. As of 2013 June, the continuum emission has since dimmed by a factor of ~2 and the BALs have appeared to weaken, with both decreased maximum and minimum velocities. Such dramatic shifts in BAL velocity are unprecedented, as BAL variability is typically confined to changes in optical depth. What is the nature of the variability in this BAL wind? The upcoming (as of the writing of this abstract) December observation should give us more insight into tackling that question, whether it be the transient response of a continuous flow to a fluctuating continuum or perhaps the continued decline of a discrete outflow event.

Current status of the thiol redox model for the regulation of hexose transport by insulin.

PubMed

Czech, M P

1976-12-01

Data obtained over the last two years pertinent to the thiol redox model for the modulation of hexose transport activity by insulin is summarized. The model proposes that activation of hexose transport in fat cells involves sulfhydryl oxidation to the disulfide form in a key protein component of the fat cell surface membrane. Theoretically, the rapid activation of transport by insulin may involve either the conversion of inactive membrane carriers to the active form as originally proposed, or the conversion of a low Vmax transport system to a high Vmax form. The present experiments showed that the percent inhibition of insulin-activated transport rates by submaximal levels of cytochalasin B was decreased compared to its effects on basal transport. Treatment of fat cells with N-ethylmaleimide inhibited cytochalasin B action but not transport activity. When insulin or the oxidant vitamin K5 was added to cells 5 minutes before the N-ethylmaleimide, the elevated transport activity was also resistant to the sulfhydryl reagent, but cytochalasin B retained its potent inhibitory effect on transport. The data demonstrate that unique properties characterize basal versus insulin-activated transport activity with respect to the sensitivity of cytochalasin B action to sulfhydryl blockade in isolated fat cells. The data are consistent with the concept that activation of transport activity reflects the conversion of a reduced (sulfhydryl) system characterized by a low Vmax to an oxidized (disulfide), high Vmax transport system.

Tunnel transport and interlayer excitons in bilayer fractional quantum Hall systems

NASA Astrophysics Data System (ADS)

Zhang, Yuhe; Jain, J. K.; Eisenstein, J. P.

2017-05-01

In a bilayer system consisting of a composite-fermion (CF) Fermi sea in each layer, the tunnel current is exponentially suppressed at zero bias, followed by a strong peak at a finite-bias voltage Vmax. This behavior, which is qualitatively different from that observed for the electron Fermi sea, provides fundamental insight into the strongly correlated non-Fermi-liquid nature of the CF Fermi sea and, in particular, offers a window into the short-distance high-energy physics of this highly nontrivial state. We identify the exciton responsible for the peak current and provide a quantitative account of the value of Vmax. The excitonic attraction is shown to be quantitatively significant, and its variation accounts for the increase of Vmax with the application of an in-plane magnetic field. We also estimate the critical Zeeman energy where transition occurs from a fully spin-polarized composite-fermion Fermi sea to a partially spin-polarized one, carefully incorporating corrections due to finite width and Landau level mixing, and find it to be in satisfactory agreement with the Zeeman energy where a qualitative change has been observed for the onset bias voltage [J. P. Eisenstein et al., Phys. Rev. B 94, 125409 (2016), 10.1103/PhysRevB.94.125409]. For fractional quantum Hall states, we predict a substantial discontinuous jump in Vmax when the system undergoes a transition from a fully spin-polarized state to a spin singlet or a partially spin-polarized state.

[Temperature sensitivity of soil organic carbon mineralization and β-glucosidase enzymekinetics in the northern temperate forests at different altitudes, China].

PubMed

Fan, Jin-juan; Li, Dan-dan; Zhang, Xin-yu; He, Nian-peng; Bu, Jin-feng; Wang, Qing; Sun, Xiao-min; Wen, Xue-fa

2016-01-01

Soil samples, which were collected from three typical forests, i.e., Betula ermanii forest, coniferous mixed broad-leaved forest, and Pinus koraiensis forest, at different altitudes along the southern slope of Laotuding Mountain of Changbai Mountain range in Liaoning Province of China, were incubated over a temperature gradient in laboratory. Soil organic carbon mineralization rates (Cmin), soil β-1,4-glucosidase (βG) kinetics and their temperature sensitivity (Q₁₀) were measured. The results showed that both altitude and temperature had significant effects on Cmin · Cmin increased with temperature and was highest in the B. ermanii forest. The temperature sensitivity of Cmin [Q₁₀(Cmin)] ranked in order of B. ermanii forest > P. koraiensis forest > coniferous mixed broad-leaved forest, but did not differ significantly among the three forests. Both the maximum activity (Vmax) and the Michaelis constant (Km) of the βG responded positively to temperature for all the forests. The temperature sensitivity of Vmax [Q₁₀(Vmax)] ranged from 1.78 to 1.90, and the temperature sensitivity of Km [Q₁₀(Km)] ranged from 1.79 to 2.00. The Q₁₀(Vmax)/Q10(Km) ratios were significantly greater in the B. ermanii soil than in the other two forest soils, suggesting that the βG kinetics-dependent impacts of the global warming or temperature increase on the decomposition of soil organic carbon were temperature sensitive for the forests at the higher altitudes.

Role of 3D Ultrasound and Doppler in Differentiating Clinically Suspected Cases of Leiomyoma and Adenomyosis of Uterus

PubMed Central

Sharma, Kaveri; Venkatesh, B.P; Barman, Partho; Roy, Sumit Kumar; Jayagurunathan, Usha; Sellamuthu, Eswaramoorthy; Moidu, Fazil

2015-01-01

Introduction Adenomyosis and Leiomyoma are common disorders affecting females in their reproductive age. They mimic each other in clinical presentation. Due to similarities in clinical symptoms and signs, missing one diagnosis in favour of the other is not very uncommon. Accurate diagnosis of these two conditions is important for their management. In this study we evaluated role of 3D Ultrasound and Doppler in differentiating clinically suspected cases of leiomyoma and adenomyosis of uterus. Materials and Methods A total of 100 patients with symptoms of abnormal uterine bleeding (with or without dysmenorrhoea), lump abdomen, chronic pelvic pain or dysparaunia who were clinically diagnosed as leiomyoma of uterus and/or adenomyosis were enrolled in to the study. These patients underwent transvaginal sonography (TVS), trans abdominal sonography (TAS) along with color and spectral Doppler sonography. Scanning was done in follicular phase of the menstrual cycle to avoid bias due high vascularity of endometrium in secretory phase. The morphology of the lesion, its vascularity, and Pulsality Index (PI), Resistive Index (RI) and Vmax (maximum velocity) were measured. Only those patients who were chosen for operative treatment were included in the study. Radiological diagnosis was then correlated with intra-operative and histopathological diagnosis. Results On imaging, while using morphological criteria and Doppler for diagnosing leiomyoma, it was found that “peripheral vascularity” was seen in 52 (89%) cases, which was the highest. Similarly while diagnosing adenomyosis it was, the criteria “central vascularity” was seen in 28 cases (93%) and “ill defined junctional zone in 3D ultrasound” was seen in 26 cases (86%), which was also observed to be highest. With the cut off values taken for PI,RI and Vmax, diagnosis of leiomyoma was found to be 93.4% sensitive, 95.6% specific and with a positive predictive value of 97.6% and negative predictive value of 88.6%. Diagnosis of adenomyosis showed a sensitivity of 95.6%, specificity of 93.4% and a positive predictive value of 88.6% and negative predictive value of 97.6%. Imaging dignosed the co-existence of both the conditions correctly in 8 (66%) cases. Conclusion The parameters of blood flow impedance (that is PI, RI, and Vmax) of arteries within or around the uterine lesions revealed a consistent and significant difference between leiomyoma and adenomyosis. So apart from morphological criteria used in 3D TAS and TVS, aid of color Doppler can more accurately differentiate and diagnose these conditions. PMID:26023602

Three-dimensional quantitative structure-activity relationship studies on UGT1A9-mediated 3-O-glucuronidation of natural flavonols using a pharmacophore-based comparative molecular field analysis model.

PubMed

Wu, Baojian; Morrow, John Kenneth; Singh, Rashim; Zhang, Shuxing; Hu, Ming

2011-02-01

Glucuronidation is often recognized as one of the rate-determining factors that limit the bioavailability of flavonols. Hence, design and synthesis of more bioavailable flavonols would benefit from the establishment of predictive models of glucuronidation using kinetic parameters [e.g., K(m), V(max), intrinsic clearance (CL(int)) = V(max)/K(m)] derived for flavonols. This article aims to construct position (3-OH)-specific comparative molecular field analysis (CoMFA) models to describe UDP-glucuronosyltransferase (UGT) 1A9-mediated glucuronidation of flavonols, which can be used to design poor UGT1A9 substrates. The kinetics of recombinant UGT1A9-mediated 3-O-glucuronidation of 30 flavonols was characterized, and kinetic parameters (K(m), V(max), CL(int)) were obtained. The observed K(m), V(max), and CL(int) values of 3-O-glucuronidation ranged from 0.04 to 0.68 μM, 0.04 to 12.95 nmol/mg/min, and 0.06 to 109.60 ml/mg/min, respectively. To model UGT1A9-mediated glucuronidation, 30 flavonols were split into the training (23 compounds) and test (7 compounds) sets. These flavonols were then aligned by mapping the flavonols to specific common feature pharmacophores, which were used to construct CoMFA models of V(max) and CL(int), respectively. The derived CoMFA models possessed good internal and external consistency and showed statistical significance and substantive predictive abilities (V(max) model: q(2) = 0.738, r(2) = 0.976, r(pred)(2) = 0.735; CL(int) model: q(2) = 0.561, r(2) = 0.938, r(pred)(2) = 0.630). The contour maps derived from CoMFA modeling clearly indicate structural characteristics associated with rapid or slow 3-O-glucuronidation. In conclusion, the approach of coupling CoMFA analysis with a pharmacophore-based structural alignment is viable for constructing a predictive model for regiospecific glucuronidation rates of flavonols by UGT1A9.

Methane and Trichloroethylene Degradation by Methylosinus trichosporium OB3b Expressing Particulate Methane Monooxygenase

PubMed Central

Lontoh, Sonny; Semrau, Jeremy D.

1998-01-01

Whole-cell assays of methane and trichloroethylene (TCE) consumption have been performed on Methylosinus trichosporium OB3b expressing particulate methane monooxygenase (pMMO). From these assays it is apparent that varying the growth concentration of copper causes a change in the kinetics of methane and TCE degradation. For M. trichosporium OB3b, increasing the copper growth concentration from 2.5 to 20 μM caused the maximal degradation rate of methane (Vmax) to decrease from 300 to 82 nmol of methane/min/mg of protein. The methane concentration at half the maximal degradation rate (Ks) also decreased from 62 to 8.3 μM. The pseudo-first-order rate constant for methane, Vmax/Ks, doubled from 4.9 × 10−3 to 9.9 × 10−3 liters/min/mg of protein, however, as the growth concentration of copper increased from 2.5 to 20 μM. TCE degradation by M. trichosporium OB3b was also examined with varying copper and formate concentrations. M. trichosporium OB3b grown with 2.5 μM copper was unable to degrade TCE in both the absence and presence of an exogenous source of reducing equivalents in the form of formate. Cells grown with 20 μM copper, however, were able to degrade TCE regardless of whether formate was provided. Without formate the Vmax for TCE was 2.5 nmol/min/mg of protein, while providing formate increased the Vmax to 4.1 nmol/min/mg of protein. The affinity for TCE also increased with increasing copper, as seen by a change in Ks from 36 to 7.9 μM. Vmax/Ks for TCE degradation by pMMO also increased from 6.9 × 10−5 to 5.2 × 10−4 liters/min/mg of protein with the addition of formate. From these whole-cell studies it is apparent that the amount of copper available is critical in determining the oxidation of substrates in methanotrophs that are expressing only pMMO. PMID:16349516

Connexin43 contributes to electrotonic conduction across scar tissue in the intact heart

NASA Astrophysics Data System (ADS)

Mahoney, Vanessa M.; Mezzano, Valeria; Mirams, Gary R.; Maass, Karen; Li, Zhen; Cerrone, Marina; Vasquez, Carolina; Bapat, Aneesh; Delmar, Mario; Morley, Gregory E.

2016-05-01

Studies have demonstrated non-myocytes, including fibroblasts, can electrically couple to myocytes in culture. However, evidence demonstrating current can passively spread across scar tissue in the intact heart remains elusive. We hypothesize electrotonic conduction occurs across non-myocyte gaps in the heart and is partly mediated by Connexin43 (Cx43). We investigated whether non-myocytes in ventricular scar tissue are electrically connected to surrounding myocardial tissue in wild type and fibroblast-specific protein-1 driven conditional Cx43 knock-out mice (Cx43fsp1KO). Electrical coupling between the scar and uninjured myocardium was demonstrated by injecting current into the myocardium and recording depolarization in the scar through optical mapping. Coupling was significantly reduced in Cx43fsp1KO hearts. Voltage signals were recorded using microelectrodes from control scars but no signals were obtained from Cx43fsp1KO hearts. Recordings showed significantly decreased amplitude, depolarized resting membrane potential, increased duration and reduced upstroke velocity compared to surrounding myocytes, suggesting that the non-excitable cells in the scar closely follow myocyte action potentials. These results were further validated by mathematical simulations. Optical mapping demonstrated that current delivered within the scar could induce activation of the surrounding myocardium. These data demonstrate non-myocytes in the scar are electrically coupled to myocytes, and coupling depends on Cx43 expression.

Optically Nonlinear Polymeric Materials.

DTIC Science & Technology

1983-01-01

distilled water . Polymer A was deposited onto two back-to-back clean glass slides on the upstroke. The subphase surface was cleaned and either behenic acid...having a water soluble head orouc I and a hydrophobic tail group such as a long chain fatty acid. The amphiphiles are spread on the water surface of a L/B...trough (Figure 1). The hydrophilic head group sits on . the water surface, while the hydrophobic alkyl tails orient themselves away from the water By

Bat flight: aerodynamics, kinematics and flight morphology.

PubMed

Hedenström, Anders; Johansson, L Christoffer

2015-03-01

Bats evolved the ability of powered flight more than 50 million years ago. The modern bat is an efficient flyer and recent research on bat flight has revealed many intriguing facts. By using particle image velocimetry to visualize wake vortices, both the magnitude and time-history of aerodynamic forces can be estimated. At most speeds the downstroke generates both lift and thrust, whereas the function of the upstroke changes with forward flight speed. At hovering and slow speed bats use a leading edge vortex to enhance the lift beyond that allowed by steady aerodynamics and an inverted wing during the upstroke to further aid weight support. The bat wing and its skeleton exhibit many features and control mechanisms that are presumed to improve flight performance. Whereas bats appear aerodynamically less efficient than birds when it comes to cruising flight, they have the edge over birds when it comes to manoeuvring. There is a direct relationship between kinematics and the aerodynamic performance, but there is still a lack of knowledge about how (and if) the bat controls the movements and shape (planform and camber) of the wing. Considering the relatively few bat species whose aerodynamic tracks have been characterized, there is scope for new discoveries and a need to study species representing more extreme positions in the bat morphospace. © 2015. Published by The Company of Biologists Ltd.

Wing kinematics measurement and aerodynamics of a dragonfly in turning flight.

PubMed

Li, Chengyu; Dong, Haibo

2017-02-03

This study integrates high-speed photogrammetry, 3D surface reconstruction, and computational fluid dynamics to explore a dragonfly (Erythemis Simplicicollis) in free flight. Asymmetric wing kinematics and the associated aerodynamic characteristics of a turning dragonfly are analyzed in detail. Quantitative measurements of wing kinematics show that compared to the outer wings, the inner wings sweep more slowly with a higher angle of attack during the downstroke, whereas they flap faster with a lower angle of attack during the upstroke. The inner-outer asymmetries of wing deviations result in an oval wingtip trajectory for the inner wings and a figure-eight wingtip trajectory for the outer wings. Unsteady aerodynamics calculations indicate significantly asymmetrical force production between the inner and outer wings, especially for the forewings. Specifically, the magnitude of the drag force on the inner forewing is approximately 2.8 times greater than that on the outer forewing during the downstroke. In the upstroke, the outer forewing generates approximately 1.9 times greater peak thrust than the inner forewing. To keep the body aloft, the forewings contribute approximately 64% of the total lift, whereas the hindwings provide 36%. The effect of forewing-hindwing interaction on the aerodynamic performance is also examined. It is found that the hindwings can benefit from this interaction by decreasing power consumption by 13% without sacrificing force generation.

In vivo recording of aerodynamic force with an aerodynamic force platform: from drones to birds.

PubMed

Lentink, David; Haselsteiner, Andreas F; Ingersoll, Rivers

2015-03-06

Flapping wings enable flying animals and biomimetic robots to generate elevated aerodynamic forces. Measurements that demonstrate this capability are based on experiments with tethered robots and animals, and indirect force calculations based on measured kinematics or airflow during free flight. Remarkably, there exists no method to measure these forces directly during free flight. Such in vivo recordings in freely behaving animals are essential to better understand the precise aerodynamic function of their flapping wings, in particular during the downstroke versus upstroke. Here, we demonstrate a new aerodynamic force platform (AFP) for non-intrusive aerodynamic force measurement in freely flying animals and robots. The platform encloses the animal or object that generates fluid force with a physical control surface, which mechanically integrates the net aerodynamic force that is transferred to the earth. Using a straightforward analytical solution of the Navier-Stokes equation, we verified that the method is accurate. We subsequently validated the method with a quadcopter that is suspended in the AFP and generates unsteady thrust profiles. These independent measurements confirm that the AFP is indeed accurate. We demonstrate the effectiveness of the AFP by studying aerodynamic weight support of a freely flying bird in vivo. These measurements confirm earlier findings based on kinematics and flow measurements, which suggest that the avian downstroke, not the upstroke, is primarily responsible for body weight support during take-off and landing.

Dynamics of the vortex wakes of flying and swimming vertebrates.

PubMed

Rayner, J M

1995-01-01

The vortex wakes of flying and swimming animals provide evidence of the history of aero- and hydrodynamic force generation during the locomotor cycle. Vortex-induced momentum flux in the wake is the reaction of forces the animal imposes on its environment, which must be in equilibrium with inertial and external forces. In flying birds and bats, the flapping wings generate lift both to provide thrust and to support the weight. Distinct wingbeat and wake movement patterns can be identified as gaits. In flow visualization experiments, only two wake patterns have been identified: a vortex ring gait with inactive upstroke, and a continuous vortex gait with active upstroke. These gaits may be modelled theoretically by free vortex and lifting line theory to predict mechanical energy consumption, aerodynamic forces and muscle activity. Longer-winged birds undergo a distinct gait change with speed, but shorter-winged species use the vortex ring gait at all speeds. In swimming fish, the situation is more complex: the wake vortices form a reversed von Kármán vortex street, but little is known about the mechanism of generation of the wake, or about how it varies with speed and acceleration or with body form and swimming mode. An unresolved complicating factor is the interaction between the drag wake of the flapping fish body and the thrusting wake from the tail.

Mechanical properties of the avian acrocoracohumeral ligament and its role in shoulder stabilization in flight.

PubMed

Baier, David Bradley

2012-02-01

Control of movement in the avian shoulder joint is fundamental to understanding the avian wingstroke. The acrocoracohumeral ligament (AHL) is thought to play a key role in stabilizing the glenoid and balancing the pectoralis in gliding flight. If the AHL has to be taut to balance the pectoralis, then it must constrain glenohumeral motion during flapping flight as well. However, birds vary wing kinematics depending on flight speed and behavior. How can a passive ligament accommodate such varying joint movements? Herein, mechanical testing and 3-D modeling are used to link the mechanical properties and morphology of the AHL to its functional role during flapping flight. The bone-ligament-bone complex of the pigeon (Columba livia) fails at a tensile loading of 141 ± 18 N (± s .D., n = 10) or 39 times body weight, which corresponds to a failure stress of 51 MPa, well above expected loads during flight. Simulated AHL length changes, comparisons to glenohumeral kinematics from the literature, and manipulations of partially dissected pigeon specimens all support the hypothesis that the AHL remains taut through downstroke and most of upstroke while becoming slack during the downstroke/upstroke transition. The digital AHL model provides a mechanism for explaining how the AHL can stabilize the shoulder joint under a broad array of humeral paths by constraining the coordination of glenohumeral degrees of freedom. © 2011 WILEY PERIODICALS, INC.

Ethanol intake and sup 3 H-serotonin uptake I: A study in Fawn-Hooded rats

DOE Office of Scientific and Technical Information (OSTI.GOV)

Daoust, M.; Compagnon, P.; Legrand, E.

1991-01-01

Ethanol intake and synaptosomal {sup 3}H-serotonin uptake were studied in male Fawn-Hooded and Sprague-Dawley rats. Fawn-Hooded rats consumed more alcohol and more water than Sprague-Dawley rats. Plasma alcohol levels of Sprague-Dawley rats were not detectable but were about 5 mg/dl in Fawn-Hooded rats. Ethanol intake increased the Vmax of serotonin uptake in Fawn-Hooded rats in hippocampus and cortex, but not in thalamus. In Fawn-Hooded rats, serotonin uptake (Vmax) was higher than in Sprague-Dawley rats cortex. Ethanol intake reduced the Vmax of serotonin uptake in Fawn-Hooded rats in hippocampus and cortex. In cortex, the carrier affinity for serotonin was increased inmore » alcoholized Fawn-Hooded rats. These results indicate that synaptosomal {sup 3}H-serotonin uptake is affected by ethanol intake. In Fawn-Hooded rats, high ethanol consumption is associated with high serotonin uptake. In rats presenting high serotonin uptake, alcoholization reduces {sup 3}H-serotonin internalization in synaptosomes, indicating a specific sensitivity to alcohol intake of serotonin uptake system.« less

Characterization of protease from bacillus sp. on medium containing FeCl3 exposed to magnetic field 0.2 mt

NASA Astrophysics Data System (ADS)

Sumardi; Agustrina, Rochmah; Nugroho Ekowati, Christina; Selvie Pasaribu, Yovita

2018-03-01

This purpose of this research is to determine the character of the protease enzymes from Bacillus sp. on media content of FeCl3 exposed to 0.2 mT magnetic field. The data obtained were analyzed descriptively. The result showed that protease enzyme without Fe resulted in the highest activity at pH 8, temperature. 30°C with the addition of activator Mn2+, and Vmax of 0.28 U/ml, and Km of 4.60 U/ml. The protease enzyme on media without magnetic field exposure and containing Fe yielded the highest activity at pH 8, temperature 30°C with the addition of activator Mn2+, and Vmax of 0.33 U/ml, and Km of 5.64 U/ml. The protease enzyme on medium with magnetic field exposure and use Fe as inductors have the highest activity at pH 9, the temperature of 55° C with the addition of activator Mn2+, and Vmax of 0.35 U/ml, and Km 10.04 U/ml.

Enzymatic carotenoid cleavage in star fruit (Averrhoa carambola).

PubMed

Fleischmann, Peter; Watanabe, Naoharu; Winterhalter, Peter

2003-05-01

This paper presents the first description of an enzyme fraction exhibiting carotenoid cleavage activity isolated from fruit skin of Averrhoa carambola. Partial purification of the enzyme could be achieved by acetone precipitation, ultrafiltration (300 kDa, 50 kDa), isoelectric focusing (pH 3-10) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (7.5%). In this way, an enzymatically active protein fraction was obtained, consisting of four proteins in the molecular weight range of between 12 and 90 kDa. Using beta-carotene as substrate, the enzyme activity was detected spectrophotometrically at 505 nm. The main reaction product, detected by GC analysis, was beta-ionone. This proves that the isolated enzymes are closely related to aroma metabolism and release of star fruit. The time constant of the reaction was 16.6 min, the Michaelis Constant K(m)=3.6 micromol 1(-1) and the maximum velocity V(max)=10.5 x 10(-3) micromol l(-1) s(-1) mg((Protein))(-1). The optimum temperature was 45 degrees C.

( sup 3 H)Dopamine uptake by platelet storage granules in schizophrenia

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rabey, J.M.; Graff, E.; Oberman, Z.

1992-01-01

({sup 3}H)Dopamine (DA) uptake by platelet storage granules was determined in 26 schizophrenic male patients, paranoid type (14 acute stage; 12 in remission) and 20 age-matched, normal controls. maximum velocity (Vmax) of DA uptake was significantly higher in acute patients, than patients in remission or controls (p>0.05). The apparent Michaelis constant (kM) of DA uptake in acute patients was also significantly different from chronic patients a substantial diminution of DA uptake, while haloperidol produced a substantial diminution of DA uptake, while haloperidol (10{sup {minus}4}, 10{sup {minus}5} M) did not affect the assay. Considering that a DA disequilibrium in schizophrenia maymore » be expressed not only in the brain, but also in the periphery and that an increased amount of DA accumulated in the vesicles, implies that an increased quantity of catecholamine is available for release, our findings suggest additional evidence for the role of DA overactivity in the pathophysiology of this disorder.« less

Transport of Phosphoenolpyruvate by Chloroplasts from Mesembryanthemum crystallinum L. Exhibiting Crassulacean Acid Metabolism 1

PubMed Central

Neuhaus, H. Ekkehard; Holtum, Joseph A. M.; Latzko, Erwin

1988-01-01

Chloroplasts from CAM-Mesembryanthemum crystallinum can transport phosphoenolpyruvate (PEP) across the envelope. The initial velocities of PEP uptake in the dark at 4°C exhibited saturation kinetics with increasing external PEP concentration. PEP uptake had a Vmax of 6.46 (±0.05) micromoles per milligram chlorophyll per hour and an apparent Kmpep of 0.148 (±0.004) millimolar. The uptake was competitively inhibited by Pi (apparent Ki = 0.19 millimolar), by glycerate 3-phosphate (apparent Ki = 0.13 millimolar), and by dihydroxyacetone phosphate, but malate and pyruvate were without effect. The chloroplasts were able to synthesize PEP when presented with pyruvate. PEP synthesis was light dependent. The prolonged synthesis and export of PEP from the chloroplasts required the presence of Pi or glycerate 3-phosphate in the external medium. It is suggested that the transport of pyruvate and PEP across the chloroplasts envelope is required during the gluconeogenic conversion of carbon from malate to storage carbohydrate in the light. PMID:16666128

Functional characteristics of pyruvate transport in Phycomyces blakesleeanus.

PubMed

Marcos, J A; de Arriaga, D; Busto, F; Soler, J

1998-12-01

A saturable and accumulative transport system for pyruvate has been detected in Phycomyces blakesleeanus NRRL 1555(-) mycelium. It was strongly inhibited by alpha-cyano-4-hydroxycinnamate. l-Lactate and acetate were competitive inhibitors of pyruvate transport. The initial pyruvate uptake velocity and accumulation ratio was dependent on the external pH. The Vmax of transport greatly decreased with increasing pH, whereas the affinity of the carrier for pyruvate was not affected. The pyruvate transport system mediated its homologous exchange, which was essentially pH independent, and efflux, which increased with increasing external pH. The uptake of pyruvate was energy dependent and was strongly inhibited by inhibitors of oxidative phosphorylation and of the formation of proton gradients. Glucose counteracted the inhibitory effect of the pyruvate transport produced by inhibitors of mitochondrial ATP synthesis. Our results are consistent with a pyruvate/proton cotransport in P. blakesleeanus probably driven by an electrochemical gradient of H+ generated by a plasma membrane H+-ATPase. Copyright 1998 Academic Press.

A fibrinolytic, alkaline and thermostable metalloprotease from the newly isolated Serratia sp RSPB11.

PubMed

Lakshmi Bhargavi, P; Prakasham, R S

2013-10-01

This study shows the purification and characterization of metalloprotease (serralysin) with fibrin and fibrinogenolytic property, from the newly isolated Serratia marcescens RSPB11. This protein macro molecule was more stable over a wide range of pH (6-10) and the temperatures up to 60 °C. It showed optimum enzyme activity at pH 9.0 and at a temperature of 37 °C. Inhibitory analysis revealed that this enzyme is metalloprotease and its enzyme activity could be regained by the addition of Co(2+), Cu(2+), Fe(2+), Mg(2+)and Zn(2+) ions after chelation of ions with EDTA. This enzyme showed the Michaelis-Menten's constant Km (1.261 mg/ml) for its substrate, casein and the observed maximum attainable velocity was Vmax (24,842 U/min). The purified enzyme showed an apparent molecular mass of approximately 50 kDa in SDS-PAGE. The results also suggested that this serralysin is having potential application thrombolytic therapy. Copyright © 2013 Elsevier B.V. All rights reserved.

Characterization of procaine metabolism as probe for the butyrylcholinesterase enzyme investigation by simultaneous determination of procaine and its metabolite using capillary electrophoresis with electrochemiluminescence detection.

PubMed

Yuan, Jipei; Yin, Jianyuan; Wang, Erkang

2007-06-22

Capillary electrophoresis with electrochemiluminescene detection was used to characterize procaine hydrolysis as a probe for butyrylcholinesterase by in vitro procaine metabolism in plasma with butyrylcholinesterase acting as bioscavenger. Procaine and its metabolite N,N-diethylethanolamine were separated at 16 kV and then detected at 1.25 V in the presence of 5.0 mM Ru(bpy)(3)2+, with the detection limits of 2.4x10(-7) and 2.0x10(-8) mol/L (S/N=3), respectively. The Michaelis constant Km value was 1.73x10(-4) mol/L and the maximum velocity Vmax was 1.62x10(-6) mol/L/min. Acetylcholine bromide and choline chloride presented inhibition effects on the enzymatic cleavage of procaine, with the 50% inhibition concentration (IC50) of 6.24x10(-3) and 2.94x10(-4) mol/L.

A normalized plot as a novel and time-saving tool in complex enzyme kinetic analysis.

PubMed

Bravo, I G; Busto, F; De Arriaga, D; Ferrero, M A; Rodríguez-Aparicio, L B; Martínez-Blanco, H; Reglero, A

2001-09-15

A new data treatment is described for designing kinetic experiments and analysing kinetic results for multi-substrate enzymes. Normalized velocities are plotted against normalized substrate concentrations. Data are grouped into n + 1 families across the range of substrate or product tested, n being the number of substrates plus products assayed. It has the following advantages over traditional methods: (1) it reduces to less than a half the amount of data necessary for a proper description of the system; (2) it introduces a self-consistency checking parameter that ensures the 'scientific reliability' of the mathematical output; (3) it eliminates the need for a prior knowledge of Vmax; (4) the normalization of data allows the use of robust and fuzzy methods suitable for managing really 'noisy' data; (5) it is appropriate for analysing complex systems, as the complete general equation is used, and the actual influence of effectors can be typified; (6) it is amenable to being implemented as a software that incorporates testing and electing among rival kinetic models.

Evaluation of the effects of sildenafil citrate (viagra) on vertebral artery blood flow in patients with vertebro-basilar insufficiency.

PubMed

Bozgeyik, Zulkif; Berilgen, Sait; Ozdemir, Huseyin; Tekatas, Aslan; Ogur, Erkin

2008-01-01

To investigate the effects of sildenafil citrate (Viagra) on the vertebral artery blood flow of patients with vertebro-basilar insufficiency (VBI) using color duplex sonography (CDS). The study included 21 patients with VBI (aged 31-76; mean 61.0 +/- 10.5 yrs). We administered a 50 mg oral dose of sildenafil citrate to all patients. Next, we measured the peak systolic velocity (Vmax), end diastolic velocity (Vmin), resistive index (RI), pulsatility index (PI), diameter, area, and flow volume (FV) of vertebral arteries using CDS before the administration of sildenafil citrate; 45 minutes after, and 75 minutes after administration. Statistical testing was performed using SPSS for windows version 11.0. The statistical test used to determine the outcome of the analysis was the repeated measures analysis of variance (ANOVA) test. Compared to the baseline values, the vertebral artery diameter, area, and FV increased significantly following the administration of sildenafil citrate. The diameter, area and FV increased from 3.39 mm at 45 minutes to 3.64 mm at 75 minutes, 9.43 cm(2) to 10.80 cm(2) at 45 minutes and 10.81 cm(2) at 75 minutes, as well as from 0.07 L/min at baseline to 0.09 L/min at 45 minutes and unchanged at 75 minutes, respectively. Sildenafil citrate elicited a significant effect on vertebral artery diameter, area and FVs.

Cloning, Expression, and Purification of Choline Dehydrogenase from the Moderate Halophile Halomonas elongata

PubMed Central

Gadda, Giovanni; McAllister-Wilkins, Elien Elizabeth

2003-01-01

Choline dehydrogenase (EC 1.1.99.1) catalyzes the four-electron oxidation of choline to glycine-betaine via a betaine-aldehyde intermediate. Such a reaction is of considerable interest for biotechnological applications in that transgenic plants engineered with bacterial glycine-betaine-synthesizing enzymes have been shown to have enhanced tolerance towards various environmental stresses, such as hypersalinity, freezing, and high temperatures. To date, choline dehydrogenase has been poorly characterized in its biochemical and kinetic properties, mainly because its purification has been hampered by instability of the enzyme in vitro. In the present report, we cloned and expressed in Escherichia coli the betA gene from the moderate halophile Halomonas elongata which codes for a hypothetical choline dehydrogenase. The recombinant enzyme was purified to more than 70% homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by treatment with 30 to 50% saturation of ammonium sulfate followed by column chromatography using DEAE-Sepharose. The purified enzyme showed similar substrate specificities with either choline or betaine-aldehyde as the substrate, as indicated by the apparent V/K values (where V is the maximal velocity and K is the Michaelis constant) of 0.9 and 0.6 μmol of O2 min−1 mg−1 mM−1 at pH 7 and 25°C, respectively. With 1 mM phenazine methosulfate as the primary electron acceptor, the apparent Vmax values for choline and betaine-aldehyde were 10.9 and 5.7 μmol of O2 min−1 mg−1, respectively. These Vmax values decreased four- to sevenfold when molecular oxygen was used as the electron acceptor. Altogether, the kinetic data are consistent with the conclusion that H. elongata betA codes for a choline dehydrogenase that can also act as an oxidase when electron acceptors other than molecular oxygen are not available. PMID:12676692

[Effect of phosphatidic acid on the reaction of linoleic acid oxidation by 5-lipooxygenase from potatoes].

PubMed

Skaterna, T D; Kharchenko, O V

2008-01-01

Influence of anionogenic phospholipid of phosphatidic acid (PA) on oxidation of linoleic acid by 5-lipoxygenase (5-LO) from Solanum tuberosum was studied. The influence of PA was studied in micellar system which consisted of mixed micelles of linolenic acid (LK), Lubrol PX and different quantity of enzyme effector PA. The reaction was initiated by addition of 5-LO. It was established that 5-LO had two pHopt. in the presence of 50 microM phosphatidic acid: pH 5.0 and 6.9. In concentration of 50 microM PA was able to activate 5-LO 15 times at pH 5.0. The reaction maximum velocity (Vmax) coincided with Vmax of lipoxygenase reaction without the effector at pH 6.9 under such conditions. It was found that 30-50 microM phospholipid in the reaction mixture decreased the concentration of half saturation by the substrate by 43-67%. The enzyme demonstrated positive cooperation in respect of the substrate, the reaction is described by the Hill equation. Hill coefficient value (h) of the substrate was 3.34 +/- 0.22 (pH 6.9) and 5.61 +/- 0.88 (pH 5.0), that is with the change of pH to acidic region the number of substrate molecules increased and they could interact with the enzyme molecule. In case of substrate insufficiency the enzyme demonstrated positive cooperation of PA, it added from 4 to 3 effectors' molecules at pH 5.0, that is the phospholipid acted as the allosteric regulator of 5-LO. A comparative analysis of the influence of 4-hydroxy-TEMPO displayed, that the level of nonenzymatic processes in the case of physiological pH values was lower by 15-50% in the presence of PA in the range of 30-80 microM than without the effector.

Renal functional reserve and renal hemodynamics in hypertensive patients.

PubMed

Gaipov, Abduzhappar; Solak, Yalcin; Zhampeissov, Nurlan; Dzholdasbekova, Aliya; Popova, Nadezhda; Molnar, Miklos Z; Tuganbekova, Saltanat; Iskandirova, Elmira

2016-10-01

The renal functional reserve (RFR) is the ability of the kidneys to increase renal plasma flow and glomerular filtration rate (GFR) in response to protein intake. It is a measure of functional and anatomic integrity of nephrons. It is not known what relation between RFR and kidney Doppler parameters. We aimed to study the relation between the RFR and renal hemodynamic parameters in hypertensive patients with and without nephropathy who had normal kidney function. Twenty-four hypertensive subjects with nephropathy (HTN-n, n = 10) and hypertension without nephropathy (HTN, n = 14) were included in the study. Control group included 11 healthy subjects. Baseline GFR (GFR1) and GFR after intake of egg protein 1 mg/kg of body weight were determined (GFR2). RFR was calculated by the following formula: (GFR2-GFR1)/GFR1 × 100%. Doppler ultrasonography was performed. Arterial blood pressure (BP), body mass index (BMI), and estimated GFR were also recorded. HTN and HTN-n groups had impaired levels of RFR compared with controls (p < 0.05), significantly decreased value of flow velocity parameters (Vmax, Vmin), and increased RRI compared with controls. There was significant negative correlation of RFR with blood pressure levels (sBP, r = -0.435, p = 0.009; dBP, r = -0.504, p = 0.002), RRI (r = -0.456, p = 0.008), micro albuminuria (MAU, r = -0.366, p = 0.031) and positive correlation with Vmax and Vmin (r = 0.556, p = 0.001 and r = 0.643, respectively, p < 0.001). Linear regression showed that RRI and MAU were independent predictors of decreased RFR. RFR is lower in hypertensive patients despite near-normal level of kidney function and is related to particular level of BP. RRI and MAU were independent predictors of decreased RFR.

THERAPEUTIC HYPOTHERMIA DECREASES PHENYTOIN ELIMINATION IN CHILDREN WITH TRAUMATIC BRAIN INJURY

PubMed Central

Empey, Philip E.; Velez de Mendizabal, Nieves; Bell, Michael J.; Bies, Robert R.; Anderson, Kacey B.; Kochanek, Patrick M.; Adelson, P. David; Poloyac, Samuel M.

2013-01-01

Objective Preclinical and clinical studies have suggested that therapeutic hypothermia, while decreasing neurological injury, may also lead to drug toxicity that may limit its benefit. Cooling decreases cytochrome p450(CYP)-mediated drug metabolism and limited clinical data suggest that drug levels are elevated. Fosphenytoin is metabolized by CYP2C, has a narrow therapeutic range, and is a commonly used antiepileptic medication. The objective of the study was to evaluate the impact of therapeutic hypothermia on phenytoin levels and pharmacokinetics in children with severe TBI. Design Pharmacokinetic analysis of subjects participating in a multicenter randomized Phase III study of therapeutic hypothermia for severe TBI. Setting Intensive care unit at the Children’s Hospital of Pittsburgh Patients Nineteen children with severe TBI. Interventions None Measurements and Main Results A total of 121 total and 114 free phenytoin levels were evaluated retrospectively in 10 hypothermia- and 9 normothermia-treated children who were randomized to 48h of cooling to 32–33°C followed by slow rewarming or controlled normothermia. Drug dosing, body temperatures, and demographics were collected during cooling, rewarming, and post-treatment periods(8 days). A trend towards elevated free phenytoin levels in the hypothermia group(p=0.051) to a median of 2.2 mg/L during rewarming was observed and was not explained by dosing differences. Nonlinear mixed effects modeling incorporating both free and total levels demonstrated that therapeutic hypothermia specifically decreased the time-variant component of the maximum velocity of phenytoin metabolism(Vmax) 4.6-fold(11.6 to 2.53 mg/h) and reduced the overall Vmax by ~50%. Simulations showed that the increased risk for drug toxicity extends many days beyond the end of the cooling period. Conclusions Therapeutic hypothermia significantly reduces phenytoin elimination in children with severe TBI leading to increased drug levels for an extended period of time after cooling. Pharmacokinetic interactions between hypothermia and medications should be considered when caring for children receiving this therapy. PMID:23896831

Presynaptic Na+-dependent transport and exocytose of GABA and glutamate in brain in hypergravity.

NASA Astrophysics Data System (ADS)

Borisova, T.; Pozdnyakova, N.; Krisanova, N.; Himmelreich, N.

γ-Aminobutyric acid (GABA) and L-glutamate are the most widespread neurotransmitter amino acids in the mammalian central nervous system. GABA is now widely recognized as the major inhibitory neurotransmitter. L-glutamate mediates the most of excitatory synaptic neurotransmission in the brain. They involved in the main aspects of normal brain function. The nerve terminals (synaptosomes) offer several advantages as a model system for the study of general mechanisms of neurosecretion. Our data allowed to conclude that exposure of animals to hypergravity (centrifugation of rats at 10G for 1 hour) had a profound effect on synaptic processes in brain. Comparative analysis of uptake and release of GABA and glutamate have demonstrated that hypergravity loading evokes oppositely directed alterations in inhibitory and excitatory signal transmission. We studied the maximal velocities of [^3H]GABA reuptake and revealed more than twofold enhancement of GABA transporter activity (Vmax rises from 1.4 |pm 0.3 nmol/min/mg of protein in the control group to 3.3 ± 0.59 nmol/min/mg of protein for animals exposed to hypergravity (P ≤ 0.05)). Recently we have also demonstrated the significant lowering of glutamate transporter activity (Vmax of glutamate reuptake decreased from 12.5 ± 3.2 nmol/min/mg of protein in the control group to 5.6 ± 0.9 nmol/min/mg of protein in the group of animals, exposed to the hypergravity stress (P ≤ 0.05)). Significant changes occurred in release of neurotransmitters induced by stimulating exocytosis with the agents, which depolarized nerve terminal plasma membrane. Depolarization-evoked Ca2+-stimulated release was more abundant for GABA (7.2 ± 0.54% and 11,74 ±1,2 % of total accumulated label for control and hypergravity, respectively (P≤0.05)) and was essentially less for glutamate (14.4 ± 0.7% and 6.2 ± 1.9%) after exposure of animals to centrifuge induced artificial gravity. Changes observed in depolarization-evoked exocytotic release seem to be in a concert with alterations of plasma membrane transporters activity studied. Perhaps, lowering of glutamate transporter activity and increase of the velocity of GABA uptake correlated with diminution and augmentation of exocytotic release of these neurotransmitters, respectively. It is possible to suggest that observed changes in the activity of the processes responsible for the uptake and release of excitatory and inhibitory neurotransmitters are likely to be physiologically important and reflect making protective mechanisms more active for neutralization of harm influence of hypergravity stress.

Differential responses of targeted lung redox enzymes to rat exposure to 60 or 85% oxygen

PubMed Central

Gan, Zhuohui; Roerig, David L.; Clough, Anne V.

2011-01-01

Rat exposure to 60% O2 (hyper-60) or 85% O2 (hyper-85) for 7 days confers susceptibility or tolerance, respectively, of the otherwise lethal effects of exposure to 100% O2. The objective of this study was to determine whether activities of the antioxidant cytosolic enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1) and mitochondrial complex III are differentially altered in hyper-60 and hyper-85 lungs. Duroquinone (DQ), an NQO1 substrate, or its hydroquinone (DQH2), a complex III substrate, was infused into the arterial inflow of isolated, perfused lungs, and the venous efflux rates of DQH2 and DQ were measured. Based on inhibitor effects and kinetic modeling, capacities of NQO1-mediated DQ reduction (Vmax1) and complex III-mediated DQH2 oxidation (Vmax2) increased by ∼140 and ∼180% in hyper-85 lungs, respectively, compared with rates in lungs of rats exposed to room air (normoxic). In hyper-60 lungs, Vmax1 increased by ∼80%, with no effect on Vmax2. Additional studies revealed that mitochondrial complex I activity in hyper-60 and hyper-85 lung tissue homogenates was ∼50% lower than in normoxic lung homogenates, whereas mitochondrial complex IV activity was ∼90% higher in only hyper-85 lung tissue homogenates. Thus NQO1 activity increased in both hyper-60 and hyper-85 lungs, whereas complex III activity increased in hyper-85 lungs only. This increase, along with the increase in complex IV activity, may counter the effects the depression in complex I activity might have on tissue mitochondrial function and/or reactive oxygen species production and may be important to the tolerance of 100% O2 observed in hyper-85 rats. PMID:21551015

Differential responses of targeted lung redox enzymes to rat exposure to 60 or 85% oxygen.

PubMed

Gan, Zhuohui; Roerig, David L; Clough, Anne V; Audi, Said H

2011-07-01

Rat exposure to 60% O(2) (hyper-60) or 85% O(2) (hyper-85) for 7 days confers susceptibility or tolerance, respectively, of the otherwise lethal effects of exposure to 100% O(2). The objective of this study was to determine whether activities of the antioxidant cytosolic enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1) and mitochondrial complex III are differentially altered in hyper-60 and hyper-85 lungs. Duroquinone (DQ), an NQO1 substrate, or its hydroquinone (DQH(2)), a complex III substrate, was infused into the arterial inflow of isolated, perfused lungs, and the venous efflux rates of DQH(2) and DQ were measured. Based on inhibitor effects and kinetic modeling, capacities of NQO1-mediated DQ reduction (V(max1)) and complex III-mediated DQH(2) oxidation (V(max2)) increased by ∼140 and ∼180% in hyper-85 lungs, respectively, compared with rates in lungs of rats exposed to room air (normoxic). In hyper-60 lungs, V(max1) increased by ∼80%, with no effect on V(max2). Additional studies revealed that mitochondrial complex I activity in hyper-60 and hyper-85 lung tissue homogenates was ∼50% lower than in normoxic lung homogenates, whereas mitochondrial complex IV activity was ∼90% higher in only hyper-85 lung tissue homogenates. Thus NQO1 activity increased in both hyper-60 and hyper-85 lungs, whereas complex III activity increased in hyper-85 lungs only. This increase, along with the increase in complex IV activity, may counter the effects the depression in complex I activity might have on tissue mitochondrial function and/or reactive oxygen species production and may be important to the tolerance of 100% O(2) observed in hyper-85 rats.

Is ADH1C genotype relevant for the cardioprotective effect of alcohol?

PubMed

Høiseth, Gudrun; Magnus, Per; Knudsen, Gun Peggy; Jansen, Mona Dverdal; Næss, Oyvind; Tambs, Kristian; Mørland, Jørg

2013-03-01

The cardioprotective effect of ethanol has been suggested to be linked to one of the ethanol metabolizing enzymes (ADH1C), which constitutes a high V(max) and a low V(max) variant. This has been demonstrated in some studies, while others have not been able to replicate the findings. The aim of the present study was to investigate the relation between the different ADH1C genotypes, death from coronary heart disease (CHD) and alcohol in a material larger than the previously published studies. Eight hundred CHD deaths as well as 1303 controls were genotyped for the high V(max) (γ1) and the low V(max) (γ2) ADH1C variant. Information of alcohol use was available for all subjects. Multiple logistic regression analyses was used to study if the decreased risk of death from CHD in alcohol consuming subjects was more pronounced in subjects homozygous for the γ2 allele (γ2γ2 subjects) compared to γ1γ1 and γ1γ2 subjects. The odds ratio (OR) for death from CHD in alcohol consumers compared to abstainers was similar in the genotype groups, i.e., 0.62 (95% CI: 0.43-0.88) in γ1γ1 subjects and 0.62 (95% CI: 0.42-0.91) in γ2γ2 subjects. Also when stratifying the results by gender and when dividing alcohol consumers into different alcohol consumption groups, there was no difference in the OR between the different genotype groups. This study, which included the largest study group published so far, failed to find any link between the ADH1C genotype and the cardioprotective effects of alcohol. Copyright © 2013 Elsevier Inc. All rights reserved.

Saccharomyces cerevisiae FLO1 Gene Demonstrates Genetic Linkage to Increased Fermentation Rate at Low Temperatures

PubMed Central

Deed, Rebecca C.; Fedrizzi, Bruno; Gardner, Richard C.

2017-01-01

Low fermentation temperatures are of importance to food and beverage industries working with Saccharomyces cerevisiae. Therefore, the identification of genes demonstrating a positive impact on fermentation kinetics is of significant interest. A set of 121 mapped F1 progeny, derived from a cross between haploid strains BY4716 (a derivative of the laboratory yeast S288C) and wine yeast RM11-1a, were fermented in New Zealand Sauvignon Blanc grape juice at 12.5°. Analyses of five key fermentation kinetic parameters among the F1 progeny identified a quantitative trait locus (QTL) on chromosome I with a significant degree of linkage to maximal fermentation rate (Vmax) at low temperature. Independent deletions of two candidate genes within the region, FLO1 and SWH1, were constructed in the parental strains (with S288C representing BY4716). Fermentation of wild-type and deletion strains at 12.5 and 25° confirmed that the genetic linkage to Vmax corresponds to the S288C version of the FLO1 allele, as the absence of this allele reduced Vmax by ∼50% at 12.5°, but not at 25°. Reciprocal hemizygosity analysis (RHA) between S288C and RM11-1a FLO1 alleles did not confirm the prediction that the S288C version of FLO1 was promoting more rapid fermentation in the opposing strain background, suggesting that the positive effect on Vmax derived from S288C FLO1 may only provide an advantage in haploids, or is dependent on strain-specific cis or trans effects. This research adds to the growing body of evidence demonstrating the role of FLO1 in providing stress tolerance to S. cerevisiae during fermentation. PMID:28143947

Allosteric modulation of semicarbazide-sensitive amine oxidase activities in vitro by imidazoline receptor ligands

PubMed Central

Holt, Andrew; Wieland, Barbara; Baker, Glen B

2004-01-01

Evidence indicates that imidazoline I2 binding sites (I2BSs) are present on monoamine oxidase (MAO) and on soluble (plasma) semicarbazide-sensitive amine oxidase enzymes. The binding site on MAO has been described as a modulatory site, although no effects on activity are thought to have been observed as a result of ligands binding to these sites. We examined the effects in vitro of several imidazoline binding site ligands on activities of bovine plasma amine oxidase (BPAO) and porcine kidney diamine oxidase (PKDAO) in a spectrophotometric protocol. While both enzymes were inhibited at high concentrations of all ligands, clonidine, cirazoline and oxymetazoline were seen, at lower concentrations, to increase activity of BPAO versus benzylamine, but not of PKDAO versus putrescine. This effect was substrate dependent, with mixed or biphasic inhibition of spermidine, methylamine, p-tyramine and β-phenylethylamine oxidation observed at cirazoline concentrations that increased benzylamine oxidation. With benzylamine as substrate, clonidine decreased KM (EC50 8.82 μM, Emax 75.1% of control) and increased Vmax (EC50 164.6 μM, Emax 154.1% of control). Cirazoline decreased Vmax (EC50 2.15 μM, Emax 91.4% of control), then decreased KM (EC50 5.63 μM, Emax 42.6% of control) and increased Vmax (EC50 49.0 μM, Emax 114.4% of decreased Vmax value). Data for clonidine fitted a mathematical model for two-site nonessential activation plus linear intersecting noncompetitive inhibition. Data for cirazoline were consistent with involvement of a fourth site. These results reveal an ability of imidazoline ligands to modulate BPAO kinetics allosterically. The derived mechanism may have functional significance with respect to modulation of MAO by I2BS ligands. PMID:15451775

High-intensity interval training increases in vivo oxidative capacity with no effect on P(i)→ATP rate in resting human muscle.

PubMed

Larsen, Ryan G; Befroy, Douglas E; Kent-Braun, Jane A

2013-03-01

Mitochondrial ATP production is vital for meeting cellular energy demand at rest and during periods of high ATP turnover. We hypothesized that high-intensity interval training (HIT) would increase ATP flux in resting muscle (VPi→ATP) in response to a single bout of exercise, whereas changes in the capacity for oxidative ATP production (Vmax) would require repeated bouts. Eight untrained men (27 ± 4 yr; peak oxygen uptake = 36 ± 4 ml·kg(-1)·min(-1)) performed six sessions of HIT (4-6 × 30-s bouts of all-out cycling with 4-min recovery). After standardized meals and a 10-h fast, VPi→ATP and Vmax of the vastus lateralis muscle were measured using phosphorus magnetic resonance spectroscopy at 4 Tesla. Measurements were obtained at baseline, 15 h after the first training session, and 15 h after completion of the sixth session. VPi→ATP was determined from the unidirectional flux between Pi and ATP, using the saturation transfer technique. The rate of phosphocreatine recovery (kPCr) following a maximal contraction was used to calculate Vmax. While kPCr and Vmax were unchanged after a single session of HIT, completion of six training sessions resulted in a ∼14% increase in muscle oxidative capacity (P ≤ 0.004). In contrast, neither a single nor six training sessions altered VPi→ATP (P = 0.74). This novel analysis of resting and maximal high-energy phosphate kinetics in vivo in response to HIT provides evidence that distinct aspects of human skeletal muscle metabolism respond differently to this type of training.

Decline in spirometric variables in grain workers from start of employment: differential effect of duration of follow up.

PubMed Central

Zejda, J E; Pahwa, P; Dosman, J A

1992-01-01

Prospective study of 164 young men from the start of employment in grain elevators showed that of those seen at the initial evaluation of respiratory state only 30% were available for a complete four year follow up. The drop out of subjects could represent a health related selection leading to the underestimation of respiratory effects of exposure to grain dust as assessed in the survivor group. This hypothesis was examined by comparisons of longitudinal changes in lung function in four groups defined by the duration of follow up involving the initial examination and periodic evaluations after one, two, and four years of work. Sixty four men were tested only on the initial examination (group I), 18 underwent two (group II), 31 underwent three (group III), and 51 (group IV) all four examinations. The groups had similar mean ages (range: 19.4-20.1 years), mean duration of previous exposure to grain dust (range: 8-13 weeks), smoking habits, lung function, and prevalences of respiratory symptoms evaluated on the initial occasion. The average decline in lung function over the first year was associated with duration of follow up. The annual decline in FVC (ml) was 58 in group II, 41 in group III and -55 (increase) in group IV; the decline in FEV1 (ml) was 224, 130, and 70 respectively. The differences for the annual declines of FEV1, FEF25-759 Vmax509 and Vmax25 were significant between groups II and IV, and the FEF25-759 Vmax509 and Vmax25 differed significantly between groups II and III. The results show that the restriction of analysis to the survivors may underestimate the relation between work and respiratory impairment. PMID:1515349

Phosphate and acidosis act synergistically to depress peak power in rat muscle fibers.

PubMed

Nelson, Cassandra R; Debold, Edward P; Fitts, Robert H

2014-11-15

Skeletal muscle fatigue is characterized by the buildup of H(+) and inorganic phosphate (Pi), metabolites that are thought to cause fatigue by inhibiting muscle force, velocity, and power. While the individual effects of elevated H(+) or Pi have been well characterized, the effects of simultaneously elevating the ions, as occurs during fatigue in vivo, are still poorly understood. To address this, we exposed slow and fast rat skinned muscle fibers to fatiguing levels of H(+) (pH 6.2) and Pi (30 mM) and determined the effects on contractile properties. At 30°C, elevated Pi and low pH depressed maximal shortening velocity (Vmax) by 15% (4.23 to 3.58 fl/s) in slow and 31% (6.24 vs. 4.55 fl/s) in fast fibers, values similar to depressions from low pH alone. Maximal isometric force dropped by 36% in slow (148 to 94 kN/m(2)) and 46% in fast fibers (148 to 80 kN/m(2)), declines substantially larger than what either ion exerted individually. The strong effect on force combined with the significant effect on velocity caused peak power to decline by over 60% in both fiber types. Force-stiffness ratios significantly decreased with pH 6.2 + 30 mM Pi in both fiber types, suggesting these ions reduced force by decreasing the force per bridge and/or increasing the number of low-force bridges. The data indicate the collective effects of elevating H(+) and Pi on maximal isometric force and peak power are stronger than what either ion exerts individually and suggest the ions act synergistically to reduce muscle function during fatigue. Copyright © 2014 the American Physiological Society.

Propulsion of the Water Flea, Daphnia magna: Experiments, Scaling, and Modelling

NASA Astrophysics Data System (ADS)

Skipper, A. N.; Murphy, D.; Webster, D. R.; Yen, J.

2016-02-01

The freshwater crustacean Daphnia magna is a widely studied zooplankton in relation to food webs, predator-prey interactions, and other biological/ecological considerations; however, their locomotion is poorly quantified and understood. These water fleas utilize a hop-and-sink mechanism that consists of making quick, impulsive jumps by beating their antennae to propel themselves forward ( 1 body length). The animals then sink for a period, during which they stretch out their antennae to increase drag and thereby reduce their sinking velocity. Time-resolved three-dimensional flow fields surrounding the animals were quantified with a unique infrared tomographic particle image velocity (tomo-PIV) system. Three-dimensional kinematics data were also extracted from the image sequences. In the current work, we compared body kinematics and flow disturbance among organisms of size in the range of 1.3 to 2.8 mm. The stroke cycle averaged 150 ms in duration, ranging from 100 to 180 ms; this period is generally evenly split between the power and recovery strokes. The range of peak hop velocity was 27.2 to 32.5 mm/s, and peak acceleration was in the range of 0.68 to 1.8 m/s2. The results showed a distinct relationship between peak hop speed (Vmax 14 BL/s) and body size; these data collapsed onto a single time-record curve during the power stroke when properly non-dimensionalized. The fluid flow induced by each antennae consisted of a viscous vortex ring that demonstrated a slow decay in the wake. The strength, size, and decay of the induced viscous vortex rings were compared as a function of organism size. Finally, the viscous vortex rings were analyzed in the context of a double Stokeslet model that consisted of two impulsively applied point forces separated by the animal width.

Risk factors for left ventricular hypertrophy: role of Na(+)-Li+ countertransport.

PubMed

Neves, P L; Faisca, M; Gomes, V; Cacodcar, S; Bernardo, I; Anunciada, A I; Viegas, E; Martins, H; da Silva, A M

1996-06-01

Left ventricular hypertrophy (LVH) is associated with an increase in cardiovascular death in essential hypertension (EH). The factors involved in LVH are multiple and complex. We looked for risk factors of LVH in a group of 28 nonobese patients with EH (mean age = 45.3 years). We analyzed the activity of several erythrocyte ion transports (Vmax of NaLi countertransport, NaKCl cotransport and NaK-pump, and the Na-leak Kp Na), the intracellular Na and the insulin sensitivity index. All these parameters were used as independent variables whereas the left ventricular mass index (LVMI) was used as the dependent variable. Variables showing a significant univariate correlation (age, time of EH, mean blood pressure and Vmax of NaLi countertransport) were introduced in a stepwise multiple regression model. Only age (P = 0.014), time of EH (P = 0.038) and Vmax of NaLi countertransport (P = 0.032) were independently associated with LVMI (R2 = 0.581, P = 0.0001). The NaLi CT, an operating mode of the NaH exchanger that facilitates cellular growth, may be a marker of LVH, and consequently a marker of increased cardiovascular risk.

Renormalization-group study of the Nagel-Schreckenberg model

NASA Astrophysics Data System (ADS)

Teoh, Han Kheng; Yong, Ee Hou

2018-03-01

We study the phase transition from free flow to congested phases in the Nagel-Schreckenberg (NS) model by using the dynamically driven renormalization group (DDRG). The breaking probability p that governs the driving strategy is investigated. For the deterministic case p =0 , the dynamics remain invariant in each renormalization-group (RG) transformation. Two fully attractive fixed points, ρc*=0 and 1, and one unstable fixed point, ρc*=1 /(vmax+1 ) , are obtained. The critical exponent ν which is related to the correlation length is calculated for various vmax. The critical exponent appears to decrease weakly with vmax from ν =1.62 to the asymptotical value of 1.00. For the random case p >0 , the transition rules in the coarse-grained scale are found to be different from the NS specification. To have a qualitative understanding of the effect of stochasticity, the case p →0 is studied with simulation, and the RG flow in the ρ -p plane is obtained. The fixed points p =0 and 1 that govern the driving strategy of the NS model are found. A short discussion on the extension of the DDRG method to the NS model with the open-boundary condition is outlined.

Chemical mechanism of D-amino acid oxidase from Rhodotorula gracilis: pH dependence of kinetic parameters.

PubMed Central

Ramón, F; Castillón, M; De La Mata, I; Acebal, C

1998-01-01

The variation of kinetic parameters of d-amino acid oxidase from Rhodotorula gracilis with pH was used to gain information about the chemical mechanism of the oxidation of D-amino acids catalysed by this flavoenzyme. d-Alanine was the substrate used. The pH dependence of Vmax and Vmax/Km for alanine as substrate showed that a group with a pK value of 6.26-7.95 (pK1) must be unprotonated and a group with a pK of 10.8-9.90 (pK2) must be protonated for activity. The lower pK value corresponded to a group on the enzyme involved in catalysis and whose protonation state was not important for binding. The higher pK value was assumed to be the amino group of the substrate. Profiles of pKi for D-aspartate as competitive inhibitor showed that binding is prevented when a group on the enzyme with a pK value of 8.4 becomes unprotonated; this basic group was not detected in Vmax/Km profiles suggesting its involvement in binding of the beta-carboxylic group of the inhibitor. PMID:9461524

Small intestinal sulphoxidation of albendazole.

PubMed

Villaverde, C; Alvarez, A I; Redondo, P; Voces, J; Del Estal, J L; Prieto, J G

1995-05-01

1. The in vitro sulphoxidation of Albendazole (ABZ) by rat intestinal microsomes has been examined. The results revealed intestinal sulphoxidation of ABZ by intestinal microsomes in a NADPH-dependent enzymatic system. The kinetic constants for sulphoxidase activity were Vmax = 46 pmol/min/mg protein and Michaelis constant Km = 6.8 microM. 2. The possible effect of inducers (Arochlor 1254 and ABZ pretreatment) and inhibitors (erythromycin, methimazole, carbon monoxide and fenbendazole), was also studied. In rat pretreated with Arochlor 1254, Vmax was 52 pmol/min/mg protein, whereas oral administration of ABZ increased the intestinal sulphoxidation of the drug, Vmax being 103 pmol/min/mg protein. 3. Erythromycin did not change the enzymatic bioconversion of ABZ, but methimazole and carbon monoxide inhibited the enzyme activity by approximately 60 and 30% respectively. Fenbendazole (a structural analogue of ABZ) was a competitive inhibitor of the sulphoxidation process, characterized by a Ki or 69 microM. 4. These data demonstrate that the intestinal enzymes contributing to the initial sulphoxidation of ABZ may be similar to the hepatic enzymes involved in the biotransformation process by the P450 and FMO systems, a conclusion that needs to be further established.

Near- and far-field aerodynamics in insect hovering flight: an integrated computational study.

PubMed

Aono, Hikaru; Liang, Fuyou; Liu, Hao

2008-01-01

We present the first integrative computational fluid dynamics (CFD) study of near- and far-field aerodynamics in insect hovering flight using a biology-inspired, dynamic flight simulator. This simulator, which has been built to encompass multiple mechanisms and principles related to insect flight, is capable of 'flying' an insect on the basis of realistic wing-body morphologies and kinematics. Our CFD study integrates near- and far-field wake dynamics and shows the detailed three-dimensional (3D) near- and far-field vortex flows: a horseshoe-shaped vortex is generated and wraps around the wing in the early down- and upstroke; subsequently, the horseshoe-shaped vortex grows into a doughnut-shaped vortex ring, with an intense jet-stream present in its core, forming the downwash; and eventually, the doughnut-shaped vortex rings of the wing pair break up into two circular vortex rings in the wake. The computed aerodynamic forces show reasonable agreement with experimental results in terms of both the mean force (vertical, horizontal and sideslip forces) and the time course over one stroke cycle (lift and drag forces). A large amount of lift force (approximately 62% of total lift force generated over a full wingbeat cycle) is generated during the upstroke, most likely due to the presence of intensive and stable, leading-edge vortices (LEVs) and wing tip vortices (TVs); and correspondingly, a much stronger downwash is observed compared to the downstroke. We also estimated hovering energetics based on the computed aerodynamic and inertial torques, and powers.

Systolic time interval data acquisition system. Specialized cardiovascular studies

NASA Technical Reports Server (NTRS)

Baker, J. T.

1976-01-01

The development of a data acquisition system for noninvasive measurement of systolic time intervals is described. R-R interval from the ECG determines instantaneous heart rate prior to the beat to be measured. Total electromechanical systole (Q-S2) is measured from the onset of the ECG Q-wave to the onset of the second heart sound (S2). Ejection time (ET or LVET) is measured from the onset of carotid upstroke to the incisure. Pre-ejection period (PEP) is computed by subtracting ET from Q-S2. PEP/ET ratio is computed directly.

Evaluation of the Effects of Sildenafil Citrate (Viagra) on Vertebral Artery Blood Flow in Patients with Vertebro-Basilar Insufficiency

PubMed Central

Berilgen, Sait; Ozdemir, Huseyin; Tekatas, Aslan; Ogur, Erkin

2008-01-01

Objective To investigate the effects of sildenafil citrate (Viagra) on the vertebral artery blood flow of patients with vertebro-basilar insufficiency (VBI) using color duplex sonography (CDS). Materials and Methods The study included 21 patients with VBI (aged 31-76; mean 61.0 ± 10.5 yrs). We administered a 50 mg oral dose of sildenafil citrate to all patients. Next, we measured the peak systolic velocity (Vmax), end diastolic velocity (Vmin), resistive index (RI), pulsatility index (PI), diameter, area, and flow volume (FV) of vertebral arteries using CDS before the administration of sildenafil citrate; 45 minutes after, and 75 minutes after administration. Statistical testing was performed using SPSS for windows version 11.0. The statistical test used to determine the outcome of the analysis was the repeated measures analysis of variance (ANOVA) test. Results Compared to the baseline values, the vertebral artery diameter, area, and FV increased significantly following the administration of sildenafil citrate. The diameter, area and FV increased from 3.39 mm at 45 minutes to 3.64 mm at 75 minutes, 9.43 cm2 to 10.80 cm2 at 45 minutes and 10.81 cm2 at 75 minutes, as well as from 0.07 L/min at baseline to 0.09 L/min at 45 minutes and unchanged at 75 minutes, respectively. Conclusion Sildenafil citrate elicited a significant effect on vertebral artery diameter, area and FVs. PMID:19039262

Optimization of terrestrial ecosystem model parameters using atmospheric CO2 concentration data with a global carbon assimilation system (GCAS)

NASA Astrophysics Data System (ADS)

Chen, Z.; Chen, J.; Zhang, S.; Zheng, X.; Shangguan, W.

2016-12-01

A global carbon assimilation system (GCAS) that assimilates ground-based atmospheric CO2 data is used to estimate several key parameters in a terrestrial ecosystem model for the purpose of improving carbon cycle simulation. The optimized parameters are the leaf maximum carboxylation rate at 25° (Vmax25 ), the temperature sensitivity of ecosystem respiration (Q10), and the soil carbon pool size. The optimization is performed at the global scale at 1°resolution for the period from 2002 to 2008. Optimized multi-year average Vmax25 values range from 49 to 51 μmol m-2 s-1 over most regions of world. Vegetation from tropical zones has relatively lower values than vegetation in temperate regions. Optimized multi-year average Q10 values varied from 1.95 to 2.05 over most regions of the world. Relatively high values of Q10 are derived over high/mid latitude regions. Both Vmax25 and Q10 exhibit pronounced seasonal variations at mid-high latitudes. The maximum in occurs during the growing season, while the minima appear during non-growing seasons. Q10 values decreases with increasing temperature. The seasonal variabilities of and Q10 are larger at higher latitudes with tropical or low latitude regions showing little seasonal variabilities.

Effects of CYP2C19 Variants on Fluoxetine Metabolism in vitro.

PubMed

Fang, Ping; He, Jia-Yang; Han, Ai-Xia; Lan, Tian; Dai, Da-Peng; Cai, Jian-Ping; Hu, Guo-Xin

2017-01-01

CYP2C19 is an important member of the cytochrome P450 enzyme superfamily. We recently identified 31 CYP2C19 alleles in the Han Chinese population. The aim of this study was to assess the catalytic activities of these allelic isoforms and their effects on the metabolism of fluoxetine in vitro. The wild-type and 30 CYP2C19 variants were expressed in insect cells and each variant was characterized using fluoxetine as the substrate. Reactions were performed at 37°C with 20-1,000 µmol/L substrate for 30 min. By using ultra-high performance liquid chromatography-mass spectrometry to detect the products, the kinetic parameters Km, Vmax, and intrinsic clearance (Vmax/Km) of norfluoxetine were determined. Among the CYP2C19 variants tested, T130M showed similar intrinsic clearance (Vmax/Km) values with CYP2C19*1, while the intrinsic clearance values of other variants were significantly decreased (from 9.56 to 77.77%). In addition, CYP2C19*3 and *35FS could not be detected because they have no detectable enzyme activity. In China, the assessment of CYP2C19 variants in vitro offers valuable information relevant to the personalized medicine for CYP2C19-metabolized drug. © 2017 S. Karger AG, Basel.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tam, Tsz Kin; Chen, Baowei; Lei, Chenghong

NAD/NADH is a coenzyme found in all living cells, carrying electrons from one reaction to another. We report on characterizations of in situ regeneration of NADH via lipoamide dehydrogenase (LD)-catalyzed electron transfer reaction to regenerate NADH using UV-vis spectroelectrochemistry. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) of NADH regeneration were measured as 0.80 {+-} 0.15 mM and 1.91 {+-} 0.09 {micro}M s-1 in a 1-mm thin-layer spectroelectrochemical cell using gold gauze as the working electrode at the applied potential -0.75 V (vs. Ag/AgCl). The electrocatalytic reduction of the NAD system was further coupled with the enzymatic conversion of pyruvatemore » to lactate by lactate dehydrogenase to examine the coenzymatic activity of the regenerated NADH. Although the reproducible electrocatalytic reduction of NAD into NADH is known to be difficult compared to the electrocatalytic oxidation of NADH, our spectroelectrochemical results indicate that the in situ regeneration of NADH via LD-catalyzed electron transfer reaction is fast and sustainable and can be potentially applied to many NAD/NADH-dependent enzyme systems.« less

Morphological and metabolic adjustments in the small intestine to energy demands of growth, storage, and fasting in the first annual cycle of a hibernating lizard (Tupinambis merianae).

PubMed

do Nascimento, Lucas Francisco R; da Silveira, Lilian Cristina; Nisembaum, Laura Gabriela; Colquhoun, Alison; Abe, Agusto S; Mandarim-de-Lacerda, Carlos Alberto; de Souza, Silvia Cristina R

2016-05-01

Seasonal plasticity in the small intestine of neonatal tegu lizards was investigated using morphometry and analysis of enzymes involved in supplying energy to the intestinal tissue. In the autumn, the intestinal mass (Mi) was 1.0% of body mass and the scaling exponent b=0.92 indicated that Mi was larger in smaller neonates. During arousal from dormancy Mi was 23% smaller; later in spring, Mi increased 60% in relation to the autumn and the exponent b=0.14 indicated that the recovery was disproportionate in smaller tegus. During the autumn, the intestinal villi were greatly elongated; by midwinter, the Hv, SvEp, and VvEp were smaller than during the autumn (59%, 54%, 29%) and were restored to autumn levels during spring. In the active tegus, the maximum activity (Vmax) of enzymes indicated that the enterocytes can obtain energy from different sources, and possess gluconeogenic capacity. During winter, the Vmax of CS, HOAD, GDH, PEPCK was 40-50% lower in relation to the autumn and spring, while the Vmax of HK, PK, LDH, AST was unchanged. The hypoglycemia and the mucosal atrophy/ischemia during winter would prevent the enterocytes from using glucose, whereas they could slowly oxidize fatty acids released from body stores and amino acids from the tissue proteolysis to satisfy their needs of energy. Contrastingly, starvation during spring caused severe mass loss (50%); the tissue protein and the VvEp and VvLP did not change while the thickness of the muscular layer increased 51%, which suggested different effects along the length of the organ. In addition, the Vmax of the glycolytic enzymes was lower, indicating that a regulatory mechanism would spare blood glucose for vital organs during unanticipated food restriction. Copyright © 2016 Elsevier Inc. All rights reserved.

In vivo and in vitro kinetics of ethylene oxide metabolism in rats and mice.

PubMed

Brown, C D; Wong, B A; Fennell, T R

1996-01-01

Ethylene oxide (EO) is a direct-acting mutagen and animal carcinogen used as an industrial intermediate and sterilant with a high potential for human exposure. Kinetics of EO metabolism in rodents can be used to develop human EO dosimetry models. This study examined the kinetics of EO metabolism in vivo and in vitro in male and female F-344 rats and B6C3F1 mice. In vivo studies measured blood and tissue EO levels during and 2-20 min following whole-body inhalation exposure (4 hr, 100 or 330 ppm EO). At 100 ppm EO, the half-life of elimination (t1/2) in rats was 13.8 +/- 0.3 (mean +/- SD) and 10.8 +/- 2.4 min for males and females, respectively, compared to a t1/2 in mice of 3.12 +/- 0.2 and 2.4 +/- 0.2 min in males and females, respectively. On exposure to 330 ppm EO, the t1/2 in mice increased approx twofold, while no change in t1/2 was observed in rats. In vitro kinetic parameters (Vmax and KM) of EO metabolism were determined using tissue cytosol and microsomes. EO metabolism in vitro occurred primarily via cytosolic glutathione S-transferase-mediated EO-GSH conjugation (cGST-EO), with highest activity in the liver. Liver cGST-EO activity (Vmax) was 258 +/- 86.9 and 287 +/- 49.0 nmol/mg protein/min (mean +/- SD) in male and female mice, respectively, compared to 52.7 +/- 10.8 and 29.3 +/- 4.9 in male and female rats, respectively. In rats, but not mice, there was a statistically significant (p < 0.05) gender difference in the Vmax for liver cGST. The KM for liver cGST-EO was approximately 10 mM in both species. The higher Vmax values observed in mice are consistent with the more rapid elimination of EO observed for this species in vivo compared to rats.

Microscale Measurements of Michaelis–Menten Constants of Neuraminidase with Nanogel Capillary Electrophoresis for the Determination of the Sialic Acid Linkage

PubMed Central

2016-01-01

Phospholipid nanogels enhance the stability and performance of the exoglycosidase enzyme neuraminidase and are used to create a fixed zone of enzyme within a capillary. With nanogels, there is no need to covalently immobilize the enzyme, as it is physically constrained. This enables rapid quantification of Michaelis–Menten constants (KM) for different substrates and ultimately provides a means to quantify the linkage (i.e., 2-3 versus 2-6) of sialic acids. The fixed zone of enzyme is inexpensive and easily positioned in the capillary to support electrophoresis mediated microanalysis using neuraminidase to analyze sialic acid linkages. To circumvent the limitations of diffusion during static incubation, the incubation period is reproducibly achieved by varying the number of forward and reverse passes the substrate makes through the stationary fixed zone using in-capillary electrophoretic mixing. A KM value of 3.3 ± 0.8 mM (Vmax, 2100 ± 200 μM/min) was obtained for 3′-sialyllactose labeled with 2-aminobenzoic acid using neuraminidase from Clostridium perfringens that cleaves sialic acid monomers with an α2-3,6,8,9 linkage, which is similar to values reported in the literature that required benchtop analyses. The enzyme cleaves the 2-3 linkage faster than the 2-6, and a KM of 2 ± 1 mM (Vmax, 400 ± 100 μM/min) was obtained for the 6′-sialyllactose substrate. An alternative neuraminidase selective for 2-3 sialic acid linkages generated a KM value of 3 ± 2 mM (Vmax, 900 ± 300 μM/min) for 3′-sialyllactose. With a knowledge of Vmax, the method was applied to a mixture of 2-3 and 2-6 sialyllactose as well as 2-3 and 2-6 sialylated triantennary glycan. Nanogel electrophoresis is an inexpensive, rapid, and simple alternative to current technologies used to distinguish the composition of 3′ and 6′ sialic acid linkages. PMID:27936604

Hindlimb motion during steady flight of the lesser dog-faced fruit bat, Cynopterus brachyotis.

PubMed

Cheney, Jorn A; Ton, Daniel; Konow, Nicolai; Riskin, Daniel K; Breuer, Kenneth S; Swartz, Sharon M

2014-01-01

In bats, the wing membrane is anchored not only to the body and forelimb, but also to the hindlimb. This attachment configuration gives bats the potential to modulate wing shape by moving the hindlimb, such as by joint movement at the hip or knee. Such movements could modulate lift, drag, or the pitching moment. In this study we address: 1) how the ankle translates through space during the wingbeat cycle; 2) whether amplitude of ankle motion is dependent upon flight speed; 3) how tension in the wing membrane pulls the ankle; and 4) whether wing membrane tension is responsible for driving ankle motion. We flew five individuals of the lesser dog-faced fruit bat, Cynopterus brachyotis (Family: Pteropodidae), in a wind tunnel and documented kinematics of the forelimb, hip, ankle, and trailing edge of the wing membrane. Based on kinematic analysis of hindlimb and forelimb movements, we found that: 1) during downstroke, the ankle moved ventrally and during upstroke the ankle moved dorsally; 2) there was considerable variation in amplitude of ankle motion, but amplitude did not correlate significantly with flight speed; 3) during downstroke, tension generated by the wing membrane acted to pull the ankle dorsally, and during upstroke, the wing membrane pulled laterally when taut and dorsally when relatively slack; and 4) wing membrane tension generally opposed dorsoventral ankle motion. We conclude that during forward flight in C. brachyotis, wing membrane tension does not power hindlimb motion; instead, we propose that hindlimb movements arise from muscle activity and/or inertial effects.

Enhanced thrust and speed revealed in the forward flight of a butterfly with transient body translation.

PubMed

Fei, Yueh-Han John; Yang, Jing-Tang

2015-09-01

A butterfly with broad wings, flapping at a small frequency, flies an erratic trajectory at an inconstant speed. A large variation of speed within a cycle is observed in the forward flight of a butterfly. A self-propulsion model to simulate a butterfly is thus created to investigate the transient translation of the body; the results, which are in accordance with experimental data, show that the shape of the variation of the flight speed is similar to a sinusoidal wave with a maximum (J=0.89) at the beginning of the downstroke, and a decrease to a minimum (J=0.17) during a transition from downstroke to upstroke; the difference between the extrema of the flight speed is enormous in a flapping cycle. At a high speed, a clapping motion of the butterfly wings decreases the generation of drag. At a small speed, a butterfly is able to capture the induced wakes generated in a downstroke, and effectively generates a thrust at the beginning of an upstroke. The wing motion of a butterfly skillfully interacts with its speed so as to enable an increased speed with the same motion. Considering a butterfly to fly in a constant inflow leads to either an underestimate of its speed or an overestimate of its generated lift, which yields an inaccurate interpretation of the insect's flight. Our results reveal the effect of transient translation on a butterfly in forward flight, which is especially important for an insect with a small flapping frequency.

Control for small-speed lateral flight in a model insect.

PubMed

Zhang, Yan Lai; Sun, Mao

2011-09-01

Controls required for small-speed lateral flight of a model insect were studied using techniques based on the linear theories of stability and control (the stability and control derivatives were computed by the method of computational fluid dynamics). The main results are as follows. (1) Two steady-state lateral motions can exist: one is a horizontal side translation with the body rolling to the same side of the translation by a small angle, and the other is a constant-rate yaw rotation (rotation about the vertical axis). (2) The side translation requires an anti-symmetrical change in the stroke amplitudes of the contralateral wings, and/or an anti-symmetrical change in the angles of attack of the contralateral wings, with the down- and upstroke angles of attack of a wing having equal change. The constant-rate yaw rotation requires an anti-symmetrical change in the angles of attack of the contralateral wings, with the down- and upstroke angles of attack of a wing having differential change. (3) For the control of the horizontal side translation, control input required for the steady-state motion has an opposite sign to that needed for initiating the motion. For example, to have a steady-state left side-translation, the insect needs to increase the stroke amplitude of the left wing and decrease that of the right wing to maintain the steady-state flight, but it needs an opposite change in stroke amplitude (decreasing the stroke amplitude of the left wing and increasing that of the right wing) to enter the flight.

Enhanced thrust and speed revealed in the forward flight of a butterfly with transient body translation

NASA Astrophysics Data System (ADS)

Fei, Yueh-Han John; Yang, Jing-Tang

2015-09-01

A butterfly with broad wings, flapping at a small frequency, flies an erratic trajectory at an inconstant speed. A large variation of speed within a cycle is observed in the forward flight of a butterfly. A self-propulsion model to simulate a butterfly is thus created to investigate the transient translation of the body; the results, which are in accordance with experimental data, show that the shape of the variation of the flight speed is similar to a sinusoidal wave with a maximum (J =0.89 ) at the beginning of the downstroke, and a decrease to a minimum (J =0.17 ) during a transition from downstroke to upstroke; the difference between the extrema of the flight speed is enormous in a flapping cycle. At a high speed, a clapping motion of the butterfly wings decreases the generation of drag. At a small speed, a butterfly is able to capture the induced wakes generated in a downstroke, and effectively generates a thrust at the beginning of an upstroke. The wing motion of a butterfly skillfully interacts with its speed so as to enable an increased speed with the same motion. Considering a butterfly to fly in a constant inflow leads to either an underestimate of its speed or an overestimate of its generated lift, which yields an inaccurate interpretation of the insect's flight. Our results reveal the effect of transient translation on a butterfly in forward flight, which is especially important for an insect with a small flapping frequency.

Intracellular Calcium Dynamics and the Acceleration of Sinus Rhythm by β-Adrenergic Stimulation

PubMed Central

Joung, Boyoung; Tang, Liang; Maruyama, Mitsunori; Han, Seongwook; Chen, Zhenhui; Stucky, Marcelle; Jones, Larry R.; Fishbein, Michael C.; Weiss, James N.; Chen, Peng-Sheng; Lin, Shien-Fong

2009-01-01

Background Recent evidence indicates that membrane voltage and Ca2+ clocks jointly regulate sinoatrial node (SAN) automaticity. Here we test the hypothesis that sinus rate acceleration by β-adrenergic stimulation involves synergistic interactions between these clock mechanisms. Methods and Results We simultaneously mapped intracellular calcium (Cai) and membrane potential (Vm) in 25 isolated canine right atrium (RA), using previously described criteria of the timing of late diastolic Cai elevation (LDCAE) relative to the action potential (AP) upstroke to detect the Ca2+ clock. Before isoproterenol, the earliest pacemaking site occurred in the inferior SAN, and LDCAE was observed in only 4/25 preparations. Isoproterenol (1 μmol/L) increased sinus rate and shifted pacemaking site to superior SAN, concomitant with the appearance of LDCAE preceding the AP upstroke by 98 ± 31 ms. Caffeine had similar effects, while SR Ca2+ depletion with ryanodine and thapsigargin prevented isoproterenol-induced LDCAE and blunted sinus rate acceleration. Cai transient relaxation time during ISO was shorter in superior SAN (124 ± 34 ms) than inferior SAN (138 ± 24 ms, p = 0.01) or RA (164 ± 33 ms, p = 0.001), and was associated with a lower SR Ca2+ ATPase pump to phospholamban protein ratio in SAN than in RA. If current blockade with ZD 7288 modestly blunted, but did not prevent LDCAE or sinus rate acceleration by isoproterenol. Conclusions Acceleration of the Ca2+ clock in the superior SAN plays an important role in sinus acceleration during β-adrenergic stimulation, interacting synergistically with the voltage clock to increase sinus rate. PMID:19188501

Effect of quantifying peptide release on ruminal protein degradation determined using the inhibitor in vitro system.

PubMed

Colombini, S; Broderick, G A; Clayton, M K

2011-04-01

The aim of this work was to compare use of an o-phthaldialdehyde (OPA) colorimetric assay (OPA-C), which responds to both free AA and peptides, with an OPA fluorimetric assay (OPA-F), which is insensitive to peptides, to quantify rates of ruminal protein degradation in the inhibitor in vitro system using Michaelis-Menten saturation kinetics. Four protein concentrates (expeller-extracted soybean meal, ESBM; 2 solvent-extracted soybean meals, SSBM1 and SSBM2; and casein) were incubated in a ruminal in vitro system treated with hydrazine and chloramphenicol to inhibit microbial uptake of protein degradation products. Proteins were weighed to give a range of N concentrations (from 0.15 to 3 mg of N/mL of inoculum) and incubated with 10 mL of ruminal inoculum and 5 mL of buffer; fermentations were stopped after 2 h by adding trichloroacetic acid (TCA). Proteins were analyzed for buffer-soluble N and buffer extracts were treated with TCA to determine N degraded at t=0 (FD0). The TCA supernatants were analyzed for ammonia (phenol-hypochlorite assay), total AA (TAA; OPA-F), and TAA plus oligopeptides (OPA-C) by flow injection analysis. Velocity of protein degradation was computed from extent of release of 1) ammonia plus free TAA or 2) ammonia plus free TAA and peptides. Rate of degradation (kd) was quantified using nonlinear regression of the integrated Michaelis-Menten equation. The parameters Km (Michaelis constant) and kd (Vmax/Km), where Vmax=maximum velocity, were estimated directly; kd values were adjusted (Akd) for the fraction FD0 using the equation Akd=kd-FD0/2. The OPA-C assay yielded faster degradation rates due to the contribution of peptides to the fraction degraded (overall mean=0.280/h by OPA-C and 0.219/h by OPA-F). Degradation rates for SSBM samples (0.231/h and 0.181/h) and ESBM (0.086/h) obtained by the OPA-C assay were more rapid than rates reported by the National Research Council (NRC). Both assays indicated that the 2 SSBM differed in rumen-undegradable protein (RUP) content; the more slowly degraded SSBM had RUP content (35% by OPA-C) similar to that reported by the NRC. The RUP content of ESBM (42% by OPA-C) was lower than the NRC value. Preliminary studies with 4 additional protein concentrates confirmed that accounting for peptide formation increased degradation rate; however, a trend for an interaction between assay and protein source suggested that peptide release made a smaller contribution to rate for more slowly degraded proteins. The OPA-C assay is a simple and reliable method to quantify formation of small peptides. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The Inhibition of Hepatic and Renal Glucuronidation of p-Nitrophenol and 4-Methylumbelliferone by Oil Palm Empty Fruit Bunch Lignin and Its Main Oxidation Compounds

PubMed Central

Salleh, Norliyana Mohamad; Ismail, Sabariah; Ibrahim, Mohamad Nasir Mohamad

2017-01-01

Background: In order to develop oil palm empty fruit bunch (EFB) lignin as a nutraceutical and health supplement, the investigation of its potential in interacting with other drugs via inhibition of drug-metabolizing enzymes (DMEs) would ensure product safety. Objective: The study was aimed to investigate the in vitro effect of oil palm EFB lignin and its main oxidation compounds on phase II DME UDP-glucuronosyltransferases (UGTs) in rat liver and kidney microsomes. Materials and Methods: The p-nitrophenol (p-NP) and 4-methylumbelliferone (4-MU) were employed as probe substrates in glucuronidation assays. The effect of soda oil palm EFB lignin on Vmax, Km, CLint, Ki, and mode of inhibition of 4-MU glucuronidation in RLM was also determined. Results: The inhibitory potency of oil palm EFB lignin for both p-NP and 4-MU glucuronidation in rat liver microsome (RLM) and rat kidneys microsomes (RKM) was found to be in the rank order of soda > kraft > organosolv. However, the inhibitory potency of its main oxidation compounds were in the rank order of vanillin > syringaldehyde > p-hydroxybenzaldehyde. Soda oil palm EFB lignin exhibited mixed-type inhibition against 4-MU glucuronidation in RLM, showing the change in apparent Vmax and with only a minor effect on Km compared with control. Conclusions: The findings showed that effect of oil palm EFB lignin on both p-NP and 4-MU glucuronidation in RLM and RKM was enhanced by the presence of vanillin as well as flavonoids. Kinetic study showed that soda oil palm EFB lignin exhibited strong inhibition on UGT activity in RLM with mixed-type inhibition mode. SUMMARY The inhibitory potential of oil palm EFB lignin extracts for p-NP and 4-MU glucuronidation in RLM and RKM can be listed in the following rank order: soda > kraft > organosolvThe inhibitory potential of oil palm EFB lignin main oxidation compounds for p-NP and 4-MU glucuronidation in RLM and RKM can be listed in the following rank order: vanillin > syringaldehyde > p-hydroxybenzaldehydeResults suggested that the effect of oil palm EFB lignin on p-NP and 4-MU glucuronidation activity in both RLM and RKM was enhanced by the presence of vanillin as well as total flavonoid contentResults also suggested that oil palm EFB lignin may inhibit glucuronidation of substrate by UGT enzymes, especially UGT1A6, particularly in rat liver Abbreviations used: p-NP: p-Nitrophenol, 4-MU: 4-Methylumbelliferone, EFB: Empty fruit bunch, DME: Drug-metabolizing enzymes, UGT: UDPglucuronosyltransferase, Vmax: Maximal reaction velocity, Km: Michaelis-Menten constant, CLint: Intrinsic clearance, Ki: Dissociation constant of an inhibitor enzyme complex, 4-MUG: 4-Methylumbelliferone glucuronide, DMSO: Dimethyl sulfoxide, IC50: Half maximal inhibitory concentration, p-NPG: p-Nitrophenol glucuronide, RKM: Rat kidneys microsomes, RLM: Rat liver microsome, UDPGA: UDPglucuronic acid, TCA: trichloroacetic acid, MPA: mycophenolic acid PMID:28479734

Kinetic and crystallographic studies of Escherichia coli UDP-N-acetylmuramate:L-alanine ligase.

PubMed Central

Emanuele, J. J.; Jin, H.; Jacobson, B. L.; Chang, C. Y.; Einspahr, H. M.; Villafranca, J. J.

1996-01-01

Uridine diphosphate-N-acetylmuramate:L-alanine ligase (EC 6.3.2.8, UNAM:L-Ala ligase or MurC gene product) catalyzes the ATP-dependent ligation of the first amino acid to the sugar moiety of the peptidoglycan precursor. This is an essential step in cell wall biosynthesis for both gram-positive and gram-negative bacteria. Optimal assay conditions for initial velocity studies have been established. Steady-state assays were carried out to determine the effect of various parameters on enzyme activity. Factors studies included: cation specificity, ionic strength, buffer composition and pH. At 37 degrees C and pH 8.0, kcat was equal to 980 +/- 40 min-1, while K(m) values for ATP, UNAM, and L-alanine were, 130 +/- 10, 44 +/- 3, and 48 +/- 6 microM, respectively. Of the metals tested only Mn, Mg, and Co were able to support activity. Sodium chloride, potassium chloride, ammonium chloride, and ammonium sulfate had no effect on activity up to 75 mM levels. The enzyme, in appropriate buffer, was stable enough to be assayed over the pH range of 5.6 to 10.1. pH profiles of Vmax/K(m) for the three substrates and of Vmax were obtained. Crystallization experiments with the enzyme produced two crystal forms. One of these has been characterized by X-ray diffraction as monoclinic, space group C2, with cell dimensions a = 189.6, b = 92.1, c = 75.2 A, beta = 105 degrees, and two 54 kDa molecules per asymmetric unit. It was discovered that the enzyme will hydrolyze ATP in the absence of L-alanine. This L-alanine independent activity is dependent upon the concentrations of both ATP and UNAM; kcat for this activity is less than 4% of the biosynthetic activity measured in the presence of saturating levels of L-alanine. Numerous L-alanine analogs tested were shown to stimulate ATP hydrolysis. A number of these L-alanine analogs produced novel products as accessed by HPLC and mass spectral analysis. All of the L-alanine analogs tested as inhibitors were competitive versus L-alanine. PMID:8976565

Evaluation of NT-proBNP concentrations during exercise in asymptomatic patients with severe high-gradient aortic stenosis.

PubMed

Dobrowolski, Piotr; Lech, Agnieszka; Klisiewicz, Anna; Hoffman, Piotr

2016-08-11

INTRODUCTION The effect of asymptomatic severe aortic stenosis (ASAS) on N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels ar rest and during exercise, as well as their relevance for clinical practice remain controversial.  OBJECTIVES The aim of this study was to test the hypothesis of whether the evaluation of NT-proBNP concentrations during exercise provides additional information about the severity of aortic stenosis and left ventricular remodeling in patients with ASAS. PATIENTS AND METHODS A total of 50 patients with ASAS (mean age, 38.4 ±18.1 years) and 21 healthy subjects (mean age, 43.4 ±10.6 years) were enrolled. Rest and exercise echocardiography was performed to evaluate maximum velocity (Vmax), mean aortic gradient (AG), and aortic valve area (AVA). The left ventricular mass index (LVMI) was calculated. NT-proBNP concentrations at rest and during exercise were assessed, and the difference between the 2 values was calculated (ΔNT-proBNP). RESULTS NT-proBNP and ΔNT-proBNP levels at rest and during exercise were significantly higher in the ASAS group compared with the control group. In the ASAS group, NT-proBNP levels at rest significantly correlated with LVMI (r = 0.432; P <0.0001), AVA (r = -0.408; P <0.0001), Vmax (r = 0.375; P = 0.002), and mean AG (r = 0.257; P = 0.03). NT-proBNP levels during exercise significantly correlated with LVMI (r = 0.432; P <0.0001), mean AG (r = 0.401; P = 0.001), and AVA (r = -0.375; P = 0.001). In the multivariate logistic regression model, the factors independently associated with NT-proBNP both at rest and during exercise were age, AVA, and LVMI. CONCLUSIONS NT-proBNP levels at rest provide valuable information for identifying patients with more advanced left ventricular hypertrophy secondary to severe aortic stenosis. NT-proBNP levels during exercise do not provide new information on the severity of AS.

Purification and properties of trimethylamine oxide reductase from Salmonella typhimurium.

PubMed Central

Kwan, H S; Barrett, E L

1983-01-01

The major inducible trimethylamine oxide reductase was purified from Salmonella typhimurium LT2. The molecular weights of the native enzyme were estimated to be 332,000 by gel filtration and 170,000 by nondenaturing disc gel electrophoresis. In sodium dodecyl sulfate-gel electrophoresis, the enzyme formed a single band of molecular weight 84,000. The isoelectric point was 4.28. Maximum activity was at pH 5.65 and 45 degrees C. Reduced flavin mononucleotide, but not reduced flavin adenine dinucleotide, served as an electron donor. The Km for trimethylamine oxide was 0.89 mM and Vmax was 1,450 U/mg of protein. The enzyme reduced chlorate with a Km of 2.2 mM and a Vmax of 350 U/mg of protein. Images PMID:6350272

POPDC1S201F causes muscular dystrophy and arrhythmia by affecting protein trafficking

PubMed Central

Schindler, Roland F.R.; Scotton, Chiara; Zhang, Jianguo; Passarelli, Chiara; Ortiz-Bonnin, Beatriz; Simrick, Subreena; Schwerte, Thorsten; Poon, Kar-Lai; Fang, Mingyan; Rinné, Susanne; Froese, Alexander; Nikolaev, Viacheslav O.; Grunert, Christiane; Müller, Thomas; Tasca, Giorgio; Sarathchandra, Padmini; Drago, Fabrizio; Dallapiccola, Bruno; Rapezzi, Claudio; Arbustini, Eloisa; Di Raimo, Francesca Romana; Neri, Marcella; Selvatici, Rita; Gualandi, Francesca; Fattori, Fabiana; Pietrangelo, Antonello; Li, Wenyan; Jiang, Hui; Xu, Xun; Bertini, Enrico; Decher, Niels; Wang, Jun; Brand, Thomas; Ferlini, Alessandra

2015-01-01

The Popeye domain–containing 1 (POPDC1) gene encodes a plasma membrane–localized cAMP-binding protein that is abundantly expressed in striated muscle. In animal models, POPDC1 is an essential regulator of structure and function of cardiac and skeletal muscle; however, POPDC1 mutations have not been associated with human cardiac and muscular diseases. Here, we have described a homozygous missense variant (c.602C>T, p.S201F) in POPDC1, identified by whole-exome sequencing, in a family of 4 with cardiac arrhythmia and limb-girdle muscular dystrophy (LGMD). This allele was absent in known databases and segregated with the pathological phenotype in this family. We did not find the allele in a further screen of 104 patients with a similar phenotype, suggesting this mutation to be family specific. Compared with WT protein, POPDC1S201F displayed a 50% reduction in cAMP affinity, and in skeletal muscle from patients, both POPDC1S201F and WT POPDC2 displayed impaired membrane trafficking. Forced expression of POPDC1S201F in a murine cardiac muscle cell line (HL-1) increased hyperpolarization and upstroke velocity of the action potential. In zebrafish, expression of the homologous mutation (popdc1S191F) caused heart and skeletal muscle phenotypes that resembled those observed in patients. Our study therefore identifies POPDC1 as a disease gene causing a very rare autosomal recessive cardiac arrhythmia and LGMD, expanding the genetic causes of this heterogeneous group of inherited rare diseases. PMID:26642364

Voltage-dependent inward currents in smooth muscle cells of skeletal muscle arterioles

PubMed Central

Shirokov, Roman E.

2018-01-01

Voltage-dependent inward currents responsible for the depolarizing phase of action potentials were characterized in smooth muscle cells of 4th order arterioles in mouse skeletal muscle. Currents through L-type Ca2+ channels were expected to be dominant; however, action potentials were not eliminated in nominally Ca2+-free bathing solution or by addition of L-type Ca2+ channel blocker nifedipine (10 μM). Instead, Na+ channel blocker tetrodotoxin (TTX, 1 μM) reduced the maximal velocity of the upstroke at low, but not at normal (2 mM), Ca2+ in the bath. The magnitude of TTX-sensitive currents recorded with 140 mM Na+ was about 20 pA/pF. TTX-sensitive currents decreased five-fold when Ca2+ increased from 2 to 10 mM. The currents reduced three-fold in the presence of 10 mM caffeine, but remained unaltered by 1 mM of isobutylmethylxanthine (IBMX). In addition to L-type Ca2+ currents (15 pA/pF in 20 mM Ca2+), we also found Ca2+ currents that are resistant to 10 μM nifedipine (5 pA/pF in 20 mM Ca2+). Based on their biophysical properties, these Ca2+ currents are likely to be through voltage-gated T-type Ca2+ channels. Our results suggest that Na+ and at least two types (T- and L-) of Ca2+ voltage-gated channels contribute to depolarization of smooth muscle cells in skeletal muscle arterioles. Voltage-gated Na+ channels appear to be under a tight control by Ca2+ signaling. PMID:29694371

ParamAP: Standardized Parameterization of Sinoatrial Node Myocyte Action Potentials.

PubMed

Rickert, Christian; Proenza, Catherine

2017-08-22

Sinoatrial node myocytes act as cardiac pacemaker cells by generating spontaneous action potentials (APs). Much information is encoded in sinoatrial AP waveforms, but both the analysis and the comparison of AP parameters between studies is hindered by the lack of standardized parameter definitions and the absence of automated analysis tools. Here we introduce ParamAP, a standalone cross-platform computational tool that uses a template-free detection algorithm to automatically identify and parameterize APs from text input files. ParamAP employs a graphic user interface with automatic and user-customizable input modes, and it outputs data files in text and PDF formats. ParamAP returns a total of 16 AP waveform parameters including time intervals such as the AP duration, membrane potentials such as the maximum diastolic potential, and rates of change of the membrane potential such as the diastolic depolarization rate. ParamAP provides a robust AP detection algorithm in combination with a standardized AP parameter analysis over a wide range of AP waveforms and firing rates, owing in part to the use of an iterative algorithm for the determination of the threshold potential and the diastolic depolarization rate that is independent of the maximum upstroke velocity, a parameter that can vary significantly among sinoatrial APs. Because ParamAP is implemented in Python 3, it is also highly customizable and extensible. In conclusion, ParamAP is a powerful computational tool that facilitates quantitative analysis and enables comparison of sinoatrial APs by standardizing parameter definitions and providing an automated work flow. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Responses of soil hydrolase kinetics to nitrogen and phosphorus additions in Chinese fir plantations of subtropical China

NASA Astrophysics Data System (ADS)

Zhang, X.; Zhang, C.; Yang, Y.; Wang, H.; Chen, F.; Fu, X.; Fang, X.; Sun, X.

2016-12-01

Nitrogen (N) deposition and low soil phosphorus (P) content aggravate the P limitation in subtropical forest soils. However, the responses of soil organic matter related hydrolyase kinetics to N and P additions in subtropical plantations are still not clear. We tested the hypothesis that P application can improve the potential maximum activities of soil carbon (C) and N related hydrolayase but substrate demand (Km) may tradeoff the catalytic efficiency of the enzymes. Thirty 20m×20m plots were established in November 2011 and six different treatments were randomly distributed with five replicates in the Chinese fir plantations in subtropical China. The ongoing treatments are control (CK, no N and P application), low N addition (N1:50 kg N ha-1 yr-1), high N addition (N2: 100 kg N ha-1 yr-1), P addition (P: 50 kg P ha-1 yr-1), low N andP addition (N1P: 50 kg N ha-1 yr-1 and 50 kg P ha-1 yr-1) and high N and P addition (N2P: 100 kg N ha-1yr-1and 50 kg P ha-1 yr-1). Soil enzyme kinetic parameters for b-1,4-glucosidase (βG), β-1,4-N-acetylglucosaminidase (NAG), and acid phosphatase (aP) were measured in November 2015. The substrate affinities (Km) of βG and NAG were not affected by N or /and P additions. However, the substrate affinities of aP were decreased by N additions (N1, N2) with higher Km values than the other treatments. N additions (N1, N2) or higher N combined P additions (N2P) increased Vmax and catalytic efficiencies for βG, while with P addition treatments (N1P, N2P, and P) decreased Vmax and catalytic efficiencies for aP. The effects of N combined P treatments (N1P and N2P) on kinetic parameters (Vmax, Km) and catalytic efficiencies for AP were similar to P treatment, indicating that P had stronger effects on organic phosphorus hydrolysis than N in the research site. The N additions (N1 and N2) did not affect the catalytic efficiencies for NAG despite of their positive responses to Vmax for NAG compared with CK. The catalytic efficiencies of aP and NAG were negatively correlated with soil TP and available P contents, and both the enzyme kinetics for aP exhibited strong negative correlations with TP and available P contents. However, the Vmax for BG and NAG were positively correlated with SOC contents, but were negatively correlated with soil pH.

Enhancement of force generated by individual myosin heads in skinned rabbit psoas muscle fibers at low ionic strength.

PubMed

Sugi, Haruo; Abe, Takahiro; Kobayashi, Takakazu; Chaen, Shigeru; Ohnuki, Yoshiki; Saeki, Yasutake; Sugiura, Seiryo

2013-01-01

Although evidence has been presented that, at low ionic strength, myosin heads in relaxed skeletal muscle fibers form linkages with actin filaments, the effect of low ionic strength on contraction characteristics of Ca(2+)-activated muscle fibers has not yet been studied in detail. To give information about the mechanism of muscle contraction, we have examined the effect of low ionic strength on the mechanical properties and the contraction characteristics of skinned rabbit psoas muscle fibers in both relaxed and maximally Ca(2+)-activated states. By progressively decreasing KCl concentration from 125 mM to 0 mM (corresponding to a decrease in ionic strength μ from 170 mM to 50 mM), relaxed fibers showed changes in mechanical response to sinusoidal length changes and ramp stretches, which are consistent with the idea of actin-myosin linkage formation at low ionic strength. In maximally Ca(2+)-activated fibers, on the other hand, the maximum isometric force increased about twofold by reducing KCl concentration from 125 to 0 mM. Unexpectedly, determination of the force-velocity curves indicated that, the maximum unloaded shortening velocity Vmax, remained unchanged at low ionic strength. This finding indicates that the actin-myosin linkages, which has been detected in relaxed fibers at low ionic strength, are broken quickly on Ca(2+) activation, so that the linkages in relaxed fibers no longer provide any internal resistance against fiber shortening. The force-velocity curves, obtained at various levels of steady Ca(2+)-activated isometric force, were found to be identical if they are normalized with respect to the maximum isometric force. The MgATPase activity of muscle fibers during isometric force generation was found not to change appreciably at low ionic strength despite the two-fold increase in Ca(2+)-activated isometric force. These results can be explained in terms of enhancement of force generated by individual myosin heads, but not by any changes in kinetic properties of cyclic actin-myosin interaction.

[Positive inotropic and lusitropic effect of RP 62719, a new class III antiarrhythmia agent].

PubMed

Beregi, J P; Escande, D; Coudray, N; Chemla, D; Mestre, M; Péry, N; Lecarpentier, Y

1994-02-01

Antiarrhythmic drugs, especially the Class I family, exert a negative inotropic effect on the myocardium which is particularly undesirable in patients with depressed left ventricular function. Therefore, research has been directed to the development of new, more specific molecules of the Class III family. The authors studies the mechanical effects of RP 62719 on guinea pig left ventricular papillary muscle. This new molecule is a pure Class III antiarrhythmic, known to lengthen the duration of the cardiac action potential by selectively blocking the potassium current iK1 (inward rectifier K+ current). The mechanical parameters were determined during the phases of contraction and relaxation under isotonic and isometric conditions. At 0.2 and 2 microM concentrations, RP 62719 improved cardiac contraction under both isotonic and isometric conditions with an increase of about 30% of Vmax (p < 0.001), the maximum unloaded shortening velocity delta 1 (p < 0.001), the peak isometric active force normalized per cross-sectional area [AF/S (p < 0.001)]. At these two concentrations, a positive lusitropic effect (improved relaxation) was demonstrated by an increase in negative peak of derivative per mm2-dF/s and maximum lengthening velocity VR max (p < 0.01). At higher concentrations (20 microM), the inotropic and lusitropic effects were less marked with a bell-shaped form of the dose-effect curve. This study indicates that RP 62719 has moderate but significant positive inotropic and lusitropic effects. These actions could provide significant therapeutic advantages especially in patients cardiac failure.

Looping tracks associated with tropical cyclones approaching an isolated mountain. Part I: Essential parameters

NASA Astrophysics Data System (ADS)

Huang, Yi-Chih; Lin, Yuh-Lang

2018-06-01

Essential parameters for making a looping track when a westward-moving tropical cyclone (TC) approaches a mesoscale mountain are investigated by examining several key nondimensional control parameters with a series of systematic, idealized numerical experiments, such as U/ Nh, V max/ Nh, U/ fL x , V max/ fR, h/ L x , and R/ L y . Here U is the uniform zonal wind velocity, N the Brunt-Vaisala frequency, h the mountain height, f the Coriolis parameter, V max the maximum tangential velocity at a radius of R from the cyclone center and L x is the halfwidth of the mountain in the east-west direction. It is found that looping tracks (a) tend to occur under small U/ Nh and U/ fL x , moderate h/ L x , and large V max/ Nh, which correspond to slow movement (leading to subgeostrophic flow associated with strong orographic blocking), moderate steepness, and strong tangential wind associated with TC vortex; (b) are often accompanied by an area of perturbation high pressure to the northeast of the mountain, which lasts for only a short period; and (c) do not require the existence of a northerly jet. The nondimensional control parameters are consolidated into a TC looping index (LI), {U2 R2 }/{V_{max 2 hLy }} , which is tested by several historical looping and non-looping typhoons approaching Taiwan's Central Mountain Range (CMR) from east or southeast. It is found that LI < 0.0125 may serve as a criterion for looping track to occur.

Improved lower extremity pedaling mechanics in individuals with stroke under maximal workloads.

PubMed

Linder, Susan M; Rosenfeldt, Anson B; Bazyk, Andrew S; Koop, Mandy Miller; Ozinga, Sarah; Alberts, Jay L

2018-05-01

Background Individuals with stroke present with motor control deficits resulting in the abnormal activation and timing of agonist and antagonist muscles and inefficient movement patterns. The analysis of pedaling biomechanics provides a window into understanding motor control deficits, which vary as a function of workload. Understanding the relationship between workload and motor control is critical when considering exercise prescription during stroke rehabilitation. Objectives To characterize pedaling kinematics and motor control processes under conditions in which workload was systematically increased to an eventual patient-specific maximum. Methods A cohort study was conducted in which 18 individuals with chronic stroke underwent a maximal exertion cardiopulmonary exercise test on a stationary cycle ergometer, during which pedaling torque was continuously recorded. Measures of force production, pedaling symmetry, and pedaling smoothness were obtained. Results Mean Torque increased significantly (p < 0.05) for both legs from initial to terminal workloads. Mean torque Symmetry Index, calculated for down and upstroke portions of the pedaling action, improved from 0.37(0.29) to 0.29(0.35) during downstroke (p = 0.007), and worsened during the upstroke: -0.37(0.38) to -0.62(0.46) (p < 0.001) from initial to terminal workloads. Low Torque Duration improved from initial to terminal workloads, decreasing from 121.1(52.9) to 58.1(39.6) degrees (p < 0.001), respectively. Smoothness of pedaling improved significantly from initial to terminal workloads (p < 0.001). Conclusions Improved pedaling kinematics at terminal workloads indicate that individuals with stroke demonstrate improved motor control with respect to the timing, sequencing, and activation of hemiparetic lower extremity musculature compared to lower workloads. Therapeutic prescription involving higher resistance may be necessary to sufficiently engage and activate the paretic lower extremity.

Wing motion measurement and aerodynamics of hovering true hoverflies.

PubMed

Mou, Xiao Lei; Liu, Yan Peng; Sun, Mao

2011-09-01

Most hovering insects flap their wings in a horizontal plane (body having a large angle from the horizontal), called `normal hovering'. But some of the best hoverers, e.g. true hoverflies, hover with an inclined stroke plane (body being approximately horizontal). In the present paper, wing and body kinematics of four freely hovering true hoverflies were measured using three-dimensional high-speed video. The measured wing kinematics was used in a Navier-Stokes solver to compute the aerodynamic forces of the insects. The stroke amplitude of the hoverflies was relatively small, ranging from 65 to 85 deg, compared with that of normal hovering. The angle of attack in the downstroke (∼50 deg) was much larger that in the upstroke (∼20 deg), unlike normal-hovering insects, whose downstroke and upstroke angles of attack are not very different. The major part of the weight-supporting force (approximately 86%) was produced in the downstroke and it was contributed by both the lift and the drag of the wing, unlike the normal-hovering case in which the weight-supporting force is approximately equally contributed by the two half-strokes and the lift principle is mainly used to produce the force. The mass-specific power was 38.59-46.3 and 27.5-35.4 W kg(-1) in the cases of 0 and 100% elastic energy storage, respectively. Comparisons with previously published results of a normal-hovering true hoverfly and with results obtained by artificially making the insects' stroke planes horizontal show that for the true hoverflies, the power requirement for inclined stroke-plane hover is only a little (<10%) larger than that of normal hovering.

Aerodynamics and flow features of a damselfly in takeoff flight.

PubMed

Bode-Oke, Ayodeji T; Zeyghami, Samane; Dong, Haibo

2017-09-26

Flight initiation is fundamental for survival, escape from predators and lifting payload from one place to another in biological fliers and can be broadly classified into jumping and non-jumping takeoffs. During jumping takeoffs, the legs generate most of the initial impulse. Whereas the wings generate most of the forces in non-jumping takeoffs, which are usually voluntary, slow, and stable. It is of great interest to understand how these non-jumping takeoffs occur and what strategies insects use to generate large amount of forces required for this highly demanding flight initiation mode. Here, for the first time, we report accurate wing and body kinematics measurements of a damselfly during a non-jumping takeoff. Furthermore, using a high fidelity computational fluid dynamics simulation, we identify the 3D flow features and compute the wing aerodynamics forces to unravel the key mechanisms responsible for generating large flight forces. Our numerical results show that a damselfly generates about three times its body weight during the first half-stroke for liftoff. In generating these forces, the wings flap through a steeply inclined stroke plane with respect to the horizon, slicing through the air at high angles of attack (45°-50°). Consequently, a leading edge vortex (LEV) is formed during both the downstroke and upstroke on all the four wings. The formation of the LEV, however, is inhibited in the subsequent upstrokes following takeoff. Accordingly, we observe a drastic reduction in the magnitude of the aerodynamic force, signifying the importance of LEV in augmenting force production. Our analysis also shows that forewing-hindwing interaction plays a favorable role in enhancing both lift and thrust production during takeoff.

Electrocardiogram changes and atrial arrhythmias in individuals carrying sodium channel SCN5A D1275N mutation.

PubMed

Vanninen, Sari U M; Nikus, Kjell; Aalto-Setälä, Katriina

2017-09-01

The cardiac sodium channel SCN5A regulates atrioventricular and ventricular depolarization as well as cardiac conduction. Patients with cardiac electrical abnormalities have an increased risk of sudden cardiac death (SCD) and cardio-embolic stroke. Optimal management of cardiac disease includes the understanding of association between the causative mutations and the clinical phenotype. A 12-lead electrocardiogram (ECG) is an easy and inexpensive tool for finding risk patients. A blood sample for DNA extraction was obtained in a Finnish family with 43 members; systematic 12-lead ECG analysis was performed in 13 of the family members carrying an SCN5A D1275N mutation. Conduction defects and supraventricular arrhythmias, including atrial fibrillation/flutter, atrioventricular nodal re-entry tachycardia (AVNRT) and junctional rhythm were searched for. Five (38%) mutation carriers had fascicular or bundle branch block, 10 had atrial arrhythmias; no ventricular arrhythmias were found. Notching of the R- and S waves - including initial QRS fragmentation - and prolonged S-wave upstroke were present in all the affected family members. Notably, four (31%) affected family members had a stroke before the age of 31 and two experienced premature death. A 12-lead ECG can be used to predict arrhythmias in SCN5A D1275N mutation carriers. Key messages The 12-lead ECG may reveal cardiac abnormalities even before clinical symptoms occur. Specific ECG findings - initial QRS fragmentation, prolonged S-wave upstroke as well as supraventricular arrhythmias - were frequently encountered in all SCN5A D1257N mutation carriers. ECG follow-up is recommended for all SCN5A D1275N mutation carriers.

Cellular assessment of muscle in COPD: case studies of two males.

PubMed

Green, Howard J; Bombardier, Eric; Burnett, Margaret E; D'Arsigny, Christine L; Iqbal, Sobia; Webb, Katherine A; Ouyang, Jing; O'Donnell, Denis E

2009-12-29

The objective of this paper is to provide an overview of the recent developments in muscle physiology and biochemistry in general, and with respect to chronic obstructive pulmonary disease (COPD) specifically. As a way of illustration, we have presented data on the remodeling that occurs in vastus lateralis in two patients with COPD (COPD #1, forced expiratory volume in one second/forced vital capacity [FEV(1)/FVC] = 63%; COPD #2, FEV(1)/FVC = 41%) exhibiting differences in muscle wasting as compared to healthy controls (CON; FEV(1)/FVC = 111 +/- 2.2%, n = 4). Type I fibers percentages were lower in both COPD #1 (16.7) and COPD #2 (24.9) compared to CON (57.3 +/- 5.2). Cross sectional area of the type I fibers of the patients ranged between 65%-68% of CON and for the type II subtypes (IIA, IIAX, IIX) between 74% and 89% (COPD #1) and 17%-32% (COPD #2). A lower number of capillary contacts were observed for all fiber types in COPD #1 but not COPD #2. Lower concentrations of adenosine triphosphate (ATP) (24%-26%) and phosphocreatine (18%-20%), but not lactate occurred in COPD. In contrast to COPD #1, who displayed normal glucose transporter content, GLUT1 and GLUT4 were only 71% and 54%, respectively of CON in COPD #2. Lower monocarboxylate contents were found for MCT1 in both COPD #1 (63%) and COPD #2 (41%) and for MCT4 (78%) in COPD #1. Maximal oxidative enzyme activities (V(max)) for COPD #2 ranged between 37% (succinic dehydrogenase) and 70% (cytochrome C oxidase) of CON. For the cytosolic enzymes, V(max) ranged between 89% (hexokinase) to 31% (pyruvate kinase) of CON. Depressions were also observed in V(max) of the Na(+)-K(+)-ATPase for COPD #1 (66% of CON) but not COPD #2 (92% of CON) while V(max) of the Ca(2+)-ATPase was near normal in COPD #1 (84% CON). It is concluded that disturbances can occur in muscle to a wide range of excitation, contraction and metabolic processes in COPD.

Pyruvate and ketone-body transport across the mitochondrial membrane. Exchange properties, pH-dependence and mechanism of the carrier.

PubMed

Halestrap, A P

1978-06-15

The effects of exchangeable ions and pH on the efflux of pyruvate from preloaded mitochondria are reported. Efflux obeys first-order kinetics, and the stimulation of efflux by exchangeable ions such as acetoacetate and lactate obeys Michaelis--Menten kinetics. The apparent Km value +/- S.E. for acetoacetate was 0.56 +/- 0.14 mM (n = 5) and that for lactate 12.3 +/- 2.3 mM (n = 6). The Vmax. values +/- S.E. at 0 degrees C were 16.2 +/- 2.0 and 21.9 +/- 2.7 nmol/min per mg of protein. The exchange of a variety of other substituted monocarboxylates was also studied. Efflux was also stimulated by increasing the external pH. The data gave a pK for the transport process of 8.35 and a Vmax. of 3.31 +/- 0.14 nmol/min per mg. The similarity of the Vmax. values for various exchangeable ions but the difference of this from the Vmax. in the absence of exchangeable ions may indicate that transport of pyruvate occurs with H+ and not in exchange for an OH- ion. The inhibition of transport by alpha-cyano-4-hydroxycinnamate took several seconds to reach completion at 0 degrees C. It is proposed that inhibition occurs by binding to the substrate site and subsequent reaction with an -SH group on the inside of the membrane. The inhibitor can be displaced by substrates that can also enter the mitochondria independently of the carrier and so compete with the inhibitor for the substrate-binding site on the inside of the membrane. A mechanism for transport is proposed that invokes a transition state of pyruvate involving addition of an -SH group to the 2-carbon of pyruvate. Evidence is presented that suggests that ketone bodies may cross the mitochondrial membrane either on the carrier or by free diffusion. The physiological involvement of the carrier in ketone-body metabolism is discussed. The role of ketone bodies and pH in the physiological regulation of pyruvate transport is considered.

Catalytical Properties of Free and Immobilized Aspergillus niger Tannase.

PubMed

Flores-Maltos, Abril; Rodríguez-Durán, Luis V; Renovato, Jacqueline; Contreras, Juan C; Rodríguez, Raúl; Aguilar, Cristóbal N

2011-01-01

A fungal tannase was produced, recovered, and immobilized by entrapment in calcium alginate beads. Catalytical properties of the immobilized enzyme were compared with those of the free one. Tannase was produced intracellularly by the xerophilic fungus Aspergillus niger GH1 in a submerged fermentation system. Enzyme was recovered by cell disruption and the crude extract was partially purified. The catalytical properties of free and immobilized tannase were evaluated using tannic acid and methyl gallate as substrates. K(M) and V(max) values for free enzyme were very similar for both substrates. But, after immobilization, K(M) and V(max) values increased drastically using tannic acid as substrate. These results indicated that immobilized tannase is a better biocatalyst than free enzyme for applications on liquid systems with high tannin content, such as bioremediation of tannery or olive-mill wastewater.

Purification, characterization and kinetic properties of extracellular L-asparaginase produced by Cladosporium sp.

PubMed

Mohan Kumar, N S; Manonmani, H K

2013-04-01

L-asparaginase from Cladosporium sp. grown on wheat bran by SSF was purified. Enzyme appeared to be a trimer with homodimer of 37 kDa and another 47 kDa amounting to total mass of 121 kDa as estimated by SDS-PAGE and 120 kDa on gel filtration column. The optimum temperature and pH of the enzyme were 30 °C and 6.3, respectively with Vmax of 4.44 μmol/mL/min and Km of 0.1 M. Substrate specificity studies indicated that, L-asparaginase has greater affinity towards L-asparagine with substrate hydrolysis efficiency (Vmax/Km ratio) eightfold higher than that of L-glutamine. L-asparaginase activity in presence of thiols studied showed decrease in Vmax and increase in Km, indicating nonessential mode of inactivation. Among the thiols tested, β-mercaptomethanol, exerted inhibitory effect, suggesting a critical role of disulphide linkages in maintaining a suitable conformation of the enzyme. Metal ions such as Ca(2+), Co(2+), Cu(2+), Mg(2+), Na(+), K(+) and Zn(2+) significantly affected enzyme activity whereas presence of Fe(3+), Pb(2+) and KI stimulated the activity. Detergents studied also enhanced L-asparaginase activity. In-vitro half-life of purified L-asparaginase in mammalian blood serum was 93.69 h. The enzyme inhibited acrylamide formation in potato chips by 96 % making it a potential candidate for food industry to reduce acrylamide content in starchy fried food commodities.

[Contrast enhanced power Doppler and color Doppler ultrasound in breast masses: Efficiency in diagnosis and contributions to differential diagnosis].

PubMed

Algül, Ali; Balci, Pinar; Seçil, Mustafa; Canda, Tülay

2003-06-01

To compare ability of detection of vascular structures by utilizing ultrasonographic contrast agent (Levovist) prior to and following power Doppler ultrasound (PDUS) and colour Doppler ultrasound (CDUS) and to determine useful parameters in the differentiation of malignant and benign breast masses by means of verified data. Vascularisation characteristics of 38 breast masses (22 malignant, 16 benign) which were confirmed by mammography and B-mode sonography were evaluated by both CDUS and PDUS following and prior to intravenous contrast application. In addition, Vmax and RI values of vascular structures were calculated by Doppler spectral evaluation. Malignant lesions showed more vascularity than benign lesions both with and without contrast enhancement. With both methods, by utilizing contrast agent, central, penetrating and tortuous vascular structures became more significant in malignant lesions when compared with benign lesions. PDUS was able to detect vascular structures better than CDUS; however, the difference was not statistically significant. Presence of peripheral vascularity was not useful in differentiating malignant from benign lesions. Vmax and RI values were higher in malignant lesions and the difference was statistically significant. In both methods, Vmax > 15 cm/sec and RI > 0.80 (CDUS), and RI > 0.70 (PDUS) were accepted as malignancy parameters. Vascular patterns of breast masses as determined with PDUS and CDUS with contrast enhancement and Doppler spectral examinations enabled differentiation of malignant and benign breast lesions. Thus, it is possible to decrease the number of unnecessary surgical interventions.

Adaptation of skeletal muscle energy metabolism to repeated hypoxic-normoxic exposures and drug treatment.

PubMed

Pastoris, O; Dossena, M; Gorini, A; Vercesi, L; Benzi, G

1985-03-01

Muscular glycolytic fuels, intermediates and end-products (glycogen, glucose, glucose-6-phosphate, pyruvate, lactate), Krebs cycle intermediates (citrate, alpha-ketoglutarate, succinate, malate), related free amino acids (glutamate, alanine), ammonia, energy store (creatine phosphate), energy mediators (ATP, ADP, AMP) and energy charge potential were evaluated. Furthermore the maximum rate (Vmax) of the following muscular enzyme activities was evaluated in the crude extract and/or mitochondrial fraction: for the anaerobic glycolytic pathway: hexokinase, phosphofructokinase, pyruvate kinase, lactate dehydrogenase; for the tricarboxylic acid cycle: citrate synthase, malate dehydrogenase; for the electron transfer chain: total NADH cytochrome c reductase, cytochrome oxidase. The rat gastrocnemius muscles were analyzed in normoxia and after repeated, alternate hypoxic and normoxic exposures (12 hours of hypoxia daily; for 5 days). Naftidrofuryl was administered daily at three different doses: 10, 15 and 22.5 mg/kg i.m., 30 min before the beginning of the experimental hypoxia. The biochemical adaptation to intermittent normobaric hypoxic-normoxic exposures was characterized by the decrease of the muscular contents of creatine phosphate, citrate, alpha-ketoglutarate and glutamate. This adaptation occurred in absence of significant changes in the Vmax of the muscle enzymes tested. By naftidrofuryl treatment, in gastrocnemius muscle from hypoxic rats both alpha-ketoglutarate and creatine phosphate contents maintained normal values, while glutamate concentration remained reduced to subnormal values. With the exception of hexokinase, naftidrofuryl treatment did not modify the Vmax of marker enzymes related to energy transduction.

Characterizing Isozymes of Chlorite Dismutase for Water Treatment

PubMed Central

Mobilia, Kellen C.; Hutchison, Justin M.; Zilles, Julie L.

2017-01-01

This work investigated the potential for biocatalytic degradation of micropollutants, focusing on chlorine oxyanions as model contaminants, by mining biology to identify promising biocatalysts. Existing isozymes of chlorite dismutase (Cld) were characterized with respect to parameters relevant to this high volume, low-value product application: kinetic parameters, resistance to catalytic inactivation, and stability. Maximum reaction velocities (Vmax) were typically on the order of 104 μmol min-1 (μmol heme)-1. Substrate affinity (Km) values were on the order of 100 μM, except for the Cld from Candidatus Nitrospira defluvii (NdCld), which showed a significantly lower affinity for chlorite. NdCld also had the highest susceptibility to catalytic inactivation. In contrast, the Cld from Ideonella dechloratans was least susceptible to catalytic inactivation, with a maximum turnover number of approximately 150,000, more than sevenfold higher than other tested isozymes. Under non-reactive conditions, Cld was quite stable, retaining over 50% of activity after 30 days, and most samples retained activity even after 90–100 days. Overall, Cld from I. dechloratans was the most promising candidate for environmental applications, having high affinity and activity, a relatively low propensity for catalytic inactivation, and excellent stability. PMID:29312158

The mechanism of zinc uptake by cultured rat liver cells.

PubMed Central

Taylor, J A; Simons, T J

1994-01-01

1. The initial rate of 65Zn uptake into cultured rat hepatocytes has been measured over a range of Zn2+ concentrations from 3 x 10(-10) M to 5 x 10(-6) M. Histidine and albumin were used to buffer Zn2+ ions at concentrations below 1 x 10(-6) M. 2. The results suggest there are two mechanisms for Zn2+ uptake; a high-affinity, saturable pathway, with a maximum velocity (Vmax) of 20-30 pmol (mg protein)-1 min-1 and a Michaelis-Menten constant (Km) of about 2 x 10(-9) M Zn2+ (with histidine), and a low-affinity, linear pathway, that only makes a significant contribution to Zn2+ uptake at Zn2+ concentrations above 1 x 10(-6) M. 3. Transport via the high-affinity pathway is dependent on the concentration of Zn2+ ions and not on the concentrations of Zn(2+)-ligand complexes, suggesting that Zn2+ is the transported species. 4. The affinity of the saturable pathway for Zn2+ is slightly lower in the presence of albumin, with a Km of about 1.3 x 10(-8) M. The reason for this is uncertain. PMID:8014898

Purified reconstituted lac carrier protein from Escherichia coli is fully functional.

PubMed

Viitanen, P; Garcia, M L; Kaback, H R

1984-03-01

Proteoliposomes reconstituted with lac carrier protein purified from the plasma membrane of Escherichia coli catalyze each of the translocation reactions typical of the beta-galactoside transport system (i.e., active transport, counterflow, facilitated influx and efflux) with turnover numbers and apparent Km values comparable to those observed in right-side-out membrane vesicles. Furthermore, detailed kinetic studies show that the reconstituted system exhibits properties analogous to those observed in membrane vesicles. Imposition of a membrane potential (delta psi, interior negative) causes a marked decrease in apparent Km (by a factor of 7 to 10) with a smaller increase in Vmax (approximately equal to 3-fold). At submaximal values of delta psi, the reconstituted carrier exhibits biphasic kinetics, with one component manifesting the kinetic parameters of active transport and the other exhibiting the characteristics of facilitated diffusion. Finally, at low lactose concentrations, the initial velocity of influx varies linearly with the square of the proton electro-chemical gradient. The results provide quantitative support for the contention that a single polypeptide species, the product of the lac y gene, is responsible for each of the transport reactions typical of the beta-galactoside transport system.

The dark side of galaxy colour

NASA Astrophysics Data System (ADS)

Hearin, Andrew P.; Watson, Douglas F.

2013-10-01

We present age distribution matching, a theoretical formalism for predicting how galaxies of luminosity L and colour C occupy dark matter haloes. Our model supposes that there are just two fundamental properties of a halo that determine the colour and brightness of the galaxy it hosts: the maximum circular velocity Vmax and the redshift zstarve that correlates with the epoch at which the star formation in the galaxy ceases. The halo property zstarve is intended to encompass physical characteristics of halo mass assembly that may deprive the galaxy of its cold gas supply and, ultimately, quench its star formation. The new, defining feature of the model is that, at fixed luminosity, galaxy colour is in monotonic correspondence with zstarve, with the larger values of zstarve being assigned redder colours. We populate an N-body simulation with a mock galaxy catalogue based on age distribution matching and show that the resulting mock galaxy distribution accurately describes a variety of galaxy statistics. Our model suggests that halo and galaxy assembly are indeed correlated. We make publicly available our low-redshift, Sloan Digital Sky Survey Mr < -19 mock galaxy catalogue, and main progenitor histories of all z = 0 haloes, at http://logrus.uchicago.edu/~aphearin

Importance of Xanthobacter autotrophicus in toluene biodegradation within a contaminated stream.

PubMed

Tay, S T; Hemond, H F; Polz, M F; Cavanaugh, C M; Krumholz, L R

1999-02-01

Toluene-degrading strains T101 and T102 were isolated from rock surface biomass in a toluene-contaminated freshwater stream. These organisms were present at a density of 5.5 x 10(6) cells/g of rock surface biomass. Both are aerobic, rod-shaped, Gram-negative, non-motile, catalase-positive, oxidase-positive, with yellow pigments, and can grow on benzene. Phylogenetic analyses show that strains T101 and T102 have 16S rDNA sequences identical to Xanthobacter autotrophicus. Fatty acid analyses indicate that they are different strains of the same species Xanthobacter autotrophicus, and that they have high levels of cis-11-octadecenoic acid and cis-9-hexadecenoic acid; 3-hydroxyhexadecanoic acid is the major hydroxy fatty acid present. Strains T101 and T102 had maximal velocities (Vmax) for toluene biodegradation of 3.8 +/- 0.5 and 28.3 +/- 2.2 mumoles toluene/mgprotein-hr, and half-saturation constants (Ks) of 0.8 +/- 0.5 and 11.5 +/- 2.4 microM, respectively. Strain T102 has a higher capacity than strain T101 to degrade toluene, and kinetic calculations suggest that strain T102 may be a major contributor to toluene biodegradation in the stream.

Lysine and glutamate transport in the erythrocytes of common brushtail possum, Tammar Wallaby and eastern grey, kangaroo.

PubMed

Ogawa, E; Kuchel, P W; Agar, N S

1998-04-01

It was recently coincidentally discovered, using 1H NMR spectroscopy, that the erythrocytes of two species of Australian marsupials, Tammar Wallaby (Macropus eugenii) and Bettong (Bettongia penicillata), contain relatively high concentrations of the essential amino acid lysine (Agar NS, Rae CD, Chapman BE, Kuchel PW. Comp Biochem Physiol 1991;99B:575-97). Hence, in the present work the rates of transport of lysine into the erythrocytes from the Common Brushtail Possum (Dactylopsilia trivirgata) and Eastern Grey Kangaroo (Macropus giganteus) (which both have low lysine concentrations), and Tammar Wallaby were studied, to explore the mechanistic basis of this finding. The concentration-dependence of the uptake was studied with lysine alone and in the presence of arginine, which may be a competitor of the transport in some species. In relation to GSH metabolism, glutamate uptake was determined in the presence and absence of Na+. The data was analysed to yield estimates of the maximal velocity (Vmax) and the Km in each of the species. Erythrocytes from Tammar Wallaby lacked saturable lysine transport in contrast to the other two species. The glutamate uptake was normal in all three animals for adequate GSH biosynthesis.

The inertial power and inertial force of robotic and natural bat wing

NASA Astrophysics Data System (ADS)

Yin, Dongfu; Zhang, Zhisheng

2016-03-01

Based on the acquired length and angle data of bat skeletons, a four-degree freedom robotic bat wing and an identical computational model with flap, sweep, elbow and wrist motions were presented. By considering the digits motions, a biomimetic bat skeleton model with seven-degree freedom was established as well. The effects of frequency, amplitude and downstroke ratio, as well as the components of inertial power and force on different directions, were studied. The experimental and computational results indicated that the inertial power and force accounted for the largest part on flap direction, the wing fold during upstroke could reduce the inertial power and force.

Scaling Factor Variability and Toxicokinetic Outcomes in Children

EPA Science Inventory

Abstract title: Scaling Factor Variability and Toxicokinetic Outcomes in ChildrenBackgroundBiotransformation rates (Vmax) extrapolated from in vitro data are used increasingly in human physiologically based pharmacokinetic (PBPK) models. PBPK models are widely used in human hea...

Hidden Risks of Erectile Dysfunction "Treatments" Sold Online

MedlinePlus

... screen and stop these shipments from entering U.S. commerce," says Huascar Batista, team leader of OOC's Import- ... Free Rhino V Max V.Max True Man Energy Max HS Joy of Love NaturalUp Blue Steel ...

Influence of rete testis fluid deprivation on the kinetic parameters of goat epididymal 5 alpha-reductase.

PubMed

Kelce, W R; Lubis, A M; Braun, W F; Youngquist, R S; Ganjam, V K

1990-01-01

A surgical technique to cannulate the rete testis of the goat was utilized to examine the effects of rete testis fluid (RTF) deprivation on the enzymatic activity of epididymal 5 alpha-reductase. Kinetic techniques were used to determine whether the regional enzymatic effect of RTF deprivation is to decrease the apparent number of 5 alpha-reductase active sites or the catalytic activity of each active site within the epididymal epithelium. Paired comparisons of (Vmax)app and (Km)app values between control and RTF-deprived epididymides indicated that RTF deprivation affected the value of (Vmax)app with no apparent change in the values of (Km)app in caput, corpus, and cauda epididymal regions. We conclude that RTF deprivation in the goat epididymis for 7 days results in a decreased number of apparent 5 alpha-reductase active sites within the epididymal epithelium.

Kinetics of 3H-serotonin uptake by platelets in infantile autism and developmental language disorder (including five pairs of twins)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Katsui, T.; Okuda, M.; Usuda, S.

The kinetics of 5-HT uptake by platelets was studied in cases of infantile autism and developmental language disorder (DLD) and normal subjects. Two patients of the autism group were twins, and the seven patients of the DLD group were members of four pairs of twins. The Vmax values (means +/- SD) for autism and DLD were 6.46 +/- .90 pmol 5-HT/10(7) cells/min and 4.85 +/- 1.50 pmol 5-HT/10(7) cells/min, respectively. These values were both significantly higher than that of 2.25 +/- .97 pmole 5-HT/10(7) cells/min for normal children. The Km values of the three groups were not significantly different. Datamore » on the five pairs of twins examined suggested that the elevated Vmax of 5-HT uptake by platelets was determined genetically.« less

Influence of cimetidine and diethyldithiocarbamate on the metabolism of halothane and methoxyflurane in vitro.

PubMed

Loesch, J; Siegers, C P; Younes, M

1987-06-01

The metabolism of halothane and methoxyflurane was measured in vitro by the vial equilibration method using the S-9-fraction from rat liver as source of enzymes. Kinetic values were measured for halothane: Vmax = 11.6 nmol/g.min, KM = 19.6 mumol/l and methoxyflurane: Vmax = 12.0 nmol/g.min, KM = 17.5 mumol/l. Dithiocarb showed strong inhibitory activity on halothane and methoxyflurane metabolism; inhibition constants were calculated as Ki = 0.051 mmol/l and Ki = 0.004 mmol/l, respectively. Cimetidine inhibited the metabolism of both anesthetics to a lesser extent. Inhibition constants were calculated as Ki = 16.2 mmol/l and Ki = 8.2 mmol/l for halothane and methoxyflurane, respectively. The observed inhibitory properties of dithiocarb and cimetidine on the metabolism of halothane and methoxyflurane may be of interest in connection with the problem of toxic liver and kidney injury after anesthesia with these agents.

Toxicity and kinetic parameters of the aerobic biodegradation of the phenol and alkylphenols by a mixed culture.

PubMed

Acuña-Argüelles, M E; Olguin-Lora, P; Razo-Flores, E

2003-04-01

A mixed culture aerobically metabolized phenol, cresol isomers (o-,m-,p-), 2-ethylphenol and xylenol isomers (2,5-DMP and 3,4-DMP) as the sole carbon and energy source. This culture had a high tolerance towards phenol with values of maximum degradation rate (Vmax) of 47 microM phenol mg-1 protein h-1 and inhibition substrate constant (Ki) of 10 mM. These kinetic parameters were considerably diminished and the toxicity increased with the alkylphenols. For example with 2,5-xylenol, Vmax and Ki values of 0.8 microM 2,5-xylenol mg-1 protein h-1 and 1.3 mM, respectively, were obtained. The cresols were 5-fold more toxic than phenol, whereas 2-ethylphenol and 3,4-xylenol were 11-fold more toxic, and 2,5-xylenol was 34-fold more toxic than phenol.

Cloning of Arabidopsis serotonin N-acetyltransferase and its role with caffeic acid O-methyltransferase in the biosynthesis of melatonin in vitro despite their different subcellular localizations.

PubMed

Lee, Hyoung Yool; Byeon, Yeong; Lee, Kyungjin; Lee, Hye-Jung; Back, Kyoungwhan

2014-11-01

Serotonin N-acetyltransferase (SNAT) is the penultimate enzyme in melatonin biosynthesis. We cloned SNAT from Arabidopsis thaliana (AtSNAT) and functionally characterized this enzyme for the first time from dicotyledonous plants. Similar to rice SNAT, AtSNAT was found to localize to chloroplasts with peak enzyme activity at 45 °C (Km , 309 μm; Vmax , 1400 pmol/min/mg protein). AtSNAT also catalyzed 5-methoxytryptamine (5-MT) into melatonin with high catalytic activity (Km , 51 μm; Vmax , 5300 pmol/min/mg protein). In contrast, Arabidopsis caffeic acid O-methyltransferase (AtCOMT) localized to the cytoplasm. Interestingly, AtCOMT can methylate serotonin into 5-MT with low catalytic activity (Km , 3.396 mm; Vmax , 528 pmol/min/mg protein). These data suggest that serotonin can be converted into either N-acetylserotonin by SNAT or into 5-MT by COMT, after which it is metabolized into melatonin by COMT or SNAT, respectively. To support this hypothesis, serotonin was incubated in the presence of both AtSNAT and AtCOMT enzymes. In addition to melatonin production, the production of major intermediates depended on incubation temperatures; N-acetylserotonin was predominantly produced at high temperatures (45 °C), while low temperatures (37 °C) favored the production of 5-MT. Our results provide biochemical evidence for the presence of a serotonin O-methylation pathway in plant melatonin biosynthesis. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Characterization of Two Distinct Glycosyl Hydrolase Family 78 α-l-Rhamnosidases from Pediococcus acidilactici▿†

PubMed Central

Michlmayr, Herbert; Brandes, Walter; Eder, Reinhard; Schümann, Christina; del Hierro, Andrés M.; Kulbe, Klaus D.

2011-01-01

α-l-Rhamnosidases play an important role in the hydrolysis of glycosylated aroma compounds (especially terpenes) from wine. Although several authors have demonstrated the enological importance of fungal rhamnosidases, the information on bacterial enzymes in this context is still limited. In order to fill this important gap, two putative rhamnosidase genes (ram and ram2) from Pediococcus acidilactici DSM 20284 were heterologously expressed, and the respective gene products were characterized. In combination with a bacterial β-glucosidase, both enzymes released the monoterpenes linalool and cis-linalool oxide from a muscat wine extract under ideal conditions. Additionally, Ram could release significant amounts of geraniol and citronellol/nerol. Nevertheless, the potential enological value of these enzymes is limited by the strong negative effects of acidity and ethanol on the activities of Ram and Ram2. Therefore, a direct application in winemaking seems unlikely. Although both enzymes are members of the same glycosyl hydrolase family (GH 78), our results clearly suggest the distinct functionalities of Ram and Ram2, probably representing two subclasses within GH 78: Ram could efficiently hydrolyze only the synthetic substrate p-nitrophenyl-α-l-rhamnopyranoside (Vmax = 243 U mg−1). In contrast, Ram2 displayed considerable specificity toward hesperidin (Vmax = 34 U mg−1) and, especially, rutinose (Vmax = 1,200 U mg−1), a disaccharide composed of glucose and rhamnose. Both enzymes were unable to hydrolyze the flavanone glycoside naringin. Interestingly, both enzymes displayed indications of positive substrate cooperativity. This study presents detailed kinetic data on two novel rhamnosidases, which could be relevant for the further study of bacterial glycosidases. PMID:21784921

Metabolism of ethylbenzene by human liver microsomes and recombinant human cytochrome P450s (CYP).

PubMed

Sams, Craig; Loizou, George D; Cocker, John; Lennard, Martin S

2004-03-07

The enzyme kinetics of the initial hydroxylation of ethylbenzene to form 1-phenylethanol were determined in human liver microsomes. The individual cytochrome P450 (CYP) forms catalysing this reaction were identified using selective inhibitors and recombinant preparations of hepatic CYPs. Production of 1-phenylethanol in hepatic microsomes exhibited biphasic kinetics with a high affinity, low Km, component (mean Km = 8 microM; V(max) = 689 pmol/min/mg protein; n = 6 livers) and a low affinity, high Km, component (Km = 391 microM; V(max) = 3039 pmol/min/mg protein; n = 6). The high-affinity component was inhibited 79%-95% (mean 86%) by diethyldithiocarbamate, and recombinant CYP2E1 was shown to metabolise ethylbenzene with low Km (35 microM), but also low (max) (7 pmol/min/pmol P450), indicating that this isoform catalysed the high-affinity component. Recombinant CYP1A2 and CYP2B6 exhibited high V(max) (88 and 71 pmol/min/pmol P450, respectively) and high Km (502 and 219 microM, respectively), suggesting their involvement in catalysing the low-affinity component. This study has demonstrated that CYP2E1 is the major enzyme responsible for high-affinity side chain hydroxylation of ethylbenzene in human liver microsomes. Activity of this enzyme in the population is highly variable due to induction or inhibition by physiological factors, chemicals in the diet or some pharmaceuticals. This variability can be incorporated into the risk assessment process to improve the setting of occupational exposure limits and guidance values for biological monitoring.

Hopantenate interference on the adaptation of muscular energy metabolism to intermittent hypoxia.

PubMed

Pastoris, O; Vercesi, L; Mazzocchi, A; Dossena, M; Benzi, G

1986-06-01

In rat gastrocnemius muscle, the concentrations of glycolytic fuels, intermediates and end-products; Krebs cycle intermediates and related free amino acids; ammonia; energy store and mediators; and the energy charge potential were evaluated in normoxia or after repeated, alternate hypoxic and normoxic exposures (12 hr of hypoxia daily; for 5 days) with or without treatment with hopantenate (HOPA). Furthermore, in the crude extract and/or mitochondrial fraction the maximum rate (Vmax) of some muscular enzymes related to the anaerobic glycolytic pathway; the tricarboxylic acid cycle; and the electron transfer chain were evaluated. Hopantenate was administered daily at the dose of 250 mg.kg-1 i.p., for 5 days, 30 min before the beginning of the experimental normobaric hypoxia. The biochemical adaptation to intermittent normobaric hypoxic-normoxic exposures was characterized by the decrease of the muscular concentrations of citrate, alpha-ketoglutarate and glutamate, in absence of changes in the Vmax of the muscle enzymes related to energy transduction. In gastrocnemius muscle from hypoxic rats, by HOPA treatment, both citrate and alpha-ketoglutarate maintained normal values, aspartate decreased, while glutamate remained reduced to subnormal values. In the muscle from hypoxic animals, by hopantenate treatment the Vmax of the mitochondrial enzymes tested (citrate synthase, malate dehydrogenase, total NADH cytochrome c reductase, cytochrome oxidase) decreased in comparison with both hypoxic and normoxic untreated animals. This behaviour could be tentatively related to a mitochondrial sparing action concomitant with an intervention of the glutamate group of amino acids, even if the results do not allow a clear interpretation of the mechanism of HOPA action.

Warming rate drives microbial limitation and enzyme expression during peat decomposition

NASA Astrophysics Data System (ADS)

Inglett, P.; Sihi, D.; Inglett, K. S.

2015-12-01

Recent developments of enzyme-based decomposition models highlight the importance of enzyme kinetics with warming, but most modeling exercises are based on studies with a step-wise warming. This approach may mask the effect of temperature in controlling in-situ activities as in most ecosystems soil temperature change more gradually than air temperature. We conducted an experiment to test the effects of contrasting warming rates on the kinetics of C, N, and P degradation enzymes in subtropical peat soils. We also wanted to evaluate if the stoichiometry of enzyme kinetics shifts under contrasting warming rates and if so, how does it relate to the stoichiometry in microbial biomass. Contrasting warming rates altered microbial biomass stoichiometry leading to differing patterns of enzyme expression and microbial nutrient limitation. Activity (higher Vmax) and efficiency (lower Km) of C acquisition enzymes were greater in the step treatment; however, expressions of nutrient (N and P) acquiring enzymes were enhanced in the ramp treatment at the end of the experiment. In the step treatment, there was a typical pattern of an initial peak in the Vmax and drop in the Km for all enzyme groups followed by later adjustments. On the other hand, a consistent increase in Vmax and decline in Km of all enzyme groups were observed in the slow warming treatment. These changes were sufficient to alter microbial identity (as indicated by enzyme Km and biomass stoichiometry) with two apparently stable endpoints under contrasting warming rates. This observation resembles the concept of alternate stable states and highlights a need for improved representation of warming in models.

Positive inotropic effects of RP 62719, a new pure class III antiarrhythmic agent, on guinea pig myocardium.

PubMed

Beregi, J P; Escande, D; Coudray, N; Mery, P; Mestre, M; Chemla, D; Lecarpentier, Y

1992-12-01

The mechanical effects of RP 62719 [(-)1-[-2-(3,4-dihydro-2H-1- benzopyran-4-yl)ethyl]-4-(3,4-dimethoxyphenyl)-piperidine] were tested in vitro on guinea pig left ventricular papillary muscle. RP 62719 is a novel pure class III antiarrhythmic agent known to prolong the cardiac action potential duration by selectively blocking the inward rectifying K+ current. Mechanical parameters were determined from contraction and relaxation phases under isotonic and isometric conditions. At a concentration of 0.02 microM, RP 62719 did not produce significant effects on inotropy or lusitropy. At 0.2 and 2 microM, the drug improved contraction under both heavy and low loading conditions, as evidenced by a 30% increase in maximum unloaded shortening velocity (Vmax, P < .001), peak amplitude of shortening (delta L, P < .001), peak isometric active force normalized per cross-sectional area (AF/s, P < .001) and positive peak of the force derivative per mm2 (+dF/s, P < .001). At the same concentrations, positive lusitropic effects were evidenced by an increase in maximum lengthening velocity (maxVr) and negative peak of force derivative per mm2 (-dF/s, P < .001). At a higher concentration (20 microM), effects of RP 62719 on inotropy and lusitropy were less marked, thus accounting for the bell-shaped form of the dose-response curve. An increase in the extracellular Ca++ concentration from 2.5 to 3.75 mM improved inotropy to a similar extent (+30-50%) as did 2 microM RP 62719. However, lusitropy and mechanical coupling between contraction and relaxation were not modified in the same proportion under RP 62719 and under 3.75 mM Ca++.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of methomyl on acetylcholinesterase in erythrocyte membrane and various brain areas].

PubMed

Zhao, Fei; Li, Tao; Zhang, Changchun; Xu, Yiping; Xu, Hangong; Shi, Nian

2015-06-01

To study the toxicity of methomyl to acetylcholinesterase (AChE) in different regions. The optimal temperature and time for measurement of AChE activity were determined in vitro. The dose- and time-response relationships of methomyl with AChE activity in human erythrocyte membrane, rat erythrocyte membrane, cortical synapses, cerebellar synapses, hippocampal synapses, and striatal synapses were evaluated. The half maximal inhibitory concentration (IC50) and bimolecular rate constant (K) of methomyl for AChE activity in different regions were calculated, and the type of inhibition of AChE activity by methomyl was determined. AChE achieved the maximum activity at 370 °C, and the optimal time to determine initial reaction velocity was 0-17 min. There were dose- and time-response relationships between methomyl and AChE activity in the erythrocyte membrane and various brain areas. The IC50 value of methomyl for AChE activity in human erythrocyte membrane was higher than that in rat erythrocyte membrane, while the Ki value of methomyl for AChE activity in rat erythrocyte membrane was higher than that in human erythrocyte membrane. Among synapses in various brain areas, the striatum had the highest IC50 value, followed by the cerebellum, cerebral cortex, and hippocampus, while the cerebral cortex had the highest Ki value, followed by the hippocampus, striatum, and cerebellum. Lineweaver-Burk diagram demonstrated that with increasing concentration of methomyl, the maximum reaction velocity (Vmax) of AChE decreased, and the Michaelis constant (Km) remained the same. Methomyl is a reversible non-competitive inhibitor of AChE. AChE of rat erythrocyte membrane is more sensitive to methomyl than that of human erythrocyte membrane; the cerebral cortical synapses have the most sensitive AChE to methomyl among synapses in various brain areas.

Haloes gone MAD: The Halo-Finder Comparison Project

NASA Astrophysics Data System (ADS)

Knebe, Alexander; Knollmann, Steffen R.; Muldrew, Stuart I.; Pearce, Frazer R.; Aragon-Calvo, Miguel Angel; Ascasibar, Yago; Behroozi, Peter S.; Ceverino, Daniel; Colombi, Stephane; Diemand, Juerg; Dolag, Klaus; Falck, Bridget L.; Fasel, Patricia; Gardner, Jeff; Gottlöber, Stefan; Hsu, Chung-Hsing; Iannuzzi, Francesca; Klypin, Anatoly; Lukić, Zarija; Maciejewski, Michal; McBride, Cameron; Neyrinck, Mark C.; Planelles, Susana; Potter, Doug; Quilis, Vicent; Rasera, Yann; Read, Justin I.; Ricker, Paul M.; Roy, Fabrice; Springel, Volker; Stadel, Joachim; Stinson, Greg; Sutter, P. M.; Turchaninov, Victor; Tweed, Dylan; Yepes, Gustavo; Zemp, Marcel

2011-08-01

We present a detailed comparison of fundamental dark matter halo properties retrieved by a substantial number of different halo finders. These codes span a wide range of techniques including friends-of-friends, spherical-overdensity and phase-space-based algorithms. We further introduce a robust (and publicly available) suite of test scenarios that allow halo finder developers to compare the performance of their codes against those presented here. This set includes mock haloes containing various levels and distributions of substructure at a range of resolutions as well as a cosmological simulation of the large-scale structure of the universe. All the halo-finding codes tested could successfully recover the spatial location of our mock haloes. They further returned lists of particles (potentially) belonging to the object that led to coinciding values for the maximum of the circular velocity profile and the radius where it is reached. All the finders based in configuration space struggled to recover substructure that was located close to the centre of the host halo, and the radial dependence of the mass recovered varies from finder to finder. Those finders based in phase space could resolve central substructure although they found difficulties in accurately recovering its properties. Through a resolution study we found that most of the finders could not reliably recover substructure containing fewer than 30-40 particles. However, also here the phase-space finders excelled by resolving substructure down to 10-20 particles. By comparing the halo finders using a high-resolution cosmological volume, we found that they agree remarkably well on fundamental properties of astrophysical significance (e.g. mass, position, velocity and peak of the rotation curve). We further suggest to utilize the peak of the rotation curve, vmax, as a proxy for mass, given the arbitrariness in defining a proper halo edge. Airport code for Madrid, Spain

Cardiohemodynamic and electrophysiological effects of a selective EP4 receptor agonist ONO--AE1--329 in the halothane-anesthetized dogs.

PubMed

Nomura, Hiroaki; Nakamura, Yuji; Cao, Xin; Honda, Atsushi; Katagi, Jun; Ohara, Hiroshi; Izumi-Nakaseko, Hiroko; Satoh, Yoshioki; Ando, Kentaro; Sugiyama, Atsushi

2015-08-15

Cardiovascular effects of a highly selective prostaglandin E2 type 4 (EP4) receptor agonist ONO-AE1-329 were assessed with the halothane-anesthetized dogs (n=6). ONO-AE1-329 was intravenously infused in three escalating doses of 0.3, 1 and 3ng/kg/min for 10min with a pause of 20min between the doses. The low dose of 0.3ng/kg/min significantly increased maximum upstroke velocity of left ventricular pressure by 18% at 20min, indicating increase of ventricular contractility. The middle dose of 1ng/kg/min significantly decreased total peripheral resistance by 24% and left ventricular end-diastolic pressure by 32% at 10min, indicating dilation of arteriolar resistance vessels and venous capacitance ones, respectively; and increased cardiac output by 25% at 10min in addition to the change induced by the low dose. The high dose of 3ng/kg/min increased heart rate by 34% at 10min; decreased mean blood pressure by 14% at 10min and atrioventricular nodal conduction time by 13% at 5min; and shortened left ventricular systolic period by 8% at 10min and electromechanical coupling defined as an interval from completion of repolarization to the start of ventricular diastole by 39% at 10min in addition to the changes induced by the middle dose. No significant change was detected in a ventricular repolarization period. These results indicate that ONO-AE1-329 may possess a similar cardiovascular profile to typical phosphodiesterase 3 inhibitors as an inodilator, and suggest that EP4 receptor stimulation can become an alternative strategy for the treatment of congestive heart failure. Copyright © 2015 Elsevier B.V. All rights reserved.

Acute heat tolerance of cardiac excitation in the brown trout (Salmo trutta fario).

PubMed

Vornanen, Matti; Haverinen, Jaakko; Egginton, Stuart

2014-01-15

The upper thermal tolerance and mechanisms of heat-induced cardiac failure in the brown trout (Salmo trutta fario) was examined. The point above which ion channel function and sinoatrial contractility in vitro, and electrocardiogram (ECG) in vivo, started to fail (break point temperature, BPT) was determined by acute temperature increases. In general, electrical excitation of the heart was most sensitive to heat in the intact animal (electrocardiogram, ECG) and least sensitive in isolated cardiac myocytes (ion currents). BPTs of Ca(2+) and K(+) currents of cardiac myocytes were much higher (>28°C) than BPT of in vivo heart rate (23.5 ± 0.6°C) (P<0.05). A striking exception among sarcolemmal ion conductances was the Na(+) current (INa), which was the most heat-sensitive molecular function, with a BPT of 20.9 ± 0.5°C. The low heat tolerance of INa was reflected as a low BPT for the rate of action potential upstroke in vitro (21.7 ± 1.2°C) and the velocity of impulse transmission in vivo (21.9 ± 2.2°C). These findings from different levels of biological organization strongly suggest that heat-dependent deterioration of Na(+) channel function disturbs normal spread of electrical excitation over the heart, leading to progressive variability of cardiac rhythmicity (missed beats, bursts of fast beating), reduction of heart rate and finally cessation of the normal heartbeat. Among the cardiac ion currents INa is 'the weakest link' and possibly a limiting factor for upper thermal tolerance of electrical excitation in the brown trout heart. Heat sensitivity of INa may result from functional requirements for very high flux rates and fast gating kinetics of the Na(+) channels, i.e. a trade-off between high catalytic activity and thermal stability.

A computational study on the influence of insect wing geometry on bee flight mechanics

PubMed Central

Feaster, Jeffrey; Bayandor, Javid

2017-01-01

ABSTRACT Two-dimensional computational fluid dynamics (CFD) is applied to better understand the effects of wing cross-sectional morphology on flow field and force production. This study investigates the influence of wing cross-section on insect scale flapping flight performance, for the first time, using a morphologically representative model of a bee (Bombus pensylvanicus) wing. The bee wing cross-section was determined using a micro-computed tomography scanner. The results of the bee wing are compared with flat and elliptical cross-sections, representative of those used in modern literature, to determine the impact of profile variation on aerodynamic performance. The flow field surrounding each cross-section and the resulting forces are resolved using CFD for a flight speed range of 1 to 5 m/s. A significant variation in vortex formation is found when comparing the ellipse and flat plate with the true bee wing. During the upstroke, the bee and approximate wing cross-sections have a much shorter wake structure than the flat plate or ellipse. During the downstroke, the flat plate and elliptical cross-sections generate a single leading edge vortex, while the approximate and bee wings generate numerous, smaller structures that are shed throughout the stroke. Comparing the instantaneous aerodynamic forces on the wing, the ellipse and flat plate sections deviate progressively with velocity from the true bee wing. Based on the present findings, a simplified cross-section of an insect wing can misrepresent the flow field and force production. We present the first aerodynamic study using a true insect wing cross-section and show that the wing corrugation increases the leading edge vortex formation frequency for a given set of kinematics. PMID:29061734

Altered profile of mRNA expression in atrioventricular node of streptozotocin-induced diabetic rats

PubMed Central

Howarth, Frank Christopher; Parekh, Khatija; Jayaprakash, Petrilla; Inbaraj, Edward Samuel; Oz, Murat; Dobrzynski, Halina; Adrian, Thomas Edward

2017-01-01

Prolonged action potential duration, reduced action potential firing rate, upstroke velocity and rate of diastolic depolarization have been demonstrated in atrioventricular node (AVN) cells from streptozotocin (STZ)-induced diabetic rats. To further clarify the molecular basis of these electrical disturbances, the mRNA profiles encoding a variety of proteins associated with the generation and conduction of electrical activity in the AVN, were evaluated in the STZ-induced diabetic rat heart. Expression of mRNA was measured in AVN biopsies using reverse transcription-quantitative polymerase chain reaction techniques. Notable differences in mRNA expression included upregulation of genes encoding membrane and intracellular Ca2+ transport, including solute carrier family 8 member A1, transient receptor potential channel 1, ryanodine receptor 2/3, hyperpolarization-activated cyclic-nucleotide 2 and 3, calcium channel voltage-dependent, β2 subunit and sodium channels 3a, 4a, 7a and 3b. In addition to this, potassium channels potassium voltage-gated channel subfamily A member 4, potassium channel calcium activated intermediate/small conductance subfamily N α member 2, potassium voltage-gated channel subfamily J members 3, 5, and 11, potassium channel subfamily K members 1, 2, 3 and natriuretic peptide B (BNP) were upregulated in AVN of STZ heart, compared with controls. Alterations in gene expression were associated with upregulation of various proteins including the inwardly rectifying, potassium channel Kir3.4, NCX1 and BNP. The present study demonstrated notable differences in the profile of mRNA encoding proteins associated with the generation, conduction and regulation of electrical signals in the AVN of the STZ-induced diabetic rat heart. These data will provide a basis for a substantial range of future studies to investigate whether variations in mRNA translate into alterations in electrophysiological function. PMID:28731153

The Interaction between Adult Cardiac Fibroblasts and Embryonic Stem Cell-Derived Cardiomyocytes Leads to Proarrhythmic Changes in In Vitro Cocultures

PubMed Central

Trieschmann, Jan; Bettin, Daniel; Haustein, Moritz; Köster, Annette; Molcanyi, Marek; Halbach, Marcel; Hanna, Mira; Fouad, Mariam; Brockmeier, Konrad; Hescheler, Jürgen; Pfannkuche, Kurt; Hannes, Tobias

2016-01-01

Transplantation of stem cell-derived cardiomyocytes is one of the most promising therapeutic approaches after myocardial infarction, as loss of cardiomyocytes is virtually irreversible by endogenous repair mechanisms. In myocardial scars, transplanted cardiomyocytes will be in immediate contact with cardiac fibroblasts. While it is well documented how the electrophysiology of neonatal cardiomyocytes is modulated by cardiac fibroblasts of the same developmental stage, it is unknown how adult cardiac fibroblasts (aCFs) affect the function of embryonic stem cell-derived cardiomyocytes (ESC-CMs). To investigate the effects of aCFs on ESC-CM electrophysiology, we performed extra- and intracellular recordings of murine aCF-ESC-CM cocultures. We observed that spontaneous beating behaviour was highly irregular in aCF-ESC-CM cocultures compared to cocultures with mesenchymal stem cells (coefficient of variation of the interspike interval: 40.5 ± 15.2% versus 9.3 ± 2.0%, p = 0.008) and that action potential amplitude and maximal upstroke velocity (V max) were reduced (amplitude: 52.3 ± 1.7 mV versus 65.1 ± 1.5 mV, V max: 7.0 ± 1.0 V/s versus 36.5 ± 5.3 V/s), while action potential duration (APD) was prolonged (APD50: 25.6 ± 1.0 ms versus 16.8 ± 1.9 ms, p < 0.001; APD90: 52.2 ± 1.5 ms versus 43.3 ± 3.3 ms, p < 0.01) compared to controls. Similar changes could be induced by aCF-conditioned medium. We conclude that the presence of aCFs changes automaticity and induces potentially proarrhythmic changes of ESC-CM electrophysiology. PMID:26880949

Electrophysiological characteristics of embryonic stem cell-derived cardiomyocytes are cell line-dependent.

PubMed

Hannes, Tobias; Wolff, Marie; Doss, Michael Xavier; Pfannkuche, Kurt; Haustein, Moritz; Müller-Ehmsen, Jochen; Sachinidis, Agapios; Hescheler, Jürgen; Khalil, Markus; Halbach, Marcel

2015-01-01

Modelling of cardiac development, physiology and pharmacology by differentiation of embryonic stem cells (ESCs) requires comparability of cardiac differentiation between different ESC lines. To investigate whether the outcome of cardiac differentiation is consistent between different ESC lines, we compared electrophysiological properties of ESC-derived cardiomyocytes (ESC-CMs) of different murine ESC lines. Two wild-type (D3 and R1) and two transgenic ESC lines (D3/aPIG44 and CGR8/AMPIGX-7) were differentiated under identical culture conditions. The transgenic cell lines expressed enhanced green fluorescent protein (eGFP) and puromycin-N-acetyltransferase under control of the cardiac specific α-myosin heavy chain (αMHC) promoter. Action potentials (APs) were recorded using sharp electrodes and multielectrode arrays in beating clusters of ESC-CMs. Spontaneous AP frequency and AP duration (APD) as well as maximal upstroke velocity differed markedly between unpurified CMs of the four ESC lines. APD heterogeneity was negligible in D3/aPIG44, moderate in D3 and R1 and extensive in CGR8/AMPIGX-7. Interspike intervals calculated from long-term recordings showed a high degree of variability within and between recordings in CGR8/AMPIGX-7, but not in D3/aPIG44. Purification of the αMHC+ population by puromycin treatment posed only minor changes to APD in D3/aPIG44, but significantly shortened APD in CGR8/AMPIGX-7. Electrophysiological properties of ESC-CMs are strongly cell line-dependent and can be influenced by purification of cardiomyocytes by antibiotic selection. Thus, conclusions on cardiac development, physiology and pharmacology derived from single stem cell lines have to be interpreted carefully. © 2015 S. Karger AG, Basel.

The Interaction between Adult Cardiac Fibroblasts and Embryonic Stem Cell-Derived Cardiomyocytes Leads to Proarrhythmic Changes in In Vitro Cocultures.

PubMed

Trieschmann, Jan; Bettin, Daniel; Haustein, Moritz; Köster, Annette; Molcanyi, Marek; Halbach, Marcel; Hanna, Mira; Fouad, Mariam; Brockmeier, Konrad; Hescheler, Jürgen; Pfannkuche, Kurt; Hannes, Tobias

2016-01-01

Transplantation of stem cell-derived cardiomyocytes is one of the most promising therapeutic approaches after myocardial infarction, as loss of cardiomyocytes is virtually irreversible by endogenous repair mechanisms. In myocardial scars, transplanted cardiomyocytes will be in immediate contact with cardiac fibroblasts. While it is well documented how the electrophysiology of neonatal cardiomyocytes is modulated by cardiac fibroblasts of the same developmental stage, it is unknown how adult cardiac fibroblasts (aCFs) affect the function of embryonic stem cell-derived cardiomyocytes (ESC-CMs). To investigate the effects of aCFs on ESC-CM electrophysiology, we performed extra- and intracellular recordings of murine aCF-ESC-CM cocultures. We observed that spontaneous beating behaviour was highly irregular in aCF-ESC-CM cocultures compared to cocultures with mesenchymal stem cells (coefficient of variation of the interspike interval: 40.5 ± 15.2% versus 9.3 ± 2.0%, p = 0.008) and that action potential amplitude and maximal upstroke velocity (V max) were reduced (amplitude: 52.3 ± 1.7 mV versus 65.1 ± 1.5 mV, V max: 7.0 ± 1.0 V/s versus 36.5 ± 5.3 V/s), while action potential duration (APD) was prolonged (APD50: 25.6 ± 1.0 ms versus 16.8 ± 1.9 ms, p < 0.001; APD90: 52.2 ± 1.5 ms versus 43.3 ± 3.3 ms, p < 0.01) compared to controls. Similar changes could be induced by aCF-conditioned medium. We conclude that the presence of aCFs changes automaticity and induces potentially proarrhythmic changes of ESC-CM electrophysiology.

Matrigel Mattress: A Method for the Generation of Single Contracting Human-Induced Pluripotent Stem Cell-Derived Cardiomyocytes.

PubMed

Feaster, Tromondae K; Cadar, Adrian G; Wang, Lili; Williams, Charles H; Chun, Young Wook; Hempel, Jonathan E; Bloodworth, Nathaniel; Merryman, W David; Lim, Chee Chew; Wu, Joseph C; Knollmann, Björn C; Hong, Charles C

2015-12-04

The lack of measurable single-cell contractility of human-induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs) currently limits the utility of hiPSC-CMs for evaluating contractile performance for both basic research and drug discovery. To develop a culture method that rapidly generates contracting single hiPSC-CMs and allows quantification of cell shortening with standard equipment used for studying adult CMs. Single hiPSC-CMs were cultured for 5 to 7 days on a 0.4- to 0.8-mm thick mattress of undiluted Matrigel (mattress hiPSC-CMs) and compared with hiPSC-CMs maintained on a control substrate (<0.1-mm thick 1:60 diluted Matrigel, control hiPSC-CMs). Compared with control hiPSC-CMs, mattress hiPSC-CMs had more rod-shape morphology and significantly increased sarcomere length. Contractile parameters of mattress hiPSC-CMs measured with video-based edge detection were comparable with those of freshly isolated adult rabbit ventricular CMs. Morphological and contractile properties of mattress hiPSC-CMs were consistent across cryopreserved hiPSC-CMs generated independently at another institution. Unlike control hiPSC-CMs, mattress hiPSC-CMs display robust contractile responses to positive inotropic agents, such as myofilament calcium sensitizers. Mattress hiPSC-CMs exhibit molecular changes that include increased expression of the maturation marker cardiac troponin I and significantly increased action potential upstroke velocity because of a 2-fold increase in sodium current (INa). The Matrigel mattress method enables the rapid generation of robustly contracting hiPSC-CMs and enhances maturation. This new method allows quantification of contractile performance at the single-cell level, which should be valuable to disease modeling, drug discovery, and preclinical cardiotoxicity testing. © 2015 American Heart Association, Inc.

A computational study on the influence of insect wing geometry on bee flight mechanics.

PubMed

Feaster, Jeffrey; Battaglia, Francine; Bayandor, Javid

2017-12-15

Two-dimensional computational fluid dynamics (CFD) is applied to better understand the effects of wing cross-sectional morphology on flow field and force production. This study investigates the influence of wing cross-section on insect scale flapping flight performance, for the first time, using a morphologically representative model of a bee ( Bombus pensylvanicus ) wing. The bee wing cross-section was determined using a micro-computed tomography scanner. The results of the bee wing are compared with flat and elliptical cross-sections, representative of those used in modern literature, to determine the impact of profile variation on aerodynamic performance. The flow field surrounding each cross-section and the resulting forces are resolved using CFD for a flight speed range of 1 to 5 m/s. A significant variation in vortex formation is found when comparing the ellipse and flat plate with the true bee wing. During the upstroke, the bee and approximate wing cross-sections have a much shorter wake structure than the flat plate or ellipse. During the downstroke, the flat plate and elliptical cross-sections generate a single leading edge vortex, while the approximate and bee wings generate numerous, smaller structures that are shed throughout the stroke. Comparing the instantaneous aerodynamic forces on the wing, the ellipse and flat plate sections deviate progressively with velocity from the true bee wing. Based on the present findings, a simplified cross-section of an insect wing can misrepresent the flow field and force production. We present the first aerodynamic study using a true insect wing cross-section and show that the wing corrugation increases the leading edge vortex formation frequency for a given set of kinematics. © 2017. Published by The Company of Biologists Ltd.

High-speed digital imaging of cytosolic Ca2+ and contraction in single cardiomyocytes.

PubMed

O'Rourke, B; Reibel, D K; Thomas, A P

1990-07-01

A charge-coupled device (CCD) camera, with the capacity for simultaneous spatially resolved photon counting and rapid frame transfer, was utilized for high-speed digital image collection from an inverted epifluorescence microscope. The unique properties of the CCD detector were applied to an analysis of cell shortening and the Ca2+ transient from fluorescence images of fura-2-loaded [corrected] cardiomyocytes. On electrical stimulation of the cell, a series of sequential subimages was collected and used to create images of Ca2+ within the cell during contraction. The high photosensitivity of the camera, combined with a detector-based frame storage technique, permitted collection of fluorescence images 10 ms apart. This rate of image collection was sufficient to resolve the rapid events of contraction, e.g., the upstroke of the Ca2+ transient (less than 40 ms) and the time to peak shortening (less than 80 ms). The technique was used to examine the effects of beta-adrenoceptor activation, fura-2 load, and stimulus frequency on cytosolic Ca2+ transients and contractions of single cardiomyocytes. beta-Adrenoceptor stimulation resulted in pronounced increases in peak Ca2+, maximal rates of rise and decay of Ca2+, extent of shortening, and maximal velocities of shortening and relaxation. Raising the intracellular load of fura-2 had little effect on the rising phase of Ca2+ or the extent of shortening but extended the duration of the Ca2+ transient and contraction. In related experiments utilizing differential-interference contrast microscopy, the same technique was applied to visualize sarcomere dynamics in contracting cells. This newly developed technique is a versatile tool for analyzing the Ca2+ transient and mechanical events in studies of excitation-contraction coupling in cardiomyocytes.

Aquatic wing flapping at low Reynolds numbers: swimming kinematics of the Antarctic pteropod, Clione antarctica.

PubMed

Borrell, Brendan J; Goldbogen, Jeremy A; Dudley, Robert

2005-08-01

We studied swimming kinematics of the Antarctic pteropod, Clione antarctica, to investigate how propulsive forces are generated by flexible oscillating appendages operating at low Reynolds numbers (1025) exhibited gliding during the recovery phase of each half-stroke. Maximum translational and rotational accelerations of the body occurred at the initiation of each power phase, suggesting that rotational circulation, the acceleration reaction, and wake recapture may all potentially contribute to vertical force production. Individual contributions of these mechanisms cannot, however, be assessed from these kinematic data alone. During recovery phases of each half-stroke, C. antarctica minimized adverse drag forces by orienting the wings parallel to flow and by moving them along the body surface, possibly taking advantage of boundary layer effects. Vertical force production was altered through changes in the hydrodynamic angle of attack of the wing that augmented drag during the power phase of each half-stroke. At higher translational velocities of the body, the inclination of the power phase also became more nearly vertical. These results indicate that, in addition to serotonin-mediated modulation of wingbeat frequency reported previously in Clione, geometric alteration of wingbeat kinematics offers a precise means of controlling swimming forces.

Performance assessment of Pulse Wave Imaging using conventional ultrasound in canine aortas ex vivo and normal human arteries in vivo

PubMed Central

Li, Ronny X.; Qaqish, William; Konofagou, Elisa. E.

2015-01-01

The propagation behavior of the arterial pulse wave may provide valuable diagnostic information for cardiovascular pathology. Pulse Wave Imaging (PWI) is a noninvasive, ultrasound imaging-based technique capable of mapping multiple wall motion waveforms along a short arterial segment over a single cardiac cycle, allowing for the regional pulse wave velocity (PWV) and propagation uniformity to be evaluated. The purpose of this study was to improve the clinical utility of PWI using a conventional ultrasound system. The tradeoff between PWI spatial and temporal resolution was evaluated using an ex vivo canine aorta (n = 2) setup to assess the effects of varying image acquisition and signal processing parameters on the measurement of the PWV and the pulse wave propagation uniformity r2. PWI was also performed on the carotid arteries and abdominal aortas of 10 healthy volunteers (24.8 ± 3.3 y.o.) to determine the waveform tracking feature that would yield the most precise PWV measurements and highest r2 values in vivo. The ex vivo results indicated that the highest precision for measuring PWVs ~ 2.5 – 3.5 m/s was achieved using 24–48 scan lines within a 38 mm image plane width (i.e. 0.63 – 1.26 lines/mm). The in vivo results indicated that tracking the 50% upstroke of the waveform would consistently yield the most precise PWV measurements and minimize the error in the propagation uniformity measurement. Such findings may help establish the optimal image acquisition and signal processing parameters that may improve the reliability of PWI as a clinical measurement tool. PMID:26640603

Electrophysiological and mechanical effects of caffeic acid phenethyl ester, a novel cardioprotective agent with antiarrhythmic activity, in guinea-pig heart.

PubMed

Chang, Gwo-Jyh; Chang, Chi-Jen; Chen, Wei-Jan; Yeh, Yung-Hsin; Lee, Hsiao-Yu

2013-02-28

Caffeic acid phenethyl ester (CAPE) is an active component of propolis that exhibits cardioprotective and antiarrhythmic effects. The detailed mechanisms underlying these effects, however, are not entirely understood. The aim of this study was to elucidate the electromechanical effects of CAPE in guinea-pig cardiac preparations. Intracardiac electrograms, left ventricular (LV) pressure, and the anti-arrhythmic efficacy were determined using isolated hearts. Action potentials of papillary muscles were assessed with microelectrodes, Ca(2+) transients were measured by fluorescence, and ion fluxes were measured by patch-clamp techniques. In a perfused heart model, CAPE prolonged the atrio-ventricular conduction interval, the Wenckebach cycle length, and the refractory periods of the AV node and His-Purkinje system, while shortening the QT interval. CAPE reduced the occurrence of reperfusion-induced ventricular fibrillation and decreased LV pressure in isolated hearts. In papillary muscles, CAPE shortened the action potential duration and reduced both the maximum upstroke velocity and contractile force. In fura-2-loaded single ventricular myocytes, CAPE decreased cell shortening and the Ca(2+) transient amplitude. Patch-clamp experiments revealed that CAPE produced a use-dependent decrease in L-type Ca(2+) current (ICa,L) (IC50=1.1 μM) and Na(+) current (INa) (IC50=0.43 μM), caused a negative-shift of the voltage-dependent inactivation and a delay of recovery from inactivation. CAPE decreased the delayed outward K(+) current (IK) slightly, without affecting the inward rectifier K(+) current (IK1). These results suggest that the preferential inhibition of Ca(2+) inward and Na(+) inward currents by CAPE may induce major electromechanical alterations in guinea-pig cardiac preparations, which may underlie its antiarrhythmic action. Copyright © 2013 Elsevier B.V. All rights reserved.

Role of suppression of the inward rectifier current in terminal action potential repolarization in the failing heart.

PubMed

Klein, Michael G; Shou, Matie; Stohlman, Jayna; Solhjoo, Soroosh; Haigney, Myles; Tidwell, Richard R; Goldstein, Robert E; Flagg, Thomas P; Haigney, Mark C

2017-08-01

The failing heart exhibits an increased arrhythmia susceptibility that is often attributed to action potential (AP) prolongation due to significant ion channel remodeling. The inwardly rectifying K + current (I K1 ) has been reported to be reduced, but its contribution to shaping the AP waveform and cell excitability in the failing heart remains unclear. The purpose of this study was to define the effect of I K1 suppression on the cardiac AP and excitability in the normal and failing hearts. We used electrophysiological and pharmacological approaches to investigate I K1 function in a swine tachy-pacing model of heart failure (HF). Terminal repolarization of the AP (TRAP; the time constant of the exponential fit to terminal repolarization) was markedly prolonged in both myocytes and arterially perfused wedges from animals with HF. TRAP was increased by 54.1% in HF myocytes (P < .001) and 26.2% in HF wedges (P = .014). The increase in TRAP was recapitulated by the potent and specific I K1 inhibitor, PA-6 (pentamidine analog 6), indicating that I K1 is the primary determinant of the final phase of repolarization. Moreover, we find that I K1 suppression reduced the ratio of effective refractory period to AP duration at 90% of repolarization, permitting re-excitation before full repolarization, reduction of AP upstroke velocity, and likely promotion of slow conduction. Using an objective measure of terminal repolarization, we conclude that I K1 is the major determinant of the terminal repolarization time course. Moreover, suppression of I K1 prolongs repolarization and reduces postrepolarization refractoriness without marked effects on the overall AP duration. Collectively, these findings demonstrate how I K1 suppression may contribute to arrhythmogenesis in the failing heart. Published by Elsevier Inc.

A spectral model for signal elements isolated from zebrafish photopic electroretinogram

PubMed Central

Nelson, Ralph; Singla, Nirmish

2009-01-01

The zebrafish photopic ERG sums isolatable elements. In each element red, blue, green and UV (r, g, b, u) cone signals combine in a way that reflects retinal organization. ERG responses to monochromatic stimuli of different wavelengths and irradiances were recorded on a white, rod suppressing background using superfused eyecups. Onset elements were isolated with glutamatergic blockers and response subtractions. CNQX blocked ionotropic (AMPA/kainate) glutamate receptors; L-AP4 or CPPG blocked metabotropic (mGluR6) glutamate receptors; TBOA blocked glutamate transporters; and L-Aspartate inactivated all glutamatergic mechanisms. Seven elements emerged: photopic PIII, the L-Aspartate-isolated cone response; b1, a CNQX-sensitive early b-wave element of inner retinal origin; PII, a photopic, CNQX-insensitive, composite b-wave element from ON bipolar cells; PIIm, an L-AP4/CPPG-sensitive, CNQX-insensitive metabotropic sub-element of PII; PIInm, an L-AP4/CPPG/CNQX-insensitive, non-metabotropic sub-element of PII; a1nm, a TBOA-sensitive, CNQX/L-AP4/CPPG-insensitive, non-metabotropic, post-photoreceptor a-wave element; and a2, a CNQX-sensitive a-wave element linked to OFF bipolar cells. The first five elements were fit with a spectral model that demonstrates independence of cone color pathways. From this Vmax and half-saturation values (k) for the contributing r- g- b- and u-cone signals were calculated. Two signal patterns emerged. For PIII or PIInm the Vmax order was Vr > Vg ≫ Vb ≈ Vu. For b1, PII, and PIIm the Vmax order was Vr ≈ Vb > Vg > Vu. In either pattern u-cone amplitude (Vu) was smallest, but u-cone sensitivity (ku362) was greatest, some 10-30 times greater than r-cone (kr570). The spectra of b1/PII/PIIm elements peaked near b-cone and u-cone absorbance maxima regardless of criteria, but the spectra of PIII/PIInm elements shifted from b- towards r-cone absorbance maxima as criterion levels increased. The greatest gains in Vmax relative to PIII occurred for the b- and u-cone signals in the b1/PII/PIIm b-wave elements. This suggests a high-gain, prolific metabotropic circuitry for b- and u-cone bipolar cells. PMID:19723365

Dependence of renal (Na+ + k+)-adenosine triphosphatase activity on thyroid status.

PubMed

Lo, S C; August, T R; Liberman, U A; Edelman, I S

1976-12-25

In thyroidectomized rats, a single injection of L-2,,5,2'-triiodothyronine (T3) (50mug/100 g body weight) elicited at 45% increase in (Na+ + k+)-dependent adenosine triphosphatase (NaK-ATPase) activity of the membrane-rich fraction of renal cortex at the optimal time of response, 48 h after injection. Three successive doses of T3 (50 mug/100 g body weight), given on alternate days, increased NaK-ATPase by 67% in the renal cortex but had no significant effect on the outer medulla or the papilla. Moreover, T3 had no effect on Mg2+-dependent adenosine trisphatase (MgATPase) in cortex, cedulla, or papilla. Three doses of T3 (50 mug/100 g body weight) given on alternate days to thyroidectomized rats elecited a 134, 79, and 46% increase in Vmax for ATP, Na4, and K+, respectively. There were no changes in the Km for ATP or the K1/2 values for Na+ and K+. Two methods were used to estimate the effect of T3 on the number of NaK-ATPase units (assumed to represent the number of Na+ pump sites); rat renal plasma membrane fractions were incubated with [gamma-32P]ATP, Mg2+, and Na+; the 32P-labeled membrane protein yeild was quantitatively dependent on Na+ and was hydrolyzed on addition of K+. There was a linear correlation between the specific activity of NaK-ATPase (Vmax) and the amount of phosphorylated intermediate formed, in renal cortical membrane fractions from thyroidectomized rats given T3 or the diluent. There was also a linear correlation between the specific activity of NaK-ATPase (Vmax) and the amount of [3H]ouabain specifically bound (Na+-, Mg2+-, APT-dependent) to the NaK-ATPase preparation. Injection of T3 resulted in a 70% increase in NaK-ATPase activity, a 79% increase in formation of the phosphorylated intermediate, and a 65% increase in the [H]ouabain specifically bound to the NaK-ATPase system. The T3-dependent increases in Vmax for ATP, Na+, and K+ and the proportionate increases in the phosphorylated intermediate and in the amount of [3H]ouabain bound indicate that T3 increases the number of NaK-ATPase units and that this increase accounts for the increase in NaK-ATPase activity.

Characterization of deltamethrin metabolism by rat plasma and liver microsomes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anand, Sathanandam S.; Bruckner, James V.; Haines, Wendy T.

2006-04-15

Deltamethrin, a widely used type II pyrethroid insecticide, is a relatively potent neurotoxicant. While the toxicity has been extensively examined, toxicokinetic studies of deltamethrin and most other pyrethroids are very limited. The aims of this study were to identify, characterize, and assess the relative contributions of esterases and cytochrome P450s (CYP450s) responsible for deltamethrin metabolism by measuring deltamethrin disappearance following incubation of various concentrations (2 to 400 {mu}M) in plasma (esterases) and liver microsomes (esterases and CYP450s) prepared from adult male rats. While the carboxylesterase metabolism in plasma and liver was characterized using an inhibitor, tetra isopropyl pyrophosphoramide (isoOMPA), CYP450more » metabolism was characterized using the cofactor, NADPH. Michaelis-Menten rate constants were calculated using linear and nonlinear regression as applicable. The metabolic efficiency of these pathways was estimated by calculating intrinsic clearance (Vmax/Km). In plasma, isoOMPA completely inhibited deltamethrin biotransformation at concentrations (2 and 20 {mu}M of deltamethrin) that are 2- to 10-fold higher than previously reported peak blood levels in deltamethrin-poisoned rats. For carboxylesterase-mediated deltamethrin metabolism in plasma, Vmax = 325.3 {+-} 53.4 nmol/h/ml and Km = 165.4 {+-} 41.9 {mu}M. Calcium chelation by EGTA did not inhibit deltamethrin metabolism in plasma or liver microsomes, indicating that A-esterases do not metabolize deltamethrin. In liver microsomes, esterase-mediated deltamethrin metabolism was completely inhibited by isoOMPA, confirming the role of carboxylesterases. The rate constants for liver carboxylesterases were Vmax = 1981.8 {+-} 132.3 nmol/h/g liver and Km = 172.5 {+-} 22.5 {mu}M. Liver microsomal CYP450-mediated biotransformation of deltamethrin was a higher capacity (Vmax = 2611.3 {+-} 134.1 nmol/h/g liver) and higher affinity (Km = 74.9 {+-} 5.9 {mu}M) process than carboxylesterase (plasma or liver) detoxification. Genetically engineered individual rat CYP450s (Supersomes) were used to identify specific CYP450 isozyme(s) involved in the deltamethrin metabolism. CYP1A2, CYP1A1, and CYP2C11 in decreasing order of importance quantitatively, metabolized deltamethrin. Intrinsic clearance by liver CYP450s (35.5) was more efficient than that by liver (12.0) or plasma carboxylesterases (2.4)« less

Investigation of micropump mechanism for medical application (blood transport application)

NASA Astrophysics Data System (ADS)

Piterah, N. S. M.; Ong, N. R.; Aziz, M. H. A.; Alcain, J. B.; Haimi, W. M. W. N.; Sauli, Z.

2017-09-01

A microfluidic device is a beneficial device in transporting and controling the flow of fluid in microfluidic system especially in biomedical research and application. This study proposed a valveless micropump design with reciprocating micropumping concept. This micropump mechanism model was specifically designed to overcome hydrodynamic reversibility effectively at low Reynolds number and work on finite pressure loads. The transportation of microfluidic especially biological material such as blood was presented clearly in this micropumping mechanism. The transportation of fluid throughout microchannel with low Reynolds number 16 produced 7.5 m3 maximum net volume of blood pumped from left to right and configured upstroke and downstroke situation during 0.74 seconds and 0.24 seconds respectively.

Rational mutagenesis by engineering disulphide bonds improves Kluyveromyces lactis beta-galactosidase for high-temperature industrial applications

PubMed Central

Rico-Díaz, Agustín; Álvarez-Cao, María-Efigenia; Escuder-Rodríguez, Juan-José; González-Siso, María-Isabel; Cerdán, M. Esperanza; Becerra, Manuel

2017-01-01

Kluyveromyces lactis β-galactosidase (Kl-β-Gal) is one of the most important enzymes in the dairy industry. The poor stability of this enzyme limits its use in the synthesis of galactooligosaccharides (GOS) and other applications requiring high operational temperature. To obtain thermoresistant variants, a rational mutagenesis strategy by introducing disulphide bonds in the interface between the enzyme subunits was used. Two improved mutants, R116C/T270C and R116C/T270C/G818C, had increased half-lives at 45 °C compared to Kl-β-Gal (2.2 and 6.8 fold increases, respectively). Likewise, Tm values of R116C/T270C and R116C/T270C/G818C were 2.4 and 8.5 °C, respectively, higher than Kl-β-Gal Tm. Enrichment in enzymatically active oligomeric forms in these mutant variants also increased their catalytic efficiency, due to the reinforcement of the interface contacts. In this way, using an artificial substrate (p-nitrophenyl-β-D-galactopyranoside), the Vmax values of the mutants were ~1.4 (R116C/T270C) and 2 (R116C/T270C/G818C) fold higher than that of native Kl-β-Gal. Using the natural substrate (lactose) the Vmax for R116C/T270C/G818C almost doubled the Vmax for Kl-β-Gal. Validation of these mutant variants of the enzyme for their use in applications that depend on prolonged incubations at high temperatures was achieved at the laboratory scale by monitoring their catalytic activity in GOS synthesis. PMID:28361909

Evaluation of possible reasons for the low phenylalanine ammonia lyase activity in cellulose nitrate membrane microcapsules.

PubMed

Habibi-Moini, S; D'mello, A P

2001-03-14

Microencapsulated phenylalanine ammonia lyase (PAL) exhibits a marked reduction in activity compared to the activity of the free enzyme in pH 8.5 Tris buffer. The purpose of this investigation was to evaluate the contribution of incomplete entrapment, the internal environment of cellulose nitrate membrane microcapsules, the diffusional barrier of the membrane and the microcapsulation process to the low activity of encapsulated PAL. A solution of PAL and 10% w/v hemoglobin was incorporated into cellulose nitrate membrane microcapsules. Hemoglobin incorporation was used as a surrogate marker of PAL entrapment. Using 14C hemoglobin, the encapsulation efficiency was determined to be 70% and suggested that incomplete entrapment might partially account for the low activity of encapsulated PAL. The effect of the internal environment of the microcapsule (10% hemoglobin solution) on PAL activity was evaluated by comparing enzyme activity in 10% w/v hemoglobin solution and pH 8.5 Tris buffer. Similar K(M) and V(max) values of PAL in the two media indicated that the internal environment of the microcapsule did not contribute to the reduction in activity of the encapsulated enzyme. The contribution of a membrane diffusional barrier was determined by breaking the putative barrier and measuring PAL activity in intact and broken microcapsules. Similar activity of PAL in these two conditions is evidence for the lack of a diffusional barrier. The effect of the microencapsulation process on PAL activity was evaluated by comparing K(M) and V(max) of free and encapsulated PAL. Similar K(M) values in these two media suggested that the process did not affect the conformation of PAL. However, encapsulated PAL had a 50% lower V(max) value compared to free PAL, which showed that the microencapsulation process deactivated a substantial proportion of the enzyme.

Comparative study of invertases of Streptococcus mutans.

PubMed

Tanzer, J M; Brown, A T; McInerney, M F; Woodiel, F N

1977-04-01

Sucrase activity was studied in 13 strains of Streptococcus mutans representing the five Bratthall serotypes. Sucrose-adapted cells have sucrase activity in the 37,000 x g-soluble fraction of all strains. The enzyme was identified as invertase (beta-d-fructofuranoside fructohydrolase; EC 3.2.1.26) because it hydrolyzed the beta-fructofuranoside trisaccharide raffinose, giving fructose and melibiose as its products, and because it hydrolyzed the beta-fructofuranoside dissacharide sucrose, giving equimolar glucose and fructose as its products. Invertases of c and e strains exhibit two activity peaks by molecular exclusion chromatography with molecular weights of 45,000 to 50,000 and about 180,000; those of serotypes a, b, and d strains exhibit only a single component of 45,000 to 50,000 molecular weight. The electrophoretic mobility of invertases is different between the serotypes and the same within them. Inorganic orthophosphate (P(i)) has a weak positive effect on the V(max) of invertases of serotypes c and e cells but a strong positive effect on the invertases of serotype b cells; P(i) has a strong positive effect on the apparent K(m) of the invertases of serotype d cells, but has no effect on the V(max); P(i) has a strong positive effect on both the apparent K(m) and V(max) of the invertases of serotype a cells. Thus, the invertases were different between all of the serotypes but similar within the serotypes. These findings support the taxonomic schemes of Coykendall and of Bratthall. It was additionally noted that 37,000 x g-soluble fractions of only serotypes b and c but not serotypes a, d, and e cells have melibiase activity, and it could be deduced that serotype d cells lack an intact raffinose permease system.

Permeability, transport, and metabolism of solutes in Caco-2 cell monolayers: a theoretical study.

PubMed

Sun, Huadong; Pang, K Sandy

2008-01-01

We explored the properties of a catenary model that includes the basolateral (B), apical (A), and cellular compartments via simulations under linear and nonlinear conditions to understand the asymmetric observations arising from transporters, enzymes, and permeability in Caco-2 cells. The efflux ratio (EfR; P(app,B-->A)/P(app,A-->B)), obtained from the effective permeability from the A-->B and B-->A direction under linear conditions, was unity for passively permeable drugs whose transport does not involve transporters; the value was unaffected by cellular binding or metabolism, but increased with apical efflux. Metabolism was asymmetric, showing lesser metabolite accrual for the B-->A than A-->B direction because of inherent differences in the volumes for A and B. Moreover, the net flux (total - passive permeation) due to saturable apical efflux, absorption, or metabolism showed nonconformity to simple Michaelis-Menten kinetics against C(D,0), the loading donor concentration. EfR values differed with saturable apical efflux and metabolism (>1), as well as apical absorption (EfRs <1), but approached unity with high passive diffusive clearance (CL(d)) and increasing C(D,0) at a higher degree of saturation of the process. The J(max) (apparent V(max) estimated for the carrier system) and K(m)(') [or the K(m)('') based on a modified equation with the Hill coefficient (beta)] estimates from the Eadie-Hofstee plot revealed spurious correlations with the assigned V(max) and K(m). The sampling time, CL(d), and parameter space of K(m) and V(max) strongly influenced both the correlation and accuracy of estimates. Improved correlation was found for compounds with high CL(d). These observations showed that the catenary model is appropriate in the description of transport and metabolic data in Caco-2 cells.

Effect of chronic pre-treatment with angiotensin converting enzyme inhibition on skeletal muscle mitochondrial recovery after ischemia/reperfusion.

PubMed

Thaveau, Fabien; Zoll, Joffrey; Bouitbir, Jamal; N'guessan, Benoît; Plobner, Philippe; Chakfe, Nabil; Kretz, Jean-Georges; Richard, Ruddy; Piquard, François; Geny, Bernard

2010-06-01

Impaired skeletal muscle energetic participates in peripheral arterial disease (PAD) patient's morbidity and mortality. Angiotensin converting enzyme inhibition (ACEi), cornerstone for pharmacologic risk factor management in PAD patients, might also be interesting by protecting skeletal muscle energetic. We therefore determined whether chronic ACEi might reduce ischemia-induced mitochondrial respiratory chain dysfunction in the frequent setting of hindlimb ischemia-reperfusion. Ischemic legs of rats submitted to 5 h ischemia induced by a rubber band tourniquet applied on the root of the hindlimb followed by reperfusion without (IR, n = 11) or after ACEi (n = 14; captopril 40 mg/kg per day during 28 days before surgery) were studied and compared to that of sham-operated animals (n = 11). The effect of ACEi on the non-ischemic contralateral leg was also determined in the ACEi group. Maximal oxidative capacities (V(max)) and complexes I, II and IV activities of the mitochondrial respiratory chain of the gastrocnemius muscle were determined using glutamate-malate, succinate and TMPD-ascorbate substrates. Arterial blood pressure was significantly decreased after ACEi (124 +/- 2.8 vs. 108 +/- 4.19 mmHg; P = 0.01). Ischemia-reperfusion reduced V(max) (4.4 +/- 0.4 vs. 8.7 +/- 0.5 micromol O2/min/g dry weight, -49%, P < 0.001), affecting mitochondrial complexes I, II and IV activities. ACEi failed to modulate ischemia-induced dysfunction (V(max) 5.1 +/- 0.7 micromol O2/min/g dry weight) or the non-ischemic contralateral muscle respiratory rate. Ischemia-reperfusion significantly impaired the mitochondrial respiratory chain I, II and IV complexes of skeletal muscle. Pharmacologic pre-treatment with ACEi did not prevent or increase such alterations. Further studies might be useful to improve the pharmacologic conditioning of PAD patients needing arterial revascularization.

Organization of metabolic pathways in vastus lateralis of patients with chronic obstructive pulmonary disease.

PubMed

Green, Howard J; Bombardier, Eric; Burnett, Margaret; Iqbal, Sobia; D'Arsigny, Christine L; O'Donnell, Dennis E; Ouyang, Jing; Webb, Katherine A

2008-09-01

The objective of this study was to determine whether patients with chronic obstructive lung disease (COPD) display differences in organization of the metabolic pathways and segments involved in energy supply compared with healthy control subjects. Metabolic pathway potential, based on the measurement of the maximal activity (V(max)) of representative enzymes, was assessed in tissue extracted from the vastus lateralis in seven patients with COPD (age 67 +/- 4 yr; FEV(1)/FVC = 44 +/- 3%, where FEV(1) is forced expiratory volume in 1 s and FVC is forced vital capacity; means +/- SE) and nine healthy age-matched controls (age 68 +/- 2 yr; FEV(1)/FVC = 75 +/- 2%). Compared with control, the COPD patients displayed lower (P < 0.05) V(max) (mol.kg protein(-1).h(-1)) for cytochrome c oxidase (COX; 21.2 +/- 2.0 vs. 28.7 +/- 2.2) and 3-hydroxyacyl-CoA dehydrogenase (HADH; 2.54 +/- 0.14 vs. 3.74 +/- 0.12) but not citrate synthase (CS; 2.20 +/- 0.16 vs. 3.19 +/- 0.5). While no differences between groups were observed in V(max) for creatine phosphokinase, phosphorylase (PHOSPH), phosphofructokinase (PFK), pyruvate kinase, and lactate dehydrogenase, hexokinase (HEX) was elevated in COPD (P < 0.05). Enzyme activity ratios were higher (P < 0.05) for HEX/CS, HEX/COX, PHOSPH/HADH and PFK/HADH in COPD compared with control. It is concluded that COPD patients exhibit a reduced potential for both the electron transport system and fat oxidation and an increased potential for glucose phosphorylation while the potential for glycogenolysis and glycolysis remains normal. A comparison of enzyme ratios indicated greater potentials for glucose phosphorylation relative to the citric acid cycle and the electron transport chain and glycogenolysis and glycolysis relative to beta-oxidation.

Cellulose binding domain assisted immobilization of lipase (GSlip-CBD) onto cellulosic nanogel: characterization and application in organic medium.

PubMed

Kumar, Ashok; Zhang, Shaowei; Wu, Gaobing; Wu, Cheng Chao; Chen, JunPeng; Baskaran, R; Liu, Ziduo

2015-12-01

A cbd gene was cloned into the C-terminal region of a lip gene from Geobacillus stearothermophilus. The native lipase (43.5 kDa) and CBD-Lip fusion protein (60.2 kDa) were purified to homogeneity by SDS-PAGE. A highly stable cellulosic nanogel was prepared by controlled hydrolysis of microcrystalline cellulose onto which the CBD-lip fusion protein was immobilized through bio-affinity based binding. The nanogel-bound lipase showed optimum activity at 55 °C, and it remains stable and active at pH 10-10.5. Furthermore, the immobilized lipase showed an over two-fold increase of relative activity in the presence of DMSO, isopropanol, isoamyl alcohol and n-butanol, but a mild activity decrease at a low concentration of methanol and ethanol. The immobilized biocatalyst retained ~50% activity after eight repetitive hydrolytic cycles. Enzyme kinetic studies of the immobilized lipase showed a 1.24 fold increase in Vmax and 5.25 fold increase in kcat towards p-NPP hydrolysis. Additionally, the nanogel bound lipase was tested to synthesize a biodiesel ester, ethyl oleate in DMSO. Kinetic analysis showed the km 100.5 ± 4.3 mmol and Vmax 0.19 ± 0.015 mmolmin(-1) at varied oleic acid concentration. Also, the values of km and Vmax at varying concentration of ethanol were observed to be 95.9 ± 13.9 mmol and 0.22 ± 0.013 mmolmin(-1) respectively. The maximum yield of ethyl oleate 111.2 ± 1.24 mM was obtained under optimized reaction conditions in organic medium. These results suggest that this immobilized biocatalyst can be used as an efficient tool for the biotransformation reactions on an industrial scale. Copyright © 2015 Elsevier B.V. All rights reserved.

Rational mutagenesis by engineering disulphide bonds improves Kluyveromyces lactis beta-galactosidase for high-temperature industrial applications.

PubMed

Rico-Díaz, Agustín; Álvarez-Cao, María-Efigenia; Escuder-Rodríguez, Juan-José; González-Siso, María-Isabel; Cerdán, M Esperanza; Becerra, Manuel

2017-03-31

Kluyveromyces lactis β-galactosidase (Kl-β-Gal) is one of the most important enzymes in the dairy industry. The poor stability of this enzyme limits its use in the synthesis of galactooligosaccharides (GOS) and other applications requiring high operational temperature. To obtain thermoresistant variants, a rational mutagenesis strategy by introducing disulphide bonds in the interface between the enzyme subunits was used. Two improved mutants, R116C/T270C and R116C/T270C/G818C, had increased half-lives at 45 °C compared to Kl-β-Gal (2.2 and 6.8 fold increases, respectively). Likewise, Tm values of R116C/T270C and R116C/T270C/G818C were 2.4 and 8.5 °C, respectively, higher than Kl-β-Gal Tm. Enrichment in enzymatically active oligomeric forms in these mutant variants also increased their catalytic efficiency, due to the reinforcement of the interface contacts. In this way, using an artificial substrate (p-nitrophenyl-β-D-galactopyranoside), the Vmax values of the mutants were ~1.4 (R116C/T270C) and 2 (R116C/T270C/G818C) fold higher than that of native Kl-β-Gal. Using the natural substrate (lactose) the Vmax for R116C/T270C/G818C almost doubled the Vmax for Kl-β-Gal. Validation of these mutant variants of the enzyme for their use in applications that depend on prolonged incubations at high temperatures was achieved at the laboratory scale by monitoring their catalytic activity in GOS synthesis.

Substrate preferences and catalytic parameters determined by structural characteristics of sterol 14alpha-demethylase (CYP51) from Leishmania infantum.

PubMed

Hargrove, Tatiana Y; Wawrzak, Zdzislaw; Liu, Jialin; Nes, W David; Waterman, Michael R; Lepesheva, Galina I

2011-07-29

Leishmaniasis is a major health problem that affects populations of ∼90 countries worldwide, with no vaccine and only a few moderately effective drugs. Here we report the structure/function characterization of sterol 14α-demethylase (CYP51) from Leishmania infantum. The enzyme catalyzes removal of the 14α-methyl group from sterol precursors. The reaction is essential for membrane biogenesis and therefore has great potential to become a target for antileishmanial chemotherapy. Although L. infantum CYP51 prefers C4-monomethylated sterol substrates such as C4-norlanosterol and obtusifoliol (V(max) of ∼10 and 8 min(-1), respectively), it is also found to 14α-demethylate C4-dimethylated lanosterol (V(max) = 0.9 min(-1)) and C4-desmethylated 14α-methylzymosterol (V(max) = 1.9 min(-1)). Binding parameters with six sterols were tested, with K(d) values ranging from 0.25 to 1.4 μM. Thus, L. infantum CYP51 is the first example of a plant-like sterol 14α-demethylase, where requirements toward the composition of the C4 atom substituents are not strict, indicative of possible branching in the postsqualene portion of sterol biosynthesis in the parasite. Comparative analysis of three CYP51 substrate binding cavities (Trypanosoma brucei, Trypanosoma cruzi, and L. infantum) suggests that substrate preferences of plant- and fungal-like protozoan CYP51s largely depend on the differences in the enzyme active site topology. These minor structural differences are also likely to underlie CYP51 catalytic rates and drug susceptibility and can be used to design potent and specific inhibitors.

Kinetics of phosphate uptake, growth, and accumulation of cyclic diphosphoglycerate in a phosphate-limited continuous culture of Methanobacterium thermoautotrophicum.

PubMed Central

Krueger, R D; Harper, S H; Campbell, J W; Fahrney, D E

1986-01-01

The archaebacterium Methanobacterium thermoautotrophicum was grown in continuous culture at 65 degrees C in a phosphate-limited medium at specific growth rates from 0.06 to 0.28 h-1 (maximum growth rate [mu max] = 0.36 h-1). Cyclic-2,3-diphosphoglycerate (cyclic DPG) levels ranged from 2 to 20 mM in Pi-limited cells, compared with about 30 mM in batch-grown cells. The Monod constant for Pi-limited growth was 5 nM. Pi uptake rates were determined by following the disappearance of 32Pi from the medium. Interrupting the H2 supply stopped the uptake of Pi and the release of organic phosphates. Little or no efflux of Pi occurred in the presence or absence of H2. Pi uptake of cells adapted to nanomolar Pi concentrations could be accounted for by the operation of one uptake system with an apparent Km of about 25 nM and a Vmax of 58 nmol of Pi per min per g (dry weight). Uptake curves at 30 microM Pi or above were biphasic due to a sevenfold decrease in Vmax after an initial phase of rapid movement of Pi into the cell. Under these conditions the growth rate slowed to zero and the cyclic DPG pool expanded before growth resumed. Thus, three properties of M. thermoautotrophicum make it well adapted to live in a low-P environment: the presence of a low-Km, high-Vmax uptake system for Pi; the ability to accumulate cyclic DPG rapidly; and a growth strategy in which accumulation of Pi and cyclic DPG takes precedence over a shift-up in growth rate when excess Pi becomes available. PMID:3722128

Kinetics of phosphate uptake, growth, and accumulation of cyclic diphosphoglycerate in a phosphate-limited continuous culture of Methanobacterium thermoautotrophicum.

PubMed

Krueger, R D; Harper, S H; Campbell, J W; Fahrney, D E

1986-07-01

The archaebacterium Methanobacterium thermoautotrophicum was grown in continuous culture at 65 degrees C in a phosphate-limited medium at specific growth rates from 0.06 to 0.28 h-1 (maximum growth rate [mu max] = 0.36 h-1). Cyclic-2,3-diphosphoglycerate (cyclic DPG) levels ranged from 2 to 20 mM in Pi-limited cells, compared with about 30 mM in batch-grown cells. The Monod constant for Pi-limited growth was 5 nM. Pi uptake rates were determined by following the disappearance of 32Pi from the medium. Interrupting the H2 supply stopped the uptake of Pi and the release of organic phosphates. Little or no efflux of Pi occurred in the presence or absence of H2. Pi uptake of cells adapted to nanomolar Pi concentrations could be accounted for by the operation of one uptake system with an apparent Km of about 25 nM and a Vmax of 58 nmol of Pi per min per g (dry weight). Uptake curves at 30 microM Pi or above were biphasic due to a sevenfold decrease in Vmax after an initial phase of rapid movement of Pi into the cell. Under these conditions the growth rate slowed to zero and the cyclic DPG pool expanded before growth resumed. Thus, three properties of M. thermoautotrophicum make it well adapted to live in a low-P environment: the presence of a low-Km, high-Vmax uptake system for Pi; the ability to accumulate cyclic DPG rapidly; and a growth strategy in which accumulation of Pi and cyclic DPG takes precedence over a shift-up in growth rate when excess Pi becomes available.

Enhancement of Force Generated by Individual Myosin Heads in Skinned Rabbit Psoas Muscle Fibers at Low Ionic Strength

PubMed Central

Sugi, Haruo; Abe, Takahiro; Kobayashi, Takakazu; Chaen, Shigeru; Ohnuki, Yoshiki; Saeki, Yasutake; Sugiura, Seiryo

2013-01-01

Although evidence has been presented that, at low ionic strength, myosin heads in relaxed skeletal muscle fibers form linkages with actin filaments, the effect of low ionic strength on contraction characteristics of Ca2+-activated muscle fibers has not yet been studied in detail. To give information about the mechanism of muscle contraction, we have examined the effect of low ionic strength on the mechanical properties and the contraction characteristics of skinned rabbit psoas muscle fibers in both relaxed and maximally Ca2+-activated states. By progressively decreasing KCl concentration from 125 mM to 0 mM (corresponding to a decrease in ionic strength μ from 170 mM to 50 mM), relaxed fibers showed changes in mechanical response to sinusoidal length changes and ramp stretches, which are consistent with the idea of actin-myosin linkage formation at low ionic strength. In maximally Ca2+-activated fibers, on the other hand, the maximum isometric force increased about twofold by reducing KCl concentration from 125 to 0 mM. Unexpectedly, determination of the force-velocity curves indicated that, the maximum unloaded shortening velocity Vmax, remained unchanged at low ionic strength. This finding indicates that the actin-myosin linkages, which has been detected in relaxed fibers at low ionic strength, are broken quickly on Ca2+ activation, so that the linkages in relaxed fibers no longer provide any internal resistance against fiber shortening. The force-velocity curves, obtained at various levels of steady Ca2+-activated isometric force, were found to be identical if they are normalized with respect to the maximum isometric force. The MgATPase activity of muscle fibers during isometric force generation was found not to change appreciably at low ionic strength despite the two-fold increase in Ca2+-activated isometric force. These results can be explained in terms of enhancement of force generated by individual myosin heads, but not by any changes in kinetic properties of cyclic actin-myosin interaction. PMID:23691080

Comparison of shuttle and straight repeated-sprint ability tests and their relationship to anthropometrics and explosive muscular performance of lower limb in elite handball players.

PubMed

Hermassi, Souhail; Schwesig, René; Wollny, Rainer; Fieseler, Georg; van den Tillaar, Roland; Fernandez-Fernandez, Jaime; Shephard, Roy J; Chelly, Mohamed S

2017-06-21

This study assessed inter-relationships between shuttle and straight repeated-sprint ability tests and the relationship of each measure to anthropometric and explosive lower limb performance data in elite handball players. Twenty-two elite male handball players (18.9 ± 0.2 years; body mass: 83.3 ± 1.1 kg, height: 1.79 ± 2.30 m, body fat: 12.8 ± 0.2%) completed tests that included a lower limb force-velocity test of peak power (Wpeak), jumping ability (squat and counter-movement jumps; SJ, CMJ), 1 repetition maximum (1-RM) half back squat, average sprint velocity over 5 m (V-5 m), peak velocity between 25-30 m (Vmax), and Yo-Yo Intermittent Recovery Test level 1 (Yo-Yo IR1), and anthropometric determinations of cross-sectional areas and muscle volumes for leg and thigh muscles. Data were compared with performance on 2 repeated sprint tests; the straight test [6 × 30 m (RSA30)] and the intermittent test [6 x (2 x 15 m) shuttle sprints (RSA15)]. V-5 m; 1-RM; and thigh and leg muscle cross-sectional areas and volumes showed relevant relationships to the shuttle RSA test (r2≥0.5). The Yo-Yo IR1 explained the largest percentage of variance in RSA15 total time (60%), and V-5 m also explained 56% of the variance in RSA15 total time. RSA15% fatigue was only related to 1-RM (explained variance: 58%), and the RSA30 test showed no useful correlations with any of the investigated predictors. Multiple-regression analyses using all physical tests explained 91% of the variance in RSA15 total time (p<0.001). The volume of the leg muscles seems important to power production and repeated sprints over distances of 15 m or less. RSA15 is related to muscular power, maximal strength, speed and endurance performance; however, the straight repeated sprint ability test is not, and thus does not seem relevant for the testing of handball players.

Measurement of nucleotide exchange rate constants in single rabbit soleus myofibrils during shortening and lengthening using a fluorescent ATP analog.

PubMed Central

Shirakawa, I; Chaen, S; Bagshaw, C R; Sugi, H

2000-01-01

The kinetics of displacement of a fluorescent nucleotide, 2'(3')-O-[N[2-[[Cy3]amido]ethyl]carbamoyl]-adenosine 5'-triphosphate (Cy3-EDA-ATP), bound to rabbit soleus muscle myofibrils were studied using flash photolysis of caged ATP. Use of myofibrils from this slow twitch muscle allowed better resolution of the kinetics of nucleotide exchange than previous studies with psoas muscle myofibrils (, Biophys. J. 73:2033-2042). Soleus myofibrils in the presence of Cy3-EDA-nucleotides (Cy3-EDA-ATP or Cy3-EDA-ADP) showed selective fluorescence staining of the A-band. The K(m) for Cy3-EDA-ATP and the K(d) for Cy3-EDA-ADP binding to the myofibril A-band were 1.9 microM and 3.8 microM, respectively, indicating stronger binding of nucleotide to soleus cross-bridges compared to psoas cross-bridges (2.6 microM and 50 microM, respectively). After flash photolysis of caged ATP, the A-band fluorescence of the myofibril in the Cy3-EDA-ATP solution under isometric conditions decayed exponentially with a rate constant of 0.045 +/- 0.007 s(-1) (n = 32) at 10 degrees C, which was about seven times slower than that for psoas myofibrils. When a myofibril was allowed to shorten with a constant velocity, the nucleotide displacement rate constant increased from 0.066 s(-1) (isometric) to 0.14 s(-1) at 20 degrees C with increasing shortening velocity up to 0.1 myofibril length/s (V(max), the shortening velocity under no load was approximately 0. 2 myofibril lengths/s). The rate constant was not significantly affected by an isovelocity stretch of up to 0.1 myofibril lengths/s. These results suggest that the cross-bridge kinetics are not significantly affected at higher strain during lengthening but depend on the lower strain during shortening. These data also indicate that the interaction distance between a cross-bridge and the actin filament is at least 16 nm for a single cycle of the ATPase. PMID:10653804

Purification and functional characterisation of the pyruvate (monocarboxylate) carrier from baker's yeast mitochondria (Saccharomyces cerevisiae).

PubMed

Nałecz, M J; Nałecz, K A; Azzi, A

1991-08-09

Isolated yeast mitochondria were subjected to solubilization by Triton X-114 and the detergent extract was subsequently chromatrographed on dry hydroxyapatite. Purification of the yeast monocarboxylate (pyruvate) carrier was achieved by affinity chromatography on immobilized 2-cyano-4-hydroxycinnamate, as described previously for bovine heart mitochondria (Bolli, R., Nałecz K.A. and Azzi, A. (1989) J. Biol. Chem. 264 18024-18030). The final preparation contained two polypeptides of apparent molecular mass 26 and 50 kDa. The yeast carrier appeared to be less abundant, but more active, than the analogous protein from higher eukaryotes. The carrier was able to catalyse the pyruvate / pyruvate and pyruvate / acetoacetate exchange reactions, both reactions being sensitive to cyanocinnamate and its derivatives, to phenylpyruvate and to mersalyl and p-chloromercuribenzoate. In the pyruvate / acetoacetate exchange reaction (200 mM internal acetoacetate, enzymatic assay), the Km value for external pyruvate was found to be 0.8 mM and the Vmax 135 mumol/min per mg protein. Among other substrates of the yeast carrier, all transported with similar affinity and identical maximal velocity against acetoacetate, we identified 2-oxoisocaproate, 2-oxoisovalerate and 2-oxo-3-methylvalerate. Lactate was not translocated by this carrier with a measurable rate, neither were di- or tricarboxylates.

Reaction kinetics and inhibition of adenosine kinase from Leishmania donovani.

PubMed

Bhaumik, D; Datta, A K

1988-04-01

The reaction kinetics and the inhibitor specificity of adenosine kinase (ATP:adenosine 5'-phosphotransferase, EC 2.7.1.20) from Leishmania donovani, have been analysed using homogeneous preparation of the enzyme. The reaction proceeds with equimolar stoichiometry of each reactant. Double reciprocal plots of initial velocity studies in the absence of products yielded intersecting lines for both adenosine and Mg2+-ATP. AMP is a competitive inhibitor of the enzyme with respect to adenosine and noncompetitive inhibitor with respect to ATP. In contrast, ADP was a noncompetitive inhibitor with respect to both adenosine and ATP, with inhibition by ADP becoming uncompetitive at very high concentration of ATP. Parallel equilibrium dialysis experiments against [3H]adenosine and [gamma-32P]ATP resulted in binding of adenosine to fre enzyme. Tubercidin (7-deazaadenosine) and 6-methyl-mercaptopurine riboside acted as substrates for the enzyme and were found to inhibit adenosine phosphorylation competitively in vitro. 'Substrate efficiency (Vmax/Km)' and 'turnover numbers (Kcat)' of the enzyme with respect to specific analogs were determined. Taken together the results suggest that (a) the kinetic mechanism of adenosine kinase is sequential Bi-Bi, (b) AMP and ADP may regulate enzyme activity in vivo and (c) tubercidin and 6-methylmercaptopurine riboside are monophosphorylated by the parasite enzyme.

Optimal immobilization of β-galactosidase onto κ-carrageenan gel beads using response surface methodology and its applications.

PubMed

Elnashar, Magdy M; Awad, Ghada E; Hassan, Mohamed E; Mohy Eldin, Mohamed S; Haroun, Bakry M; El-Diwany, Ahmed I

2014-01-01

β-Galactosidase (β-gal) was immobilized by covalent binding on novel κ-carrageenan gel beads activated by two-step method; the gel beads were soaked in polyethyleneimine followed by glutaraldehyde. 2(2) full-factorial central composite experiment designs were employed to optimize the conditions for the maximum enzyme loading efficiency. 11.443 U of enzyme/g gel beads was achieved by soaking 40 units of enzyme with the gel beads for eight hours. Immobilization process increased the pH from 4.5 to 5.5 and operational temperature from 50 to 55 °C compared to the free enzyme. The apparent K(m) after immobilization was 61.6 mM compared to 22.9 mM for free enzyme. Maximum velocity Vmax was 131.2 μ mol · min(-1) while it was 177.1 μ mol · min(-1) for free enzyme. The full conversion experiment showed that the immobilized enzyme form is active as that of the free enzyme as both of them reached their maximum 100% relative hydrolysis at 4 h. The reusability test proved the durability of the κ-carrageenan beads loaded with β -galactosidase for 20 cycles with retention of 60% of the immobilized enzyme activity to be more convenient for industrial uses.

Degradation kinetics of chlorpyrifos and 3,5,6-trichloro-2-pyridinol (TCP) by fungal communities.

PubMed

Maya, K; Upadhyay, S N; Singh, R S; Dubey, Suresh K

2012-12-01

Fungal isolates obtained from soil were used for degrading chlorpyrifos (CP) and TCP. The percentage degradation ranged from 69.4 to 89.8 for CP and 62.2 to 92.6 for TCP after one week. The values of K(s) and V(max) were different for different isolates. The K(s) ranged from 66.66 to 169.5mg/L and V(max) from 6.56 to 40.4 mg/L/d for CP and from 53.19 to 163.9 mg/L and 3.41 to 40.40 mg/L/d, respectively, for TCP. Fungal community showed high affinity for both CP and TCP. The genetic relatedness of isolate F1 to Aspergillus sp., F2 and F3 to Penicillium sp., F4 to Eurotium sp. and F5 to Emericella sp. were confirmed. The degradation potential was in the order: F1>F2=F3>F4>F5. Copyright © 2012 Elsevier Ltd. All rights reserved.

Explicit solution of integrated 1 - exp equation for predicting accumulation and decline of concentrations for drugs obeying nonlinear saturation kinetics.

PubMed

Keller, Frieder; Hartmann, Bertram; Czock, David

2009-12-01

To describe nonlinear, saturable pharmacokinetics, the Michaelis-Menten equation is frequently used. However, the Michaelis-Menten equation has no integrated solution for concentrations but only for the time factor. Application of the Lambert W function was proposed recently to obtain an integrated solution of the Michaelis-Menten equation. As an alternative to the Michaelis-Menten equation, a 1 - exp equation has been used to describe saturable kinetics, with the advantage that the integrated 1 - exp equation has an explicit solution for concentrations. We used the integrated 1 - exp equation to predict the accumulation kinetics and the nonlinear concentration decline for a proposed fictive drug. In agreement with the recently proposed method, we found that for the integrated 1 - exp equation no steady state is obtained if the maximum rate of change in concentrations (Vmax) within interval (Tau) is less than the difference between peak and trough concentrations (Vmax x Tau < C peak - C trough).

Beyond Vmax and Km: How details of enzyme function influence geochemical cycles

NASA Astrophysics Data System (ADS)

Steen, A. D.

2015-12-01

Enzymes catalyze the vast majority of chemical reactions relevant to geomicrobiology. Studies of the activities of enzymes in environmental systems often report Vmax (the maximum possible rate of reaction; often proportional to the concentration of enzymes in the system) and sometimes Km (a measure of the affinity between enzymes and their substrates). However, enzyme studies - particularly those related to enzymes involved in organic carbon oxidation - are often limited to only those parameters, and a relatively limited and mixed set of enzymes. Here I will discuss some novel methods to assay and characterize the specific sets of enzymes that may be important to the carbon cycle in aquatic environments. First, kinetic experiments revealed the collective properties of the complex mixtures of extracellular peptidases that occur where microbial communities are diverse. Crystal structures combined with biochemical characterization of specific enzymes can yield more detailed information about key steps in organic carbon transformations. These new techniques have the potential to provide mechanistic grounding to geomicrobiological models.

Influence of lifestyle habits, nutritional status and insulin resistance in NAFLD.

PubMed

Malavolti, Marcella; Battistini, Nino Carlo; Miglioli, Lucia; Bagni, Ilaria; Borelli, Luca; Marino, Mariano; Scaglioni, Federica; Bellentani, Stefano

2012-01-01

Non alcoholic fatty liver disease (NAFLD) is associated with obesity, diabetes and insulin resistance (IR). The aim of our study was to assess the relationship between IR, anthropometry, lifestyle habits, resting energy expenditure (REE) and degree of fatty liver at ultrasound in 48 overweight patients with NAFLD as compared to 24 controls without fatty liver, matched for age. Nutritional status, alcohol intake and physical activity were assessed by skinfold thickness measurements, a 7-day diary, and SenseWear armband (SWA). REE was assessed by both SWA (REE-SWA) and a Vmax metabolic cart (REE-Vmax). Fatty liver was measured by US and the Doppler Power Index was calculated. IR was assessed using the HOMA index. There was significant correlation between waist circumference, HOMA, Doppler power index and fatty liver grade at US. Multivariate analysis showed that alteration of waist circumference, Doppler power index, and HOMA were the major significant predictors of fatty liver. Our data demonstrated a significant association between NAFLD and central adiposity and IR.

Effect of swim exercise training on human muscle fiber function

NASA Technical Reports Server (NTRS)

Fitts, R. H.; Costill, D. L.; Gardetto, P. R.

1989-01-01

The effect of swim exercise training on the human muscle fiber function was investigated in swimmers trained in a typical collegiate swim-training program followed by an intensified 10-day training period. The measured parameters included the peak tension (P0), negative log molar Ca(2+) concentration (pCa)-force, and maximal shortening speed (Vmax) of the slow-twitch type I and fast-twitch type II fibers obtained by biopsy from the deltoid muscle. The P0 values were found to be not altered after either the training or the 10-day intensive program. The type I fibers from the trained swimmers showed pCa-force curves shifted to the right, such that higher free Ca(2+) levels were required to elicit a given percent of P0. The training program significantly increased the Vmax in the type I fibers and decreased that of the type II fibers, and the 10-day intensive training produced a further significant decrease of the type II fibers.

Enzyme Activity Experiments Using a Simple Spectrophotometer

ERIC Educational Resources Information Center

Hurlbut, Jeffrey A.; And Others

1977-01-01

Experimental procedures for studying enzyme activity using a Spectronic 20 spectrophotometer are described. The experiments demonstrate the effect of pH, temperature, and inhibitors on enzyme activity and allow the determination of Km, Vmax, and Kcat. These procedures are designed for teaching large lower-level biochemistry classes. (MR)

Functional characterisation of a tropine-forming reductase gene from Brugmansia arborea, a woody plant species producing tropane alkaloids.

PubMed

Qiang, Wei; Xia, Ke; Zhang, Qiaozhuo; Zeng, Junlan; Huang, Yuanshe; Yang, Chunxian; Chen, Min; Liu, Xiaoqiang; Lan, Xiaozhong; Liao, Zhihua

2016-07-01

Brugmansia arborea is a woody plant species that produces tropane alkaloids (TAs). The gene encoding tropine-forming reductase or tropinone reductase I (BaTRI) in this plant species was functionally characterised. The full-length cDNA of BaTRI encoded a 272-amino-acid polypeptide that was highly similar to tropinone reductase I from TAs-producing herbal plant species. The purified 29kDa recombinant BaTRI exhibited maximum reduction activity at pH 6.8-8.0 when tropinone was used as substrate; it also exhibited maximum oxidation activity at pH 9.6 when tropine was used as substrate. The Km, Vmax and Kcat values of BaTRI for tropinone were 2.65mM, 88.3nkatmg(-1) and 2.93S(-1), respectively, at pH 6.4; the Km, Vmax and Kcat values of TRI from Datura stramonium (DsTRI) for tropinone were respectively 4.18mM, 81.20nkatmg(-1) and 2.40S(-1) at pH 6.4. At pH 6.4, 6.8 and 7.0, BaTRI had a significantly higher activity than DsTRI. Analogues of tropinone, 4-methylcyclohexanone and 3-quinuclidinone hydrochloride, were also used to investigate the enzymatic kinetics of BaTRI. The Km, Vmax and Kcat values of BaTRI for tropine were 0.56mM, 171.62nkat.mg(-1) and 5.69S(-1), respectively, at pH 9.6; the Km, Vmax and Kcat values of DsTRI for tropine were 0.34mM, 111.90nkatmg(-1) and 3.30S(-1), respectively, at pH 9.6. The tissue profiles of BaTRI differed from those in TAs-producing herbal plant species. BaTRI was expressed in all examined organs but was most abundant in secondary roots. Finally, tropane alkaloids, including hyoscyamine, anisodamine and scopolamine, were detected in various organs of B. arborea by HPLC. Interestingly, scopolamine constituted most of the tropane alkaloids content in B. arborea, which suggests that B. arborea is a scopolamine-rich plant species. The scopolamine content was much higher in the leaves and stems than in other organs. The gene expression and TAs accumulation suggest that the biosynthesis of hyoscyamine, especially scopolamine, occurred not only in the roots but also in the aerial parts of B. arborea. Copyright © 2016 Elsevier Ltd. All rights reserved.

Study of Flapping Flight Using Discrete Vortex Method Based Simulations

NASA Astrophysics Data System (ADS)

Devranjan, S.; Jalikop, Shreyas V.; Sreenivas, K. R.

2013-12-01

In recent times, research in the area of flapping flight has attracted renewed interest with an endeavor to use this mechanism in Micro Air vehicles (MAVs). For a sustained and high-endurance flight, having larger payload carrying capacity we need to identify a simple and efficient flapping-kinematics. In this paper, we have used flow visualizations and Discrete Vortex Method (DVM) based simulations for the study of flapping flight. Our results highlight that simple flapping kinematics with down-stroke period (tD) shorter than the upstroke period (tU) would produce a sustained lift. We have identified optimal asymmetry ratio (Ar = tD/tU), for which flapping-wings will produce maximum lift and find that introducing optimal wing flexibility will further enhances the lift.

Detoxification of Acetylcholinesterase Inhibitors.

DTIC Science & Technology

1987-02-19

IIB:HB-o Mvse three machanisms can be distinguished by appropriate labelling experiments in oxygen-18 H.O. If mechanism C is operating, hydrolysis...enzyue reveals that there is a single break at pa 6.2 relative to both Vmax and Vfax/Km. This would appear to represent the titration of the basic residue

Satellite retrievals of leaf chlorophyll and photosynthetic capacity for improved modeling of GPP

USDA-ARS?s Scientific Manuscript database

This study investigates the utility of in-situ and satellite-based leaf chlorophyll (Chl) estimates for quantifying leaf photosynthetic capacity and for constraining model simulations of Gross Primary Productivity (GPP) over a corn field in Maryland, U.S.A. The maximum rate of carboxylation (Vmax) r...

Left Atrial Mechanical Functions in Professional Soccer Players: A Pilot Study

ERIC Educational Resources Information Center

Kartal, Alper; Güngör, Hasan; Kartal, Resat; Ergin, Esin

2017-01-01

Long-term regular exercise is associated with physiologic and morphologic alterations in the heart chambers. The aim of this study to evaluate left atrium (LA) phasic functions in professional football players and compare with control subjects. Left atrial volume was calculated at end-systole (Vmax), end-diastole and pre-atrial contraction by…

Inhibition, by 2-oxo acids that accumulate in maple-syrup-urine disease, of lactate, pyruvate, and 3-hydroxybutyrate transport across the blood-brain barrier.

PubMed

Cremer, J E; Teal, H M; Cunningham, V J

1982-09-01

Data are presented in support of the transport of (-)-D-3-hydroxybutyrate across the blood-brain barrier (BBB) being a carrier-mediated process. The kinetic parameters in 21-day-old pentobarbital-anaesthetized rats were Vmax 2.0 mumol.g-1.min-1, Km 29 mM, and KD 0.024 ml.g-1.min-1. The value for Vmax was the same as that for L-lactate and pyruvate transport in animals of the same age. The transport of all three substrates was sensitive to inhibition by low concentrations of either 2-oxo-3-methylbutanoate or 2-oxo-4-methylpentanoate, the 2-oxo acids that can accumulate in patients with maple-syrup-urine disease. The Ki values for the 2-oxo acids were severalfold lower than the respective Km values. 2-Oxo-3-phenylpropionate was a poor inhibitor. The relative affinities of the various monocarboxylic acids for the transport system of the BBB distinguished it from similar systems described in brain, heart, and liver mitochondria; human erythrocytes; and Ehrlich ascites-tumour cells.

Influence of carbon source and inoculum type on anaerobic biomass adhesion on polyurethane foam in reactors fed with acid mine drainage.

PubMed

Rodriguez, Renata P; Zaiat, Marcelo

2011-04-01

This paper analyzes the influence of carbon source and inoculum origin on the dynamics of biomass adhesion to an inert support in anaerobic reactors fed with acid mine drainage. Formic acid, lactic acid and ethanol were used as carbon sources. Two different inocula were evaluated: one taken from an UASB reactor and other from the sediment of a uranium mine. The values of average colonization rates and the maximum biomass concentration (C(max)) were inversely proportional to the number of carbon atoms in each substrate. The highest C(max) value (0.35 g TVS g(-1) foam) was observed with formic acid and anaerobic sludge as inoculum. Maximum colonization rates (v(max)) were strongly influenced by the type of inoculum when ethanol and lactic acid were used. For both carbon sources, the use of mine sediment as inoculum resulted in a v(max) of 0.013 g TVS g(-1) foam day(-1), whereas 0.024 g TVS g(-1) foam day(-1) was achieved with anaerobic sludge. Copyright © 2011 Elsevier Ltd. All rights reserved.

Phosphate-dependent glutaminase in enterocyte mitochondria and its regulation by ammonium and other ions.

PubMed

Masola, B; Zvinavashe, E

2003-06-01

The effects of ammonium and other ions on phosphate dependent glutaminase (PDG) activity in intact rat enterocyte mitochondria were investigated. Sulphate and bicarbonate activated the enzyme in absence and presence of added phosphate. In presence of 10 mM phosphate, ammonium at concentrations <1 mM inhibited the enzyme. This inhibition was reversed by increased concentration of phosphate or sulphate. The inhibition of PDG by ammonium in presence of 10 mM phosphate was biphasic with respect to glutamine concentration, its effect being through a lowering of V(max) at glutamine concentration of

Observation of the wing deformation and the CFD study of cicada

NASA Astrophysics Data System (ADS)

Dai, Hu; Mohd Adam Das, Shahrizan; Luo, Haoxiang

2011-11-01

We studied the wing properties and kinematics of cicada when the 13-year species emerged in amazingly large numbers in middle Tennessee during May 2011. Using a high-speed camera, we recorded the wing motion of the insect and then reconstructed the three-dimensional wing kinematics using a video digitization software. Like many other insects, the deformation of the cicada wing is asymmetric between the downstroke and upstroke half cycles, and this particular deformation pattern would benefit production of the lift and propulsive forces. Both two-dimensional and three-dimensional CFD studies are carried out based on the reconstructed wing motion. The implication of the study on the role of the aerodynamic force in the wing deformation will be discussed. This work is sponsored by the NSF.

Asymptomatic Wolff–Parkinson–White syndrome: incidental ECG diagnosis and a review of literature regarding current treatment

PubMed Central

Liu, Alexander; Pusalkar, Pawan

2011-01-01

A 19-year-old male presented with periorbital cellulitis responsive to intravenous antibiotics. A routine ECG on admission showed slurred upstroke of the QRS complexes consistent with Wolff–Parkinson–White syndrome (WPW). He has never experienced any cardiac-related symptoms. Once the periorbital cellulitis resolved, he was referred to the specialist cardiology ablation clinic. He was counselled on the arrythmogenic risks of untreated WPW and the potential complications of radiofrequency catheter ablation (RFCA). He decided to go ahead with electrophysiological studies and RFCA, which took place successfully. This case highlights the importance of routine ECG in the diagnosis of asymptomatic WPW. The use of prophylactic ablation of asymptomatic WPW is controversial and should be considered on a case-specific basis. PMID:22693197

The Core of Allosteric Motion in Thermus caldophilus l-Lactate Dehydrogenase*

PubMed Central

Ikehara, Yoko; Arai, Kazuhito; Furukawa, Nayuta; Ohno, Tadashi; Miyake, Tatsuya; Fushinobu, Shinya; Nakajima, Masahiro; Miyanaga, Akimasa; Taguchi, Hayao

2014-01-01

For Thermus caldophilus l-lactate dehydrogenase (TcLDH), fructose 1,6-bisphosphate (FBP) reduced the pyruvate S0.5 value 103-fold and increased the Vmax value 4-fold at 30 °C and pH 7.0, indicating that TcLDH has a much more T state-sided allosteric equilibrium than Thermus thermophilus l-lactate dehydrogenase, which has only two amino acid replacements, A154G and H179Y. The inactive (T) and active (R) state structures of TcLDH were determined at 1.8 and 2.0 Å resolution, respectively. The structures indicated that two mobile regions, MR1 (positions 172–185) and MR2 (positions 211–221), form a compact core for allosteric motion, and His179 of MR1 forms constitutive hydrogen bonds with MR2. The Q4(R) mutation, which comprises the L67E, H68D, E178K, and A235R replacements, increased Vmax 4-fold but reduced pyruvate S0.5 only 5-fold in the reaction without FBP. In contrast, the P2 mutation, comprising the R173Q and R216L replacements, did not markedly increase Vmax, but 102-reduced pyruvate S0.5, and additively increased the FBP-independent activity of the Q4(R) enzyme. The two types of mutation consistently increased the thermal stability of the enzyme. The MR1-MR2 area is a positively charged cluster, and its center approaches another positively charged cluster (N domain cluster) across the Q-axis subunit interface by 5 Å, when the enzyme undergoes the T to R transition. Structural and kinetic analyses thus revealed the simple and unique allosteric machinery of TcLDH, where the MR1-MR2 area pivotally moves during the allosteric motion and mediates the allosteric equilibrium through electrostatic repulsion within the protein molecule. PMID:25258319

Temporal and spatial variations in kinetics of alkaline phosphatase in sediments of a shallow Chinese eutrophic lake (Lake Donghu).

PubMed

Yiyong, Zhou; Jianqiu, Li; Min, Zhang

2002-04-01

Monthly sediment and interstitial water samples were collected in a shallow Chinese freshwater lake (Lake Donghu) from three areas to determine if alkaline phosphatase activity (APA) plays an important role, in phosphorus cycling in sediment. The seasonal variability in the kinetics of APA and other relevant parameters were investigated from 1995-1996. The phosphatase hydrolyzable phosphorus (PHP) fluctuated seasonally in interstitial water, peaking in the spring. A synchronous pattern was observed in chlorophyll a contents in surface water in general. The orthophosphate (o-P) concentrations in the interstitial water increased during the spring. An expected negative relationship between PHP and Vmax of APA is not evident in interstitial water. The most striking feature of the two variables is their co-occurring, which can be explained in terms of an induction mechanism. It is argued that phosphatase activity mainly contributes to the driving force of o-P regeneration from PHP in interstitial water, supporting the development of phytoplankton biomass in spring. The Vmax values in sediment increased during the summer, in conjunction with lower Km values in interstitial water that suggest a higher affinity for the substrate. The accumulation of organic matter in the sediment could be traced back to the breakdown of the algal spring bloom, which may stimulate APA with higher kinetic efficiency, by a combination of the higher Vmax in sediments plus lower Km values in interstitial water, in summer. In summary, a focus on phosphatase and its substrate in annual scale may provide a useful framework for the development of novel P cycling, possible explanations for the absence of a clear relationship between PHP and APA were PHP released from the sediment which induced APA, and the presence of kinetically higher APA both in sediment and interstitial water which permitted summer mineralization of organic matter derived from the spring bloom to occur. The study highlighted the need for distinguishing functionally distinct extracellular enzymes between the sediment and interstitial water of lakes.

Kinetics of Ethylene and Ethylene Oxide in Subcellular Fractions of Lungs and Livers of Male B6C3F1 Mice and Male Fischer 344 Rats and of Human Livers

PubMed Central

Csanády, György András; Kessler, Winfried; Klein, Dominik; Pankratz, Helmut; Pütz, Christian; Richter, Nadine; Filser, Johannes Georg

2011-01-01

Ethylene (ET) is metabolized in mammals to the carcinogenic ethylene oxide (EO). Although both gases are of high industrial relevance, only limited data exist on the toxicokinetics of ET in mice and of EO in humans. Metabolism of ET is related to cytochrome P450-dependent mono-oxygenase (CYP) and of EO to epoxide hydrolase (EH) and glutathione S-transferase (GST). Kinetics of ET metabolism to EO and of elimination of EO were investigated in headspace vessels containing incubations of subcellular fractions of mouse, rat, or human liver or of mouse or rat lung. CYP-associated metabolism of ET and GST-related metabolism of EO were found in microsomes and cytosol, respectively, of each species. EH-related metabolism of EO was not detectable in hepatic microsomes of rats and mice but obeyed saturation kinetics in hepatic microsomes of humans. In ET-exposed liver microsomes, metabolism of ET to EO followed Michaelis-Menten-like kinetics. Mean values of Vmax [nmol/(min·mg protein)] and of the apparent Michaelis constant (Km [mmol/l ET in microsomal suspension]) were 0.567 and 0.0093 (mouse), 0.401 and 0.031 (rat), and 0.219 and 0.013 (human). In lung microsomes, Vmax values were 0.073 (mouse) and 0.055 (rat). During ET exposure, the rate of EO production decreased rapidly. By modeling a suicide inhibition mechanism, rate constants for CYP-mediated catalysis and CYP inactivation were estimated. In liver cytosol, mean GST activities to EO expressed as Vmax/Km [μl/(min·mg protein)] were 27.90 (mouse), 5.30 (rat), and 1.14 (human). The parameters are most relevant for reducing uncertainties in the risk assessment of ET and EO. PMID:21785163

Kinetics and spatial distribution of enzymes of carbon, nitrogen and phosphorus cycles in earthworm biopores

NASA Astrophysics Data System (ADS)

Hoang Thi Thu, Duyen; Razavi, Bahar S.

2016-04-01

Earthworms boost microbial activities and consequently form hotspots in soil. The distribution of enzyme activities inside the earthworm biopores is completely unknown. For the first time, we analyzed enzyme kinetics and visualized enzyme distribution inside and outside biopores by in situ soil zymography. Kinetic parameters (Vmax and Km) of 6 enzymes β-glucosidase (GLU), cellobiohydrolase (CBH), xylanase (XYL), chitinase (NAG), leucine aminopeptidase (LAP) and acid phosphatase (APT) were determined in biopores formed by Lumbricus terrestris L.. The spatial distributions of GLU, NAG and APT become visible via zymograms in comparison between earthworm-inhabited and earthworm-free soil. Zymography showed heterogeneous distribution of hotspots in the rhizosphere and biopores. The hotspot areas were 2.4 to 14 times larger in the biopores than in soil without earthworms. The significantly higher Vmax values for GLU, CBH, XYL, NAG and APT in biopores confirmed the stimulation of enzyme activities by earthworms. For CBH, XYL and NAG, the 2- to 3-fold higher Km values in biopores indicated different enzyme systems with lower substrate affinity compared to control soil. The positive effects of earthworms on Vmax were cancelled by the Km increase for CBH, XYL and NAG at a substrate concentration below 20 μmol g-1 soil. The change of enzyme systems reflected a shift in dominant microbial populations toward species with lower affinity to holo-celluloses and to N-acetylglucosamine, and with higher affinity to proteins as compared to the biopores-free soil. We conclude that earthworm biopores are microbial hotspots with much higher and dense distribution of enzyme activities compared to bulk soil. References Spohn M, Kuzyakov Y. (2014) Spatial and temporal dynamics of hotspots of enzyme activity in soil as affected by living and dead roots - a soil zymography analysis, Plant Soil 379: 67-77. Blagodatskaya, E., Kuzyakov, Y., 2013. Review paper: Active microorganisms in soil: Critical review of estimation criteria and approaches. Soil Biology & Biochemistry 67, 192-211.

Effect of salt acclimatization on 3 beta-hydroxysteroid dehydrogenase/isomerase activity in the interrenal of Bufo arenarum.

PubMed

Pozzi, Andrea G; Lantos, Carlos P; Ceballos, Nora R

2002-03-01

In amphibians, aldosterone (Aldo) is particularly important in the regulation of Na(+) exchange by skin and urinary bladder. In previous works we studied a key enzyme in Aldo biosynthesis, the 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta HSD/I), in the interrenals of Bufo arenarum. In those works a dual localization of the 3 beta HSD/I in both microsomes and mitochondria was described. The mitochondrial, but not the microsomal, enzyme prefers the immediate Aldo precursor, 3 beta-analogue of aldosterone, as substrate. In this order, the enzyme 3 beta HSD/I would be not only a key enzyme for the synthesis of Aldo but additionally, due to its microsomal and mitochondrial localization, a possible target for the regulation of Aldo biosynthesis. With this rationale in mind, we have used in vivo and in vitro approaches to study Aldo regulation. In the present investigation the levels of Aldo were determined in plasma of winter (W) and summer (S) toads subjected to different saline concentrations (0.125 and 0.15 M) or kept on wet land. Saline hyperosmotically treated toads had significantly lower levels than isoosmotically treated toads. These results are consistent with the response in mammals, in which salt loading provokes a reduction in Aldo secretion. In W toads, plasmatic corticosterone (B) concentration was higher than Aldo concentration, whereas in S toads, B/Aldo ratio approached unity. The reduction of Aldo levels after saline dehydration was due to a decline in its biosynthesis. K(m) and V(max) values for 3 beta HSD/I were calculated for mitochondrial and microsomal fractions obtained from animals acclimated to 0.15 M NaCl or kept on land. As previously described, V(max) differs between W and S toads. However, only mitochondrial V(max) changed as a consequence of saline adaptation, suggesting that the mitochondrial enzyme could be involved in the regulation of Aldo biosynthesis.

GAMA/H-ATLAS: The Local Dust Mass Function and Cosmic Density as a Function of Galaxy Type - A Benchmark for Models of Galaxy Evolution

NASA Astrophysics Data System (ADS)

Beeston, R. A.; Wright, A. H.; Maddox, S.; Gomez, H. L.; Dunne, L.; Driver, S. P.; Robotham, A.; Clark, C. J. R.; Vinsen, K.; Takeuchi, T. T.; Popping, G.; Bourne, N.; Bremer, M. N.; Phillipps, S.; Moffett, A. J.; Baes, M.; Bland-Hawthorn, J.; Brough, S.; De Vis, P.; Eales, S. A.; Holwerda, B. W.; Loveday, J.; Liske, J.; Smith, M. W. L.; Smith, D. J. B.; Valiante, E.; Vlahakis, C.; Wang, L.

2018-06-01

We present the dust mass function (DMF) of 15,750 galaxies with redshift z < 0.1, drawn from the overlapping area of the GAMA and H-ATLAS surveys. The DMF is derived using the density corrected Vmax method, where we estimate Vmax using: (i) the normal photometric selection limit (pVmax) and (ii) a bivariate brightness distribution (BBD) technique, which accounts for two selection effects. We fit the data with a Schechter function, and find M^{*}=(4.65 ± 0.18)× 107 h^2_{70} M_{⊙ }, α = ( - 1.22 ± 0.01), φ ^{*}=(6.26 ± 0.28)× 10^{-3} h^3_{70} Mpc^{-3} dex^{-1}. The resulting dust mass density parameter integrated down to 104 M⊙ is Ωd = (1.11 ± 0.02) × 10-6 which implies the mass fraction of baryons in dust is f_{m_b}=(2.40± 0.04)× 10^{-5}; cosmic variance adds an extra 7-17 per cent uncertainty to the quoted statistical errors. Our measurements have fewer galaxies with high dust mass than predicted by semi-analytic models. This is because the models include too much dust in high stellar mass galaxies. Conversely, our measurements find more galaxies with high dust mass than predicted by hydrodynamical cosmological simulations. This is likely to be from the long timescales for grain growth assumed in the models. We calculate DMFs split by galaxy type and find dust mass densities of Ωd = (0.88 ± 0.03) × 10-6 and Ωd = (0.060 ± 0.005) × 10-6 for late-types and early-types respectively. Comparing to the equivalent galaxy stellar mass functions (GSMF) we find that the DMF for late-types is well matched by the GMSF scaled by (8.07 ± 0.35) × 10-4.

Biosynthesis of agmatine in isolated mitochondria and perfused rat liver: studies with 15N-labelled arginine

PubMed Central

2005-01-01

An important but unresolved question is whether mammalian mitochondria metabolize arginine to agmatine by the ADC (arginine decarboxylase) reaction. 15N-labelled arginine was used as a precursor to address this question and to determine the flux through the ADC reaction in isolated mitochondria obtained from rat liver. In addition, liver perfusion system was used to examine a possible action of insulin, glucagon or cAMP on a flux through the ADC reaction. In mitochondria and liver perfusion, 15N-labelled agmatine was generated from external 15N-labelled arginine. The production of 15N-labelled agmatine was time- and dose-dependent. The time-course of [U-15N4]agmatine formation from 2 mM [U-15N4]arginine was best fitted to a one-phase exponential curve with a production rate of approx. 29 pmol·min−1·(mg of protein)−1. Experiments with an increasing concentration (0– 40 mM) of [guanidino-15N2]arginine showed a Michaelis constant Km for arginine of 46 mM and a Vmax of 3.7 nmol·min−1·(mg of protein)−1 for flux through the ADC reaction. Experiments with broken mitochondria showed little changes in Vmax or Km values, suggesting that mitochondrial arginine uptake had little effect on the observed Vmax or Km values. Experiments with liver perfusion demonstrated that over 95% of the effluent agmatine was derived from perfusate [guanidino-15N2]arginine regardless of the experimental condition. However, the output of 15N-labelled agmatine (nmol·min−1·g−1) increased by approx. 2-fold (P<0.05) in perfusions with cAMP. The findings of the present study provide compelling evidence that mitochondrial ADC is present in the rat liver, and suggest that cAMP may stimulate flux through this pathway. PMID:15656789

A study of the temporal effect of alcohol on human erythrocyte sodium-lithium countertransport in relation to membrane cholesterol and phospholipids.

PubMed

Adebayo, G I; Gaffney, P; Feely, J

1996-01-01

The effect of a single dose of alcohol (0.8 g/kg), given with "diet coke," on erythrocyte sodium-lithium countertransport (SLC) in relation to membrane cholesterol and phospholipids was assessed over 24 h in 10 healthy volunteers. Baseline passive lithium efflux (0.168 +/- 0.008 mmol l-1 Cell H-1) was increased 1 h (0.202 +/- 0.014 mmol l-1 cell h-1; p < 0.030), and 4 h (0.200 +/- 0.014 mmol l-1 cell h-1; p < 0.020), but similar to that at 24 h postalcohol (0.173 +/- 0.011 mmol l-1 cell h-1). These changes were not associated with any change in intracellular lithium. Control SLC VMAX of 0.387 +/- 0.054 mmol l-1 cell h-1 fell at 1 h (0.328 +/- 0.050 mmol l-1 cell h-1; p = 0.0012) and 4 h (0.312 +/- 0.048 mmol l-1 cell h-1; p < 0.0005). Its value 24 h postalcohol (0.371 +/- 0.047 mmol l-1 cell h-1) was comparable to that at baseline. There was no significant change in the affinity of the transporter for external sodium throughout the experimental period, suggesting that the reduction in VMAX 1 and 4 h after alcohol ingestion resulted from a noncompetitive inhibition. Intracellular sodium 4 h after alcohol was lower than at baseline, but returned to the control value within 24 h. In a control group (n = 5), pretreatment with "diet coke" alone did not alter any of the measured parameters. It is concluded that alcohol pretreatment increases passive lithium efflux and decreases SLC Vmax. Both effects are evident up to at least 4 h postdosing, but recover within 24 h in the absence of further alcohol intake.

Process optimization for production and purification of a thermostable, organic solvent tolerant lipase from Acinetobacter sp. AU07.

PubMed

Gururaj, P; Ramalingam, Subramanian; Nandhini Devi, Ganesan; Gautam, Pennathur

2016-01-01

The purpose of this study was to isolate, purify and optimize the production conditions of an organic solvent tolerant and thermostable lipase from Acinetobacter sp. AU07 isolated from distillery waste. The lipase production was optimized by response surface methodology, and a maximum production of 14.5U/mL was observed at 30°C and pH 7, using a 0.5% (v/v) inoculum, 2% (v/v) castor oil (inducer), and agitation 150rpm. The optimized conditions from the shake flask experiments were validated in a 3L lab scale bioreactor, and the lipase production increased to 48U/mL. The enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography and the overall yield was 36%. SDS-PAGE indicated a molecular weight of 45kDa for the purified protein, and Matrix assisted laser desorption/ionization time of flight analysis of the purified lipase showed sequence similarity with GDSL family of lipases. The optimum temperature and pH for activity of the enzyme was found to be 50°C and 8.0, respectively. The lipase was completely inhibited by phenylmethylsulfonyl fluoride but minimal inhibition was observed when incubated with ethylenediaminetetraacetic acid and dithiothreitol. The enzyme was stable in the presence of non-polar hydrophobic solvents. Detergents like SDS inhibited enzyme activity; however, there was minimal loss of enzyme activity when incubated with hydrogen peroxide, Tween 80 and Triton X-100. The kinetic constants (Km and Vmax) revealed that the hydrolytic activity of the lipase was specific to moderate chain fatty acid esters. The Vmax, Km and Vmax/Km ratio of the enzyme were 16.98U/mg, 0.51mM, and 33.29, respectively when 4-nitrophenyl palmitate was used as a substrate. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Metabolism of endosulfan-alpha by human liver microsomes and its utility as a simultaneous in vitro probe for CYP2B6 and CYP3A4.

PubMed

Casabar, Richard C T; Wallace, Andrew D; Hodgson, Ernest; Rose, Randy L

2006-10-01

Endosulfan-alpha is metabolized to a single metabolite, endosulfan sulfate, in pooled human liver microsomes (Km = 9.8 microM, Vmax = 178.5 pmol/mg/min). With the use of recombinant cytochrome P450 (P450) isoforms, we identified CYP2B6 (Km = 16.2 microM, Vmax = 11.4 nmol/nmol P450/min) and CYP3A4 (Km = 14.4 microM, Vmax = 1.3 nmol/nmol P450/min) as the primary enzymes catalyzing the metabolism of endosulfan-alpha, although CYP2B6 had an 8-fold higher intrinsic clearance rate (CL(int) = 0.70 microl/min/pmol P450) than CYP3A4 (CL(int) = 0.09 microl/min/pmol P450). Using 16 individual human liver microsomes (HLMs), a strong correlation was observed with endosulfan sulfate formation and S-mephenytoin N-demethylase activity of CYP2B6 (r(2) = 0.79), whereas a moderate correlation with testosterone 6 beta-hydroxylase activity of CYP3A4 (r(2) = 0.54) was observed. Ticlopidine (5 microM), a potent CYP2B6 inhibitor, and ketoconazole (10 microM), a selective CYP3A4 inhibitor, together inhibited approximately 90% of endosulfan-alpha metabolism in HLMs. Using six HLM samples, the percentage total normalized rate (% TNR) was calculated to estimate the contribution of each P450 in the total metabolism of endosulfan-alpha. In five of the six HLMs used, the percentage inhibition with ticlopidine and ketoconazole in the same incubation correlated with the combined % TNRs for CYP2B6 and CYP3A4. This study shows that endosulfan-alpha is metabolized by HLMs to a single metabolite, endosulfan sulfate, and that it has potential use, in combination with inhibitors, as an in vitro probe for CYP2B6 and 3A4 catalytic activities.

Demethylation of the pesticide methoxychlor in liver and intestine from untreated, methoxychlor-treated, and 3-methylcholanthrene-treated channel catfish (Ictalurus punctatus): evidence for roles of CYP1 and CYP3A family isozymes.

PubMed

Stuchal, Leah D; Kleinow, Kevin M; Stegeman, John J; James, Margaret O

2006-06-01

Exposure to the organochlorine pesticide methoxychlor (MXC) is associated with endocrine disruption in several species through biotransformation to mono-desmethyl-MXC (OH-MXC) and bis-desmethyl-MXC (HPTE), which interact with estrogen receptors. The biotransformation of [14C]methoxychlor was examined in channel catfish (Ictalurus punctatus), a freshwater species found in the southern United States. Hepatic microsomes formed OH-MXC and HPTE, assessed by comigration with authentic standards. The Km for OH-MXC formation by control liver microsomes was 3.8 +/- 1.3 microM (mean +/- S.D., n = 4), and Vmax was 131 +/- 53 pmol/min/mg protein. These values were similar to those of catfish pretreated with 2 mg/kg methoxychlor i.p. for 6 days (Km 3.3 +/- 0.8 microM and Vmax 99 +/- 17 pmol/min/mg) but less (p < 0.05) than the kinetic parameters for catfish treated with 3-methylcholanthrene (3-MC), which had Km of 6.0 +/- 1.1 microM and Vmax of 246 +/- 6 pmol/min/mg protein. Liver microsomes from 3-MC-treated fish produced significantly more of the secondary metabolite and more potent estrogen, HPTE. Intestinal microsomes formed OH-MXC at lower rates than liver. Methoxychlor pretreatment significantly reduced intestinal metabolite formation from 32 +/- 4 to 15 +/- 6 pmol/min/mg (mean +/- S.D., n = 4), whereas 3-MC treatment significantly increased OH-MXC production to 72 +/- 22 pmol/min/mg. Ketoconazole, clotrimazole, and alpha-naphthoflavone all decreased the production of OH-MXC in liver microsomes, whereas alpha-naphthoflavone stimulated HPTE formation, suggesting that CYP1 and CYP3 family isozymes demethylated methoxychlor. The results suggest that the formation of estrogenic metabolites from methoxychlor would be more rapid in catfish coexposed to CYP1 inducers.

Inhibition of Sunn pest, Eurygaster integriceps, α-amylases by α-amylase inhibitors (T-αAI) from Triticale.

PubMed

Mehrabadi, Mohammad; Bandani, Ali R; Saadati, Fatemeh

2010-01-01

The effect of triticale α-amylases inhibitors on starch hydrolysis catalyzed by the Sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae) midgut amylases was examined. Biochemical studgawies showed that inhibitors from Triticale (a hybrid of wheat and rye) had inhibitiory effects on E. integriceps α-amylases. The effects of the triticale α-amylase inhibitor (T-αAI) on α-amylase of E. integriceps showed a dose dependent manner of inhibition, e.g. less inhibition of enzyme activity (around 10%) with a lower dose (0.25 mg protein) and high inhibition of enzyme activity (around 80%) when a high dose of inhibitor was used (1.5 mg protein). The enzyme kinetic studies using Michaelis-Menten and Lineweaver-Burk equations showed the K(m) remained constant (0.58%) but the maximum velocity (V(max)) decreased in the presence of a crude extract of Triticale inhibitors, indicating mixed inhibition. The temperature giving 50% inactivation of enzyme (T(50)) during a 30-min incubation at pH 7.0 was 73° C. The maximum inhibitory activity was achieved at 35° C and pH 5.0. Gel assays showed the meaningful inhibition of E. integriceps α-amylases by various concentrations of Triticale inhibitors. Based on the data presented in this study, it could be said that the T-αAI has good inhibitory activity on E. integriceps gut α-amylase.

Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of α-Amylase.

PubMed

Tian, Mei-ling; Fang, Ting; Du, Mu-ying; Zhang, Fu-sheng

2016-04-01

To explore an efficient, safe, and speedy application of pulsed electric field (PEF) technology for enzymatic modification, effects of PEF treatment on the enzymatic activity, property and kinetic parameters of α-amylase were investigated. Conformational transitions were also studied with the aid of circular dichroism (CD) and fluorescence spectra. The maximum enzymatic activity of α-amylase was obtained under 15 kV/cm electric field intensity and 100 mL/min flow velocity PEF treatment, in which the enzymatic activity increased by 22.13 ± 1.14% compared with control. The activation effect could last for 18 h at 4 °C. PEF treatment could widen the range of optimum temperature for α-amylase, however, it barely exerted any effect on the optimum pH. On the other hand, α-amylase treated by PEF showed an increase of Vmax, t1/2 and ΔG, whereas a decrease of Km and k were observed. Furthermore, it can be observed from fluorescence and CD spectra that PEF treatment had increased the number of amino acid residues, especially that of tryptophan, on α-amylase surface with enhanced α-helices by 34.76% and decreased random coil by 12.04% on α-amylase when compared with that of untreated. These changes in structure had positive effect on enhancing α-amylase activity and property.

Heterologous Expression and Characterization of a Thermostable Exo-β-D-Glucosaminidase from Aspergillus oryzae.

PubMed

Wu, Dingxin; Wang, Linchun; Li, Yuwei; Zhao, Shumiao; Peng, Nan; Liang, Yunxiang

2016-02-01

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50°C; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50°C than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of Vmax and kcat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.

Radiochromatographic assay of N-acyl-phosphatidylethanolamine-specific phospholipase D activity.

PubMed

Fezza, Filomena; Gasperi, Valeria; Mazzei, Cinzia; Maccarrone, Mauro

2005-04-01

A radiochromatographic method has been set up to assay the activity of N-acyl-phosphatidylethanolamine-specific phospholipase D (NAPE-PLD), based on reversed-phase high-performance liquid chromatography (HPLC) and online scintillation counting. The anandamide (N-arachidonoylethanolamine, AEA), product released by NAPE-PLD from the N-arachidonoyl-phosphatidylethanolamine (NArPE) substrate, was separated using a C18 column eluted with methanol-water-acetic acid and was quantified with an external standard method. Baseline separation of AEA and NArPE was completed in less than 15 min, with a detection limit of 0.5 fmol AEA at a signal-to-noise ratio of 4:1. The sensitivity and accuracy of the radiochromatographic procedure allowed detection and characterization of NAPE-PLD activity in very tiny tissue samples or in samples where the enzymatic activity is very low. With this method, we could determine the kinetic constants (i.e., apparent Michaelis-Menten constant (Km) of 40.0+/-5.6 microM and maximum velocity (Vmax) of 22.2+/-3.5 pmol/min per milligram protein toward NArPE) and the distribution of NAPE-PLD activity in brain areas and peripheral tissues of mouse. In addition, we could collect unprecedented evidence that compounds widely used in studies of the endocannabinoid system (e.g., AEA and congeners, receptor a(nta)gonists and inhibitors of AEA degradation) can also affect NAPE-PLD activity.

Inhibition of Sunn Pest, Eurygaster integriceps, α-Amylases by α-Amylase Inhibitors (T-αAI) from Triticale

PubMed Central

Mehrabadi, Mohammad; Bandani, Ali R.; Saadati, Fatemeh

2010-01-01

The effect of triticale α-amylases inhibitors on starch hydrolysis catalyzed by the Sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae) midgut amylases was examined. Biochemical studgawies showed that inhibitors from Triticale (a hybrid of wheat and rye) had inhibitiory effects on E. integriceps α-amylases. The effects of the triticale α-amylase inhibitor (T-αAI) on α-amylase of E. integriceps showed a dose dependent manner of inhibition, e.g. less inhibition of enzyme activity (around 10%) with a lower dose (0.25 mg protein) and high inhibition of enzyme activity (around 80%) when a high dose of inhibitor was used (1.5 mg protein). The enzyme kinetic studies using Michaelis-Menten and Lineweaver-Burk equations showed the Km remained constant (0.58%) but the maximum velocity (Vmax) decreased in the presence of a crude extract of Triticale inhibitors, indicating mixed inhibition. The temperature giving 50% inactivation of enzyme (T50) during a 30-min incubation at pH 7.0 was 73° C. The maximum inhibitory activity was achieved at 35° C and pH 5.0. Gel assays showed the meaningful inhibition of E. integriceps α-amylases by various concentrations of Triticale inhibitors. Based on the data presented in this study, it could be said that the T-αAI has good inhibitory activity on E. integriceps gut α-amylase. PMID:21062146

Secretory Expression and Characterization of an Acidic Endo-Polygalacturonase Gene from Aspergillus niger SC323 in Saccharomyces cerevisiae.

PubMed

Zhou, Huoxiang; Li, Xi; Guo, Mingyue; Xu, Qingrui; Cao, Yu; Qiao, Dairong; Cao, Yi; Xu, Hui

2015-07-01

The endo-polygalacturonase gene (endo-pgaA) was cloned from DNA of Aspergillus niger SC323 using the cDNA synthesized by overlapping PCR, and successfully expressed in Saccharomyces cerevisiae EBY100 through fusing the α-factor signal peptide of yeast. The full-length cDNA consists of 1,113 bp and encodes a protein of 370 amino acids with a calculated molecular mass of 38.8 kDa. After induction by galactose for 48 h, the activity of recombinant endo-PgaA in the culture supernatant can reach up to 1,448.48 U/mg. The recombinant protein was purified to homogeneity by ammonium sulfate precipitation and gel filtration column chromatography and subsequently characterized. The optimal pH and temperature of the purified recombinant enzyme were 5.0 and 50°C, respectively. The Michaelis-Menten constant (Km) and maximal velocity (Vmax) of the enzyme for pectin were 88.54 μmol/ml and 175.44 μmol/mg/min, respectively. The enzyme activity was enhanced by Ca(2+), Cu(2+), and Na(+), and strongly inhibited by Pb(2+) and Mn(2+). The pectin hydrolysates were mainly galacturonic acid and other oligo-galacturonates. Therefore, these characteristics suggest that the recombinant endo-PgaA may be of potential use in the food and feed industries.

Purification and characterization of polyphenol oxidase from rape flower.

PubMed

Sun, Han-Ju; Wang, Jing; Tao, Xue-Ming; Shi, Juan; Huang, Mei-Ying; Chen, Zhe

2012-01-25

The purification and partial enzymology characteristics of polyphenol oxidase (PPO) from rape flower were studied. After preliminary treatments, the crude enzyme solution was in turn purified with ammonium sulfate, dialysis, and Sephadex G-75 gel chromatography. The optimal conditions and stability of PPO were examined at different pH values and temperatures. Subsequently, PPO was also characterized by substrate (catechol) concentrations, inhibitors, kinetic parameters, and molecular weight. Results showed that the optimal pH for PPO activity was 5.5 in the presence of catechol and that PPO was relatively stable at pH 3.5-5.5. PPO was moderately stable at temperatures from 60 to 70 °C, whereas it was easily denatured at 80-90 °C. Ethylenediaminetetraacetic acid, sodium chloride, and calcium chloride had little inhibitive effects on PPO, whereas citric acid, sodium sulfite, and ascorbic acid had strongly inhibitive effects. The Michaelis-Menten constant (K(m)) and maximal reaction velocity (V(max)) of PPO were 0.767 mol/L and 0.519 Ab/min/mL of the crude PPO solution, respectively. PPO was finally purified to homogeneity with a purification factor of 4.41-fold and a recovery of 12.41%. Its molecular weight was 60.4 kDa, indicating that the PPO is a dimer. The data obtained in this research may help to prevent the enzymatic browning of rape flower during its storage and processing.

Screening of traditional antidiabetic medicinal plants of Mauritius for possible alpha-amylase inhibitory effects in vitro.

PubMed

Kotowaroo, M I; Mahomoodally, M F; Gurib-Fakim, A; Subratty, A H

2006-03-01

In this study, seven exotic/indigenous medicinal plants of Mauritius, namely Coix lacryma-jobi (Poaceae), Aegle marmelos (Rutaceae), Artocarpus heterophyllus (Moraceae), Vangueria madagascariensis (Rubiaceae), Azadirachta indica (Meliaceae), Eriobotrya japonica (Rosaceae) and Syzigium cumini (Myrtaceae) were studied for possible effects on starch breakdown by alpha-amylase in vitro. The results showed that only Artocarpus heterophyllus significantly (p < 0.05) inhibited alpha-amylase activity in vitro. To confirm the observed effects, a further biochemical assay was undertaken to investigate the effects of Artocarpus heterophyllus on alpha-amylase activity using rat plasma in vitro. It was found that the aqueous leaf extract significantly (p < 0.05) inhibited alpha-amylase activity in rat plasma. The highest inhibitory activity (27.20 +/- 5.00%) was observed at a concentration of 1000 microg/mL. However, in both cases dose dependency was not observed. Enzyme kinetic studies using the Michaelis-Menten and Lineweaver-Burk equations were performed to establish the type of inhibition involved. In the presence of the plant extract the maximal velocity (Vmax) remained constant (1/150 g / L/s) whereas the Michaelis-Menten constant (Km) increased by 5.79 g / L, indicating that the aqueous leaf extract of Artocarpus heterophyllus behaved as a competitive inhibitor. Results from the present study tend to indicate that Artocarpus heterophyllus could act as a 'starch blocker' thereby reducing post-prandial glucose peaks. Copyright 2006 John Wiley & Sons, Ltd.

Hydrolysis of penicillins and related compounds by the cell-bound penicillin acylase of Escherichia coli

PubMed Central

Cole, M.

1969-01-01

1. A method is given for the preparation of penicillin acylase by using Escherichia coli N.C.I.B. 8743 and a strain selected for higher yield. The enzyme is associated with the bacterial cells and removes the side chains of penicillins to give 6-amino-penicillanic acid and a carboxylic acid. 2. The rates of penicillin deacylation indicated that p-hydroxybenzylpenicillin was the best substrate, followed in diminishing order by benzyl-, dl-α-hydroxybenzyl-, 2-furylmethyl-, 2-thienylmethyl-, d-α-aminobenzyl-, n-propoxymethyl- and isobutoxymethyl-penicillin. Phenylpenicillin and dl-α-carboxybenzylpenicillin were not substrates and phenoxymethyl-penicillin was very poor. 3. Amides and esters of the above penicillins were also substrates for the deacylation reaction, as were cephalosporins with a thienylmethyl side chain. 4. For the deacylation of 2-furylmethylpenicillin at 21° the optimum pH was 8·2. The optimum temperature was 60° at pH7. 5. By using selection A of N.C.I.B. 8743 and determining reaction velocities by assaying yields of 6-amino-penicillanic acid in a 10min. reaction at 50° and pH8·2, the Km for benzylpenicillin was found to be about 30mm and the Km for 2-furylmethylpenicillin, about 10mm. The Vmax. values were 0·6 and 0·24μmole/min./mg. of bacterial cells respectively. PMID:4982417

Effects of 3G cell phone exposure on the structure and function of the human cytochrome P450 reductase.

PubMed

Tanvir, Shazia; Thuróczy, György; Selmaoui, Brahim; Silva Pires Antonietti, Viviane; Sonnet, Pascal; Arnaud-Cormos, Delia; Lévêque, Philippe; Pulvin, Sylviane; de Seze, René

2016-10-01

Cell phones increase exposure to radiofrequency (RF) electromagnetic fields (EMFs). Whether EMFs exert specific effects on biological systems remains debatable. This study investigated the effect of cell phone exposure on the structure and function of human NADPH-cytochrome P450 reductase (CPR). CPR plays a key role in the electron transfer to cytochrome P450, which takes part in a wide range of oxidative metabolic reactions in various organisms from microbes to humans. Human CPR was exposed for 60min to 1966-MHz RF inside a transverse electromagnetic cell (TEM-cell) placed in an incubator. The specific absorption rate (SAR) was 5W·kg(-1). Conformation changes have been detected through fluorescent spectroscopy of flavin and tryptophan residues, and investigated through circular dichroism, dynamic light scattering and microelectrophoresis. These showed that CPR was narrowed. By using cytochrome C reductase activity to assess the electron flux through the CPR, the Michaelis Menten constant (Km) and the maximum initial velocity (Vmax) decreased by 22% as compared with controls. This change was due to small changes in the tertiary and secondary structures of the protein at 37°C. The relevance of these findings to an actual RF exposure scenario demands further biochemical and in-vivo confirmation. Copyright © 2016 Elsevier B.V. All rights reserved.

Biphasic Kinetic Behavior of E. coli WrbA, an FMN-Dependent NAD(P)H:Quinone Oxidoreductase

PubMed Central

Kishko, Iryna; Harish, Balasubramanian; Zayats, Vasilina; Reha, David; Tenner, Brian; Beri, Dhananjay; Gustavsson, Tobias; Ettrich, Rüdiger; Carey, Jannette

2012-01-01

The E. coli protein WrbA is an FMN-dependent NAD(P)H:quinone oxidoreductase that has been implicated in oxidative defense. Three subunits of the tetrameric enzyme contribute to each of four identical, cavernous active sites that appear to accommodate NAD(P)H or various quinones, but not simultaneously, suggesting an obligate tetramer with a ping-pong mechanism in which NAD departs before oxidized quinone binds. The present work was undertaken to evaluate these suggestions and to characterize the kinetic behavior of WrbA. Steady-state kinetics results reveal that WrbA conforms to a ping-pong mechanism with respect to the constancy of the apparent Vmax to Km ratio with substrate concentration. However, the competitive/non-competitive patterns of product inhibition, though consistent with the general class of bi-substrate reactions, do not exclude a minor contribution from additional forms of the enzyme. NMR results support the presence of additional enzyme forms. Docking and energy calculations find that electron-transfer-competent binding sites for NADH and benzoquinone present severe steric overlap, consistent with the ping-pong mechanism. Unexpectedly, plots of initial velocity as a function of either NADH or benzoquinone concentration present one or two Michaelis-Menten phases depending on the temperature at which the enzyme is held prior to assay. The effect of temperature is reversible, suggesting an intramolecular conformational process. WrbA shares these and other details of its kinetic behavior with mammalian DT-diaphorase, an FAD-dependent NAD(P)H:quinone oxidoreductase. An extensive literature review reveals several other enzymes with two-plateau kinetic plots, but in no case has a molecular explanation been elucidated. Preliminary sedimentation velocity analysis of WrbA indicates a large shift in size of the multimer with temperature, suggesting that subunit assembly coupled to substrate binding may underlie the two-plateau behavior. An additional aim of this report is to bring under wider attention the apparently widespread phenomenon of two-plateau Michaelis-Menten plots. PMID:22952804

Real-time tracking of tumor motions and deformations along the leaf travel direction with the aid of a synchronized dynamic MLC leaf sequencer.

PubMed

Tacke, Martin; Nill, Simeon; Oelfke, Uwe

2007-11-21

Advanced radiotherapeutical techniques like intensity-modulated radiation therapy (IMRT) are based on an accurate knowledge of the location of the radiation target. An accurate dose delivery, therefore, requires a method to account for the inter- and intrafractional target motion and the target deformation occurring during the course of treatment. A method to compensate in real time for changes in the position and shape of the target is the use of a dynamic multileaf collimator (MLC) technique which can be devised to automatically arrange the treatment field according to real-time image information. So far, various approaches proposed for leaf sequencers have had to rely on a priori known target motion data and have aimed to optimize the overall treatment time. Since for a real-time dose delivery the target motion is not known a priori, the velocity range of the leading leaves is restricted by a safety margin to c x v(max) while the following leaves can travel with an additional maximum speed to compensate for the respective target movements. Another aspect to be considered is the tongue and groove effect. A uniform radiation field can only be achieved if the leaf movements are synchronized. The method presented in this note is the first to combine a synchronizing sequencer and real-time tracking with a dynamic MLC. The newly developed algorithm is capable of online optimizing the leaf velocities by minimizing the overall treatment time while at the same time it synchronizes the leaf trajectories in order to avoid the tongue and groove effect. The simultaneous synchronization is performed with the help of an online-calculated mid-time leaf trajectory which is common for all leaf pairs and which takes into account the real-time target motion and deformation information.

Assessment of renal dopaminergic system activity during cyclosporine A administration in the rat.

PubMed Central

Pestana, M.; Vieira-Coelho, M. A.; Pinto-do-O, P. C.; Fernandes, M. H.; Soares-da-Silva, P.

1995-01-01

1. Administration of cyclosporine A (CsA; 50 mg kg-1 day-1, s.c.) for 14 days produced an increase in both systolic (SBP) and diastolic (DBP) blood pressure by 60 and 25 mmHg, respectively. The urinary excretion of dopamine, DOPAC and HVA was reduced from day 5-6 of CsA administration onwards (dopamine from 19 to 46%, DOPAC from 16 to 48%; HVA from 18 to 42%). In vehicle-treated rats, the urinary excretion of dopamine and DOPAC increased (from 7 to 60%) from day 5 onwards; by contrast, the urinary excretion of HVA was reduced (from 27 to 60%) during the second week. 2. No significant difference was observed between the Vmax and Km values of renal aromatic L-amino acid decarboxylase (AAAD) in rats treated with CsA for 7 and 14 days or with vehicle. 3. Km and Vmax of monoamine oxidase types A and B did not differ significantly between rats treated with CsA for 7 and 14 days or with vehicle. 4. Maximal catechol-O-methyltransferase activity (Vmax) in homogenates of renal tissues obtained from rats treated with CsA for 7 or 14 days was significantly higher than that in vehicle-treated rats; Km (22.3 +/- 1.5 microM) values for COMT did not differ between the three groups of rats. 5. The accumulation of newly-formed dopamine and DOPAC in cortical tissues of rats treated with CsA for 14 days was three to four times higher than in controls. The outflow of both dopamine and DOPAC declined progressively with time and reflected the amine and amine metabolite tissue contents. No significant difference was observed between the DOPAC/dopamine ratios in the perifusate of renal tissues obtained from CsA- and vehicle-treated rats. In addition, no significant differences were observed in k values or in the slope of decline of both DA and DOPAC between experiments performed with CsA and vehicle-treated animals. 6. The Vmax for the saturable component of L-3,4-dihydroxyphenylalanine (L-DOPA) uptake in renal tubules from rats treated with CsA was twice that of vehicle-treated animals. Km in CsA- and vehicle-treated rats did not differ. 7. The decrease in the urinary excretion of sodium and an increase in blood pressure during CsA treatment was accompanied by a reduction in daily urinary excretion of dopamine. This appears to result from a reduction in the amount of L-DOPA made available to the kidney and does not involve changes in tubular AAAD, the availability of dopamine to leave the renal cells and dopamine metabolism.(ABSTRACT TRUNCATED AT 400 WORDS) PMID:8564191

Estimation of ground motion for Bhuj (26 January 2001; Mw 7.6 and for future earthquakes in India

USGS Publications Warehouse

Singh, S.K.; Bansal, B.K.; Bhattacharya, S.N.; Pacheco, J.F.; Dattatrayam, R.S.; Ordaz, M.; Suresh, G.; ,; Hough, S.E.

2003-01-01

Only five moderate and large earthquakes (Mw ???5.7) in India-three in the Indian shield region and two in the Himalayan arc region-have given rise to multiple strong ground-motion recordings. Near-source data are available for only two of these events. The Bhuj earthquake (Mw 7.6), which occurred in the shield region, gave rise to useful recordings at distances exceeding 550 km. Because of the scarcity of the data, we use the stochastic method to estimate ground motions. We assume that (1) S waves dominate at R < 100 km and Lg waves at R ??? 100 km, (2) Q = 508f0.48 is valid for the Indian shield as well as the Himalayan arc region, (3) the effective duration is given by fc-1 + 0.05R, where fc is the corner frequency, and R is the hypocentral distance in kilometer, and (4) the acceleration spectra are sharply cut off beyond 35 Hz. We use two finite-source stochastic models. One is an approximate model that reduces to the ??2-source model at distances greater that about twice the source dimension. This model has the advantage that the ground motion is controlled by the familiar stress parameter, ????. In the other finite-source model, which is more reliable for near-source ground-motion estimation, the high-frequency radiation is controlled by the strength factor, sfact, a quantity that is physically related to the maximum slip rate on the fault. We estimate ???? needed to fit the observed Amax and Vmax data of each earthquake (which are mostly in the far field). The corresponding sfact is obtained by requiring that the predicted curves from the two models match each other in the far field up to a distance of about 500 km. The results show: (1) The ???? that explains Amax data for shield events may be a function of depth, increasing from ???50 bars at 10 km to ???400 bars at 36 km. The corresponding sfact values range from 1.0-2.0. The ???? values for the two Himalayan arc events are 75 and 150 bars (sfact = 1.0 and 1.4). (2) The ???? required to explain Vmax data is, roughly, half the corresponding value for Amax, while the same sfact explains both sets of data. (3) The available far-field Amax and Vmax data for the Bhuj mainshock are well explained by ???? = 200 and 100 bars, respectively, or, equivalently, by sfact = 1.4. The predicted Amax and Vmax in the epicentral region of this earthquake are 0.80 to 0.95 g and 40 to 55 cm/sec, respectively.

Hymenolepis diminuta (Cestoda): uptake of cycloleucine by metacestodes.

PubMed

Jeffs, S A; Arme, C

1985-01-01

Cycloleucine uptake by metacestodes of H. diminuta of various ages was investigated. Absorption occurs by active mediated transport, mean Kt = 0.28 mM. Vmax values are age-related, and can be correlated to developmental changes. Cycloleucine uptake in the metacestode is very similar to that in the adult worm and the implications of this are discussed.

Intelligent Hybrid Vehicle Power Control - Part 1: Machine Learning of Optimal Vehicle Power

DTIC Science & Technology

2012-06-30

time window ),[ tWt DT : vave, vmax, vmin, ac, vst and vend, where the first four parameters are, respectively, the average speed, maximum speed...minimum speed and average acceleration, during the time period ),[ tWt DT , vst is the vehicle speed at )( DTWt  , and vend is the vehicle

Effects of a prostagrandin EP4-receptor agonist ONO-AE1-329 on the left ventricular pressure-volume relationship in the halothane-anesthetized dogs.

PubMed

Honda, Atsushi; Nakamura, Yuji; Ohara, Hiroshi; Cao, Xin; Nomura, Hiroaki; Katagi, Jun; Wada, Takeshi; Izumi-Nakaseko, Hiroko; Ando, Kentaro; Sugiyama, Atsushi

2016-03-15

Cardiac effects of a prostagrandin EP4-receptor agonist ONO-AE1-329 were assessed in the halothane-anesthetized dogs under the monitoring of left ventricular pressure-volume relationship, which were compared with those of clinically recommended doses of dopamine, dobutamine and milrinone (n=4-5 for each treatment). ONO-AE1-329 was intravenously administered in doses of 0.3, 1 and 3 ng/kg/min for 10 min with a pause of 20 min. Dopamine in a dose of 3 µg/kg/min for 10 min, dobutamine in a dose of 1 µg/kg/min for 10 min and milrinone in a dose of 5 µg/kg/min for 10 min followed by 0.5 µg/kg/min for 10 min were intravenously administered. Low dose of ONO-AE1-329 increased the stroke volume. Middle dose of ONO-AE1-329 increased the cardiac output, left ventricular end-diastolic volume, ejection fraction, maximum upstroke/downstroke velocities of the left ventricular pressure and external work, but decreased the end-systolic pressure and internal work besides the change by the low dose. High dose of ONO-AE1-329 increased the heart rate and maximum elastance, but decreased the end-systolic volume besides the changes by the middle dose. Dopamine, dobutamine and milrinone exerted essentially similar cardiac effects to ONO-AE1-329, but they did not significantly change the end-diastolic volume, end-systolic volume, stroke volume, ejection fraction, end-systolic pressure, maximum elastance, external work or internal work. Thus, EP4-receptor stimulation by ONO-AE1-329 may have potential to better promote the passive ventricular filling than the conventional cardiotonic drugs, which could become a candidate of novel therapeutic strategy for the treatment of heart failure with preserved ejection fraction. Copyright © 2016 Elsevier B.V. All rights reserved.

Cross-correlation analysis of pulse wave propagation in arteries: in vitro validation and in vivo feasibility.

PubMed

Nauleau, Pierre; Apostolakis, Iason; McGarry, Matthew; Konofagou, Elisa

2018-05-29

The stiffness of the arteries is known to be an indicator of the progression of various cardiovascular diseases. Clinically, the pulse wave velocity (PWV) is used as a surrogate for arterial stiffness. Pulse wave imaging (PWI) is a non-invasive, ultrasound-based imaging technique capable of mapping the motion of the vessel walls, allowing the local assessment of arterial properties. Conventionally, a distinctive feature of the displacement wave (e.g. the 50% upstroke) is tracked across the map to estimate the PWV. However, the presence of reflections, such as those generated at the carotid bifurcation, can bias the PWV estimation. In this paper, we propose a two-step cross-correlation based method to characterize arteries using the information available in the PWI spatio-temporal map. First, the area under the cross-correlation curve is proposed as an index for locating the regions of different properties. Second, a local peak of the cross-correlation function is tracked to obtain a less biased estimate of the PWV. Three series of experiments were conducted in phantoms to evaluate the capabilities of the proposed method compared with the conventional method. In the ideal case of a homogeneous phantom, the two methods performed similarly and correctly estimated the PWV. In the presence of reflections, the proposed method provided a more accurate estimate than conventional processing: e.g. for the soft phantom, biases of  -0.27 and -0.71 m · s -1 were observed. In a third series of experiments, the correlation-based method was able to locate two regions of different properties with an error smaller than 1 mm. It also provided more accurate PWV estimates than conventional processing (biases:  -0.12 versus -0.26 m · s -1 ). Finally, the in vivo feasibility of the proposed method was demonstrated in eleven healthy subjects. The results indicate that the correlation-based method might be less precise in vivo but more accurate than the conventional method.

Attraction of Rotors to the Pulmonary Veins in Paroxysmal Atrial Fibrillation: A Modeling Study

PubMed Central

Calvo, Conrado J.; Deo, Makarand; Zlochiver, Sharon; Millet, José; Berenfeld, Omer

2014-01-01

Maintenance of paroxysmal atrial fibrillation (AF) by fast rotors in the left atrium (LA) or at the pulmonary veins (PVs) is not fully understood. To gain insight into this dynamic and complex process, we studied the role of the heterogeneous distribution of transmembrane currents in the PVs and LA junction (PV-LAJ) in the localization of rotors in the PVs. We also investigated whether simple pacing protocols could be used to predict rotor drift in the PV-LAJ. Experimentally observed heterogeneities in IK1, IKs, IKr, Ito, and ICaL in the PV-LAJ were incorporated into two- and pseudo three-dimensional models of Courtemanche-Ramirez-Nattel-Kneller human atrial kinetics to simulate various conditions and investigate rotor drifting mechanisms. Spatial gradients in the currents resulted in shorter action potential duration, minimum diastolic potential that was less negative, and slower upstroke and conduction velocity for rotors in the PV region than in the LA. Rotors under such conditions drifted toward the PV and stabilized at the shortest action potential duration and less-excitable region, consistent with drift direction under intercellular coupling heterogeneities and regardless of the geometrical constraint in the PVs. Simulations with various IK1 gradient conditions and current-voltage relationships substantiated its major role in the rotor drift. In our 1:1 pacing protocol, we found that among various action potential properties, only the minimum diastolic potential gradient was a rate-independent predictor of rotor drift direction. Consistent with experimental and clinical AF studies, simulations in an electrophysiologically heterogeneous model of the PV-LAJ showed rotor attraction toward the PV. Our simulations suggest that IK1 heterogeneity is dominant compared to other currents in determining the drift direction through its impact on the excitability gradient. These results provide a believed novel framework for understanding the complex dynamics of rotors in AF. PMID:24739180

Action potential-based MEA platform for in vitro screening of drug-induced cardiotoxicity using human iPSCs and rat neonatal myocytes.

PubMed

Jans, Danny; Callewaert, Geert; Krylychkina, Olga; Hoffman, Luis; Gullo, Francesco; Prodanov, Dimiter; Braeken, Dries

2017-09-01

Drug-induced cardiotoxicity poses a negative impact on public health and drug development. Cardiac safety pharmacology issues urged for the preclinical assessment of drug-induced ventricular arrhythmia leading to the design of several in vitro electrophysiological screening assays. In general, patch clamp systems allow for intracellular recordings, while multi-electrode array (MEA) technology detect extracellular activity. Here, we demonstrate a complementary metal oxide semiconductor (CMOS)-based MEA system as a reliable platform for non-invasive, long-term intracellular recording of cardiac action potentials at high resolution. Quinidine (8 concentrations from 10 -7 to 2.10 -5 M) and verapamil (7 concentrations from 10 -11 to 10 -5 M) were tested for dose-dependent responses in a network of cardiomyocytes. Electrophysiological parameters, such as the action potential duration (APD), rates of depolarization and repolarization and beating frequency were assessed. In hiPSC, quinidine prolonged APD with EC 50 of 2.2·10 -6 M. Further analysis indicated a multifactorial action potential prolongation by quinidine: (1) decreasing fast repolarization with IC 50 of 1.1·10 -6 M; (2) reducing maximum upstroke velocity with IC 50 of 2.6·10 -6 M; and (3) suppressing spontaneous activity with EC 50 of 3.8·10 -6 M. In rat neonatal cardiomyocytes, verapamil blocked spontaneous activity with EC 50 of 5.3·10 -8 M and prolonged the APD with EC 50 of 2.5·10 -8 M. Verapamil reduced rates of fast depolarization and repolarization with IC 50 s of 1.8 and 2.2·10 -7 M, respectively. In conclusion, the proposed action potential-based MEA platform offers high quality and stable long-term recordings with high information content allowing to characterize multi-ion channel blocking drugs. We anticipate application of the system as a screening platform to efficiently and cost-effectively test drugs for cardiac safety. Copyright © 2017 Elsevier Inc. All rights reserved.

Structural and Functional Maturation of Cardiomyocytes Derived from Human Pluripotent Stem Cells

PubMed Central

Lundy, Scott D.; Zhu, Wei-Zhong

2013-01-01

Despite preclinical studies demonstrating the functional benefit of transplanting human pluripotent stem cell-derived cardiomyocytes (PSC-CMs) into damaged myocardium, the ability of these immature cells to adopt a more adult-like cardiomyocyte (CM) phenotype remains uncertain. To address this issue, we tested the hypothesis that prolonged in vitro culture of human embryonic stem cell (hESC)- and human induced pluripotent stem cell (hiPSC)-derived CMs would result in the maturation of their structural and contractile properties to a more adult-like phenotype. Compared to their early-stage counterparts (PSC-CMs after 20–40 days of in vitro differentiation and culture), late-stage hESC-CMs and hiPSC-CMs (80–120 days) showed dramatic differences in morphology, including increased cell size and anisotropy, greater myofibril density and alignment, sarcomeres visible by bright-field microscopy, and a 10-fold increase in the fraction of multinucleated CMs. Ultrastructural analysis confirmed improvements in the myofibrillar density, alignment, and morphology. We measured the contractile performance of late-stage hESC-CMs and hiPSC-CMs and noted a doubling in shortening magnitude with slowed contraction kinetics compared to the early-stage cells. We then examined changes in the calcium-handling properties of these matured CMs and found an increase in calcium release and reuptake rates with no change in the maximum amplitude. Finally, we performed electrophysiological assessments in hESC-CMs and found that late-stage myocytes have hyperpolarized maximum diastolic potentials, increased action potential amplitudes, and faster upstroke velocities. To correlate these functional changes with gene expression, we performed qPCR and found a robust induction of the key cardiac structural markers, including β-myosin heavy chain and connexin-43, in late-stage hESC-CMs and hiPSC-CMs. These findings suggest that PSC-CMs are capable of slowly maturing to more closely resemble the phenotype of adult CMs and may eventually possess the potential to regenerate the lost myocardium with robust de novo force-producing tissue. PMID:23461462

Attraction of rotors to the pulmonary veins in paroxysmal atrial fibrillation: a modeling study.

PubMed

Calvo, Conrado J; Deo, Makarand; Zlochiver, Sharon; Millet, José; Berenfeld, Omer

2014-04-15

Maintenance of paroxysmal atrial fibrillation (AF) by fast rotors in the left atrium (LA) or at the pulmonary veins (PVs) is not fully understood. To gain insight into this dynamic and complex process, we studied the role of the heterogeneous distribution of transmembrane currents in the PVs and LA junction (PV-LAJ) in the localization of rotors in the PVs. We also investigated whether simple pacing protocols could be used to predict rotor drift in the PV-LAJ. Experimentally observed heterogeneities in IK1, IKs, IKr, Ito, and ICaL in the PV-LAJ were incorporated into two- and pseudo three-dimensional models of Courtemanche-Ramirez-Nattel-Kneller human atrial kinetics to simulate various conditions and investigate rotor drifting mechanisms. Spatial gradients in the currents resulted in shorter action potential duration, minimum diastolic potential that was less negative, and slower upstroke and conduction velocity for rotors in the PV region than in the LA. Rotors under such conditions drifted toward the PV and stabilized at the shortest action potential duration and less-excitable region, consistent with drift direction under intercellular coupling heterogeneities and regardless of the geometrical constraint in the PVs. Simulations with various IK1 gradient conditions and current-voltage relationships substantiated its major role in the rotor drift. In our 1:1 pacing protocol, we found that among various action potential properties, only the minimum diastolic potential gradient was a rate-independent predictor of rotor drift direction. Consistent with experimental and clinical AF studies, simulations in an electrophysiologically heterogeneous model of the PV-LAJ showed rotor attraction toward the PV. Our simulations suggest that IK1 heterogeneity is dominant compared to other currents in determining the drift direction through its impact on the excitability gradient. These results provide a believed novel framework for understanding the complex dynamics of rotors in AF. Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Dynamic reciprocity of sodium and potassium channel expression in a macromolecular complex controls cardiac excitability and arrhythmia.

PubMed

Milstein, Michelle L; Musa, Hassan; Balbuena, Daniela Ponce; Anumonwo, Justus M B; Auerbach, David S; Furspan, Philip B; Hou, Luqia; Hu, Bin; Schumacher, Sarah M; Vaidyanathan, Ravi; Martens, Jeffrey R; Jalife, José

2012-07-31

The cardiac electrical impulse depends on an orchestrated interplay of transmembrane ionic currents in myocardial cells. Two critical ionic current mechanisms are the inwardly rectifying potassium current (I(K1)), which is important for maintenance of the cell resting membrane potential, and the sodium current (I(Na)), which provides a rapid depolarizing current during the upstroke of the action potential. By controlling the resting membrane potential, I(K1) modifies sodium channel availability and therefore, cell excitability, action potential duration, and velocity of impulse propagation. Additionally, I(K1)-I(Na) interactions are key determinants of electrical rotor frequency responsible for abnormal, often lethal, cardiac reentrant activity. Here, we have used a multidisciplinary approach based on molecular and biochemical techniques, acute gene transfer or silencing, and electrophysiology to show that I(K1)-I(Na) interactions involve a reciprocal modulation of expression of their respective channel proteins (Kir2.1 and Na(V)1.5) within a macromolecular complex. Thus, an increase in functional expression of one channel reciprocally modulates the other to enhance cardiac excitability. The modulation is model-independent; it is demonstrable in myocytes isolated from mouse and rat hearts and with transgenic and adenoviral-mediated overexpression/silencing. We also show that the post synaptic density, discs large, and zonula occludens-1 (PDZ) domain protein SAP97 is a component of this macromolecular complex. We show that the interplay between Na(v)1.5 and Kir2.1 has electrophysiological consequences on the myocardium and that SAP97 may affect the integrity of this complex or the nature of Na(v)1.5-Kir2.1 interactions. The reciprocal modulation between Na(v)1.5 and Kir2.1 and the respective ionic currents should be important in the ability of the heart to undergo self-sustaining cardiac rhythm disturbances.

Wolff-Parkinson-White (WPW) syndrome: the detection of delta wave in an electrocardiogram (ECG).

PubMed

Mahamat, Hassan Adam; Jacquir, Sabir; Khalil, Cliff; Laurent, Gabriel; Binczak, Stephane

2016-08-01

The delta wave remains an important indicator to diagnose the WPW syndrome. In this paper, a new method of detection of delta wave in an ECG signal is proposed. Firstly, using the continuous wavelet transform, the P wave, the QRS complex and the T wave are detected, then their durations are computed after determination of the boundary location (onsets and offsets of the P, QRS and T waves). Secondly, the PR duration, the QRS duration and the upstroke of the QRS complex are used to determine the presence or absence of the delta wave. This algorithm has been tested on the Physionel database (ptbdb) in order to evaluate its robustness. It has been applied to clinical signals from patients affected by WPW syndrome. This method can provide assistance to practitioners in order to detect the WPW syndrome.

Estimating Tropical Cyclone Surface Wind Field Parameters with the CYGNSS Constellation

NASA Astrophysics Data System (ADS)

Morris, M.; Ruf, C. S.

2016-12-01

A variety of parameters can be used to describe the wind field of a tropical cyclone (TC). Of particular interest to the TC forecasting and research community are the maximum sustained wind speed (VMAX), radius of maximum wind (RMW), 34-, 50-, and 64-kt wind radii, and integrated kinetic energy (IKE). The RMW is the distance separating the storm center and the VMAX position. IKE integrates the square of surface wind speed over the entire storm. These wind field parameters can be estimated from observations made by the Cyclone Global Navigation Satellite System (CYGNSS) constellation. The CYGNSS constellation consists of eight small satellites in a 35-degree inclination circular orbit. These satellites will be operating in standard science mode by the 2017 Atlantic TC season. CYGNSS will provide estimates of ocean surface wind speed under all precipitating conditions with high temporal and spatial sampling in the tropics. TC wind field data products can be derived from the level-2 CYGNSS wind speed product. CYGNSS-based TC wind field science data products are developed and tested in this paper. Performance of these products is validated using a mission simulator prelaunch.

Mechanisms and time course of impaired skeletal muscle glucose transport activity in streptozocin diabetic rats.

PubMed Central

Napoli, R; Hirshman, M F; Horton, E S

1995-01-01

Skeletal muscle glucose transport is altered in diabetes in humans, as well as in rats. To investigate the mechanisms of this abnormality, we measured glucose transport Vmax, the total transporter number, their average intrinsic activity, GLUT4 and GLUT1 contents in skeletal muscle plasma membrane vesicles from basal or insulin-stimulated streptozocin diabetic rats with different duration of diabetes, treated or not with phlorizin. The glucose transport Vmax progressively decreased with the duration of diabetes. In the basal state, this decrease was primarily associated with the reduction of transporter intrinsic activity, which appeared earlier than any change in transporter number or GLUT4 and GLUT1 content. In the insulin-stimulated state, the decrease of transport was mainly associated with severe defects in transporter translocation. Phlorizin treatment partially increased the insulin-stimulated glucose transport by improving the transporter translocation defects. In conclusion, in streptozocin diabetes (a) reduction of intrinsic activity plays a major and early role in the impairment of basal glucose transport; (b) a defect in transporter translocation is the mechanism responsible for the decrease in insulin-stimulated glucose transport; and (c) hyperglycemia per se affects the insulin-stimulated glucose transport by altering the transporter translocation. PMID:7615815

Lethal effect of electric fields on isolated ventricular myocytes.

PubMed

de Oliveira, Pedro Xavier; Bassani, Rosana Almada; Bassani, José Wilson Magalhães

2008-11-01

Defibrillator-type shocks may cause electric and contractile dysfunction. In this study, we determined the relationship between probability of lethal injury and electric field intensity (E in isolated rat ventricular myocytes, with emphasis on field orientation and stimulus waveform. This relationship was sigmoidal with irreversible injury for E > 50 V/cm . During both threshold and lethal stimulation, cells were twofold more sensitive to the field when it was applied longitudinally (versus transversally) to the cell major axis. For a given E, the estimated maximum variation of transmembrane potential (Delta V(max)) was greater for longitudinal stimuli, which might account for the greater sensitivity to the field. Cell death, however, occurred at lower maximum Delta V(max) values for transversal shocks. This might be explained by a less steep spatial decay of transmembrane potential predicted for transversal stimulation, which would possibly result in occurrence of electroporation in a larger membrane area. For the same stimulus duration, cells were less sensitive to field-induced injury when shocks were biphasic (versus monophasic). Ours results indicate that, although significant myocyte death may occur in the E range expected during clinical defibrillation, biphasic shocks are less likely to produce irreversible cell injury.

Calcium pump kinetics determined in single erythrocyte ghosts by microphotolysis and confocal imaging.

PubMed

Kubitscheck, U; Pratsch, L; Passow, H; Peters, R

1995-07-01

The activity of the plasma membrane calcium pump was measured in single cells. Human red blood cell ghosts were loaded with a fluorescent calcium indicator and either caged calcium and ATP (protocol A) or caged ATP and calcium (protocol B). In a suitably modified laser scanning microscope either calcium or ATP were released by a short UV light pulse. The time-dependent fluorescence intensity of the calcium indicator was then followed in single ghosts by repetitive confocal imaging. The fluorescence intensity was converted into calcium concentration, which in turn was used to derive the kinetic parameters of the calcium pump, the Michaelis-Menten constant Km, and the maximal transport rate vmax. Km and vmax values derived in this manner were 24 +/- 14 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol A, and 4 +/- 3 microM and 1.0 +/- 0.6 microM/(ghost s) for protocol B, respectively. The difference between A and B is presumably caused by calmodulin, which is inactive in the experiments with protocol A. The possibilities to extend the new method to living nucleus-containing cells transiently transfected with mutants of the plasma membrane calcium pump are discussed.

A mutation Ser213/Asn in the hexokinase 1 from Schizosaccharomyces pombe increases its affinity for glucose.

PubMed

Petit, T; Herrero, P; Gancedo, C

1998-10-29

Alignment of amino acids of the region implicated in glucose binding from a series of hexokinases showed that Schizosaccharomyces pombe hexokinase 1 had a Ser residue in a place where all other kinases had an Asn. We changed an AGT codon to AAT to place an Asn in the Ser213 position. This mutation decreased Km for glucose from 9.4 mM to 1.6 mM and the ratio Vmax (Fructose)/Vmax (Glucose) from 5 to 2.5. Also the Km for 2-deoxyglucose decreased from 2.7 mM to 0.8 mM. A mutation in the similar position of S. pombe hexokinase 2 (Asn196/Ser) increased the Km for glucose from 0.16 mM to 0.56 mM. Fermentation of glucose is not detectable in a S. pombe mutant with only hexokinase 1 activity but expression of the hxk1(S213/N) gene conferred ability to ferment the sugar. While the mutated hexokinase 1 partially mimicked S. cerevisiae hexokinase II in catabolite repression of invertase, the wild type one could not substitute for it. Copyright 1998 Academic Press.

Assessment of microbial methane oxidation above a petroleum-contaminated aquifer using a combination of in situ techniques

NASA Astrophysics Data System (ADS)

Urmann, Karina; Schroth, Martin H.; Noll, Matthias; Gonzalez-Gil, Graciela; Zeyer, Josef

2008-06-01

Emissions of the greenhouse gas CH4, which is often produced in contaminated aquifers, are reduced or eliminated by microbial CH4 oxidation in the overlying vadose zone. The aim of this field study was to estimate kinetic parameters and isotope fractionation factors for CH4 oxidation in situ in the vadose zone above a methanogenic aquifer in Studen, Switzerland, and to characterize the involved methanotrophic communities. To quantify kinetic parameters, several field tests, so-called gas push-pull tests (GPPTs), with CH4 injection concentrations ranging from 17 to 80 mL L-1 were performed. An apparent Vmax of 0.70 ± 0.15 mmol CH4 (L soil air)-1 h-1 and an apparent Km of 0.28 ± 0.09 mmol CH4 (L soil air)-1 was estimated for CH4 oxidation at 2.7 m depth, close to the groundwater table. At 1.1 m depth, Km (0.13 ± 0.02 mmol CH4 (L soil air)-1) was in a similar range, but Vmax (0.076 ± 0.006 mmol CH4 (L soil air)-1 h-1) was an order of magnitude lower. At 2.7 m, apparent first-order rate constants determined from a CH4 gas profile (1.9 h-1) and from a single GPPT (2.0 ± 0.03 h-1) were in good agreement. Above the groundwater table, a Vmax much higher than the in situ CH4 oxidation rate prior to GPPTs indicated a high buffer capacity for CH4. At both depths, known methanotrophic species affiliated with Methylosarcina and Methylocystis were detected by cloning and sequencing. Apparent stable carbon isotope fractionation factors α for CH4 oxidation determined during GPPTs ranged from 1.006 to 1.032. Variability was likely due to differences in methanotrophic activity and CH4 availability leading to different degrees of mass transfer limitation. This complicates the use of stable isotopes as an independent quantification method.

Transport of the placental estriol precursor 16α-hydroxy-dehydroepiandrosterone sulfate (16α-OH-DHEAS) by stably transfected OAT4-, SOAT-, and NTCP-HEK293 cells.

PubMed

Schweigmann, H; Sánchez-Guijo, A; Ugele, B; Hartmann, K; Hartmann, M F; Bergmann, M; Pfarrer, C; Döring, B; Wudy, S A; Petzinger, E; Geyer, J; Grosser, G

2014-09-01

16α-Hydroxy-dehydroepiandrosterone sulfate (16α-OH-DHEAS) mainly originates from the fetus and serves as precursor for placental estriol biosynthesis. For conversion of 16α-OH-DHEAS to estriol several intracellular enzymes are required. However, prior to enzymatic conversion, 16α-OH-DHEAS must enter the cells by carrier mediated transport. To identify these carriers, uptake of 16α-OH-DHEAS by the candidate carriers organic anion transporter OAT4, sodium-dependent organic anion transporter SOAT, Na(+)-taurocholate cotransporting polypeptide NTCP, and organic anion transporting polypeptide OATP2B1 was measured in stably transfected HEK293 cells by LC-MS-MS. Furthermore, the study aimed to localize SOAT in the human placenta. Stably transfected OAT4-HEK293 cells revealed a partly sodium-dependent transport for 16α-OH-DHEAS with an apparent Km of 23.1 ± 5.1 μM and Vmax of 485.0 ± 39.1 pmol/mg protein/min, while stably transfected SOAT- and NTCP-HEK293 cells showed uptake only under sodium conditions with Km of 319.0 ± 59.5 μM and Vmax of 1465.8 ± 118.8 pmol/mg protein/min for SOAT and Km of 51.4 ± 9.9 μM and Vmax of 1423.3 ± 109.6 pmol/mg protein/min for NTCP. In contrast, stably transfected OATP2B1-HEK293 cells did not transport 16α-OH-DHEAS at all. Immunohistochemical studies and in situ hybridization of formalin fixed and paraffin embedded sections of human late term placenta showed expression of SOAT in syncytiotrophoblasts, predominantly at the apical membrane as well as in the vessel endothelium. In conclusion, OAT4, SOAT, and NTCP were identified as carriers for the estriol precursor 16α-OH-DHEAS. At least SOAT and OAT4 seem to play a functional role for the placental estriol synthesis as both are expressed in the syncytiotrophoblast of human placenta. Copyright © 2014 Elsevier Ltd. All rights reserved.

Number of Players and Relative Pitch Area per Player: Comparing Their Influence on Heart Rate and Physical Demands in Under-12 and Under-13 Football Players.

PubMed

Castellano, Julen; Puente, Asier; Echeazarra, Ibon; Usabiaga, Oidui; Casamichana, David

2016-01-01

The aim of the present study is to analyse the influence of different large-sided games (LSGs) on the physical and physiological variables in under-12s (U12) and -13s (U13) soccer players. The effects of the combination of different number of players per team, 7, 9, and 11 (P7, P9, and P11, respectively) with three relative pitch areas, 100, 200, and 300 m(2) (A100, A200, and A300, respectively), were analysed in this study. The variables analysed were: 1) global indicator such as total distance (TD); work:rest ratio (W:R); player-load (PL) and maximal speed (Vmax); 2) heart rate (HR) mean and time spent in different intensity zones of HR (<75%, 75-84%, 84-90% and >90%), and; 3) five absolute (<8, 8-13, 13-16 and >16 Km h(-1)) and three relative speed categories (<40%, 40-60% and >60% Vmax). The results support the theory that a change in format (player number and pitch dimensions) affects no similarly in the two players categories. Although it can seem that U13 players are more demanded in this kind of LSG, when the work load is assessed from a relative point of view, great pitch dimensions and/or high number of player per team are involved in the training task to the U12 players. The results of this study could alert to the coaches to avoid some types of LSGs for the U12 players such as: P11 played in A100, A200 or A300, P9 played in A200 or A300 and P7 played in A300 due to that U13>U12 in several physical and physiological variables (W:R, time spent in 84-90%HRmax, distance in 8-13 and 13-16 Km h(-1) and time spent in 40-60%Vmax). These results may help youth soccer coaches to plan the progressive introduction of LSGs so that task demands are adapted to the physiological and physical development of participants.

Effects of genetic polymorphisms on the OCT1 and OCT2-mediated uptake of ranitidine.

PubMed

Meyer, Marleen Julia; Seitz, Tina; Brockmöller, Jürgen; Tzvetkov, Mladen Vassilev

2017-01-01

Ranitidine (Zantac®) is a H2-receptor antagonist commonly used for the treatment of acid-related gastrointestinal diseases. Ranitidine was reported to be a substrate of the organic cation transporters OCT1 and OCT2. The hepatic transporter OCT1 is highly genetically variable. Twelve major alleles confer partial or complete loss of OCT1 activity. The effects of these polymorphisms are highly substrate-specific and therefore difficult to predict. The renal transporter OCT2 has a common polymorphism, Ala270Ser, which was reported to affect OCT2 activity. In this study we analyzed the effects of genetic polymorphisms in OCT1 and OCT2 on the uptake of ranitidine and on its potency to inhibit uptake of other drugs. We characterized ranitidine uptake using HEK293 and CHO cells stably transfected to overexpress wild type OCT1, OCT2, or their naturally occurring allelic variants. Ranitidine was transported by wild-type OCT1 with a Km of 62.9 μM and a vmax of 1125 pmol/min/mg protein. Alleles OCT1*5, *6, *12, and *13 completely lacked ranitidine uptake. Alleles OCT1*2, *3, *4, and *10 had vmax values decreased by more than 50%. In contrast, OCT1*8 showed an increase of vmax by 25%. The effects of OCT1 alleles on ranitidine uptake strongly correlated with the effects on morphine uptake suggesting common interaction mechanisms of both drugs with OCT1. Ranitidine inhibited the OCT1-mediated uptake of metformin and morphine at clinically relevant concentrations. The inhibitory potency for morphine uptake was affected by the OCT1*2 allele. OCT2 showed only a limited uptake of ranitidine that was not significantly affected by the Ala270Ser polymorphism. We confirmed ranitidine as an OCT1 substrate and demonstrated that common genetic polymorphisms in OCT1 strongly affect ranitidine uptake and modulate ranitidine's potential to cause drug-drug interactions. The effects of the frequent OCT1 polymorphisms on ranitidine pharmacokinetics in humans remain to be analyzed.

Effects of genetic polymorphisms on the OCT1 and OCT2-mediated uptake of ranitidine

PubMed Central

Meyer, Marleen Julia; Seitz, Tina; Brockmöller, Jürgen

2017-01-01

Background Ranitidine (Zantac®) is a H2-receptor antagonist commonly used for the treatment of acid-related gastrointestinal diseases. Ranitidine was reported to be a substrate of the organic cation transporters OCT1 and OCT2. The hepatic transporter OCT1 is highly genetically variable. Twelve major alleles confer partial or complete loss of OCT1 activity. The effects of these polymorphisms are highly substrate-specific and therefore difficult to predict. The renal transporter OCT2 has a common polymorphism, Ala270Ser, which was reported to affect OCT2 activity. Aim In this study we analyzed the effects of genetic polymorphisms in OCT1 and OCT2 on the uptake of ranitidine and on its potency to inhibit uptake of other drugs. Methods and results We characterized ranitidine uptake using HEK293 and CHO cells stably transfected to overexpress wild type OCT1, OCT2, or their naturally occurring allelic variants. Ranitidine was transported by wild-type OCT1 with a Km of 62.9 μM and a vmax of 1125 pmol/min/mg protein. Alleles OCT1*5, *6, *12, and *13 completely lacked ranitidine uptake. Alleles OCT1*2, *3, *4, and *10 had vmax values decreased by more than 50%. In contrast, OCT1*8 showed an increase of vmax by 25%. The effects of OCT1 alleles on ranitidine uptake strongly correlated with the effects on morphine uptake suggesting common interaction mechanisms of both drugs with OCT1. Ranitidine inhibited the OCT1-mediated uptake of metformin and morphine at clinically relevant concentrations. The inhibitory potency for morphine uptake was affected by the OCT1*2 allele. OCT2 showed only a limited uptake of ranitidine that was not significantly affected by the Ala270Ser polymorphism. Conclusions We confirmed ranitidine as an OCT1 substrate and demonstrated that common genetic polymorphisms in OCT1 strongly affect ranitidine uptake and modulate ranitidine’s potential to cause drug-drug interactions. The effects of the frequent OCT1 polymorphisms on ranitidine pharmacokinetics in humans remain to be analyzed. PMID:29236753

Is bacteriostatic saline superior to normal saline as an echocardiographic contrast agent?

PubMed

Cardozo, Shaun; Gunasekaran, Prasad; Patel, Hena; McGorisk, Timothy; Toosi, Mehrdad; Faraz, Haroon; Zalawadiya, Sandip; Alesh, Issa; Kottam, Anupama; Afonso, Luis

2014-12-01

Objective data on the performance characteristics and physical properties of commercially available saline formulations [normal saline (NS) vs. bacteriostatic normal saline (bNS)] are sparse. This study sought to compare the in vitro physical properties and in vivo characteristics of two commonly employed echocardiographic saline contrast agents in an attempt to assess superiority. Nineteen patients undergoing transesophageal echocardiograms were each administered agitated regular NS and bNS injections in random order and in a blinded manner according to a standardized protocol. Video time-intensity (TI) curves were constructed from a representative region of interest, placed paraseptally within the right atrium, in the bicaval view. TI curves were analyzed for maximal plateau acoustic intensity (Vmax, dB) and dwell time (DT, s), defined as time duration between onset of Vmax and decay of video intensity below clinically useful levels, reflecting the duration of homogenous opacification of the right atrium. To further characterize the physical properties of the bubbles in vitro, fixed aliquots of similarly agitated saline were injected into a glass well slide-cover slip assembly and examined using an optical microscope to determine bubble diameter in microns (µm) and concentration [bubble count/high power field (hpf)]. A higher acoustic intensity (a less negative dB level), higher bubble concentration and longer DT were considered properties of a superior contrast agent. For statistical analysis, a paired t test was conducted to evaluate the differences in means of Vmax and DT. Compared to NS, bNS administration was associated with superior opacification (video intensity -8.69 ± 4.7 vs. -10.46 ± 4.1 dB, P = 0.002), longer DT (17.3 ± 6.1 vs. 10.2 ± 3.7 s) in vivo and smaller mean bubble size (43.4 vs. 58.6 μm) and higher bubble concentration (1,002 vs. 298 bubble/hpf) in vitro. bNS provides higher intensity and more sustained opacification of the right atrium compared to NS. Higher bubble concentration and stability appear to be additional desirable rheological characteristics favoring bNS as a contrast agent.

The Original Michaelis Constant: Translation of the 1913 Michaelis-Menten Paper

PubMed Central

Johnson, Kenneth A.; Goody, Roger S.

2011-01-01

Nearly 100 years ago Michaelis and Menten published their now classic paper (Michaelis, L., and Menten, M. L. (1913) Die Kinetik der Invertinwirkung, Biochemische Zeitschrift 49, 333–369), in which they show that the rate of an enzyme-catalyzed reaction is proportional to the concentration of enzyme-substrate complex predicted by the Michaelis-Menten equation. Because the original text was written in German, yet is often quoted by English speaking authors, we undertook a complete translation of the 1913 publication, which we provide as an online supplement (http://pubs.acs.org). Here we introduce the translation, describe the historical context of the work, and show a new analysis of the original data. In doing so, we uncovered several surprises that reveal an interesting glimpse into the early history of enzymology. In particular, our re-analysis of Michaelis and Menten’s data using modern computational methods revealed an unanticipated rigor and precision in the original publication and uncovered a sophisticated, comprehensive analysis that has been overlooked in the century since their work was published. Michaelis and Menten not only analyzed initial velocity measurements, but they also fit their full time course data to the integrated form of the rate equations, including product inhibition, and derived a single global constant to represent all of their data. That constant was not the Michaelis constant, but rather, Vmax/Km, the specificity constant times the enzyme concentration (kcat/Km*E0). PMID:21888353

The transport and metabolism of the uridine mononucleotides by rat jejunum in vitro.

PubMed Central

Bronk, J R; Hastewell, J G

1989-01-01

1. Both uridine 3'-monophosphate (3'-UMP) and uridine 5'-monophosphate (5'-UMP) when perfused through the lumen of isolated rat jejunum gave rise to uracil as the only transported pyrimidine appearing in the serosal medium; neither the nucleotide nor the nucleoside could be detected in the serosal fluid. 2. There was a low level of the nucleoside, uridine, in the luminal fluid after the nucleotide had passed through the jejunal segment. Luminal nucleoside appearance was more marked from the 3' form of the nucleotide. 3. The hydrolysis of the nucleotides to the nucleoside form occurred via a brush-border membrane enzyme, which had the same maximal velocity (Vmax) for the two nucleotides (699 +/- 35 and 747 +/- 10 nmol min-1 (mg protein)-1 for 3'-UMP and 5'-UMP, respectively) but a different Michaelis constant (Km) so that 3'-UMP (Km = 58 +/- 3 microM) hydrolysis is favoured over 5'-UMP hydrolysis (Km = 108 +/- microM) at lower concentrations. 4. At 0.05 mM, luminal 3'-UMP gave rise to a higher rate of serosal uracil appearance than luminal 5'-UMP, but at higher luminal concentrations (0.1-0.2 mM) the rate of serosal uracil appearance was the same from both nucleotides. 5. The transmural transport of uracil from the uridine mononucleotides is discussed with reference to the metabolism and compartmentalization of the small intestine responsible for the appearance of the free pyrimidine in the serosal fluid. PMID:2778724

Amyloglucosidase enzymatic reactivity inside lipid vesicles

PubMed Central

Li, Mian; Hanford, Michael J; Kim, Jin-Woo; Peeples, Tonya L

2007-01-01

Efficient functioning of enzymes inside liposomes would open new avenues for applications in biocatalysis and bioanalytical tools. In this study, the entrapment of amyloglucosidase (AMG) (EC 3.2.1.3) from Aspergillus niger into dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles (MLVs) and large unilamellar vesicles (LUVs) was investigated. Negative-stain, freeze-fracture, and cryo-transmission electron microscopy images verified vesicle formation in the presence of AMG. Vesicles with entrapped AMG were isolated from the solution by centrifugation, and vesicle lamellarity was identified using fluorescence laser confocal microscopy. The kinetics of starch hydrolysis by AMG was modeled for two different systems, free enzyme in aqueous solution and entrapped enzyme within vesicles in aqueous suspension. For the free enzyme system, intrinsic kinetics were described by a Michaelis-Menten kinetic model with product inhibition. The kinetic constants, Vmax and Km, were determined by initial velocity measurements, and Ki was obtained by fitting the model to experimental data of glucose concentration-time curves. Predicted concentration-time curves using these kinetic constants were in good agreement with experimental measurements. In the case of the vesicles, the time-dependence of product (glucose) formation was experimentally determined and simulated by considering the kinetic behavior of the enzyme and the permeation of substrate into the vesicle. Experimental results demonstrated that entrapped enzymes were much more stable than free enyzme. The entrapped enzyme could be recycled with retention of 60% activity after 3 cycles. These methodologies can be useful in evaluating other liposomal catalysis operations. PMID:18271982

Molecular modeling of the inhibition of enzyme PLA2 from snake venom by dipyrone and 1-phenyl-3-methyl-5-pyrazolone

NASA Astrophysics Data System (ADS)

Silva, S. L. Da; Comar, M., Jr.; Oliveira, K. M. T.; Chaar, J. S.; Bezerra, E. R. M.; Calgarotto, A. K.; Baldasso, P. A.; Veber, C. L.; Villar, J. A. F. P.; Oliveira, A. R. M.; Marangoni, S.

Phospholipases A2 (PLA2) are enzymes that trigger the degradation cascade of the arachidonic acid, leading to the formation of pro-inflammatory eicosanoids. The selective inhibition of PLA2s is crucial in the search for a more efficient anti-inflammatory drug with fewer side effects than the drugs currently used. Hence, we studied the influences caused by two pyrazolonic inhibitors: dipyrone (DIP) and 1-phenyl-3-methyl-5-pyrazolone (PMP) on the kinetic behavior of PLA2 from Crotalus adamanteus venom. Molecular modeling results, by DFT and MM approaches, showed that DIP is strongly associated to the active site of PLA2 through three hydrogen bonds, whereas PMP is associated to the enzyme just through hydrophobic interactions. In addition, only PMP presents an intramolecular hydrogen bond that make difficult the formation of more efficient interactions with PLA2. These results help in the understanding of the experimental observations. Experimentally, the results showed that PLA2 from C. adamanteus present a typical Michaelian behavior. In addition, the calculated kinetic parameters showed that, in the presence of DIP or PMP, the maximum enzymatic velocity (VMAX) value was kept constant, whereas the Michaelis constant (KM) values increased and the inhibition constant (KI) decreased, indicating competitive inhibition. These results show that the phenyl-pyrazolonic structures might help in the development and design of new drugs able to selectively inhibit PLA2.

A novel bifunctional endo-/exo-type cellulase from an anaerobic ruminal bacterium.

PubMed

Ko, Kyong-Cheol; Han, Yunjon; Choi, Jong Hyun; Kim, Geun-Joong; Lee, Seung-Goo; Song, Jae Jun

2011-03-01

An anaerobic microorganism termed AN-C16-KBRB was isolated from the bovine rumen and demonstrated cellulolytic activity on a NB agar plate containing azo-carboxymethyl cellulose. The 16S rRNA gene of the strain was 98% similar to that of Clostridiaceae bacterium SK082 (AB298754) as the highest homology. A novel celEdx16 gene encoding a bifunctional endo-/exocellulase (CelEdx16) was cloned by the shotgun method from AN-C16-KBRB, and the enzyme was characterized. The celEdx16 gene had an open reading frame of 1,104-base pairs, which encoded 367 amino acids to yield a protein of molecular mass 40.4 kDa. The amino acid sequence was 53% identical to that of an endoglucanase from Clostridium thermocellum. CelEdx16 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The specific endocellulase and exocellulase activities of CelEdx16 were 15.9 and 3.6 x 10⁻² U mg⁻¹, respectively. The Michaelis-Menten constant (K (m) values) and the maximal reaction velocities (V(max) values) of CelEdx16 were 47.1 μM and 9.6 x 10⁻³ μmole min⁻¹ when endocellulase activity was measured and 106.3 μM and 2.1 x 10⁻⁵ μmol min⁻¹ when exocellulase activity was assessed. CelEdx16 was optimally active at pH 5.0 and 40 °C.

Hydrodynamical simulations of coupled and uncoupled quintessence models - I. Halo properties and the cosmic web

NASA Astrophysics Data System (ADS)

Carlesi, Edoardo; Knebe, Alexander; Lewis, Geraint F.; Wales, Scott; Yepes, Gustavo

2014-04-01

We present the results of a series of adiabatic hydrodynamical simulations of several quintessence models (both with a free and an interacting scalar field) in comparison to a standard Λ cold dark matter cosmology. For each we use 2 × 10243 particles in a 250 h-1 Mpc periodic box assuming 7-year Wilkinson Microwave Anisotropy Probe cosmology. In this work we focus on the properties of haloes in the cosmic web at z = 0. The web is classified into voids, sheets, filaments and knots depending on the eigenvalues of the velocity shear tensor, which are an excellent proxy for the underlying overdensity distribution. We find that the properties of objects classified according to their surrounding environment show a substantial dependence on the underlying cosmology; for example, while Vmax shows average deviations of ≈5 per cent across the different models when considering the full halo sample, comparing objects classified according to their environment, the size of the deviation can be as large as 20 per cent. We also find that halo spin parameters are positively correlated to the coupling, whereas halo concentrations show the opposite behaviour. Furthermore, when studying the concentration-mass relation in different environments, we find that in all cosmologies underdense regions have a larger normalization and a shallower slope. While this behaviour is found to characterize all the models, differences in the best-fitting relations are enhanced in (coupled) dark energy models, thus providing a clearer prediction for this class of models.

Aerodynamic characteristics of the ventilated design for flapping wing micro air vehicle.

PubMed

Zhang, G Q; Yu, S C M

2014-01-01

Inspired by superior flight performance of natural flight masters like birds and insects and based on the ventilating flaps that can be opened and closed by the changing air pressure around the wing, a new flapping wing type has been proposed. It is known that the net lift force generated by a solid wing in a flapping cycle is nearly zero. However, for the case of the ventilated wing, results for the net lift force are positive which is due to the effect created by the "ventilation" in reducing negative lift force during the upstroke. The presence of moving flaps can serve as the variable in which, through careful control of the areas, a correlation with the decrease in negative lift can be generated. The corresponding aerodynamic characteristics have been investigated numerically by using different flapping frequencies and forward flight speeds.

Noninvasive stress testing - Methodology for elimination of the phonocardiogram

NASA Technical Reports Server (NTRS)

Spodick, D. H.; Lance, V. Q.

1976-01-01

Measurement by systolic time intervals (STI) of cardiac responses requires extremely careful recording during actual stress test performance. Previous work indicated no significant changes in the pulse transmission time (PTT) during exercise and other challenges. Since external STI depend on the carotid pulse offset by the PTT as an aortic curve equivalent, stable PTT implies that timing of the carotid upstroke and the carotid incisura would respectively track the pre-ejection period and the aortic incisura. In ten subjects, STIs were recorded at supine rest, sitting, standing, during prompt and sustained squatting and during isometric and dynamic exercise. The results demonstrated the tracking of both points. Coefficients of correlation and of determination were uniformly high for all challenges except isometric handgrip (IHG). Since left ventricular ejection time is obtained directly from the pulse curve, with the exception of IHG, STI responses during stress testing can be measured without a phonocardiogram.

Aerodynamic Characteristics of the Ventilated Design for Flapping Wing Micro Air Vehicle

PubMed Central

Zhang, G. Q.; Yu, S. C. M.

2014-01-01

Inspired by superior flight performance of natural flight masters like birds and insects and based on the ventilating flaps that can be opened and closed by the changing air pressure around the wing, a new flapping wing type has been proposed. It is known that the net lift force generated by a solid wing in a flapping cycle is nearly zero. However, for the case of the ventilated wing, results for the net lift force are positive which is due to the effect created by the “ventilation” in reducing negative lift force during the upstroke. The presence of moving flaps can serve as the variable in which, through careful control of the areas, a correlation with the decrease in negative lift can be generated. The corresponding aerodynamic characteristics have been investigated numerically by using different flapping frequencies and forward flight speeds. PMID:24683339

Comparing the aerodynamic forces produced by dragonfly forewings during inverted and non-inverted flight

NASA Astrophysics Data System (ADS)

Shumway, Nathan; Gabryszuk, Mateusz; Laurence, Stuart

2017-11-01

Experiments were conducted with live dragonflies to determine their wing kinematics during free flight. The motion of one forewing in two different tests, one where the dragonfly is inverted, is described using piecewise functions and simulated using the OVERTURNS Reynolds-averaged Navier-Stokes solver that has been used in previous work to determine trim conditions for a fruit fly model. For the inverted dragonfly the upstrokes were significantly longer than the downstrokes, pitching amplitude is lower than that for the right-side up flight and the flap amplitude is larger. Simulations of dragonfly kinematics of a single forewing are presented to determine how the forces differ for a dragonfly flying inverted and a dragonfly flying right-side up. This work was supported by the United States Army Research Laboratory's Micro Autonomous Systems and Technology Collaborative Technology Alliance Project MCE-16-17 1.2.

Kinetics of sulfate and hydrogen uptake by the thermophilic sulfate-reducing bacteria thermodesulfobacterium sp. Strain JSP and thermodesulfovibrio sp. Strain R1Ha3

PubMed

Sonne-Hansen; Westermann; Ahring

1999-03-01

Half-saturation constants (Km), maximum uptake rates (Vmax), and threshold concentrations for sulfate and hydrogen were determined for two thermophilic sulfate-reducing bacteria (SRB) in an incubation system without headspace. Km values determined for the thermophilic SRB were similar to the constants described for mesophilic SRB isolated from environments with low sulfate concentrations.

Kinetics of Sulfate and Hydrogen Uptake by the Thermophilic Sulfate-Reducing Bacteria Thermodesulfobacterium sp. Strain JSP and Thermodesulfovibrio sp. Strain R1Ha3

PubMed Central

Sonne-Hansen, Jacob; Westermann, Peter; Ahring, Birgitte K.

1999-01-01

Half-saturation constants (Km), maximum uptake rates (Vmax), and threshold concentrations for sulfate and hydrogen were determined for two thermophilic sulfate-reducing bacteria (SRB) in an incubation system without headspace. Km values determined for the thermophilic SRB were similar to the constants described for mesophilic SRB isolated from environments with low sulfate concentrations. PMID:10049897

United States Air Force Graduate Student Summer Support Program (1987). Program Technical Report. Volume 2

DTIC Science & Technology

1987-12-01

Metabolism (VMAX) Using Quantitative Structure- Activity Relationships (QSAR) 17 Directed Motion Doppler Shift Effects on Mitric Oxide (0,0) Gamma Band...chemiluminescence values were observed at characteristic times after adding glucose to the disks. We also produced virus-sized nanoparticles (Glucose...These nanoparticles were able to penetrate a .2 um filter,and they retained their enzymatic activity for weeks. They produced 20-fold greater

Effect of common NAT2 variant alleles in the acetylation of the major clonazepam metabolite, 7-aminoclonazepam.

PubMed

Olivera, M; Martínez, C; Gervasini, G; Carrillo, J A; Ramos, S; Benítez, J; García-Martin, E; Agúndez, J A G

2007-01-01

We investigated the role of NAT2 on clonazepam acetylation, using transiently expressed human NAT2 alleles. The NAT25*B and the NAT2*6A variant alleles cause a 20 and 22-fold reduction in the Vmax, respectively. We conclude that NAT2 is responsible for 7-aminoclonazepam acetylation and that NAT2 gene polymorphisms impair such metabolic pathway.

Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertases in barley (Hordeum vulgare) elongating stem tissue

NASA Technical Reports Server (NTRS)

Karuppiah, N.; Vadlamudi, B.; Kaufman, P. B.

1989-01-01

Three different isoforms of invertases have been detected in the developing internodes of barley (Hordeum vulgare). Based on substrate specificities, the isoforms have been identified to be invertases (beta-fructosidases EC 3.2.1.26). The soluble (cytosolic) invertase isoform can be purified to apparent homogeneity by diethylaminoethyl cellulose, Concanavalin-A Sepharose, organo-mercurial Sepharose, and Sephacryl S-300 chromatography. A bound (cell wall) invertase isoform can be released by 1 molar salt and purified further by the same procedures as above except omitting the organo-mercurial Sepharose affinity chromatography step. A third isoform of invertase, which is apparently tightly associated with the cell wall, cannot be isolated yet. The soluble and bound invertase isoforms were purified by factors of 60- and 7-fold, respectively. The native enzymes have an apparent molecular weight of 120 kilodaltons as estimated by gel filtration. They have been identified to be dimers under denaturing and nondenaturing conditions. The soluble enzyme has a pH optimum of 5.5, Km of 12 millimolar, and a Vmax of 80 micromole per minute per milligram of protein compared with cell wall isozyme which has a pH optimum of 4.5, Km of millimolar, and a Vmax of 9 micromole per minute per milligram of protein.

[Specific aspects of thrombocyte system of serotonin in patients with different manifestations of schizoaffective psychosis].

PubMed

Brusov, O S; Dikaia, V I; Zlobina, G P; Faktor, M I; Pavlova, O A; Bologov, P V; Korenev, A N

2000-01-01

45 women with different manifestations of schizoaffective psychosis (SAP) were examined. The diagnosis corresponded to ICD-10 (F25). According to the classification elaborated in Mental Health Research Centre of Russian Academy of Medical Sciences, groups of patients were identified with different variants of the psychoses course: a nuclear SAP type; a borderline SAP variation with phasic-recurrent course; SAP with progredient variation (schizoaffective variation of schizophrenia). The patients were examined both during the attack and remission. A rate of serotonine uptake (Vmax) in blood platelets, a specific imipramine binding (Bmax) and the level of serotonin in blood platelets were evaluated. It was found that dynamics of both Vmax and the level of serotonin in different SAP types were different, that was related to clinical and biological SAP heterogeneity. A tendency to decreasing of serotonin system functional activity was found in progredient SAP variations, especially during the remission, which was of low quality in these cases. On the contrary, in the borderline variations the indices of the decreased function of serotonin system corresponded well to those of acute psychosis. In nuclear type--a type with the most favourable course of psychosis--any significant changes weren't revealed as compared with the normal parameters.

Mode of action of family 10 and 11 endoxylanases on water-unextractable arabinoxylan.

PubMed

Vardakou, Maria; Katapodis, Petros; Samiotaki, Martina; Kekos, Dimitris; Panayotou, George; Christakopoulos, Paul

2003-11-01

Microbial endo-beta-1,4-xylanases (EXs, EC 3.2.1.8) belonging to glycanase families 10 and 11 differ in their action on water-unextractable arabinoxylan (WU-AX). WU-AX was incubated with different levels of a Thermoascus aurantiacus family 10 and a Sporotrichum thermophile family 11 endoxylanases. At 10 g l(-1) arabinoxylan, enzyme concentrations (KE values) needed to obtain half-maximal hydrolysis rates (V(max) values) were 4.4 nM for the xylanase from T. aurantiacus and 7.1 nM for the xylanase from S. thermophile. Determination of Vmax/KE revealed that the family 10 enzyme hydrolysed two times more efficiently WU-AX than the family 11 enzyme. Molecular weights of the products formed were assessed and separation of feruloyl-oligosaccharides was achieved by anion-exchange and size-exclusion chromatography (SEC). The main difference between the feruloylated products by xylanases of family 10 and 11 concerned the length of the products containing feruloyl-arabinosyl substitution. The xylanase from T. aurantiacus liberated from WU-AX a feruloyl arabinoxylodisaccharide (FAX2) as the shortest feruloylated fragment in contrast with the enzyme from S. thermophile, which liberated a feruloyl arabinoxylotrisaccharide (FAX3). These results indicated that different factors govern WU-AX breakdown by the two endoxylanases.

Temperature sensitivity and enzymatic mechanisms of soil organic matter decomposition along an altitudinal gradient on Mount Kilimanjaro

NASA Astrophysics Data System (ADS)

Blagodatskaya, Evgenia; Blagodatsky, Sergey; Khomyakov, Nikita; Myachina, Olga; Kuzyakov, Yakov

2016-02-01

Short-term acceleration of soil organic matter decomposition by increasing temperature conflicts with the thermal adaptation observed in long-term studies. Here we used the altitudinal gradient on Mt. Kilimanjaro to demonstrate the mechanisms of thermal adaptation of extra- and intracellular enzymes that hydrolyze cellulose, chitin and phytate and oxidize monomers (14C-glucose) in warm- and cold-climate soils. We revealed that no response of decomposition rate to temperature occurs because of a cancelling effect consisting in an increase in half-saturation constants (Km), which counteracts the increase in maximal reaction rates (Vmax with temperature). We used the parameters of enzyme kinetics to predict thresholds of substrate concentration (Scrit) below which decomposition rates will be insensitive to global warming. Increasing values of Scrit, and hence stronger canceling effects with increasing altitude on Mt. Kilimanjaro, explained the thermal adaptation of polymer decomposition. The reduction of the temperature sensitivity of Vmax along the altitudinal gradient contributed to thermal adaptation of both polymer and monomer degradation. Extrapolating the altitudinal gradient to the large-scale latitudinal gradient, these results show that the soils of cold climates with stronger and more frequent temperature variation are less sensitive to global warming than soils adapted to high temperatures.

Lipid changes in hepatic microsomes and its relationship to P-nitrophenol glucuronidation in an experimental model of portal hypertension.

PubMed

Ghanem, C; Ghisolfi, C; Marabotto, L; Ouviña, G; Rubio, M; Perazzo, J; Lemberg, A; Bengochea, L

1997-10-01

The liver is responsible for the most important metabolic pathway of non polar compounds. The aim of the present work was to study the p-nitrophenol glucuronidation and its relationship with lipidic composition of microsomal membrane in a model of hepatic portal hypertension and hepatocellular damage induced by monocrotaline. A global increment in liver microsomal phospholipids as well as changes in the phospholipid pattern (phosphatidylethanolamine and sphingomyelin increased up to 156 +/- 13 and 195 +/- 14% respectively) were detected in monocrotaline intoxicated rats when it were compared to control rats. The microsomal cholesterol content showed a decrease in monocrotaline intoxicated rats. (4.1 +/- 0.7 against 6.6 +/- 1.5 micrograms/mg of microsomal protein, in control rats). When p-nitrophenol activity was measured, Km from monocrotaline intoxicated rats was 0.137 mM, and Vmax was 2.9 nmol of p-nitrophenol/mg microsomal protein since in control group Km was 0.322 mM, and Vmax was 4.5 nmol of p-nitrophenol/mg microsomal protein. It is concluded that monocrotaline intoxicated rats showed a different behavior in the kinetics of p-nitrophenol UDP-glucuronyltransferase, as well as a different microsomal lipidic profile, when compared to control group.

omega-Amino acid:pyruvate transaminase from Alcaligenes denitrificans Y2k-2: a new catalyst for kinetic resolution of beta-amino acids and amines.

PubMed

Yun, Hyungdon; Lim, Seongyop; Cho, Byung-Kwan; Kim, Byung-Gee

2004-04-01

Alcaligenes denitrificans Y2k-2 was obtained by selective enrichment followed by screening from soil samples, which showed omega-amino acid:pyruvate transaminase activity, to kinetically resolve aliphatic beta-amino acid, and the corresponding structural gene (aptA) was cloned. The gene was functionally expressed in Escherichia coli BL21 by using an isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible pET expression system (9.6 U/mg), and the recombinant AptA was purified to show a specific activity of 77.2 U/mg for L-beta-amino-n-butyric acid (L-beta-ABA). The enzyme converts various beta-amino acids and amines to the corresponding beta-keto acids and ketones by using pyruvate as an amine acceptor. The apparent K(m) and V(max) for L-beta-ABA were 56 mM and 500 U/mg, respectively, in the presence of 10 mM pyruvate. In the presence of 10 mM L-beta-ABA, the apparent K(m) and V(max) for pyruvate were 11 mM and 370 U/mg, respectively. The enzyme exhibits high stereoselectivity (E > 80) in the kinetic resolution of 50 mM D,L-beta-ABA, producing optically pure D-beta-ABA (99% enantiomeric excess) with 53% conversion.

ω-Amino Acid:Pyruvate Transaminase from Alcaligenes denitrificans Y2k-2: a New Catalyst for Kinetic Resolution of β-Amino Acids and Amines

PubMed Central

Yun, Hyungdon; Lim, Seongyop; Cho, Byung-Kwan; Kim, Byung-Gee

2004-01-01

Alcaligenes denitrificans Y2k-2 was obtained by selective enrichment followed by screening from soil samples, which showed ω-amino acid:pyruvate transaminase activity, to kinetically resolve aliphatic β-amino acid, and the corresponding structural gene (aptA) was cloned. The gene was functionally expressed in Escherichia coli BL21 by using an isopropyl-β-d-thiogalactopyranoside (IPTG)-inducible pET expression system (9.6 U/mg), and the recombinant AptA was purified to show a specific activity of 77.2 U/mg for l-β-amino-n-butyric acid (l-β-ABA). The enzyme converts various β-amino acids and amines to the corresponding β-keto acids and ketones by using pyruvate as an amine acceptor. The apparent Km and Vmax for l-β-ABA were 56 mM and 500 U/mg, respectively, in the presence of 10 mM pyruvate. In the presence of 10 mM l-β-ABA, the apparent Km and Vmax for pyruvate were 11 mM and 370 U/mg, respectively. The enzyme exhibits high stereoselectivity (E > 80) in the kinetic resolution of 50 mM d,l-β-ABA, producing optically pure d-β-ABA (99% enantiomeric excess) with 53% conversion. PMID:15066855

Spectrophotometric determination of pyrrole-like substances in urine of rat and man: an assay for the evaluation of 2,5-hexanedione formed from n-hexane.

PubMed

Kessler, W; Heilmaier, H; Kreuzer, P; Shen, J H; Filser, M; Filser, J G

1990-01-01

Male Wistar rats exposed to atmospheric n-hexane excreted in their urine substances which gave rise to absorption spectra like those of pyrroles after the reaction with Ehrlich's reagent. A simple spectrophotometric assay was developed to determine these "pyrrole-like substances" in urine. Their excretion kinetics were evaluated by exposing rats for 8 h to atmospheric n-hexane concentrations between 50 and 3000 ppm. The dose-response curve revealed saturation kinetics according to Michaelis-Menten, Vmax being 1.12 [delta E526.ml urine/8 h n-hexane exposure] and "Km", the atmospheric n-hexane concentration at Vmax/2, being 250 ppm. The excretion of pyrrole-like substances closely correlated with that of 2,5-hexanedione measured by Fedtke and Bolt (1987). Pyrrole-like substances were also found in the urine of a male volunteer. When exposing the person for 3 h to atmospheric n-hexane at a concentration of 146 ppm (equivalent to 55 ppm/8 h) the excreted amount was twice the background value. Due to the sensitivity of this assay it is possible to determine pyrrole-like substances in urine according to the present German MAK or US TLV conditions for n-hexane (50 ppm/8 h).

A K-band Frequency Agile Microstrip Bandpass Filter using a Thin Film HTS/Ferroelectric/dielectric Multilayer Configuration

NASA Technical Reports Server (NTRS)

Subramanyam, Guru; VanKeuls, Fred; Miranda, Felix A.

1998-01-01

We report on YBa2Cu3O(7-delta) (YBCO) thin film/SrTiO3 (STO) thin film K-band tunable bandpass filters on LaAlO3 (LAO) dielectric substrates. The 2 pole filter has a center frequency of 19 GHz and a 4% bandwidth. Tunability is achieved through the non-linear dc electric field dependence of the relative dielectric constant of STO(epsilon(sub rSTO). A large tunability ((Delta)f/f(sub 0) = (f(sub Vmax) - f(sub 0)/f(sub 0), where f(sub 0) is the center frequency of the filter at no bias and f(sub Vmax) is the center frequency of the filter at the maximum applied bias) of greater than 10% was obtained in YBCO/STO/LAO microstrip bandpass filters operating below 77 K. A center frequency shift of 2.3 GHz (i.e., a tunability factor of approximately 15%) was obtained at a 400 V bipolar dc bias, and 30 K, with minimal degradation in the insertion loss of the filter. This paper addresses design, fabrication and testing of tunable filters based on STO ferroelectric thin films. The performance of the YBCO/STO/LAO filters is compared to that of gold/STO/LAO counterparts.

Hydrolysis is the dominating in vivo metabolism pathway for arctigenin: identification of novel metabolites of arctigenin by LC/MS/MS after oral administration in rats.

PubMed

Gao, Qiong; Zhang, Yufeng; Wo, Siukwan; Zuo, Zhong

2013-04-01

The phenylpropanoid dibenzylbutyrolactone lignan arctigenin, a key component found in Arctium lappa, or burdock, has been reported with a variety of therapeutic effects including anticancer, anti-inflammation, and antivirus effects. Using LC/MS/MS, three novel metabolites of arctigenin, namely, arctigenic acid, arctigenin-4-O'-glucuronide, and 4-O-demethylarctigenin were identified after oral administration of arctigenin in rats for the first time. Another potential metabolite of arctigenin, arctigenin-4'-O-sulfate, was identified in vitro but not in vivo. Structure of arctigenic acid, the major metabolite of arctigenin, was confirmed by 13C-NMR and 1H-NMR. Rapid hydrolysis in plasma was identified as the major metabolic pathway of arctigenin after its oral administration, with Vmax, Km, and Clint in rat plasma determined to be 2.21 ± 0.12 nmol/min/mg, 89.12 ± 9.44 µM, and 24.74 µL/min/mg, respectively. Paraoxonase 1 was further confirmed to be the enzyme responsible for arctigenin hydrolysis, with Vmax, Km, and Clint determined to be 55.39 ± 1.49 nmol/min/mg, 300.3 ± 10.86 µM, and 184.45 µL/min/mg, respectively. Georg Thieme Verlag KG Stuttgart · New York.

The maximal downstroke of epicardial potentials as an index of electrical activity in mouse hearts.

PubMed

Sohn, Kwanghyun; Sachse, Frank B; Moreno, Alonso P; Ershler, Philip R; Wende, Adam R; Abel, E Dale; Punske, Bonnie B

2011-11-01

The maximal upstroke of transmembrane voltage (dV(m)/dt(max)) has been used as an indirect measure of sodium current I(Na) upon activation in cardiac myocytes. However, sodium influx generates not only the upstroke of V(m), but also the downstroke of the extracellular potentials V(e) including epicardial surface potentials V(es). The purpose of this study was to evaluate the magnitude of the maximal downstroke of V(es) (|dV(es)/dt (min)|) as a global index of electrical activation, based on the relationship of dV(m)/dt(max) to I(Na). To fulfill this purpose, we examined |dV(es)/dt(min)| experimentally using isolated perfused mouse hearts and computationally using a 3-D cardiac tissue bidomain model. In experimental studies, a custom-made cylindrical "cage" array with 64 electrodes was slipped over mouse hearts to measure V(es) during hyperkalemia, ischemia, and hypoxia, which are conditions that decrease I(Na). Values of |dV(es)/dt(min)| from each electrode were normalized (|dV(es)/dt (min)|(n)) and averaged (|dV(es)/dt(min)|(na)). Results showed that |dV(es)/dt(min)|(na) decreased during hyperkalemia by 28, 59, and 79% at 8, 10, and 12 mM [K(+)](o), respectively. |dV(es)/dt(min)| also decreased by 54 and 84% 20 min after the onset of ischemia and hypoxia, respectively. In computational studies, |dV(es)/dt(min)| was compared to dV(m)/dt(max) at different levels of the maximum sodium conductance G(Na), extracellular potassium ion concentration [K(+)](o), and intracellular sodium ion concentration [Na(+)](i), which all influence levels of I(Na). Changes in |dV(es)/dt(min)|(n) were similar to dV(m)/dt (max) during alterations of G(Na), [K(+)](o), and [Na(+)](i). Our results demonstrate that |dV(es)/dt(min)|(na) is a robust global index of electrical activation for use in mouse hearts and, similar to dV(m)/dt(max), can be used to probe electrophysiological alterations reliably. The index can be readily measured and evaluated, which makes it attractive for characterization of, for instance, genetically modified mouse hearts and drug effects on cardiac tissue.

Incorporating Love- and Rayleigh-wave magnitudes, unequal earthquake and explosion variance assumptions and interstation complexity for improved event screening

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anderson, Dale N; Bonner, Jessie L; Stroujkova, Anastasia

Our objective is to improve seismic event screening using the properties of surface waves, We are accomplishing this through (1) the development of a Love-wave magnitude formula that is complementary to the Russell (2006) formula for Rayleigh waves and (2) quantifying differences in complexities and magnitude variances for earthquake and explosion-generated surface waves. We have applied the M{sub s} (VMAX) analysis (Bonner et al., 2006) using both Love and Rayleigh waves to events in the Middle East and Korean Peninsula, For the Middle East dataset consisting of approximately 100 events, the Love M{sub s} (VMAX) is greater than the Rayleighmore » M{sub s} (VMAX) estimated for individual stations for the majority of the events and azimuths, with the exception of the measurements for the smaller events from European stations to the northeast. It is unclear whether these smaller events suffer from magnitude bias for the Love waves or whether the paths, which include the Caspian and Mediterranean, have variable attenuation for Love and Rayleigh waves. For the Korean Peninsula, we have estimated Rayleigh- and Love-wave magnitudes for 31 earthquakes and two nuclear explosions, including the 25 May 2009 event. For 25 of the earthquakes, the network-averaged Love-wave magnitude is larger than the Rayleigh-wave estimate. For the 2009 nuclear explosion, the Love-wave M{sub s} (VMAX) was 3.1 while the Rayleigh-wave magnitude was 3.6. We are also utilizing the potential of observed variances in M{sub s} estimates that differ significantly in earthquake and explosion populations. We have considered two possible methods for incorporating unequal variances into the discrimination problem and compared the performance of various approaches on a population of 73 western United States earthquakes and 131 Nevada Test Site explosions. The approach proposes replacing the M{sub s} component by M{sub s} + a* {sigma}, where {sigma} denotes the interstation standard deviation obtained from the stations in the sample that produced the M{sub s} value. We replace the usual linear discriminant a* M{sub s}+b*{sub m{sub b}} with a* M{sub s}+b*{sub m{sub b}} + C*{sigma}. In the second approach, we estimate the optimum hybrid linear-quadratic discriminant function resulting from the unequal variance assumption. We observed slight improvement for the discriminant functions resulting from the theoretical interpretations of the unequal variance function. We have also studied the complexity of the ''magnitude spectra'' at each station. Our hypothesis is that explosion spectra should have fewer focal mechanism-produced complexities in the magnitude spectra than earthquakes. We have developed an intrastation ''complexity'' metric {Delta}M{sub s}, where {Delta}M{sub s} = M{sub s}(i)-M{sub s}(i+1) at periods, i, which are between 9 and 25 seconds. The complexity by itself has discriminating power but does not add substantially to the conditional hybrid discriminant that incorporates the differing spreads of the earthquake and explosion standard deviations.« less

Kinetic mechanism of Toxoplasma gondii adenosine kinase and the highly efficient utilization of adenosine

PubMed Central

Naguib, Fardos N. M.; Rais, Reem H.; Al Safarjalani, Omar N.; el Kouni, Mahmoud H.

2015-01-01

Toxoplasma gondii has an extraordinarily ability to utilize adenosine (Ado) as the primary source of all necessary purines in this parasite which lacks de novo purine biosynthesis. The activity of T. gondii adenosine kinase (TgAK, EC 2.7.1.20) is responsible for this efficient salvage of Ado in T. gondii. To fully understand this remarkable efficiency of TgAK in the utilization of Ado, complete kinetic parameters of this enzyme are necessary. Initial velocity and product inhibition studies of TgAK demonstrated that the basic mechanism of this enzyme is a hybrid random bi-uni ping-pong uni-bi. Initial velocity studies showed an intersecting pattern, consistent with substrate-enzyme-co-substrate complex formation and a binding pattern indicating that binding of the substrate interferes with the binding of the co-substrate and vice versa. Estimated kinetic parameters were KAdo = 0.002 ± 0.0002 mM, KATP = 0.05 ± 0.008 mM, and Vmax = 920 ± 35 μmol/min/mg protein. Ado exhibited substrate inhibition suggesting the presence of more than one binding site for Ado on the enzyme. ATP relieved substrate inhibition by Ado. Thus, Ado also binds to the ATP binding site. AMP was competitive with ATP, inferring that AMP binds to the same site as ATP. AMP, ADP and ATP were non-competitive with Ado, therefore, none of these nucleotides binds to the Ado binding site. Combining ATP with ADP was additive. Therefore, the binding of either ATP or ADP does not interfere with the binding of the other. It is concluded that for every ATP consumed, TgAK generates three new AMPs. These findings along with the fact that a wide range of nucleoside 5′-mono, di, and triphosphates could substitute for ATP as phosphate donors in this reaction may explain the efficient and central role played by TgAK in the utilization of Ado as the major source from which all other purines can be synthesized in T. gondii. PMID:26112826

Comparison of the UDP-N-Acetylmuramate:l-Alanine Ligase Enzymes from Mycobacterium tuberculosis and Mycobacterium leprae

PubMed Central

Mahapatra, Sebabrata; Crick, Dean C.; Brennan, Patrick J.

2000-01-01

In the peptidoglycan of Mycobacterium leprae, l-alanine of the side chain is replaced by glycine. When expressed in Escherichia coli, MurC (UDP-N-acetyl-muramate:l-alanine ligase) of M. leprae showed Km and Vmax for l-alanine and glycine similar to those of Mycobacterium tuberculosis MurC, suggesting that another explanation should be sought for the presence of glycine. PMID:11073931

VizieR Online Data Catalog: Luminosity and redshift of galaxies from WISE/SDSS (Toba+, 2014)

NASA Astrophysics Data System (ADS)

Toba, Y.; Oyabu, S.; Matsuhara, H.; Malkan, M. A.; Gandhi, P.; Nakagawa, T.; Isobe, N.; Shirahata, M.; Oi, N.; Ohyama, Y.; Takita, S.; Yamauchi, C.; Yano, K.

2017-07-01

We selected 12 and 22 um flux-limited galaxies based on the WISE (Cat. II/311) and SDSS (Cat. II/294) catalogs, and these galaxies were then classified into five types according to their optical spectroscopic information in the SDSS catalog. For spectroscopically classified galaxies, we constructed the luminosity functions using the 1/Vmax method, considering the detection limit of the WISE and SDSS catalogs. (1 data file).

14 CFR Appendix D to Part 23 - Wheel Spin-Up and Spring-Back Loads

Code of Federal Regulations, 2014 CFR

2014-01-01

... (0.80 may be used); F Vmax=maximum vertical force on wheel (pounds)=n j W e, where W e and n j are... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Wheel Spin-Up and Spring-Back Loads D.... D Appendix D to Part 23—Wheel Spin-Up and Spring-Back Loads D23.1 Wheel spin-up loads. (a) The...

14 CFR Appendix D to Part 23 - Wheel Spin-Up and Spring-Back Loads

Code of Federal Regulations, 2013 CFR

2013-01-01

... (0.80 may be used); F Vmax=maximum vertical force on wheel (pounds)=n j W e, where W e and n j are... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Wheel Spin-Up and Spring-Back Loads D.... D Appendix D to Part 23—Wheel Spin-Up and Spring-Back Loads D23.1 Wheel spin-up loads. (a) The...

14 CFR Appendix D to Part 23 - Wheel Spin-Up and Spring-Back Loads

Code of Federal Regulations, 2011 CFR

2011-01-01

... (0.80 may be used); F Vmax=maximum vertical force on wheel (pounds)=n j W e, where W e and n j are... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Wheel Spin-Up and Spring-Back Loads D.... D Appendix D to Part 23—Wheel Spin-Up and Spring-Back Loads D23.1 Wheel spin-up loads. (a) The...

14 CFR Appendix D to Part 23 - Wheel Spin-Up and Spring-Back Loads

Code of Federal Regulations, 2010 CFR

2010-01-01

... (0.80 may be used); F Vmax=maximum vertical force on wheel (pounds)=n j W e, where W e and n j are... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Wheel Spin-Up and Spring-Back Loads D.... D Appendix D to Part 23—Wheel Spin-Up and Spring-Back Loads D23.1 Wheel spin-up loads. (a) The...

14 CFR Appendix D to Part 23 - Wheel Spin-Up and Spring-Back Loads

Code of Federal Regulations, 2012 CFR

2012-01-01

... (0.80 may be used); F Vmax=maximum vertical force on wheel (pounds)=n j W e, where W e and n j are... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Wheel Spin-Up and Spring-Back Loads D.... D Appendix D to Part 23—Wheel Spin-Up and Spring-Back Loads D23.1 Wheel spin-up loads. (a) The...

Organic Nitrogen Utilization by Phytoplankton: The Role of Cell-Surface Deaminases

DTIC Science & Technology

1989-06-01

Pleurochrysis carterae (Coccoll-N) is a coccolithless clone isolated by the authors from Coccoll. Emiliania huxleyi (12-1) was isolated from the Sargasso Sea...another coccolithophorid, Emiliania huxleyi from the Sargasso Sea (now 12-1, CCMP) for L-amino acid oxidase activity. No activity was found under log...acid oxidase regulation. Saturated oxidase rate constants (Vmax) are shown for Pleurochrysis isolates and one Emiliania huxleyi isolate (12-1). Nlim

Benzydamine N-oxidation as an index reaction reflecting FMO activity in human liver microsomes and impact of FMO3 polymorphisms on enzyme activity

PubMed Central

Störmer, Elke; Roots, Ivar; Brockmöller, Jürgen

2000-01-01

Aims The role of flavin containing monooxygenases (FMO) on the disposition of many drugs has been insufficiently explored. In vitro and in vivo tests are required to study FMO activity in humans. Benzydamine (BZD) N-oxidation was evaluated as an index reaction for FMO as was the impact of genetic polymorphisms of FMO3 on activity. Methods BZD was incubated with human liver microsomes (HLM) and recombinant enzymes. Human liver samples were genotyped using PCR-RFLP. Results BZD N-oxide formation rates in HLM followed Michaelis-Menten kinetics (mean Km = 64.0 μm, mean Vmax = 6.9 nmol mg−1 protein min−1; n = 35). N-benzylimidazole, a nonspecific CYP inhibitor, and various CYP isoform selective inhibitors did not affect BZD N-oxidation. In contrast, formation of BZD N-oxide was almost abolished by heat treatment of microsomes in the absence of NADPH and strongly inhibited by methimazole, a competitive FMO inhibitor. Recombinant FMO3 and FMO1 (which is not expressed in human liver), but not FMO5, showed BZD N-oxidase activity. Respective Km values for FMO3 and FMO1 were 40.4 μm and 23.6 μm, and respective Vmax values for FMO3 and FMO1 were 29.1 and 40.8 nmol mg−1 protein min−1. Human liver samples (n = 35) were analysed for six known FMO3 polymorphisms. The variants I66M, P135L and E305X were not detected. Samples homozygous for the K158 variant showed significantly reduced vmax values (median 2.7 nmol mg−1 protein min−1) compared to the carriers of at least one wild type allele (median 6.2 nmol mg−1 protein min−1) (P<0.05, Mann–Whitney- U-test). The V257M and E308G substitutions had no effect on enzyme activity. Conclusions BZD N-oxidation in human liver is mainly catalysed by FMO3 and enzyme activity is affected by FMO3 genotype. BZD may be used as a model substrate for human liver FMO3 activity in vitro and may be further developed as an in vivo probe reflecting FMO3 activity. PMID:11136294

Number of Players and Relative Pitch Area per Player: Comparing Their Influence on Heart Rate and Physical Demands in Under-12 and Under-13 Football Players

PubMed Central

Castellano, Julen; Puente, Asier; Echeazarra, Ibon; Usabiaga, Oidui; Casamichana, David

2016-01-01

The aim of the present study is to analyse the influence of different large-sided games (LSGs) on the physical and physiological variables in under-12s (U12) and -13s (U13) soccer players. The effects of the combination of different number of players per team, 7, 9, and 11 (P7, P9, and P11, respectively) with three relative pitch areas, 100, 200, and 300 m2 (A100, A200, and A300, respectively), were analysed in this study. The variables analysed were: 1) global indicator such as total distance (TD); work:rest ratio (W:R); player-load (PL) and maximal speed (Vmax); 2) heart rate (HR) mean and time spent in different intensity zones of HR (<75%, 75–84%, 84–90% and >90%), and; 3) five absolute (<8, 8–13, 13–16 and >16 Km h-1) and three relative speed categories (<40%, 40–60% and >60% Vmax). The results support the theory that a change in format (player number and pitch dimensions) affects no similarly in the two players categories. Although it can seem that U13 players are more demanded in this kind of LSG, when the work load is assessed from a relative point of view, great pitch dimensions and/or high number of player per team are involved in the training task to the U12 players. The results of this study could alert to the coaches to avoid some types of LSGs for the U12 players such as: P11 played in A100, A200 or A300, P9 played in A200 or A300 and P7 played in A300 due to that U13>U12 in several physical and physiological variables (W:R, time spent in 84–90%HRmax, distance in 8–13 and 13–16 Km h-1 and time spent in 40–60%Vmax). These results may help youth soccer coaches to plan the progressive introduction of LSGs so that task demands are adapted to the physiological and physical development of participants. PMID:26752422

Electrophysiological properties of myocytes isolated from the mouse atrioventricular node: L-type ICa, IKr, If, and Na-Ca exchange

PubMed Central

Choisy, Stéphanie C; Cheng, Hongwei; Orchard, Clive H; James, Andrew F; Hancox, Jules C

2015-01-01

The atrioventricular node (AVN) is a key component of the cardiac pacemaker-conduction system. This study investigated the electrophysiology of cells isolated from the AVN region of adult mouse hearts, and compared murine ionic current magnitude with that of cells from the more extensively studied rabbit AVN. Whole-cell patch-clamp recordings of ionic currents, and perforated-patch recordings of action potentials (APs), were made at 35–37°C. Hyperpolarizing voltage commands from −40 mV elicited a Ba2+-sensitive inward rectifier current that was small at diastolic potentials. Some cells (Type 1; 33.4 ± 2.2 pF; n = 19) lacked the pacemaker current, If, whilst others (Type 2; 34.2 ± 1.5 pF; n = 21) exhibited a clear If, which was larger than in rabbit AVN cells. On depolarization from −40 mV L-type Ca2+ current, ICa,L, was elicited with a half maximal activation voltage (V0.5) of −7.6 ± 1.2 mV (n = 24). ICa,L density was smaller than in rabbit AVN cells. Rapid delayed rectifier (IKr) tail currents sensitive to E-4031 (5 μmol/L) were observed on repolarization to −40 mV, with an activation V0.5 of −10.7 ± 4.7 mV (n = 8). The IKr magnitude was similar in mouse and rabbit AVN. Under Na-Ca exchange selective conditions, mouse AVN cells exhibited 5 mmol/L Ni-sensitive exchange current that was inwardly directed negative to the holding potential (−40 mV). Spontaneous APs (5.2 ± 0.5 sec−1; n = 6) exhibited an upstroke velocity of 37.7 ± 16.2 V/s and ceased following inhibition of sarcoplasmic reticulum Ca2+ release by 1 μmol/L ryanodine, implicating intracellular Ca2+ cycling in murine AVN cell electrogenesis. PMID:26607172

Cross-correlation analysis of pulse wave propagation in arteries: in vitro validation and in vivo feasibility

NASA Astrophysics Data System (ADS)

Nauleau, Pierre; Apostolakis, Iason; McGarry, Matthew; Konofagou, Elisa

2018-06-01

The stiffness of the arteries is known to be an indicator of the progression of various cardiovascular diseases. Clinically, the pulse wave velocity (PWV) is used as a surrogate for arterial stiffness. Pulse wave imaging (PWI) is a non-invasive, ultrasound-based imaging technique capable of mapping the motion of the vessel walls, allowing the local assessment of arterial properties. Conventionally, a distinctive feature of the displacement wave (e.g. the 50% upstroke) is tracked across the map to estimate the PWV. However, the presence of reflections, such as those generated at the carotid bifurcation, can bias the PWV estimation. In this paper, we propose a two-step cross-correlation based method to characterize arteries using the information available in the PWI spatio-temporal map. First, the area under the cross-correlation curve is proposed as an index for locating the regions of different properties. Second, a local peak of the cross-correlation function is tracked to obtain a less biased estimate of the PWV. Three series of experiments were conducted in phantoms to evaluate the capabilities of the proposed method compared with the conventional method. In the ideal case of a homogeneous phantom, the two methods performed similarly and correctly estimated the PWV. In the presence of reflections, the proposed method provided a more accurate estimate than conventional processing: e.g. for the soft phantom, biases of  ‑0.27 and ‑0.71 m · s–1 were observed. In a third series of experiments, the correlation-based method was able to locate two regions of different properties with an error smaller than 1 mm. It also provided more accurate PWV estimates than conventional processing (biases:  ‑0.12 versus ‑0.26 m · s–1). Finally, the in vivo feasibility of the proposed method was demonstrated in eleven healthy subjects. The results indicate that the correlation-based method might be less precise in vivo but more accurate than the conventional method.

Developmental lead exposure causes startle response deficits in zebrafish.

PubMed

Rice, Clinton; Ghorai, Jugal K; Zalewski, Kathryn; Weber, Daniel N

2011-10-01

Lead (Pb(2+)) exposure continues to be an important concern for fish populations. Research is required to assess the long-term behavioral effects of low-level concentrations of Pb(2+) and the physiological mechanisms that control those behaviors. Newly fertilized zebrafish embryos (<2h post fertilization; hpf) were exposed to one of three concentrations of lead (as PbCl(2)): 0, 10, or 30 nM until 24 hpf. (1) Response to a mechanosensory stimulus: Individual larvae (168 hpf) were tested for response to a directional, mechanical stimulus. The tap frequency was adjusted to either 1 or 4 taps/s. Startle response was recorded at 1000 fps. Larvae responded in a concentration-dependent pattern for latency to reaction, maximum turn velocity, time to reach V(max) and escape time. With increasing exposure concentrations, a larger number of larvae failed to respond to even the initial tap and, for those that did respond, ceased responding earlier than control larvae. These differences were more pronounced at a frequency of 4 taps/s. (2) Response to a visual stimulus: Fish, exposed as embryos (2-24 hpf) to Pb(2+) (0-10 μM) were tested as adults under low light conditions (≈ 60 μW/m(2)) for visual responses to a rotating black bar. Visual responses were significantly degraded at Pb(2+) concentrations of 30 nM. These data suggest that zebrafish are viable models for short- and long-term sensorimotor deficits induced by acute, low-level developmental Pb(2+) exposures. Copyright © 2011 Elsevier B.V. All rights reserved.

Seismic displacement of gently-sloping coastal and marine sediment under multidirectional earthquake loading

USGS Publications Warehouse

Kayen, Robert E.

2017-01-01

Gentle sediment-laden slopes are typical of the onshore coastal zone and offshore continental shelf and slope. Coastal sediment are commonly young weakly consolidated materials that are well stratified, have low strength, and can mobilize shear displacements at low levels of stress. Seismically-driven plastic displacements of these sediment pose a hazard to coastal cities, buried onshore utilities, and offshore infrastructure like harbor protection and outfalls. One-dimensional rigid downslope-directed Newmark sliding block analyses have been used to predict earthquake deformations generally on steeper slopes that are modeled as frictional materials. This study probes the effect of multidirectional earthquake motions on inertial displacements of gently sloping ground of the coastal and offshore condition where soft-compliant soil is expected. Toward that objective, this investigation seeks to understand the effect on Newmark-type displacements of [1] multidirectional earthquake shaking and [2] soil compliance. In order to model multidirectional effects, the earthquake motions are rotated into the local slope strike- and dip-components. On gently sloping ground, including the strike component of motion always results in a larger and more accurate shear stress vector. Strike motions are found to contribute to downslope deformations on any declivity. Compliant response of the soil mass also influences the plastic displacements. The magnitude of seismic displacements can be estimated with a simplified model using only the estimated soil yield-acceleration (ky) and the peak ground velocity (Vmax) of the earthquake motions. Compliance effects can be effectively mapped using the concept of Plastic Displacement Response Spectra (PDRS).

Effects of Ethanol and Other Alkanols on Transport of Acetic Acid in Saccharomyces cerevisiae

PubMed Central

Casal, Margarida; Cardoso, Helena; Leão, Cecília

1998-01-01

In glucose-grown cells of Saccharomyces cerevisiae IGC 4072, acetic acid enters only by simple diffusion of the undissociated acid. In these cells, ethanol and other alkanols enhanced the passive influx of labelled acetic acid. The influx of the acid followed first-order kinetics with a rate constant that increased exponentially with the alcohol concentration, and an exponential enhancement constant for each alkanol was estimated. The intracellular concentration of labelled acetic acid was also enhanced by alkanols, and the effect increased exponentially with alcohol concentration. Acetic acid is transported across the plasma membrane of acetic acid-, lactic acid-, and ethanol-grown cells by acetate-proton symports. We found that in these cells ethanol and butanol inhibited the transport of labelled acetic acid in a noncompetitive way; the maximum transport velocity decreased with alcohol concentration, while the affinity of the system for acetate was not significantly affected by the alcohol. Semilog plots of Vmax versus alcohol concentration yielded straight lines with negative slopes from which estimates of the inhibition constant for each alkanol could be obtained. The intracellular concentration of labelled acid was significantly reduced in the presence of ethanol or butanol, and the effect increased with the alcohol concentration. We postulate that the absence of an operational carrier for acetate in glucose-grown cells of S. cerevisiae, combined with the relatively high permeability of the plasma membrane for the undissociated acid and the inability of the organism to metabolize acetic acid, could be one of the reasons why this species exhibits low tolerance to acidic environments containing ethanol. PMID:9464405

Absolute Bioavailability and Pharmacokinetics of Linezolid in Hospitalized Patients Given Enteral Feedings

PubMed Central

Beringer, Paul; Nguyen, Megan; Hoem, Nils; Louie, Stan; Gill, Mark; Gurevitch, Michael; Wong-Beringer, Annie

2005-01-01

Linezolid is a new antimicrobial agent effective against drug-resistant gram-positive pathogens which are common causes of infections in hospitalized patients. Many such patients rely on the intravenous or enteral route for nutrition and drug administration. Therefore, the bioavailability of linezolid administered enterally in the presence of enteral feedings in hospitalized patients was examined. Eighteen subjects were assessed in a randomized single-dose crossover study; 12 received continuous enteral feedings, while 6 did not (controls). Both groups received linezolid 600 mg intravenously and orally (control) or enterally, with the alternate route of administration separated by a 24-h washout period. Pharmacokinetic parameters derived from noncompartmental and compartmental analysis incorporating linear and nonlinear elimination pathways were compared between groups: F, Ka, Vs, K23, K32, Vmax, Km, and K20 (bioavailability, absorption rate constant, volume of central compartment normalized to body weight, intercompartmental rate constants, maximum velocity, Michaelis-Menten constant, and elimination rate constant, respectively). Pharmacokinetic (PK) data were available from 17 patients. The linezolid oral suspension was rapidly and completely absorbed by either the oral or enteral route of administration. Bioavailability was unaltered in the presence of enteral feedings. PK estimates remain similar regardless of the model applied. At the therapeutic dose used, only slight nonlinearity in elimination was observed. A linezolid oral suspension may be administered via the enteral route to hospitalized patients without compromise in its excellent bioavailability and rapid rate of absorption. Compartmental pharmacokinetic analysis offers a more flexible study application, since bioavailability (F) can be estimated directly with intermixed intravenous/oral doses without a need for a washout period. PMID:16127039

Isolation and characterization of mutated alcohol oxidases from the yeast Hansenula polymorpha with decreased affinity toward substrates and their use as selective elements of an amperometric biosensor

PubMed Central

Dmytruk, Kostyantyn V; Smutok, Oleh V; Ryabova, Olena B; Gayda, Galyna Z; Sibirny, Volodymyr A; Schuhmann, Wolfgang; Gonchar, Mykhailo V; Sibirny, Andriy A

2007-01-01

Background Accurate, rapid, and economic on-line analysis of ethanol is very desirable. However, available biosensors achieve saturation at very low ethanol concentrations and thus demand the time and labour consuming procedure of sample dilution. Results Hansenula polymorpha (Pichia angusta) mutant strains resistant to allyl alcohol in methanol medium were selected. Such strains possessed decreased affinity of alcohol oxidase (AOX) towards methanol: the KM values for AOX of wild type and mutant strains CA2 and CA4 are shown to be 0.62, 2.48 and 1.10 mM, respectively, whereas Vmax values are increased or remain unaffected. The mutant AOX alleles from H. polymorpha mutants CA2 and CA4 were isolated and sequenced. Several point mutations in the AOX gene, mostly different between the two mutant alleles, have been identified. Mutant AOX forms were isolated and purified, and some of their biochemical properties were studied. An amperometric biosensor based on the mutated form of AOX from the strain CA2 was constructed and revealed an extended linear response to the target analytes, ethanol and formaldehyde, as compared to the sensor based on the native AOX. Conclusion The described selection methodology opens up the possibility of isolating modified forms of AOX with further decreased affinity toward substrates without reduction of the maximal velocity of reaction. It can help in creation of improved ethanol biosensors with a prolonged linear response towards ethanol in real samples of wines, beers or fermentation liquids. PMID:17567895

Conversion of taurine into N-chlorotaurine (taurine chloramine) and sulphoacetaldehyde in response to oxidative stress.

PubMed

Cunningham, C; Tipton, K F; Dixon, H B

1998-03-01

N-Chlorotaurine (taurine chloramine), formed by treating taurine with hypochlorous acid, was shown to decompose to sulphoacetaldehyde with a first-order rate constant of 9.9+/-0.5 x 10(-4).h-1 at 37 degrees C in 0.1 M phosphate buffer, pH 7.4. Rat liver homogenates accelerated this decay in a process that was proportional to tissue-protein concentration and saturable, with maximum velocity (Vmax) and Km values of 0.28+/-0.01 nmol/min per mg of protein and 37+/-9 microM respectively. This activity was found to be lost on heat denaturation, but retained after dialysis. There was no detectable formation of sulphoacetaldehyde when taurine itself was incubated with the tissue homogenates under the same conditions. Activation of human neutrophils (1.67 x 10(6) cells/ml) with latex beads resulted in a respiratory burst of oxygen-radical production, the products of which were partially sequestered by 12.5 mM taurine. Under these conditions sulphoacetaldehyde was generated at a constant rate of 637+/-18 pmol/h per ml for over 7 h. A non-activated neutrophil suspension contained constant levels of 1.42+/-0.02 nmol/ml sulphoacetaldehyde, as did activated cells incubated in the absence of taurine, a basal level which may indicate a steady turnover of taurine in these cells. Such formation of chlorotaurine and its decay to the aldehyde may be the first steps in the metabolism of taurine to isethionate (2-hydroxyethanesulphonate) that has been demonstrated by various authors to occur in vivo.

Promotion of beta-glucan synthase activity in corn microsomal membranes by calcium and protein phosphorylation

NASA Technical Reports Server (NTRS)

Paliyath, G.; Poovaiah, B. W.

1988-01-01

Regulation of the activity of beta-glucan synthase was studied using microsomal preparations from corn coleoptiles. The specific activity as measured by the incorporation of glucose from uridine diphospho-D-[U-14C]glucose varied between 5 to 15 pmol (mg protein)-1 min-1. Calcium promoted beta-glucan synthase activity and the promotion was observed at free calcium concentrations as low as 1 micromole. Kinetic analysis of substrate-velocity curve showed an apparent Km of 1.92 x 10(-4) M for UDPG. Calcium increased the Vmax from 5.88 x 10(-7) mol liter-1 min-1 in the absence of calcium to 9.52 x 10(-7) mol liter-1 min-1 and 1.66 x 10(-6) mol liter-1 min-1 in the presence of 0.5 mM and 1 mM calcium, respectively. The Km values remained the same under these conditions. Addition of ATP further increased the activity above the calcium-promoted level. Sodium fluoride, a phosphoprotein phosphatase inhibitor, promoted glucan synthase activity indicating that phosphorylation and dephosphorylation are involved in the regulation of the enzyme activity. Increasing the concentration of sodium fluoride from 0.25 mM to 10 mM increased glucan synthase activity five-fold over the + calcium + ATP control. Phosphorylation of membrane proteins also showed a similar increase under these conditions. Calmodulin, in the presence of calcium and ATP stimulated glucan synthase activity substantially, indicating that calmodulin could be involved in the calcium-dependent phosphorylation and promotion of beta-glucan synthase activity. The role of calcium in mediating auxin action is discussed.

Effects of high-intensity interval training and moderate-intensity continuous training on glycaemic control and skeletal muscle mitochondrial function in db/db mice.

PubMed

Chavanelle, Vivien; Boisseau, Nathalie; Otero, Yolanda F; Combaret, Lydie; Dardevet, Dominique; Montaurier, Christophe; Delcros, Geoffrey; Peltier, Sébastien L; Sirvent, Pascal

2017-03-16

Physical activity is known as an effective strategy for prevention and treatment of Type 2 Diabetes. The aim of this work was to compare the effects of a traditional Moderate Intensity Continuous Training (MICT) with a High Intensity Interval Training (HIIT) on glucose metabolism and mitochondrial function in diabetic mice. Diabetic db/db male mice (N = 25) aged 6 weeks were subdivided into MICT, HIIT or control (CON) group. Animals in the training groups ran on a treadmill 5 days/week during 10 weeks. MICT group ran for 80 min (0° slope) at 50-60% of maximal speed (Vmax) reached during an incremental test. HIIT group ran thirteen times 4 minutes (20° slope) at 85-90% of Vmax separated by 2-min-rest periods. HIIT lowered fasting glycaemia and HbA1c compared with CON group (p < 0.05). In all mitochondrial function markers assessed, no differences were noted between the three groups except for total amount of electron transport chain proteins, slightly increased in the HIIT group vs CON. Western blot analysis revealed a significant increase of muscle Glut4 content (about 2 fold) and higher insulin-stimulated Akt phosphorylation ratios in HIIT group. HIIT seems to improve glucose metabolism more efficiently than MICT in diabetic mice by mechanisms independent of mitochondrial adaptations.

Neuraminidase activity of blue eye disease porcine rubulavirus: Specificity, affinity and inhibition studies.

PubMed

Santos-López, Gerardo; Borraz-Argüello, María T; Márquez-Domínguez, Luis; Flores-Alonso, Juan Carlos; Ramírez-Mendoza, Humberto; Priem, Bernard; Fort, Sébastien; Vallejo-Ruiz, Verónica; Reyes-Leyva, Julio; Herrera-Camacho, Irma

2017-10-01

Porcine rubulavirus (PorPV), also known as La Piedad Michoacan Virus (LPMV) causes encephalitis and reproductive failure in newborn and adult pigs, respectively. The hemagglutinin-neuraminidase (HN) glycoprotein is the most exposed and antigenic of the virus proteins. HN plays central roles in PorPV infection; i.e., it recognizes sialic acid-containing cell receptors that mediate virus attachment and penetration; in addition, its neuraminidase (sialic acid releasing) activity has been proposed as a virulence factor. This work describes the purification and characterization of PorPV HN protein (isolate PAC1). The specificity of neuraminidase is restricted to sialyl(α2,3)lactose (3SL). HN showed typical Michaelis-Menten kinetics with fetuin as substrate (km=0.029μM, Vmax=522.8nmolmin -1 mg -1 ). When 3SL was used as substrate, typical cooperative kinetics were found (S 50 =0.15μM, Vmax=154.3nmolmin -1 mg -1 ). The influenza inhibitor zanamivir inhibited the PorPV neuraminidase with IC 50 of 0.24μM. PorPV neuraminidase was activated by Ca 2+ and inhibited by nucleoside triphosphates with the level of inhibition depending on phosphorylation level. The present results open possibilities to study the role of neuraminidase in the pathogenicity of PorPV infection and its potential inhibitors. Copyright © 2017 Elsevier Ltd. All rights reserved.

A Search for Low-Luminosity BL Lacertae Objects

NASA Astrophysics Data System (ADS)

Rector, Travis A.; Stocke, John T.; Perlman, Eric S.

1999-05-01

Many properties of BL Lacs have become explicable in terms of the ``relativistic beaming'' hypothesis, whereby BL Lacs are FR 1 radio galaxies viewed nearly along the jet axis. However, a possible problem with this model is that a transition population between beamed BL Lacs and unbeamed FR 1 galaxies has not been detected. A transition population of ``low-luminosity BL Lacs'' was predicted to exist in abundance in X-ray-selected samples such as the Einstein Extended Medium Sensitivity Survey (EMSS) by Browne & Marcha. However, these BL Lacs may have been misidentified as clusters of galaxies. We have conducted a search for such objects in the EMSS with the ROSAT High-Resolution Imager (HRI) here we present ROSAT HRI images, optical spectra, and VLA radio maps for a small number of BL Lacs that were previously misidentified in the EMSS catalog as clusters of galaxies. While these objects are slightly lower in luminosity than other EMSS BL Lacs, their properties are too similar to the other BL Lacs in the EMSS sample to ``bridge the gap'' between BL Lacs and FR 1 radio galaxies. Also, the number of new BL Lacs found is too low to alter significantly the X-ray luminosity function or value for the X-ray-selected EMSS BL Lac sample. Thus, these observations do not explain fully the discrepancy between the X-ray- and radio-selected BL Lac samples.

Individual differences in impulsive action and dopamine transporter function in rat orbitofrontal cortex.

PubMed

Yates, J R; Darna, M; Beckmann, J S; Dwoskin, L P; Bardo, M T

2016-01-28

Impulsivity, which can be subdivided into impulsive action and impulsive choice, is implicated as a factor underlying drug abuse vulnerability. Although previous research has shown that dopamine (DA) systems in prefrontal cortex are involved in impulsivity and substance abuse, it is not known if inherent variation in DA transporter (DAT) function contributes to impulsivity. The current study determined if individual differences in either impulsive action or impulsive choice are related to DAT function in orbitofrontal (OFC) and/or medial prefrontal cortex (mPFC). Rats were first tested both for impulsive action in a cued go/no-go task and for impulsive choice in a delay-discounting task. Following behavioral evaluation, in vitro [(3)H]DA uptake assays were performed in OFC and mPFC isolated from individual rats. Vmax in OFC, but not mPFC, was correlated with performance in the cued go/no-go task, with decreased OFC DAT function being associated with high impulsive action. In contrast, Vmax in OFC and mPFC was not correlated with performance in the delay-discounting task. The current results demonstrate that impulsive behavior in cued go/no-go performance is associated with decreased DAT function in OFC, suggesting that hyperdopaminergic tone in this prefrontal subregion mediates, at least in part, increased impulsive action. Copyright © 2015 IBRO. Published by Elsevier Ltd. All rights reserved.

A Method for Multiplexed Measurement of Mitochondrial Pyruvate Carrier Activity*

PubMed Central

Gray, Lawrence R.; Rauckhorst, Adam J.; Taylor, Eric B.

2016-01-01

The discovery that the MPC1 and MPC2 genes encode the protein components of the mitochondrial pyruvate carrier (MPC) has invigorated studies of mitochondrial pyruvate transport and its regulation in normal and disease states. Indeed, recent reports have demonstrated MPC involvement in the control of cell fate in cancer and gluconeogenesis in models of type 2 diabetes. Biochemical measurements of MPC activity are foundational for understanding the role of pyruvate transport in health and disease. We developed a 96-well scaled method of [14C]pyruvate uptake that markedly decreases sample requirements and increases throughput relative to previous techniques. This method was applied to determine the mouse liver MPC Km (28.0 ± 3.9 μm) and Vmax (1.08 ± 0.05 nmol/min/mg), which have not previously been reported. Km and Vmax of the rat liver MPC were found to be 71.2 ± 17 μm and 1.42 ± 0.14 nmol/min/mg, respectively. Additionally, we performed parallel pyruvate uptake and oxidation experiments with the same biological samples and show differential results in response to fasting, demonstrating the continued importance of a direct MPC activity assay. We expect this method will be of value for understanding the contribution of the MPC activity to health and disease states where pyruvate metabolism is expected to play a prominent role. PMID:26823462

Purification and characterization of a p-coumarate decarboxylase and a vinylphenol reductase from Brettanomyces bruxellensis.

PubMed

Godoy, Liliana; Martínez, Claudio; Carrasco, Nelson; Ganga, María Angélica

2008-09-30

The presence of Brettanomyces bruxellensis has been correlated with an increase of phenolic aromas in wine. The production of these aromas results from the metabolization of cinnamic acids, present in the wine, to their ethyl derivatives. Hence, the participation of two enzymes has been proposed: a p-coumarate decarboxylase (CD) and a vinylphenol reductase (VR). Both enzymes were purified and characterized from B. bruxellensis. In denaturing conditions, the CD enzyme had a molecular mass of 21 kDa, while in native conditions its mass was 41 kDa. The optimal activity was obtained at a temperature of 40 degrees C and a pH of 6.0. For p-coumaric acid, the Km value and Vmax were 1.22+/-0.08 mM and 98+/-0.15 micromol/min mg, respectively. The VR enzyme had a molecular mass of 37 kDa in SDS-PAGE, while in natural conditions its mass was 118 kDa. The Km value was > 3.37+/-2.05 mM and its Vmax was 107.62+/-50.38 micromol/min mg for NADPH used as a cofactor. Both enzymatic activities were stable at pH 3.4, but in the presence of ethanol the CD activity decreased drastically while the VR activity was more stable. This is the first report that shows the presence of a CD and a VR enzyme in B. bruxellensis.

Kinetic studies of lipid oxidation induced by hemoglobin measured by consumption of dissolved oxygen in a liposome model system.

PubMed

Carvajal, Ana Karina; Rustad, Turid; Mozuraityte, Revilija; Storrø, Ivar

2009-09-09

The effect of hemoglobin (Hb) and lipid concentration, pH, temperature, and different antioxidants on heme-mediated lipid oxidation of liposomes from marine phospholipids was studied. The rate of lipid oxidation was measured by consumption of dissolved oxygen. Heme-mediated lipid oxidation at different Hb and lipid concentrations was modeled by Michaelis-Menten kinetics. The maximum rate (V(max)) for the reaction with cod and bovine Hb as a pro-oxidant was 66.2 +/- 3.4 and 56.6 +/- 3.4 microM/min, respectively. The Michaelis-Menten constant (K(m)) for the reaction with cod and bovine Hb was 0.67 +/- 0.09 and 1.2 +/- 0.2 microM, respectively. V(max) for the relationship between the oxygen uptake rate and lipid concentration was 43.2 +/- 1.5 microM/min, while the K(m) was 0.93 +/- 0.14 mg/mL. The effect of the temperature followed Arrhenius kinetics, and there was no significant difference in activation energy between cod and bovine Hb. The rate of lipid oxidation induced by bovine Hb was highest around pH 6. Ethylenediaminetetraacetic acid (EDTA) had no significant effect on heme-mediated lipid oxidation, but alpha-tocopherol and astaxanthin worked well as antioxidants. Kinetic differences were found between iron and Hb as pro-oxidants, and the efficacy of the antioxidants depended upon the pro-oxidant in the system.

Effects of metal ions on the catalytic degradation of dicofol by cellulase.

PubMed

Zhai, Zihan; Yang, Ting; Zhang, Boya; Zhang, Jianbo

2015-07-01

A new technique whereby cellulase immobilized on aminated silica was applied to catalyze the degradation of dicofol, an organochlorine pesticide. In order to evaluate the performance of free and immobilized cellulase, experiments were carried out to measure the degradation efficiency. The Michaelis constant, Km, of the reaction catalyzed by immobilized cellulase was 9.16 mg/L, and the maximum reaction rate, Vmax, was 0.40 mg/L/min, while that of free cellulase was Km=8.18 mg/L, and Vmax=0.79 mg/L/min, respectively. The kinetic constants of catalytic degradation were calculated to estimate substrate affinity. Considering that metal ions may affect enzyme activity, the effects of different metal ions on the catalytic degradation efficiency were explored. The results showed that the substrate affinity decreased after immobilization. Monovalent metal ions had no effect on the reaction, while divalent metal ions had either positive or inhibitory effects, including activation by Mn2+, reversible competition with Cd2+, and irreversible inhibition by Pb2+. Ca2+ promoted the catalytic degradation of dicofol at low concentrations, but inhibited it at high concentrations. Compared with free cellulase, immobilized cellulase was affected less by metal ions. This work provided a basis for further studies on the co-occurrence of endocrine-disrupting chemicals and heavy metal ions in the environment. Copyright © 2015. Published by Elsevier B.V.

Acute and chronic exposure of rat intestinal mucosa to dextran promotes SGLTI-mediated glucose transport.

PubMed

Debnam, E S; Denholm, E E; Grimble, G K

1998-08-01

The intestinal handling of dextran, an alpha-1,6-linked glucose polymer, is poor compared with starch, and some ingested dextran might therefore reach the lower small intestine. As luminal sugar up-regulates SGLT1 (sodium-dependent glucose transporter) locally, we report the effects of a dextran-enriched diet on jejunal and ileal brush border membrane (BBM) glucose uptake. Rats were maintained on a diet containing 65% maltodextrin or 32.5% maltodextrin + 32.5% dextran (10 kD or 40 kD) for 8-10 days, and the kinetics of phlorizin-sensitive [3H]-glucose uptake by purified BBM vesicles was determined. Ingestion of 40-kD but not 10-kD dextran increased Vmax for jejunal and ileal glucose uptake (+64.3% and +61.8% respectively, both P < 0.02). The transport response to 40-kD dextran was in keeping with lower levels of expired H2 at the end of the feeding period. High-performance liquid chromatography (HPLC) analysis of luminal contents indicated extensive hydrolysis of ingested dextran. Finally, 3-h jejunal exposure to 40-kD dextran in vivo increased the Vmax for glucose uptake by jejunal BBM. It is likely that increased SGLT1-mediated glucose uptake after short or longer term mucosal exposure to dextran results from luminal dextran per se or a hydrolysis product. The clinical implications of this up-regulation are discussed.

Comparison of Oxidation Kinetics of Nitrite-Oxidizing Bacteria: Nitrite Availability as a Key Factor in Niche Differentiation

PubMed Central

Nowka, Boris; Daims, Holger

2014-01-01

Nitrification has an immense impact on nitrogen cycling in natural ecosystems and in wastewater treatment plants. Mathematical models function as tools to capture the complexity of these biological systems, but kinetic parameters especially of nitrite-oxidizing bacteria (NOB) are lacking because of a limited number of pure cultures until recently. In this study, we compared the nitrite oxidation kinetics of six pure cultures and one enrichment culture representing three genera of NOB (Nitrobacter, Nitrospira, Nitrotoga). With half-saturation constants (Km) between 9 and 27 μM nitrite, Nitrospira bacteria are adapted to live under significant substrate limitation. Nitrobacter showed a wide range of lower substrate affinities, with Km values between 49 and 544 μM nitrite. However, the advantage of Nitrobacter emerged under excess nitrite supply, sustaining high maximum specific activities (Vmax) of 64 to 164 μmol nitrite/mg protein/h, contrary to the lower activities of Nitrospira of 18 to 48 μmol nitrite/mg protein/h. The Vmax (26 μmol nitrite/mg protein/h) and Km (58 μM nitrite) of “Candidatus Nitrotoga arctica” measured at a low temperature of 17°C suggest that Nitrotoga can advantageously compete with other NOB, especially in cold habitats. The kinetic parameters determined represent improved basis values for nitrifying models and will support predictions of community structure and nitrification rates in natural and engineered ecosystems. PMID:25398863

Methodological uncertainty in quantitative prediction of human hepatic clearance from in vitro experimental systems.

PubMed

Hallifax, D; Houston, J B

2009-03-01

Mechanistic prediction of unbound drug clearance from human hepatic microsomes and hepatocytes correlates with in vivo clearance but is both systematically low (10 - 20 % of in vivo clearance) and highly variable, based on detailed assessments of published studies. Metabolic capacity (Vmax) of commercially available human hepatic microsomes and cryopreserved hepatocytes is log-normally distributed within wide (30 - 150-fold) ranges; Km is also log-normally distributed and effectively independent of Vmax, implying considerable variability in intrinsic clearance. Despite wide overlap, average capacity is 2 - 20-fold (dependent on P450 enzyme) greater in microsomes than hepatocytes, when both are normalised (scaled to whole liver). The in vitro ranges contrast with relatively narrow ranges of clearance among clinical studies. The high in vitro variation probably reflects unresolved phenotypical variability among liver donors and practicalities in processing of human liver into in vitro systems. A significant contribution from the latter is supported by evidence of low reproducibility (several fold) of activity in cryopreserved hepatocytes and microsomes prepared from the same cells, between separate occasions of thawing of cells from the same liver. The large uncertainty which exists in human hepatic in vitro systems appears to dominate the overall uncertainty of in vitro-in vivo extrapolation, including uncertainties within scaling, modelling and drug dependent effects. As such, any notion of quantitative prediction of clearance appears severely challenged.

Autotrophic, hydrogen-oxidizing, denitrifying bacteria in groundwater, potential agents for bioremediation of nitrate contamination

USGS Publications Warehouse

Smith, R.L.; Ceazan, M.L.; Brooks, M.H.

1994-01-01

Addition of hydrogen or formate significantly enhanced the rate of consumption of nitrate in slurried core samples obtained from an active zone of denitrification in a nitrate-contaminated sand and gravel aquifer (Cape Cod, Mass.). Hydrogen uptake by the core material was immediate and rapid, with an apparent K(m) of 0.45 to 0.60 ??M and a V(max) of 18.7 nmol cm-3 h-1 at 30??C. Nine strains of hydrogen-oxidizing denitrifying bacteria were subsequently isolated from the aquifer. Eight of the strains grew autotrophically on hydrogen with either oxygen or nitrate as the electron acceptor. One strain grew mixotrophically. All of the isolates were capable of heterotrophic growth, but none were similar to Paracoccus denitrificans, a well-characterized hydrogen-oxidizing denitrifier. The kinetics for hydrogen uptake during denitrification were determined for each isolate with substrate depletion progress curves; the K(m)s ranged from 0.30 to 3.32 ??M, with V(max)s of 1.85 to 13.29 fmol cell-1 h-1. Because these organisms appear to be common constituents of the in situ population of the aquifer, produce innocuous end products, and could be manipulated to sequentially consume oxygen and then nitrate when both were present, these results suggest that these organisms may have significant potential for in situ bioremediation of nitrate contamination in groundwater.

Effect of complete protein 4.1R deficiency on ion transportproperties of murine erythrocytes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rivera, Alicia; De Franceschi, Lucia; Peters, Luanne L.

2006-06-02

Moderate hemolytic anemia, abnormal erythrocyte morphology(spherocytosis), and decreased membrane stability are observed in micewith complete deficiency of all erythroid protein 4.1 protein isoforms(4.1-/-; Shi TS et al., J. Clin. Invest. 103:331,1999). We have examinedthe effects of erythroid protein 4.1 (4.1R) deficiency on erythrocytecation transport and volume regulation. 4.1-/- mice exhibited erythrocytedehydration that was associated with reduced cellular K and increased Nacontent. Increased Na permeability was observed in these mice, mostlymediated by Na/H exchange with normal Na-K pump and Na-K-2Cl cotransportactivities. The Na/H exchange of 4.1-/- erythrocytes was markedlyactivated by exposure to hypertonic conditions (18.2+- 3.2 in 4.1 -/- vs.9.8 +-more » 1.3 mmol/1013 cell x h in control mice), with an abnormaldependence on osmolarity, (K0.5=417 +- 42 in 4.1 -/- vs. 460 +- 35 mOsmin control mice) suggestive of an up-regulated functional state. Whilethe affinity for internal protons was not altered (K0.5= 489.7 +- 0.7 vs.537.0+- 0.56 nM in control mice), the Vmax of the H-induced Na/H exchangeactivity was markedly elevated in 4.1-/- erythrocytes (Vmax 91.47Moderatehemolytic anemia, abnormal erythrocyte morphology (spherocytosis), anddecreased membrane stability are observed in mice with completedeficiency of all erythroid protein 4.1 protein isoforms (4.1-/-; Shi TSet al., J. Clin. Invest. 103:331,1999). We have examined the effects oferythroid protein 4.1 (4.1R) deficiency on erythrocyte cation transportand volume regulation. 4.1-/- mice exhibited erythrocyte dehydration thatwas associated with reduced cellular K and increased Na content.Increased Na permeability was observed in these mice, mostly mediated byNa/H exchange with normal Na-K pump and Na-K-2Cl cotransport activities.The Na/H exchange of 4.1-/- erythrocytes was markedly activated byexposure to hypertonic conditions (18.2 +- 3.2 in 4.1 -/- vs. 9.8 +- 1.3mmol/1013 cell x h in control mice), with an abnormal dependence onosmolarity, (K0.5=417 +- 42 in 4.1 -/- vs. 460 +- 35 mOsm in controlmice) suggestive of an up-regulated functional state. While the affinityfor internal protons was not altered (K0.5= 489.7 +- 0.7 vs. 537.0 +-0.56 nM in control mice), the Vmax of the H-induced Na/H exchangeactivity was markedly elevated in 4.1-/- erythrocytes (Vmax 91.47+-7.2compared to 46.52+-5.4 mmol/1013 cell x h in control mice). Na/H exchangeactivation by okadaic acid was absent in 4.1-/- erythrocytes. Altogether,these results suggest that erythroid protein 4.1 plays a major role involume regulation and physiologically down-regulates Na/H exchange inmouse erythrocytes. Up-regulation of the Na/H exchange is an importantcontributor to the elevated cell Na content of 4.1 -/- erythrocytes.-7.2compared to 46.52+-5.4 mmol/1013 cell x h in control mice). Na/H exchangeactivation by okadaic acid was absent in 4.1-/- erythrocytes. Altogether,these results suggest that erythroid protein 4.1 plays a major role involume regulation and physiologically down-regulates Na/H exchange inmouse erythrocytes. Up-regulation of the Na/H exchange is an importantcontributor to the elevated cell Na content of 4.1 -/-erythrocytes.« less

Expression of drug transporters and drug metabolizing enzymes in the bladder urothelium in man and affinity of the bladder spasmolytic trospium chloride to transporters likely involved in its pharmacokinetics.

PubMed

Bexten, Maria; Oswald, Stefan; Grube, Markus; Jia, Jia; Graf, Tanja; Zimmermann, Uwe; Rodewald, Kathrin; Zolk, Oliver; Schwantes, Ulrich; Siegmund, Werner; Keiser, Markus

2015-01-05

The cationic, water-soluble quaternary trospium chloride (TC) is incompletely absorbed from the gut and undergoes wide distribution but does not pass the blood-brain barrier. It is secreted by the kidneys, liver, and intestine. To evaluate potential transport mechanisms for TC, we measured affinity of the drug to the human uptake and efflux transporters known to be of pharmacokinetic relevance. Affinity of TC to the uptake transporters OATP1A2, -1B1, -1B3, -2B1, OCT1, -2, -3, OCTN2, NTCP, and ASBT and the efflux carriers P-gp, MRP2 and MRP3 transfected in HEK293 and MDCK2 cells was measured. To identify relevant pharmacokinetic mechanisms in the bladder urothelium, mRNA expression of multidrug transporters, drug metabolizing enzymes, and nuclear receptors, and the uptake of TC into primary human bladder urothelium (HBU) cells were measured. TC was shown to be a substrate of OATP1A2 (Km = 6.9 ± 1.3 μmol/L; Vmax = 41.6 ± 1.8 pmol/mg·min), OCT1 (Km = 106 ± 16 μmol/L; Vmax = 269 ± 18 pmol/mg·min), and P-gp (Km = 34.9 ± 7.5 μmol/L; Vmax = 105 ± 9.1 pmol/mg·min, lipovesicle assay). The genetic OATP1A2 variants *2 and *3 were loss-of-function transporters for TC. The mRNA expression analysis identified the following transporter proteins in the human urothelium: ABCB1 (P-gp), ABCC1-5 (MRP1-5), ABCG2 (BCRP), SLCO2B1 (OATP2B1), SLCO4A1 (OATP4A1), SLC22A1 (OCT1), SLC22A3 (OCT3), SLC22A4 (OCTN1), SLC22A5 (OCTN2), and SLC47A1 (MATE1). Immuno-reactive P-gp and OATP1A2 were localized to the apical cell layers. Drug metabolizing enzymes CYP3A5, -2B6, -2B7 -2E1, SULT1A1-4, UGT1A1-10, and UGT2B15, and nuclear receptors NR1H3 and NR1H4 were also expressed on mRNA level. TC was taken up into HBU cells (Km = 18.5 ± 4.8 μmol/L; Vmax = 106 ± 11.3 pmol/mg·min) by mechanisms that could be synergistically inhibited by naringin (IC50 = 10.8 (8.4; 13.8) μmol/L) and verapamil (IC50 = 4.6 (2.8; 7.5) μmol/L), inhibitors of OATP1A2 and OCT1, respectively. Affinity of TC to OCT1 and P-glycoprotein may be the reason for incomplete oral absorption, wide distribution into liver and kidneys, and substantial intestinal and renal secretions. Absence of brain distribution may result from affinity to P-gp and a low affinity to OATP1A2. The human urothelium expresses many drug transporters and drug metabolizing enzymes that may interact with TC and other drugs eliminated into the urine.

Leap and strike kinetics of an acoustically ‘hunting’ barn owl (Tyto alba)

PubMed Central

Usherwood, James R.; Sparkes, Emily L.; Weller, Renate

2014-01-01

Barn owls are effective hunters of small rodents. One hunting technique is a leap from the ground followed by a brief flight and a plummeting ‘strike’ onto an acoustically targeted – and potentially entirely hidden – prey. We used forceplate measurements to derive kinetics of the leap and strike. Leaping performance was similar to reported values for guinea fowl. This is likely achieved despite the owl's considerably smaller size because of its relatively long legs and use of wing upstroke. Strikes appear deliberately forceful: impulses could have been spread over larger periods during greater deflections of the centre of mass, as observed in leaping and an alighting landing measurement. The strike, despite forces around 150 times that of a mouse body weight, is not thought to be crucial to the kill; rather, forceful strikes may function primarily to enable rapid penetration of leaf litter or snow cover, allowing grasping of hidden prey. PMID:24948629

Leap and strike kinetics of an acoustically 'hunting' barn owl (Tyto alba).

PubMed

Usherwood, James R; Sparkes, Emily L; Weller, Renate

2014-09-01

Barn owls are effective hunters of small rodents. One hunting technique is a leap from the ground followed by a brief flight and a plummeting 'strike' onto an acoustically targeted - and potentially entirely hidden - prey. We used forceplate measurements to derive kinetics of the leap and strike. Leaping performance was similar to reported values for guinea fowl. This is likely achieved despite the owl's considerably smaller size because of its relatively long legs and use of wing upstroke. Strikes appear deliberately forceful: impulses could have been spread over larger periods during greater deflections of the centre of mass, as observed in leaping and an alighting landing measurement. The strike, despite forces around 150 times that of a mouse body weight, is not thought to be crucial to the kill; rather, forceful strikes may function primarily to enable rapid penetration of leaf litter or snow cover, allowing grasping of hidden prey. © 2014. Published by The Company of Biologists Ltd.

Surface pressure distributions on a delta wing undergoing large amplitude pitching oscillations. M.S. Thesis

NASA Technical Reports Server (NTRS)

Thompson, Scott A.

1989-01-01

Wind tunnel experiments were performed on a 70 deg sweep delta wing to determine the effect of a sinusoidal pitching motion on the pressure field on the suction side of the wing. Twelve pressure taps were placed from 35 to 90 percent of the chord, at 60 percent of the local semi-span. Pressure coefficients were measured as a function of Reynolds number and pitch rate. The pressure coefficient was seen to vary at approximately the same frequency as the pitching frequency. The relative pressure variation at each chord location was comparable for each case. The average pressure distribution through each periodic motion was near the static distribution for the average angle of attack. Upon comparing the upstroke and downstroke pressures for a specific angle of attack, the downstroke pressures were slightly larger. Vortex breakdown was seen to have the most significant effect at the 40 to 45 percent chord location, where a decrease in pressure was apparent.

Pressure-induced amorphization of YVO₄:Eu³⁺ nanoboxes.

PubMed

Ruiz-Fuertes, J; Gomis, O; León-Luis, S F; Schrodt, N; Manjón, F J; Ray, S; Santamaría-Pérez, D; Sans, J A; Ortiz, H M; Errandonea, D; Ferrer-Roca, C; Segura, A; Martínez-García, D; Lavín, V; Rodríguez-Mendoza, U R; Muñoz, A

2016-01-15

A structural transformation from the zircon-type structure to an amorphous phase has been found in YVO4:Eu(3+) nanoboxes at high pressures above 12.7 GPa by means of x-ray diffraction measurements. However, the pair distribution function of the high-pressure phase shows that the local structure of the amorphous phase is similar to the scheelite-type YVO4. These results are confirmed both by Raman spectroscopy and Eu(3+) photoluminescence which detect the phase transition to a scheelite-type structure at 10.1 and 9.1 GPa, respectively. The irreversibility of the phase transition is observed with the three techniques after a maximum pressure in the upstroke of around 20 GPa. The existence of two (5)D0-->(7)F0 photoluminescence peaks confirms the existence of two local environments for Eu(3+), at least for the low-pressure phase. One environment is the expected for substituting Y(3+) and the other is likely a disordered environment possibly found at the surface of the nanoboxes.

Universal ripper miner

DOEpatents

Morrell, Roger J.; Larson, David A.

1991-01-01

A universal ripper miner used to cut, collect and transfer material from an underground mine working face includes a cutter head that is vertically movable in an arcuate cutting cycle by means of drive members, such as hydraulically actuated pistons. The cutter head may support a circular cutter bit having a circular cutting edge that may be indexed to incrementally expose a fresh cutting edge. An automatic indexing system is disclosed wherein indexing occurs by means of a worm gear and indexing lever mechanism. The invention also contemplates a bi-directional bit holder enabling cutting to occur in both the upstroke and the downstroke cutting cycle. Another feature of the invention discloses multiple bits arranged in an in-line, radially staggered pattern, or a side-by-side pattern to increase the mining capacity in each cutting cycle. An on-board resharpening system is also disclosed for resharpening the cutting edge at the end of cutting stroke position. The aforementioned improvement features may be used either singly, or in any proposed combination with each other.

EVALUATION OF RIGHT AND LEFT VENTRICULAR DIASTOLIC FILLING

PubMed Central

Pasipoularides, Ares

2013-01-01

A conceptual fluid-dynamics framework for diastolic filling is developed. The convective deceleration load (CDL) is identified as an important determinant of ventricular inflow during the E-wave (A-wave) upstroke. Convective deceleration occurs as blood moves from the inflow anulus through larger-area cross-sections toward the expanding walls. Chamber dilatation underlies previously unrecognized alterations in intraventricular flow dynamics. The larger the chamber, the larger become the endocardial surface and the CDL. CDL magnitude affects strongly the attainable E-wave (A-wave) peak. This underlies the concept of diastolic ventriculoannular disproportion. Large vortices, whose strength decreases with chamber dilatation, ensue after the E-wave peak and impound inflow kinetic energy, averting an inflow-impeding, convective Bernoulli pressure-rise. This reduces the CDL by a variable extent depending on vortical intensity. Accordingly, the filling vortex facilitates filling to varying degrees, depending on chamber volume. The new framework provides stimulus for functional genomics research, aimed at new insights into ventricular remodeling. PMID:23585308

Significance of the enzymatic properties of yeast S39A enolase to the catalytic mechanism.

PubMed

Brewer, J M; Glover, C V; Holland, M J; Lebioda, L

1998-04-02

The S39A mutant of yeast enolase (isozyme 1), prepared by site-directed mutagenesis, has a relative Vmax of 0.01% and an activation constant for Mg2+ ca. 10-fold higher, compared with native enzyme. It is correctly folded. There is little effect of solvent viscosity on activity. We think that the loop Ser36-His43 fails to move to the 'closed' position upon catalytic Mg2+ binding, weakening several electrostatic interactions involved in the mechanism.

Cloning, sequencing, and expression of the gene encoding cyclic 2, 3-diphosphoglycerate synthetase, the key enzyme of cyclic 2, 3-diphosphoglycerate metabolism in Methanothermus fervidus.

PubMed

Matussek, K; Moritz, P; Brunner, N; Eckerskorn, C; Hensel, R

1998-11-01

Cyclic 2,3-diphosphoglycerate synthetase (cDPGS) catalyzes the synthesis of cyclic 2,3-diphosphoglycerate (cDPG) by formation of an intramolecular phosphoanhydride bond in 2,3-diphosphoglycerate. cDPG is known to be accumulated to high intracellular concentrations (>300 mM) as a putative thermoadapter in some hyperthermophilic methanogens. For the first time, we have purified active cDPGS from a methanogen, the hyperthermophilic archaeon Methanothermus fervidus, sequenced the coding gene, and expressed it in Escherichia coli. cDPGS purification resulted in enzyme preparations containing two isoforms differing in their electrophoretic mobility under denaturing conditions. Since both polypeptides showed the same N-terminal amino acid sequence and Southern analyses indicate the presence of only one gene coding for cDPGS in M. fervidus, the two polypeptides originate from the same gene but differ by a not yet identified modification. The native cDPGS represents a dimer with an apparent molecular mass of 112 kDa and catalyzes the reversible formation of the intramolecular phosphoanhydride bond at the expense of ATP. The enzyme shows a clear preference for the synthetic reaction: the substrate affinity and the Vmax of the synthetic reaction are a factor of 8 to 10 higher than the corresponding values for the reverse reaction. Comparison with the kinetic properties of the electrophoretically homogeneous, apparently unmodified recombinant enzyme from E. coli revealed a twofold-higher Vmax of the enzyme from M. fervidus in the synthesizing direction.

Does oxidative stress affect the activity of the sodium-proton exchanger?

PubMed

Bober, Joanna; Kedzierska, Karolina; Kwiatkowska, Ewa; Stachowska, Ewa; Gołembiewska, Edyta; Mazur, Olech; Staniewicz, Zdzisław; Ciechanowski, Kazimierz; Chlubek, Dariusz

2010-01-01

Accumulation of reactive oxygen species (ROS) takes place in patients with chronic renal failure (CRF). Oxidative stress causes disorders in the activity of the sodium-proton exchanger (NHE). Studies on NHE in CRF produced results that are discrepant and difficult to interpret. The aim of this study was to demonstrate that oxidative stress had an effect on the activity of NHE. We enrolled 87 subjects divided into 4 groups: patients with CRF treated conservatively; patients with CRF hemodialyzed without glucose--HD-g(-); patients with CRF hemodialyzed with glucose--HD-g(+); controls (C). The activity of NHE, the rate of proton efflux V(max), Michaelis constant (Km), and the concentration of thiobarbituric acid-reactive substances (TBARS, an indicator of oxidative stress) in plasma, as well as the concentration of reduced glutathione in blood were determined. The concentration of TBARS was significantly higher in hemodialyzed patients before and after dialysis and in patients with CRF on conservative treatment in comparison with group C. TBARS in plasma correlated negatively with VpH(i)6.4 in group C and with V(max) and VpH(i)6.4 after HD in group HD-g(-). We found that the concentration of creatinine correlated with TBARS (p < 0.0001; r = +0.51) in the conservatively treated group. We observed a marked oxidative stress and decreased NHE activity when dialysis was done without glucose, whereas patients dialyzed with glucose demonstrated a relatively low intensity of oxidative stress.

Effect of bio-column composed of aged refuse on methane abatement--a novel configuration of biological oxidation in refuse landfill.

PubMed

Han, Dan; Zhao, Youcai; Xue, Binjie; Chai, Xiaoli

2010-01-01

An experimental bio-column composed of aged refuse was installed around the exhaust pipe as a new way to mitigate methane in refuse landfill. One of the objectives of this work was to assess the effect of aged refuse thickness in bio-column on reducing CH4 emissions. Over the study period, methane oxidation was observed at various thicknesses, 5 cm (small size), 10 cm (middle size) and 15 cm (large size), representing one to three times of pipeline diameters. The middle and large size both showed over 90% methane conversion, and the highest methane conversion rate of above 95% occurred in the middle-size column cell. Michaelis-Menten equation addressed the methanotrophs diffusion in different layers of the bio-columns. Maximum methanotrophic activity (Vmax) measured at the three thicknesses ranged from 6.4 x 10(-3) to 15.6 x 10(-3) units, and the half-saturation value (K(M)) ranged from 0.85% to 1.67%. Both the highest Vmax and K(M) were observed at the middle-size of the bio-column, as well as the largest methanotrophs population, suggesting a significant efficiency of methane mitigation happened in the optimum zone with greatest affinity and methanotrophic bacteria activities. Therefore, bio-column is a potential style for methane abatement in landfill, and the aged refuse both naturally formed and artificially placed in the column plays a critical role in CH4 emission.

Islet transplantation under the kidney capsule fully corrects the impaired skeletal muscle glucose transport system of streptozocin diabetic rats.

PubMed Central

Napoli, R; Davalli, A M; Hirshman, M F; Weitgasser, R; Weir, G C; Horton, E S

1996-01-01

Chronic insulin therapy improves but does not restore impaired insulin-mediated muscle glucose uptake in human diabetes or muscle glucose uptake, transport, and transporter translocation in streptozocin diabetic rats. To determine whether this inability is due to inadequate insulin replacement, we studied fasted streptozocin-induced diabetic Lewis rats either untreated or after islet transplantation under the kidney capsule. Plasma glucose was increased in untreated diabetics and normalized by the islet transplantation (110 +/- 5, 452 +/- 9, and 102 +/- 3 mg/dl in controls, untreated diabetics, and transplanted diabetics, respectively). Plasma membrane and intracellular microsomal membrane vesicles were prepared from hindlimb skeletal muscle of basal and maximally insulin-stimulated rats. Islet transplantation normalized plasma membrane carrier-mediated glucose transport Vmax, plasma membrane glucose transporter content, and insulin-induced transporter translocation. There were no differences in transporter intrinsic activity (Vmax/Ro) among the three groups. Microsomal membrane GLUT4 content was reduced by 30% in untreated diabetic rats and normal in transplanted diabetics, whereas the insulin-induced changes in microsomal membrane GLUT4 content were quantitatively similar in the three groups. There were no differences in plasma membrane GLUT1 among the groups and between basal and insulin stimulated states. Microsomal membrane GLUT1 content was increased 60% in untreated diabetics and normalized by the transplantation. In conclusion, an adequate insulin delivery in the peripheral circulation, obtained by islet transplantation, fully restores the muscle glucose transport system to normal in streptozocin diabetic rats. PMID:8617870

[Metabolic kinetics of MN9202 in Beagle dog liver microsomes].

PubMed

Yang, Zhi-fu; Zhou, Si-yuan; Mei, Qi-bing; Yang, Tie-hong; Liu, Zhen-guo

2005-11-01

To study the metabolic kinetics of MN9202 in Beagle dog liver microsome. Beagle dog liver microsomes were prepared by using ultracentrifuge method. After incubating 0.4 micromol x L(-1) MN9202 with 1 g x L(-1) microsomes for 30 min at 37 degrees C, the reaction was terminated by adding 0.5 mL alkalization. The RP-HPLC was used to determine the drug in the incubation mixture. The Michaelis-Menten parameters Km, and Vmax in Beagle dog liver microsomes were initially estimated by analyzing Lineweave-Brurk plot. Various selective CYP inhibitors were used to investigate their inhibitory effect on the metabolism of MN9202. The Km, Vmax and CLint of MN9202 were (22.6 +/- 8.0) micromol x L(-1), (0.54 +/- 0.17) micromol x g(-1) x min(-1) and (0.0242 +/- 0.0009) L x g(-1) x min(-1), respectively. The metabolism of MN9202 was significantly inhibited by ketoconazole (Ket) and troleandomycin (Tro) in Beagle dog liver microsomes. Tranylcypromine (Tra) could inhibit the metabolism of drug as well. While other inhibitors showed little inhibitory effect on the metabolism of MN9202. It was shown that CYP3A and CYP2C19 were involved in MN9202 metabolism. The inhibitors of human CYP3A and CYP2C19 may have potential interaction with MN9202, and this can reduce the metabolism rate and increase the toxicity of MN9202.

Characterization of arylalkylamine N-acetyltransferase (AANAT) activities and action spectrum for suppression in the band-legged cricket, Dianemobius nigrofasciatus (Orthoptera: Gryllidae).

PubMed

Izawa, Norimitsu; Suzuki, Takeshi; Watanabe, Masakatsu; Takeda, Makio

2009-04-01

Arylalkylamine N-acetyltransferase (AANAT), constituting a large family of enzymes, catalyzes the transacetylation from acetyl-CoA to monoamine substrates, although homology among species is not very high. AANAT in vertebrates is photosensitive and mediates circadian regulation. Here, we analyzed AANAT of the cricket, Dianemobius nigrofasciatus. The central nervous system contained AANAT activity. The optimum pHs were 6.0 (a minor peak) and 10.5 (a major peak) with crude enzyme solution. We analyzed the kinetics at pH 10.5 using the sample containing collective AANAT activities, which we term AANAT. Lineweaver-Burk plot and secondary plot yielded a K(m) for tryptamine as substrate of 0.42 microM, and a V(max) of 9.39 nmol/mg protein/min. The apparent K(m) for acetyl-CoA was 59.9 microM and the V(max) was 8.14 nmol/mg protein/min. AANAT of D. nigrofasciatus was light-sensitive. The activity was higher at night-time than at day-time as in vertebrates. To investigate most effective wavelengths on AANAT activity, a series of monochromatic lights was applied (350, 400, 450, 500, 550, 600 and 650 nm). AANAT showed the highest sensitivity to around 450 nm and 550 nm. 450 nm light was more effective than 550 nm light. Therefore, the most effective light affecting AANAT activity is blue light, which corresponds to the absorption spectrum of blue wave (BW)-opsin.

Molecular genetics of the platelet serotonin system in first-degree relatives of patients with autism

PubMed Central

Cross, Sarah; Kim, Soo-Jeong; Weiss, Lauren A.; Delahanty, Ryan J.; Sutcliffe, James S.; Leventhal, Bennett L.; Cook, Edwin H.; Veenstra-VanderWeele, Jeremy

2009-01-01

Elevated platelet serotonin (5-HT) is found in a subset of children with autism and in some of their first-degree relatives. Indices of the platelet serotonin system, including whole blood serotonin (5-HT), 5-HT binding affinity for the serotonin transporter (Km), 5-HT uptake (Vmax), and lysergic acid diethylamide (LSD) receptor binding, were previously studied in twenty-four first-degree relatives of probands with autism, half of whom were selected for elevated whole blood 5-HT levels. All subjects were then genotyped for selected polymorphisms at the SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 loci. Previous studies allowed an a priori prediction of SLC6A4 haplotypes that separated the subjects into three groups that showed significantly different 5-HT binding affinity (Km, p = 0.005) and 5-HT uptake rate (Vmax, p = 0.046). Genotypes at four individual polymorphisms in SLC6A4 were not associated with platelet 5-HT indices. Haplotypes at SLC6A4 and individual genotypes of polymorphisms at SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 showed no significant association with whole blood 5-HT. Haplotype analysis of two polymorphisms in TPH1 revealed a nominally significant association with whole blood 5-HT (p = 0.046). These initial studies of indices of the 5-HT system with several SNPs at loci in this system generate hypotheses for testing in other samples. PMID:17406648

Cytochrome P-450 isoforms involved in carboxylic acid ester cleavage of Hantzsch pyridine ester of pranidipine.

PubMed

Kudo, S; Okumura, H; Miyamoto, G; Ishizaki, T

1999-02-01

Cytochrome P-450 (CYP) isoforms responsible for the cleavage of Hantzsch pyridine ester at the 3-position of pranidipine were studied in vitro using cDNA-expressed human CYP enzymes. CYP1A1, 1A2, 2D6, and 3A4 cleaved the ester with a catalytic activity of 5.5, 0. 93, 13.1, and 22.4 nmol/30 min/nmol P-450, respectively. CYP2A6, 2B6, 2C8, 2C9, 2C19, and 2E1 were not involved in the de-esterification. The Km and Vmax values for the de-esterification were 11.8 microM and 0.47 nmol/min/nmol P-450 in the CYP2D6-catalyzed reaction and 8. 7 microM and 0.84 nmol/min/nmol P-450 in the CYP3A4-catalyzed reaction. The intrinsic clearance (Vmax/Km) of the de-esterification by CYP3A4 was 2-fold greater than that by CYP2D6. Quinidine almost completely inhibited the CYP2D6-mediated de-esterification at the concentration of 1 x 10(-6) M. Ketoconazole and troleandomycin inhibited the CYP3A4-mediated reaction in a dose-related manner. The results indicate that although the multiple CYP isoforms can catalyze the de-esterification, CYP3A4 and 2D6 are the major isoforms.

The effects of a supportive knee brace on leg performance in healthy subjects.

PubMed

Veldhuizen, J W; Koene, F M; Oostvogel, H J; von Thiel, T P; Verstappen, F T

1991-12-01

Eight healthy volunteers were fitted with a supportive knee brace (Push Brace 'Heavy') to one knee for a duration of four weeks wherein they were tested before, during and after the application to establish the effect of bracing on performance. The tests consisted of isokinetic strength measurement of knee flexion and extension, 60 meter dash, vertical jump height and a progressive horizontal treadmill test until exhaustion (Vmax) with determination of oxygen uptake, heart rate and plasma lactate concentration. Wearing the brace for one day, the performance indicators showed a decline compared with the test before application (base values). Sprint time was 4% longer (p less than 0.01) and Vmax 6% slower (p less than 0.01). Peak torque of knee flexion at 60 and 240 deg.sec-1 was 6% (p less than 0.05) respectively 9% (p less than 0.05) less. Peak extension torque at 60 deg.sec-1 was 9% less (p less than 0.05). While wearing the brace for four weeks, the test performances were practically identical to their base values. After removal of the brace, all test parameters were statistically similar to the base values. Heart rate at submaximal exercise levels was even lower (p less than 0.05). In conclusion, performance in sports with test-like exercise patterns is not affected by the brace tested. Bracing does not "weaken the knee" as it is widely believed in sports practice.

Carnosine: effect on aging-induced increase in brain regional monoamine oxidase-A activity.

PubMed

Banerjee, Soumyabrata; Poddar, Mrinal K

2015-03-01

Aging is a natural biological process associated with several neurological disorders along with the biochemical changes in brain. Aim of the present investigation is to study the effect of carnosine (0.5-2.5μg/kg/day, i.t. for 21 consecutive days) on aging-induced changes in brain regional (cerebral cortex, hippocampus, hypothalamus and pons-medulla) mitochondrial monoamine oxidase-A (MAO-A) activity with its kinetic parameters. The results of the present study are: (1) The brain regional mitochondrial MAO-A activity and their kinetic parameters (except in Km of pons-medulla) were significantly increased with the increase of age (4-24 months), (2) Aging-induced increase of brain regional MAO-A activity including its Vmax were attenuated with higher dosages of carnosine (1.0-2.5μg/kg/day) and restored toward the activity that observed in young, though its lower dosage (0.5μg/kg/day) were ineffective in these brain regional MAO-A activity, (3) Carnosine at higher dosage in young rats, unlike aged rats significantly inhibited all the brain regional MAO-A activity by reducing their only Vmax excepting cerebral cortex, where Km was also significantly enhanced. These results suggest that carnosine attenuated the aging-induced increase of brain regional MAO-A activity by attenuating its kinetic parameters and restored toward the results of MAO-A activity that observed in corresponding brain regions of young rats. Copyright © 2014 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

Kinetics of DCE and VC mineralization under methanogenic and Fe(III)- reducing conditions

USGS Publications Warehouse

Bradley, P.M.; Chapelle, F.H.

1997-01-01

The kinetics of anaerobic mineralization of DCE and VC under mathanogenic and Fe(III)-reducing conditions as a function of dissolved contaminant concentration were evaluated. Microorganisms indigenous to creek bed sediments, where groundwater contaminated with chlorinated ethenes continuously discharges, demonstrated significant mineralization of DCE and VC under methanogenic and Fe(III)- reducing conditions. Over 37 days, the recovery of [1,214C]VC radioactivity as 14CO2 ranged from 5% to 44% and from 8% to 100% under methanogenic and Fe(III)-reducing conditions, respectively. The recovery of [1,2-14C]DCE radioactivity as 14CO2 ranged from 4% to 14% and did not vary significantly between methanogenic and Fe(III)reducing conditions. VC mineralization was described by Michaelis- Menten kinetics. Under methanogenic conditions, V(max) was 0.19 ?? 0.01 ??mol L-1 d-1 and the half-saturation constant, k(m), was 7.6 ?? 1.7 ??M. Under Fe(III)-reducing conditions, V(max) was 0.76 ?? 0.07 ??mol L-1 d-1 and k(m) was 1.3 ?? 0.5 ??M. In contrast, DCE mineralization could be described by first-order kinetics. The first-order degradation rate constant for DCE mineralization was 0.6 ?? 0.2% d-1 under methanogenic and Fe(III)-reducing conditions. The results indicate that the kinetics of chlorinated ethane mineralization can vary significantly with the specific contaminant and the predominant redox conditions under which mineralization occurs.

Cation transport in intact erythrocytes of hyperthyroid patients: role of the NaK-ATPase pump.

PubMed

Michels, R C; Ober, K P; Hennessy, J F

1981-11-01

Studies of erythrocyte (RBC) cation fluxes and concentrations in hyperthyroid subjects have recently been reported with the suggestion that Na-K ATPase activity was decreased. We have studied tha kinetics of total and ouabain-sensitive K+ uptake utilizing 86Rb as a tracer in the intact erythrocytes of 7 hyperthyroid subjects and compared the results of those of a healthy control population. We find total K+ transport is depressed in the RBC of hyperthyroid subjects. The Vmax for K+ transport for hyperthyroid subjects is 1.8 +/- 0.17 x 10(-4) mM K+/10(9) RBC/hour versus a control of 2.3 +/- 0.14 x 10(-4) mM K+/10(9) RBC/hour. This depression in Vmax is evident in spite of no significant differences in the Km for the system when hyperthyroid subjects (2.7 +/- 0.19 mM) are compared to controls (2.38 +/- 0.21 mM). Further, the depressed K+ transport appears to be the result of depressed ouabain--insensitive K+ transport. Although the percent of the ouabain-sensitive K+ transport is greater in the hyperthyroid subject (82.5%) versus controls (72.5%), this simply reflects a relative change in a system where total transport is dropping but the ouabain-sensitive component is remaining unchanged. None of these findings can be directly or indirectly related to thyroid hormone and it is suggested that the ion transport changes reflect factors independent of thyroid hormone.

Hydrolysis of triolein in phospholipid vesicles and microemulsions by a purified rat liver acid lipase.

PubMed

Burrier, R E; Brecher, P

1983-10-10

An acid lipase was purified from rat liver lysosomes. Lipase purification involved affinity chromatography, gel filtration, and stabilization of the purified preparation using ethylene glycol and Triton X-100. A molecular weight of 67,000-69,000 was determined independently using density gradient centrifugation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and gel filtration. To study enzyme action, model substrates were prepared by incorporating radiolabeled triolein into either unilamellar vesicles or microemulsions. Substrates were prepared by cosonicating aqueous dispersions of lecithin and triolein. Formation of vesicles or emulsions depended on the relative amount of each lipid and on sonication conditions. Vesicles were prepared at molar ratios between 70:1 and 26:1 (lecithin:triolein) and the microemulsion preparation at a molar ratio of 1:1. The substrate particles were of similar size (220-250 A) as determined by Bio-Gel A-15m chromatography. Hydrolysis of triolein contained in vesicles or emulsions was similar with respect to pH, temperature, and reaction products. Kinetic studies on vesicles with increasing triolein content showed progressively greater Vmax values (0-0.6 mumol/min/mg), and Vmax for the emulsion was 3.1 mumol/min/mg. Addition of human very low or low density lipoprotein produced a dose-dependent inhibition with both substrates. The results show that synthetically prepared microemulsions are stable and effective substrates for the acid lipase and indicate that surface-oriented triolein is hydrolyzed in both preparations.

The first evidence of cholinesterases in skin mucus of carps and its applicability as biomarker of organophosphate exposure.

PubMed

Nigam, Ashwini Kumar; Srivastava, Nidhi; Rai, Amita Kumari; Kumari, Usha; Mittal, Ajay Kumar; Mittal, Swati

2014-05-01

The presence of cholinesterase (ChE) activity in skin mucus of three carps, Cirrhinus mrigala, Labeo rohita, and Catla catla and its applicability as biomarker of the organophosphorus insecticide exposure were investigated. Biochemical characterization, using specific substrates and inhibitors, indicated that measured esterase activity in skin mucus was mainly owing to ChEs. Significant difference in the proportion of acetylcholinesterase and butyrylcholinesterase activities was observed in skin mucus of three carps. Enzyme kinetic analysis, using the substrate acetylthiocholine iodide revealed significantly high Vmax value in C. catla compared to that in L. rohita and C. mrigala. In contrast, Vmax value using the substrate butyrylthiocholine iodide was significantly high in C. mrigala than in L. rohita and C. catla. In vitro treatment of skin mucus of three carps, with the organophosphorus insecticide Nuvan®, showed strong inhibition of ChE activities. In vivo experiments conducted using C. mrigala and exposing the fish to the sublethal test concentrations (5 and 15 mg/L) of the insecticide also revealed significant inhibition of ChE activity in mucus. In C. mrigala, exposed to the sublethal test concentrations of the insecticide for 4 days and then kept for recovery for 16 days, mucus ChE activity recovered to the control level. Thus, ChE activity in skin mucus could be considered a good biomarker of the organophosphorus insecticide exposure to fish and a useful tool in monitoring environmental toxicity. Copyright © 2012 Wiley Periodicals, Inc., a Wiley company.

Competition between roots and microorganisms for nitrogen: mechanisms and ecological relevance

NASA Astrophysics Data System (ADS)

Kuzyakov, Yakov; Xu, Xingliang

2014-05-01

Demand of all living organisms on the same nutrients forms the basis for interspecific competition between plants and microorganisms in soils. This competition is especially strong in the rhizosphere. To evaluate competitive and mutualistic interactions between plants and microorganisms and to analyse ecological consequences of these interactions, we analysed 424 data pairs from 41 15N-labelling studies that investigated 15N redistribution between roots and microorganisms. Calculated Michaelis-Menten kinetics based on Km (Michaelis constant) and Vmax (maximum uptake capacity) values from 77 studies on the uptake of nitrate, ammonia, and amino acids by roots and microorganisms clearly showed that, shortly after nitrogen (N) mobilization from soil organic matter and litter, microorganisms take up most N. Lower Km values of microorganisms suggest that they are especially efficient at low N concentrations, but can also acquire more N at higher N concentrations (Vmax) compared with roots. Because of the unidirectional flow of nutrients from soil to roots, plants are the winners for N acquisition in the long run. Therefore, despite strong competition between roots and microorganisms for N, a temporal niche differentiation reflecting their generation times leads to mutualistic relationships in the rhizosphere. This temporal niche differentiation is highly relevant ecologically because it: protects ecosystems from N losses by leaching during periods of slow or no root uptake; continuously provides roots with available N according to plant demand; and contributes to the evolutionary development of mutualistic interactions between roots and microorganisms.

A designed bifunctional laccase/β-1,3-1,4-glucanase enzyme shows synergistic sugar release from milled sugarcane bagasse.

PubMed

Furtado, G P; Ribeiro, L F; Lourenzoni, M R; Ward, R J

2013-01-01

A bifunctional enzyme has been created by fusing two Bacillus subtilis enzymes: the β-1,3-1,4-glucanase (BglS, EC 3.2.1.73) that hydrolyzes plant cell wall β-glucans and the copper-dependent oxidase laccase (CotA, EC 1.10.3.2) that catalyzes the oxidation of aromatic compounds with simultaneous reduction of oxygen to water. The chimeric laccase/β-1,3-1,4-glucanase was created by insertion fusion of the bglS and cotA genes, and expressed in Escherichia coli. The affinity-purified recombinant chimeric enzyme showed both laccase and glucanase activities, with a maximum laccase activity at pH 4.5 and 75°C that showed a V(max) 30% higher than observed for the parental laccase. The maximum glucanase activity in the chimeric enzyme was at pH 6.0 and 50°C, with a slight reduction in V(max) by ∼10% compared with the parental glucanase. A decreased K(M) resulted in an overall increase in the K(cat)/K(M) value for the glucanase activity of the chimeric enzyme. The hydrolytic activity of the chimera was 20% higher against natural milled sugarcane bagasse as compared with equimolar mixtures of the separate parental enzymes. Molecular dynamics simulations indicated the approximation of the two catalytic domains in the chimeric enzyme, and the formation of an inter-domain interface may underlie the improved catalytic function.

Fast evaluation of enantioselective drug metabolism by electrophoretically mediated microanalysis: application to fluoxetine metabolism by CYP2D6.

PubMed

Asensi-Bernardi, Lucía; Martín-Biosca, Yolanda; Escuder-Gilabert, Laura; Sagrado, Salvador; Medina-Hernández, María José

2013-12-01

In this work, a capillary electrophoretic methodology for the enantioselective in vitro evaluation of drugs metabolism is applied to the evaluation of fluoxetine (FLX) metabolism by cytochrome 2D6 (CYP2D6). This methodology comprises the in-capillary enzymatic reaction and the chiral separation of FLX and its major metabolite, norfluoxetine enantiomers employing highly sulfated β-CD and the partial filling technique. The methodology employed in this work is a fast way to obtain a first approach of the enantioselective in vitro metabolism of racemic drugs, with the additional advantage of an extremely low consumption of enzymes, CDs and all the reagents involved in the process. Michaelis-Menten kinetic parameters (Km and Vmax ) for the metabolism of FLX enantiomers by CYP2D6 have been estimated by nonlinear fitting of experimental data to the Michaelis-Menten equation. Km values have been found to be 30 ± 3 μM for S-FLX and 39 ± 5 μM for R-FLX. Vmax estimations were 28.6 ± 1.2 and 34 ± 2 pmol·min(-1) ·(pmol CYP)(-1) for S- and R-FLX, respectively. Similar results were obtained using a single enantiomer (R-FLX), indicating that the use of the racemate is a good option for obtaining enantioselective estimations. The results obtained show a slight enantioselectivity in favor of R-FLX. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Active mechanical stabilization of the viscoplastic intracellular space of Dictyostelia cells by microtubule-actin crosstalk.

PubMed

Heinrich, Doris; Sackmann, Erich

2006-11-01

The micro-viscoelasticity of the intracellular space of Dictyostelium discoideum cells is studied by evaluating the intracellular transport of magnetic force probes and their viscoelastic responses to force pulses of 20-700 pN. The role of the actin cortex, the microtubule (MT) aster and their crosstalk is explored by comparing the behaviour of wild-type cells, myosin II null mutants, latrunculin A and benomyl treated cells. The MT coupled beads perform irregular local and long range directed motions which are characterized by measuring their velocity distributions (P(v)). The correlated motion of the MT and the centrosome are evaluated by microfluorescence of GFP-labelled MTs. P(v) can be represented by log-normal distributions with long tails and it is determined by random sweeping motions (v approximately 0.5 microm/s) of the MTs (caused by tangential forces on the filament ends coupled to the actin cortex) and by intermittent bead transports parallel to the MTs (v(max) approximately 1.5 microm/s). The tails are due to spontaneous filament deflections (with speeds up to 10 microm/s) attributed to pre-stressing of the MT by local cortical tensions, generated by dynactin motors generating plus-end directed forces in the MTs. The viscoelastic responses are strongly non-linear and are mostly directed opposite or perpendicular to the force, showing that the cytoplasm behaves as an active viscoplastic body with time and force dependent drag coefficients. Nano-Newton loads exerted on the soft MT are balanced by traction forces arising at the MT ends coupled to the actin cortex and the centrosome, respectively. The mechanical coupling between the soft microtubules and the viscoelastic actin cortex provides cells with high mechanical stability despite the softness of the cytoplasm.

Effects of Acute Lymphoblastic Leukemia on Ceruloplasmin Oxidase, Copper and Several Markers of Oxidative Damage, in Children.

PubMed

Mehdi, Wesen Adel; Yusof, Faridah; Mehde, Atheer Awad; Zainulabdeen, Jwan Abdulmohsin; Raus, Raha Ahmed; Abdulbari, Alaa Shawqi

2015-01-01

Acute leukaemia is characterized by fast growth of abnormal clones of haemopoietic precursor cells inside bone marrow leading to undue accumulation in the bone marrow. Acute lymphoblastic leukemia (ALL) is the most common form of childhood cancer. The study concerned 50 children diagnosed with ALL (mean age, 8.55±2.54) compared to 40 healthy controls (mean age, 8.00±1.85). The Hb, serum copper, ceruloplasmin oxidase, advanced oxidation protein products (AOPPs), total antioxidant activity (TAA) and protein were measured in all groups. One proteinous component was isolated by gel filtration chromatography from the precipitate produced by polyethylene glycol. Significantly higher levels of AOPP, copper and decrease in total antioxidant activity were noted in the cases. Statistical analysis also showed a significant increase (p<0.01) in the activity of serum ceruloplasmin oxidase in patients with ALL compared to normal subjects. The maximum velocity (Vmax) and Michaelis constant had values of 104.2 U/L and 11.7 mM, respectively. The ΔH* values for ceruloplasmin oxidase in ALL patients were positive, confirming the reaction to be endothermic. The results from this study showed a significant increase in AOPP, ceruloplasmine oxidase and decrease in total antioxidant activity .These parameters may play a role in development of DNA damage in childhood patients with acute lymphoblastic leukemia (ALL). The ΔS* and ΔG* values were negative, these refer that the reaction of ES formation is spontaneous, but needs energy in a so-called endergonic reaction. Also the negative ΔS* value of ceruloplasmin oxidase indicates that the complex [ES*] is further modulated through increasing structure arrangement.

In vitro Inhibitory Effects of Andrographis paniculata, Gynura procumbens, Ficus deltoidea, and Curcuma xanthorrhiza Extracts and Constituents on Human Liver Glucuronidation Activity.

PubMed

Husni, Zulhilmi; Ismail, Sabariah; Zulkiffli, Mohd Halimhilmi; Afandi, Atiqah; Haron, Munirah

2017-07-01

Andrographis paniculata , Gynura procumbens , Ficus deltoidea and Curcuma xanthorrhiza are commonly consumed as herbal medicines. However their effects on human liver glucuronidation activity are not yet evaluated. In this study, we evaluate the inhibitory Effects of Andrographis paniculata, Gynura procumbens, Ficus deltoidea and Curcuma xanthorrhiza extracts and their constituents on human liver glucuronidation activity. Herbal extracts (aqueous, methanolic and ethanolic extracts) and their constituents were incubated with human liver microsomes with the addition of UDPGA to initiate the reaction. Working concentrations of herbal extracts and their constituents ranged from 10 μg/mL to 1000 μg/mL and 10 μM to 300 μM respectively. IC50 was determined by monitoring the decrement of glucuronidation activity with the increment of herbal extracts or phytochemical constituent's concentrations. All herbal extracts inhibited human liver glucuronidation activity in range of 34.69 μg/mL to 398.10 μg/mL whereas for the constituents, only xanthorrhizol and curcumin (constituents of Curcuma xanthorrhiza ) inhibited human liver glucuronidation activity with IC50 of 538.50 and 32.26 μM respectively. In the present study, we have proved the capabilities of Andrographis paniculata , Gynura procumbens , Ficus deltoidea and Curcuma xanthorrhiza to interfere with in vitro glucuronidation process in human liver microsomes. This study documented the capabilities of Andrographis paniculata , Gynura procumbens , Ficus deltoidea and Curcuma xanthorrhiza to inhibit human liver glucuronidation activity which may affect the metabolism of therapeutic drugs or hazardous toxicants that follow the same glucuronidation pathway. Abbreviations used: UGT: Uridine 5'-diphospho-glucuronosyltransferase; 4-MU: 4-methylumbelliferone; IC50: Half Maximal Inhibitory Concentration; Km: Michaelis constant; Vmax: Maximum velocity.

In vitro Inhibitory Effects of Andrographis paniculata, Gynura procumbens, Ficus deltoidea, and Curcuma xanthorrhiza Extracts and Constituents on Human Liver Glucuronidation Activity

PubMed Central

Husni, Zulhilmi; Ismail, Sabariah; Zulkiffli, Mohd Halimhilmi; Afandi, Atiqah; Haron, Munirah

2017-01-01

Background: Andrographis paniculata, Gynura procumbens, Ficus deltoidea and Curcuma xanthorrhiza are commonly consumed as herbal medicines. However their effects on human liver glucuronidation activity are not yet evaluated. Objective: In this study, we evaluate the inhibitory Effects of Andrographis paniculata, Gynura procumbens, Ficus deltoidea and Curcuma xanthorrhiza extracts and their constituents on human liver glucuronidation activity. Materials and Methods: Herbal extracts (aqueous, methanolic and ethanolic extracts) and their constituents were incubated with human liver microsomes with the addition of UDPGA to initiate the reaction. Working concentrations of herbal extracts and their constituents ranged from 10 μg/mL to 1000 μg/mL and 10 μM to 300 μM respectively. IC50 was determined by monitoring the decrement of glucuronidation activity with the increment of herbal extracts or phytochemical constituent's concentrations. Results: All herbal extracts inhibited human liver glucuronidation activity in range of 34.69 μg/mL to 398.10 μg/mL whereas for the constituents, only xanthorrhizol and curcumin (constituents of Curcuma xanthorrhiza) inhibited human liver glucuronidation activity with IC50 of 538.50 and 32.26 μM respectively. Conclusion: In the present study, we have proved the capabilities of Andrographis paniculata, Gynura procumbens, Ficus deltoidea and Curcuma xanthorrhiza to interfere with in vitro glucuronidation process in human liver microsomes. SUMMARY This study documented the capabilities of Andrographis paniculata, Gynura procumbens, Ficus deltoidea and Curcuma xanthorrhiza to inhibit human liver glucuronidation activity which may affect the metabolism of therapeutic drugs or hazardous toxicants that follow the same glucuronidation pathway. Abbreviations used: UGT: Uridine 5’-diphospho-glucuronosyltransferase; 4-MU: 4-methylumbelliferone; IC50: Half Maximal Inhibitory Concentration; Km: Michaelis constant; Vmax: Maximum velocity. PMID:28808386

Administration of exercise-conditioned plasma alters muscle catalase kinetics in rat: An argument for in vivo-like Km instead of in vitro-like Vmax.

PubMed

Veskoukis, Aristidis S; Paschalis, Vassilis; Kyparos, Antonios; Nikolaidis, Michalis G

2018-05-01

Maximal velocity (V max ) is a well established biomarker for the assessment of tissue redox status. There is scarce evidence, though, that it does not probably reflect sufficiently in vivo tissue redox profile. Instead, the Michaelis constant (K m ) could more adequately image tissue oxidative stress and, thus, be a more physiologically relevant redox biomarker. Therefore, the aim of the present study was to side-by-side compare V max and K m of an antioxidant enzyme after implementing an in vivo set up that induces alterations in tissue redox status. Forty rats were divided into two groups including rats injected with blood plasma originating from rats that had previously swam until exhaustion and rats injected with blood plasma originating from sedentary rats. Tail-vein injections were performed daily for 21 days. Catalase V max and K m measured in gastrocnemius muscle were increased after administration of the exercise-conditioned plasma, denoting enhancement of the enzyme activity but impairment of its affinity for the substrate, respectively. These alterations are potential adaptations stimulated by the administered plasma pointing out that blood is an active fluid capable of regulating tissue homeostasis. Our findings suggest that K m adequately reflects in vivo modifications of skeletal muscle catalase and seems to surpass V max regarding its physiological relevance and biological interpretation. In conclusion, K m can be regarded as an in vivo-like biomarker that satisfactorily images the intracellular environment, as compared to V max that could be aptly parallelized with a biomarker that describes tissue oxidative stress in an in vitro manner. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Pharmacokinetic and pharmacodynamic interaction between nifedipine and metformin in rats: competitive inhibition for metabolism of nifedipine and metformin by each other via CYP isozymes.

PubMed

Choi, Young H; Lee, Myung G

2012-05-01

It has been reported that hypertension exponentially increases in the patients with type 2 diabetes mellitus. Thus, this study was performed to investigate the pharmacokinetic and pharmacodynamic interactions between nifedipine and metformin, since both drugs were commonly metabolized via hepatic CYP2C and 3A subfamilies in rats. Nifedipine (3 mg/kg) and metformin (100 mg/kg) were simultaneously administered intravenously or orally to rats. Concentrations (I) of each drug in the liver and intestine, maximum velocity (V(max)), Michaelis-Menten constant (K(m)), and intrinsic clearance (CL(int)) for the disappearance of each drug, apparent inhibition constant (K(i)) and [I]/K(i) ratios of each drug in liver and intestine were determined. Also the metabolism of each drug in rat and human CYPs and blood pressure were also measured. After the simultaneous single intravenous administration of both drugs together, the AUCs of each drug were significantly greater than that in each drug alone due to the competitive inhibition for the metabolism of nifedipine by metformin via hepatic CYP3A1/2 and of metformin by nifedipine via hepatic CYP2C6 and 3A1/2. After the simultaneous single oral administration of both drugs, the significantly greater AUCs of each drug than that in each drug alone could have mainly been due to the competitive inhibition for the metabolism of nifedipine and metformin by each other via intestinal CYP3A1/2 in addition to competitive inhibition for the hepatic metabolism of each drug as same as the intravenous study.

Characterization of covalent binding of N'-nitrosonornicotine in rat liver microsomes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hughes, M.F.; Brock, W.J.; Marion, L.J.

1986-01-01

The metabolism of the carcinogenic nitrosamine, N'-nitrosonornicotine (NNN), to reactive intermediates which bind covalently was assessed using male Sprague-Dawley rat liver microsomes. The NADPH-dependent covalent binding of (/sup 14/C)NNN was linear with time up to 90 min and protein concentration up to 3.0 mg/ml. The apparent Km and Vmax of the binding were determined from the initial velocities and found to be 0.91 mM and 4.7 pmol/min/mg protein, respectively. Although NNN is not a hepatocarcinogen, this amount of NADPH-dependent covalent binding is 7-fold greater than that reported for dimethylnitrosamine, a potent hepatocarcinogen. Extensive covalent binding of (/sup 14/C)NNN to livermore » and muscle microsomal protein was also present in the absence of an NADPH-generating system and in the presence of 50% methanol, indicating a non-enzymatically mediated reaction. Addition of the nucleophiles glutathione, cysteine and N-acetylcysteine significantly decreased (p less than 0.01) the non-NADPH-dependent binding, but did not affect NADPH-dependent binding. In vitro addition of the cytochrome P-450 inhibitors metyrapone, piperonyl butoxide and SKF-525A significantly decreased (p less than 0.05) NADPH-dependent binding of (14C)NNN by 27-40%. NADH did not replace NADPH in supporting covalent binding. Replacement of an air atmosphere with nitrogen or CO:O2 (8:2) significantly decreased (p less than 0.05) NADPH-dependent binding of (/sup 14/C)NNN by 40 and 27%, respectively. Aroclor 1254 pre-treatment of the rats did not enhance the NADPH-dependent binding of (/sup 14/C)NNN. These data indicate that cytochrome P-450 is at least in part responsible for the metabolic activation of the carcinogen NNN but also suggest additional mechanisms of activation.« less

The effect of Zn2+ on glucose 6-phosphate dehydrogenase activity from Bufo arenarum toad ovary and alfalfa plants.

PubMed

Fonovich de Schroeder, Teresa M

2005-02-01

The effect of Zn2+ on glucose 6-phosphate dehydrogenase (G6PD) activity was monitored in samples from Bufo arenarum toad ovary and alfalfa plants, in the search for a possible new bioindicator able to detect levels of exposure through contaminated soils, and also to elucidate possible similarities between the enzyme from animal and plant tissues. The in vivo effect was evaluated after exposure of the toads to the metal in Ringer solution during 30 days and after 10 days of treatment in 6 weeks old plants, cultured under laboratory conditions. In vitro effects were measured in different extracts from control samples and partially purified enzyme from ovarian tissue as well as in different extracts from control alfalfa plants, by addition of the metal to the reaction mixture containing the enzyme. G6PD from toad ovary was noncompetitively inhibited by zinc both in vivo and in vitro, under all the experimental conditions studied. A kinetic analysis of the enzyme activity showed that the Michaelis-Menten constant (Km) was not modified, while maximal velocity (Vmax) decreased as the consequence of treatment. It was not possible to obtain a dose-response curve for the effects of Zn2+ on G6PD from alfalfa whole plants, measured in vivo or in vitro. Only leaf extracts evidenced a possible relationship between treatment with the metal and G6PD activity alteration. The results agree with a possible role for G6PD as a biomarker of effect and exposure to Zn2+ in B. arenarum ovarian tissue but not in alfalfa plants.

Synthesis, structure, and glutathione peroxidase-like activity of amino acid containing ebselen analogues and diaryl diselenides.

PubMed

Selvakumar, Karuthapandi; Shah, Poonam; Singh, Harkesh B; Butcher, Ray J

2011-11-04

The synthesis of some ebselen analogues and diaryl diselenides, which have amino acid functions as an intramolecularly coordinating group (Se···O) has been achieved by the DCC coupling procedure. The reaction of 2,2'-diselanediylbis(5-tert-butylisophthalic acid) or the activated ester tetrakis(2,5-dioxopyrrolidin-1-yl) 2,2'-diselanediylbis(5-tert-butylisophthalate) with different C-protected amino acids (Gly, L-Phe, L-Ala, and L-Trp) afforded the corresponding ebselen analogues. The used precursor diselenides have been found to undergo facile intramolecular cyclization during the amide bond formation reaction. In contrast, the DCC coupling of 2,2'-diselanediyldibenzoic acid with C-protected amino acids (Gly, L/D-Ala and L-Phe) affords the corresponding amide derivatives and not the ebselen analogues. Some of the representative compounds have been structurally characterized by single-crystal X-ray crystallography. The glutathione peroxidase (GPx)-like activities of the ebselen analogues and the diaryl diselenides have been evaluated by using the coupled reductase assay method. Intramolecularly stabilized ebselen analogues show slightly higher maximal velocity (V(max)) than ebselen. However, they do not show any GPx-like activity at low GSH concentrations at which ebselen and related diselenides are active. This could be attributed to the peroxide-mediated intramolecular cyclization of the corresponding selenenyl sulfide and diaryl diselenide intermediates generated during the catalytic cycle. Interestingly, the diaryl diselenides with alanine (L,L or D,D) amide moieties showed excellent catalytic efficiency (k(cat)/K(M)) with low K(M) values in comparison to the other compounds. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Centrifuge-induced hypergravity: [ 3H]GABA and L-[ 14C]glutamate uptake, exocytosis and efflux mediated by high-affinity, sodium-dependent transporters

NASA Astrophysics Data System (ADS)

Borisova, T. A.; Himmelreich, N. H.

The effects of centrifuge-induced hypergravity on the presynaptic events have been investigated in order to provide further insight into regulation of glutamate and GABA neurotransmission and correlation between excitatory and inhibitory responses under artificial gravity conditions. Exposure of animals to hypergravity (centrifugation of rats at 10 G for 1 h) has been found to cause changes in the synaptic processes of brain, in particular neurotransmitter release and uptake in rat brain synaptosomes. Hypergravity loading resulted in more than two-fold enhancement of GABA transporter activity ( Vmax increased from 1.4 ± 0.3 nmol/min/mg of protein in the control group to 3.3 ± 0.59 nmol/min/mg of protein for the animals exposed to hypergravity ( P ⩽ 0.05)). The maximal velocity of L-[ 14C]glutamate uptake decreased from 12.5 ± 3.2 to 5.6 ± 0.9 nmol/min/mg of protein under artificial gravity conditions. Depolarization-evoked exocytotic release of the neurotransmitters has also changed in response to hypergravity. It increased for GABA (7.2 ± 0.54% and 11.74 ± 1.2% of total accumulated label for control and hypergravity, respectively ( P ⩽ 0.05)), but reduced for glutamate (14.4 ± 0.7% and 6.2 ± 1.9%, for control and hypergravity, respectively). Thus, comparative analysis of the neurotransmitter uptake and release has demonstrated that short-term centrifuge-induced 10 G hypergravity loading intensified inhibitory and attenuated excitatory processes in nerve terminals. The activation or reduction of neurotransmitter uptake appeared to be coupled with similarly directed alterations of the neurotransmitter release.

Structure finding in cosmological simulations: the state of affairs

NASA Astrophysics Data System (ADS)

Knebe, Alexander; Pearce, Frazer R.; Lux, Hanni; Ascasibar, Yago; Behroozi, Peter; Casado, Javier; Moran, Christine Corbett; Diemand, Juerg; Dolag, Klaus; Dominguez-Tenreiro, Rosa; Elahi, Pascal; Falck, Bridget; Gottlöber, Stefan; Han, Jiaxin; Klypin, Anatoly; Lukić, Zarija; Maciejewski, Michal; McBride, Cameron K.; Merchán, Manuel E.; Muldrew, Stuart I.; Neyrinck, Mark; Onions, Julian; Planelles, Susana; Potter, Doug; Quilis, Vicent; Rasera, Yann; Ricker, Paul M.; Roy, Fabrice; Ruiz, Andrés N.; Sgró, Mario A.; Springel, Volker; Stadel, Joachim; Sutter, P. M.; Tweed, Dylan; Zemp, Marcel

2013-10-01

The ever increasing size and complexity of data coming from simulations of cosmic structure formation demand equally sophisticated tools for their analysis. During the past decade, the art of object finding in these simulations has hence developed into an important discipline itself. A multitude of codes based upon a huge variety of methods and techniques have been spawned yet the question remained as to whether or not they will provide the same (physical) information about the structures of interest. Here we summarize and extent previous work of the `halo finder comparison project': we investigate in detail the (possible) origin of any deviations across finders. To this extent, we decipher and discuss differences in halo-finding methods, clearly separating them from the disparity in definitions of halo properties. We observe that different codes not only find different numbers of objects leading to a scatter of up to 20 per cent in the halo mass and Vmax function, but also that the particulars of those objects that are identified by all finders differ. The strength of the variation, however, depends on the property studied, e.g. the scatter in position, bulk velocity, mass and the peak value of the rotation curve is practically below a few per cent, whereas derived quantities such as spin and shape show larger deviations. Our study indicates that the prime contribution to differences in halo properties across codes stems from the distinct particle collection methods and - to a minor extent - the particular aspects of how the procedure for removing unbound particles is implemented. We close with a discussion of the relevance and implications of the scatter across different codes for other fields such as semi-analytical galaxy formation models, gravitational lensing and observables in general.

Microbial L-phenylalanine ammonia-lyase. Purification, subunit structure and kinetic properties of the enzyme from Rhizoctonia solani.

PubMed Central

Kalghatgi, K K; Subba Rao, P V

1975-01-01

1. Phenylalanine ammonia-lyase (EC 4.3.1.5) was purified to homogeneity from the acetone-dried powders of the mycelial felts of the plant pathogenic fungus Rhizoctonia solani. 2. A useful modification in protamine sulphate treatment to get substantial purification of the enzyme in a single-step is described. 3. The purified enzyme shows bisubstrate activity towards L-phenylalanine and L-tyrosine. 4. It is sensitive to carbonyl reagents and the inhibition is not reversed by gel filtration. 5. The molecular weight of the enzyme as determined by Sephadex G-200 chromatography and sucrose-density-gradient centrifugation is around 330000. 6. The enzyme is made up of two pairs of unidentical subunits, with a molecular weight of 70000 (alpha) and 90000 (beta) respectively. 7. Studies on initial velocity versus substrate concentration have shown significant deviations from Michaelis-Menten kinetics. 8. The double-reciprocal plots are biphasic (concave downwards) and Hofstee plots show a curvilinear pattern. 9. The apparent Km value increases from 0.18 mM to as high as 5.0 mM with the increase in the concentration of the substrate and during this process the Vmax, increases by 2-2.5-fold. 10. The value of Hill coefficient is 0.5. 11. Steady-state rates of phenylalanine ammonia-lyase reaction in the presence of inhibitors like D-phenylalanine, cinnamic, p-coumaric, caffeic, dihydrocaffeic and phenylpyruvic acid have shown that only one molecule of each type of inhibitor binds to a molecule of the enzyme. These observations suggest the involvement of negative homotropic interactions in phenylalanine ammonia-lyase. 12. The enzyme could not be desensitized by treatment with HgCl2, p-chloromercuribenzoic acid or by repeated freezing and thawing. PMID:1191266

Neo-deterministic seismic hazard scenarios for India—a preventive tool for disaster mitigation

NASA Astrophysics Data System (ADS)

Parvez, Imtiyaz A.; Magrin, Andrea; Vaccari, Franco; Ashish; Mir, Ramees R.; Peresan, Antonella; Panza, Giuliano Francesco

2017-11-01

Current computational resources and physical knowledge of the seismic wave generation and propagation processes allow for reliable numerical and analytical models of waveform generation and propagation. From the simulation of ground motion, it is easy to extract the desired earthquake hazard parameters. Accordingly, a scenario-based approach to seismic hazard assessment has been developed, namely the neo-deterministic seismic hazard assessment (NDSHA), which allows for a wide range of possible seismic sources to be used in the definition of reliable scenarios by means of realistic waveforms modelling. Such reliable and comprehensive characterization of expected earthquake ground motion is essential to improve building codes, particularly for the protection of critical infrastructures and for land use planning. Parvez et al. (Geophys J Int 155:489-508, 2003) published the first ever neo-deterministic seismic hazard map of India by computing synthetic seismograms with input data set consisting of structural models, seismogenic zones, focal mechanisms and earthquake catalogues. As described in Panza et al. (Adv Geophys 53:93-165, 2012), the NDSHA methodology evolved with respect to the original formulation used by Parvez et al. (Geophys J Int 155:489-508, 2003): the computer codes were improved to better fit the need of producing realistic ground shaking maps and ground shaking scenarios, at different scale levels, exploiting the most significant pertinent progresses in data acquisition and modelling. Accordingly, the present study supplies a revised NDSHA map for India. The seismic hazard, expressed in terms of maximum displacement (Dmax), maximum velocity (Vmax) and design ground acceleration (DGA), has been extracted from the synthetic signals and mapped on a regular grid over the studied territory.

The Copernicus Complexio: a high-resolution view of the small-scale Universe

NASA Astrophysics Data System (ADS)

Hellwing, Wojciech A.; Frenk, Carlos S.; Cautun, Marius; Bose, Sownak; Helly, John; Jenkins, Adrian; Sawala, Till; Cytowski, Maciej

2016-04-01

We introduce Copernicus Complexio (COCO), a high-resolution cosmological N-body simulation of structure formation in the ΛCDM model. COCO follows an approximately spherical region of radius ˜17.4 h-1 Mpc embedded in a much larger periodic cube that is followed at lower resolution. The high-resolution volume has a particle mass of 1.135 × 105 h-1 M⊙ (60 times higher than the Millennium-II simulation). COCO gives the dark matter halo mass function over eight orders of magnitude in halo mass; it forms ˜60 haloes of galactic size, each resolved with about 10 million particles. We confirm the power-law character of the subhalo mass function, overline{N}(>μ )∝ μ ^{-s}, down to a reduced subhalo mass Msub/M200 ≡ μ = 10-6, with a best-fitting power-law index, s = 0.94, for hosts of mass = 1012 h-1 M⊙. The concentration-mass relation of COCO haloes deviates from a single power law for masses M200 < afew × 108 h-1 M⊙, where it flattens, in agreement with results by Sanchez-Conde et al. The host mass invariance of the reduced maximum circular velocity function of subhaloes, ν ≡ Vmax/V200, hinted at in previous simulations, is clearly demonstrated over five orders of magnitude in host mass. Similarly, we find that the average, normalized radial distribution of subhaloes is approximately universal (I.e. independent of subhalo mass), as previously suggested by the Aquarius simulations of individual haloes. Finally, we find that at fixed physical subhalo size, subhaloes in lower mass hosts typically have lower central densities than those in higher mass hosts.

In vitro metabolic stability of moisture-sensitive rabeprazole in human liver microsomes and its modulation by pharmaceutical excipients.

PubMed

Ren, Shan; Park, Mi-Jin; Kim, Aera; Lee, Beom-Jin

2008-03-01

A reliable method to assess in vitro metabolic stability of rabeprazole and its modulation by Generally Recognized As Safe (GRAS)-listed pharmaceutical excipients was established in human liver microsomes. The metabolic stability of rabeprazole decreased as a function of incubation time, resulting in the formation of thioether rabeprazole via nonenzymatic degradation and enzymatic metabolism. Buffer type was also a determining factor for the degree of both nonenzymatic degradation and enzymatic metabolism. The net extent of enzymatic drug metabolism, obtained by calculating the difference in drug degradation between a microsome-present reaction system and a microsome-free solution, was about 9.20 +/- 0.67% in phosphate buffer and 2.27 +/- 1.76% in Tris buffer, respectively. Rabeprazole exhibited first-order kinetics in microsome-free solution but showed non-linear kinetics in the microsome-present reaction system. The maximal velocity, Vmax, in phosphate buffer was 5.07 microg mL(-1) h(-1) and the Michaelis-Menten constant, Km, was 10.39 microg mL(-1) by computer-fitting to the classical Michaelis-Menten equation for pattern of time-dependent change in the substrate concentration. The intact drug and its thioether form were well resolved and successfully identified by HPLC chromatography and liquid chromatography mass spectroscopy (LC/MS). The metabolic stability of rabeprazole was also modulated by the presence of pharmaceutical excipients. Among the five pharmaceutical excipients tested, poloxamer 188 and Gelucire 44/14 had potentially inhibitory effects on rabeprazole metabolism in human liver microsomes (p < 0.05). A greater understanding of metabolic stability and its modulation by pharmaceutical excipients would be useful for optimizing the bioavailability of rabeprazole at the early formulation stages.

Functional analysis of aldehyde oxidase using expressed chimeric enzyme between monkey and rat.

PubMed

Itoh, Kunio; Asakawa, Tasuku; Hoshino, Kouichi; Adachi, Mayuko; Fukiya, Kensuke; Watanabe, Nobuaki; Tanaka, Yorihisa

2009-01-01

Aldehyde oxidase (AO) is a homodimer with a subunit molecular mass of approximately 150 kDa. Each subunit consists of about 20 kDa 2Fe-2S cluster domain storing reducing equivalents, about 40 kDa flavine adenine dinucleotide (FAD) domain and about 85 kDa molybdenum cofactor (MoCo) domain containing a substrate binding site. In order to clarify the properties of each domain, especially substrate binding domain, chimeric cDNAs were constructed by mutual exchange of 2Fe-2S/FAD and MoCo domains between monkey and rat. Chimeric monkey/rat AO was referred to one with monkey type 2Fe-2S/FAD domains and a rat type MoCo domain. Rat/monkey AO was vice versa. AO-catalyzed 2-oxidation activities of (S)-RS-8359 were measured using the expressed enzyme in Escherichia coli. Substrate inhibition was seen in rat AO and chimeric monkey/rat AO, but not in monkey AO and chimeric rat/monkey AO, suggesting that the phenomenon might be dependent on the natures of MoCo domain of rat. A biphasic Eadie-Hofstee profile was observed in monkey AO and chimeric rat/monkey AO, but not rat AO and chimeric monkey/rat AO, indicating that the biphasic profile might be related to the properties of MoCo domain of monkey. Two-fold greater V(max) values were observed in monkey AO than in chimeric rat/monkey AO, and in chimeric monkey/rat AO than in rat AO, suggesting that monkey has the more effective electron transfer system than rat. Thus, the use of chimeric enzymes revealed that 2Fe-2S/FAD and MoCo domains affect the velocity and the quantitative profiles of AO-catalyzed (S)-RS-8359 2-oxidation, respectively.

Proton-coupled organic cation antiporter-mediated uptake of apomorphine enantiomers in human brain capillary endothelial cell line hCMEC/D3.

PubMed

Okura, Takashi; Higuchi, Kei; Kitamura, Atsushi; Deguchi, Yoshiharu

2014-01-01

R(-)-Apomorphine is a dopamine agonist used for rescue management of motor function impairment associated with levodopa therapy in Parkinson's disease patients. The aim of this study was to examine the role of proton-coupled organic cation antiporter in uptake of R(-)-apomorphine and its S-enantiomer in human brain, using human endothelial cell line hCMEC/D3 as a model. Uptake of R(-)- or S(+)-apomorphine into hCMEC/D3 cells was measured under various conditions to evaluate its time-, concentration-, energy- and ion-dependency. Inhibition by selected organic cations was also examined. Uptakes of both R(-)- and S(+)-apomorphine increased with time. The initial uptake velocities of R(-)- and S(+)-apomorphine were concentration-dependent, with similar Km and Vmax values. The cell-to-medium (C/M) ratio of R(-)-apomorphine was significantly reduced by pretreatment with sodium azide, but was not affected by replacement of extracellular sodium ion with N-methylglucamine or potassium. Intracellular alkalization markedly reduced the uptake, while intracellular acidification increased it, suggesting that the uptake is driven by an oppositely directed proton gradient. The C/M ratio was significantly decreased by amantadine, verapamil, pyrilamine and diphenhydramine (substrates or inhibitors of proton-coupled organic cation antiporter), while tetraethylammonium (substrate of organic cation transporters (OCTs)) and carnitine (substrate of carnitine/organic cation transporter 2; (OCTN2)) had no effect. R(-)-Apomorphine uptake was competitively inhibited by diphenhydramine. Our results indicate that R(-)-apomorphine transport in human blood-brain barrier (BBB) model cells is similar to S(+)-apomorphine uptake. The transport was dependent on an oppositely directed proton gradient, but was sodium- or membrane potential-independent. The transport characteristics were consistent with involvement of the previously reported proton-coupled organic cation antiporter.

Differential gene expression of CYP3A isoforms in equine liver and intestines.

PubMed

Tydén, E; Löfgren, M; Pegolo, S; Capolongo, F; Tjälve, H; Larsson, P

2012-12-01

Recently, seven CYP3A isoforms - CYP3A89, CYP3A93, CYP3A94, CYP3A95, CYP3A96, CYP3A97 and CYP129 - have been isolated from the horse genome. In this study, we have examined the hepatic and intestinal gene expression of these CYP3A isoforms using TaqMan probes. We have also studied the enzyme activity using luciferin-isopropyl acetal (LIPA) as a substrate. The results show a differential gene expression of the CYP3A isoforms in the liver and intestines in horses. In the liver, CYP3A89, CYP3A94, CYP3A96 and CYP3A97 were highly expressed, while in the intestine there were only two dominating isoforms, CYP3A93 and CYP3A96. The isoform CYP3A129 was not detected in the liver or the intestine, although this gene consists of a complete set of exons and should therefore code for a functional protein. It is possible that this gene is expressed in tissues other than the liver and intestines. In the intestine, both CYP3A96 and CYP3A93 showed the highest gene expression in the duodenum and the proximal parts of the jejunum. This correlated with a high protein expression in these tissues. Studies of the enzyme activity showed the same K(m) for the LIPA substrate in the liver and the intestine, while the maximum velocity (V(max)) in the liver was higher than in the intestine. Our finding of a differential gene expression of the CYP3A isoforms in the liver and the intestines contributes to a better understanding of drug metabolism in horses. © 2012 Blackwell Publishing Ltd.

Investigations of the Local Supercluster velocity field. III. Tracing the backside infall with distance moduli from the direct Tully-Fisher relation

NASA Astrophysics Data System (ADS)

Ekholm, T.; Lanoix, P.; Teerikorpi, P.; Fouqué, P.; Paturel, G.

2000-03-01

We have extended the discussion of Paper II (Ekholm et al. \\cite{Ekholm99a}) to cover also the backside of the Local Supercluster (LSC) by using 96 galaxies within Theta <30degr from the adopted centre of LSC and with distance moduli from the direct B-band Tully-Fisher relation. In order to minimize the influence of the Malmquist bias we required log Vmax>2.1 and sigma B_T<0.2mag. We found out that if RVirgo<20 Mpc this sample fails to follow the expected dynamical pattern from the Tolman-Bondi (TB) model. When we compared our results with the Virgo core galaxies given by Federspiel et al. (\\cite{Federspiel98}) we were able to constrain the distance to Virgo: RVirgo=20-24 Mpc. When analyzing the TB-behaviour of the sample as seen from the origin of the metric as well as that with distances from the extragalactic Cepheid PL-relation we found additional support to the estimate RVirgo= 21 Mpc given in Paper II. Using a two-component mass-model we found a Virgo mass estimate MVirgo=(1.5 - 2)x Mvirial, where Mvirial=9.375*E14Msun for RVirgo= 21 Mpc. This estimate agrees with the conclusion in Paper I (Teerikorpi et al. \\cite{Teerikorpi92}). Our results indicate that the density distribution of luminous matter is shallower than that of the total gravitating matter when q0<= 0.5. The preferred exponent in the density power law, alpha ~2.5, agrees with recent theoretical work on the universal density profile of dark matter clustering in an Einstein-deSitter universe (Tittley & Couchman \\cite{Tittley99}).

Porcine Stomach Smooth Muscle Force Depends on History-Effects

PubMed Central

Tomalka, André; Borsdorf, Mischa; Böl, Markus; Siebert, Tobias

2017-01-01

The stomach serves as food reservoir, mixing organ and absorption area for certain substances, while continually varying its position and size. Large dimensional changes during ingestion and gastric emptying of the stomach are associated with large changes in smooth muscle length. These length changes might induce history-effects, namely force depression (FD) following active muscle shortening and force enhancement (FE) following active muscle stretch. Both effects have impact on the force generating capacity of the stomach, and thus functional relevance. However, less is known about history-effects and active smooth muscle properties of stomach smooth muscle. Thus, the aim of this study was to investigate biomechanical muscle properties as force-length and force-velocity relations (FVR) of porcine stomach smooth muscle strips, extended by the analysis of history-effects on smooth muscle force. Therefore, in total n = 54 tissue strips were dissected in longitudinal direction from the ventral fundus of porcine stomachs. Different isometric, isotonic, and isokinetic contraction protocols were performed during electrical muscle stimulation. Cross-sectional areas (CSA) of smooth muscles were determined from cryo-histological sections stained with Picrosirius Red. Results revealed that maximum smooth muscle tension was 10.4 ± 2.6 N/cm2. Maximum shortening velocity (Vmax) and curvature factor (curv) of the FVR were 0.04 ± 0.01 [optimum muscle length/s] and 0.36 ± 0.15, respectively. The findings of the present study demonstrated significant (P < 0.05) FD [up to 32% maximum muscle force (Fim)] and FE (up to 16% Fim) of gastric muscle tissue, respectively. The FE- and FD-values increased with increasing ramp amplitude. This outstanding muscle behavior is not accounted for in existing models so far and strongly supports the idea of a holistic reflection of distinct stomach structure and function. For the first time this study provides a comprehensive set of stomach smooth muscle parameters including classic biomechanical muscle properties and history-dependent effects, offering the possibility for the development and validation of computational stomach models. Furthermore, this data set facilitates novel insights in gastric motility and contraction behavior based on the re-evaluation of existing contractile mechanisms. That will likely help to understand physiological functions or dysfunctions in terms of gastric accommodation and emptying. PMID:29093684

Ketopantoyl lactone reductase is a conjugated polyketone reductase.

PubMed

Hata, H; Shimizu, S; Hattori, S; Yamada, H

1989-03-01

Ketopantoyl lactone reductase (EC 1.1.1.168) of Saccharomyces cerevisiae was found to catalyze the reduction of a variety of natural and unnatural conjugated polyketone compounds and quinones, such as isatin, ninhydrin, camphorquinone and beta-naphthoquinone in the presence of NADPH. 5-Bromoisatin is the best substrate for the enzyme (Km = 3.1 mM; Vmax = 650 mumol/min/mg). The enzyme is inhibited by quercetin, and several polyketones. These results suggest that ketopantoyl lactone reductase is a carbonyl reductase which specifically catalyzes the reduction of conjugated polyketones.

Uptake of Free Choline by Isolated Perfused Rat Liver

NASA Astrophysics Data System (ADS)

Zeisel, Steven H.; Story, David L.; Wurtman, Richard J.; Brunengraber, Henri

1980-08-01

The uptake of free choline by isolated perfused rat liver was characterized. A saturable uptake mechanism [Ka=0.17± 0.07 mM (SD); Vmax=0.84± 0.16\\ μ mol/min × g dry weight] and a nonsaturable mechanism (through which uptake is proportional to choline concentration in the perfusate) were identified. Most of the choline transported into hepatocytes was converted to betaine, phosphorylcholine, or lecithin. Free choline also accumulated within the intracellular space, suggesting that choline oxidase activity does not always limit choline's uptake by the liver.

4-O-beta-D-galactopyranosyl-D-xylose: a new synthesis and application to the evaluation of intestinal lactase.

PubMed

Rivera-Sagredo, A; Fernández-Mayoralas, A; Jiménez-Barbero, J; Martín-Lomas, M; Villanueva, D; Aragón, J J

1992-04-10

4-O-beta-D-Galactopyranosyl-D-xylose (2) was prepared from benzyl 2,3-O-isopropylidene-beta-D-xylopyranoside by glycosylation with 2,3,4,6-tetra-O-benzoyl-alpha-D-galactopyranosyl bromide and subsequent deprotection. Compound 2 was hydrolyzed in vitro by intestinal lactase; the Vmax was 25% of that with lactose and the Km was 370mM (cf. 27mM for lactose). Oral administration of 2 suckling rats led to urinary excretion of D-xylose which could be estimated colorimetrically.

Single transporter for sulfate, selenate, and selenite in Escherichia coli K-12.

PubMed Central

Lindblow-Kull, C; Kull, F J; Shrift, A

1985-01-01

A Michaelis-Menten kinetic analysis of the transport of sulfate, selenate, and selenite into Escherichia coli K-12 showed that the three dianions were transported by the same carrier. Km values, used as a measure of the affinity of each ligand for the carrier, showed that sulfate was bound 5 times more tightly than selenate and 37 times more tightly than selenite. The specificity ratio, Vmax/Km, also indicated that sulfate was the preferred ligand. There was little difference in the ratios for selenate and selenite. PMID:3897189

Experimental Investigation of Aerodynamics of Feather-Covered Flapping Wing.

PubMed

Yang, Wenqing; Song, Bifeng

2017-01-01

Avian flight has an outstanding performance than the manmade flapping wing MAVs. Considering that the feather is light and strong, a new type of the flapping wing was designed and made, whose skeleton is carbon fiber rods and covered by goose feathers as the skin. Its aerodynamics is tested by experiments and can be compared with conventional artificial flapping wings made of carbon fiber rods as the skeleton and polyester membrane as the skin. The results showed that the feathered wing could generate more lift than the membrane wing in the same flapping kinematics because the feathered wing can have slots between feathers in an upstroke process, which can mainly reduce the negative lift. At the same time, the power consumption also decreased significantly, due to the decrease in the fluctuating range of the periodic lift curve, which reduced the offset consumption of lift. At the same time, the thrusts generated by the feather wing and the membrane wing are similar with each other, which increases with the increase of flapping frequency. In general, the aerodynamic performances of the feather wing are superior to that of the membrane wings.

Sea Butterfly Swimming: Time-resolved Tomographic PIV measurements

NASA Astrophysics Data System (ADS)

Murphy, David; Zheng, Lingxiao; Mittal, Rajat; Webster, Donald; Yen, Jeannette

2011-11-01

The planktonic sea butterfly Limacina helicina swims by flapping its flexible, wing-like parapodia. The appendage stroke kinematics of this shell-bearing pteropod are three-dimensional and likely contain elements of both drag-based (rowing) and lift-based (flapping) propulsion. Unsteady lift-generating mechanisms such as clap-and-fling may also be present. Upstroke and downstroke motions both propel the animal upward and roll it forwards and backwards, resulting in a sawtooth trajectory. We present time-resolved, tomographic PIV measurements of flow generated by free-swimming pteropods (Limacina helicina) moving upwards with average swimming speeds of 5 - 17 mm/s. The pteropods beat their appendages with a stroke frequency of 4 - 5 Hz. With a size range of 1 - 2 mm, the animals filmed in this study operate in a viscous environment with a Reynolds number of 5 to 20. The volumetric flow measurements provide insight into the three dimensional nature of the flow and into the relative importance of drag- and lift-based propulsion at this low Reynolds number. Preliminary results from Navier-Stokes simulations of the flow associated with the swimming of this organism will also be presented.

Force production of a hovering hummingbird

NASA Astrophysics Data System (ADS)

Luo, Haoxiang; Song, Jialei; Hedrick, Tyson

2013-11-01

A three-dimensional numerical study is performed for a hovering Ruby-throated hummingbird (Archilochus colubris) based on an immersed-boundary method. To accurately model the unsteady aerodynamics, realistic 3D wing kinematics is reconstructed from high-speed images of the wing motion filmed at 1000 frames per second, resulting in 25 frames per flapping cycle. A high-resolution grid is employed to resolve the vortices shed from the wing. The results are validated by comparing the spanwise vorticity and circulation with the previous PIV data and also by calculating the average lift. The force production shows significant asymmetry with the downstroke producing lift 2.6 times as high as the upstroke, despite a nearly horizontal stroke plane. The total power consumption is around 55 W/kg, which is twice of previous estimate. In this presentation, we will discuss several mechanisms that lead to the force asymmetry, including the drag-based lift and the leading-edge vortex behavior. We will also address the role of wing-wake interaction, which appears to be different for the hummingbird than some of the insects such as fruit flies. Supported by NSF (No. CBET-0954381).

Control of unsteady separated flow associated with the dynamic pitching of airfoils

NASA Technical Reports Server (NTRS)

Ahmed, Sajeer

1991-01-01

Although studies have been done to understand the dependence of parameters for the occurrence of deep stall, studies to control the flow for sustaining lift for a longer time has been little. To sustain the lift for a longer time, an understanding of the development of the flow over the airfoil is essential. Studies at high speed are required to study how the flow behavior is dictated by the effects of compressibility. When the airfoil is pitched up in ramp motion or during the upstroke of an oscillatory cycle, the flow development on the upper surface of the airfoil and the formation of the vortex dictates the increase in lift behavior. Vortex shedding past the training edge decreases the lift. It is not clear what is the mechanism associated with the unsteady separation and vortex formation in present unsteady environment. To develop any flow control device, to suppress the vortex formation or delay separation, it is important that this mechanism be properly understood. The research activities directed toward understanding these questions are presented and the results are summarized.

Subtype-Selective Small Molecule Inhibitors Reveal a Fundamental Role for Nav1.7 in Nociceptor Electrogenesis, Axonal Conduction and Presynaptic Release

PubMed Central

Estacion, Mark; Turner, Jamie; Mis, Malgorzata A.; Wilbrey, Anna; Payne, Elizabeth C.; Gutteridge, Alex; Cox, Peter J.; Doyle, Rachel; Printzenhoff, David; Lin, Zhixin; Marron, Brian E.; West, Christopher; Swain, Nigel A.; Storer, R. Ian; Stupple, Paul A.; Castle, Neil A.; Hounshell, James A.; Rivara, Mirko; Randall, Andrew; Dib-Hajj, Sulayman D.; Krafte, Douglas; Waxman, Stephen G.; Patel, Manoj K.; Butt, Richard P.; Stevens, Edward B.

2016-01-01

Human genetic studies show that the voltage gated sodium channel 1.7 (Nav1.7) is a key molecular determinant of pain sensation. However, defining the Nav1.7 contribution to nociceptive signalling has been hampered by a lack of selective inhibitors. Here we report two potent and selective arylsulfonamide Nav1.7 inhibitors; PF-05198007 and PF-05089771, which we have used to directly interrogate Nav1.7’s role in nociceptor physiology. We report that Nav1.7 is the predominant functional TTX-sensitive Nav in mouse and human nociceptors and contributes to the initiation and the upstroke phase of the nociceptor action potential. Moreover, we confirm a role for Nav1.7 in influencing synaptic transmission in the dorsal horn of the spinal cord as well as peripheral neuropeptide release in the skin. These findings demonstrate multiple contributions of Nav1.7 to nociceptor signalling and shed new light on the relative functional contribution of this channel to peripheral and central noxious signal transmission. PMID:27050761

A monoclonal antibody that targets a NaV1.7 channel voltage sensor for pain and itch relief

PubMed Central

Lee, Jun-Ho; Park, Chul-Kyu; Chen, Gang; Han, Qingjian; Xie, Rou-Gang; Liu, Tong; Ji, Ru-Rong; Lee, Seok-Yong

2014-01-01

Summary Voltage-gated sodium (NaV) channels control the upstroke of the action potentials in excitable cells. Multiple studies have shown distinct roles of NaV channel subtypes in human physiology and diseases, but subtype-specific therapeutics are lacking and the current efforts have been limited to small molecules. Here we present a monoclonal antibody that targets the voltage-sensor paddle of NaV1.7, the subtype critical for pain sensation. This antibody not only inhibits NaV1.7 with high selectivity but also effectively suppresses inflammatory and neuropathic pain in mice. Interestingly, the antibody inhibits acute and chronic itch, despite well-documented differences in pain and itch modulation. Using this antibody, we discovered that NaV1.7 plays a key role in spinal cord nociceptive and pruriceptive synaptic transmission. Our studies reveal that NaV1.7 is a target for itch management and the antibody has therapeutic potential for suppressing pain and itch. Our antibody strategy may have broad applications for voltage-gated cation channels. PMID:24856969

New micro waveforms firstly recorded on electrocardiogram in human.

PubMed

Liu, Renguang; Chang, Qinghua; Chen, Juan

2015-10-01

In our study, not only the P-QRS-T waves but also the micro-wavelets before QRS complex (in P wave and PR segment) and after QRS complex (ST segment and upstroke of T wave) were first to be identified on surface electrocardiogram in human by the "new electrocardiogram" machine (model PHS-A10) according to conventional 12-lead electrocardiogram connection methods. By comparison to the conventional electrocardiogram in 100 cases of healthy individuals and several patients with arrhythmias, we have found that the wavelets before P wave theoretically reflected electrical activity of sinus node and the micro-wavelets before QRS complex may be related to atrioventricular conduction system (atrioventricular node, His bundle and bundle branch) potentials. Noninvasive atrioventricular node and His bundle potential tracing will contribute to differentiation of the origin of wide QRS and the location of the atrioventricular block. We also have found that the wavelets after QRS complex may be associated with phase 2 and 3 repolarization of ventricular action potential, which will further reveal ventricular repolarization changes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Ketopantoyl-lactone reductase from Candida parapsilosis: purification and characterization as a conjugated polyketone reductase.

PubMed

Hata, H; Shimizu, S; Hattori, S; Yamada, H

1989-02-24

Ketopantoyl-lactone reductase (2-dehydropantoyl-lactone reductase, EC 1.1.1.168) was purified and crystallized from cells of Candida parapsilosis IFO 0708. The enzyme was found to be homogeneous on ultracentrifugation, high-performance gel-permeation liquid chromatography and SDS-polyacrylamide gel electrophoresis. The relative molecular mass of the native and SDS-treated enzyme is approximately 40,000. The isoelectric point of the enzyme is 6.3. The enzyme was found to catalyze specifically the reduction of a variety of natural and unnatural polyketones and quinones other than ketopantoyl lactone in the presence of NADPH. Isatin and 5-methylisatin are rapidly reduced by the enzyme, the Km and Vmax values for isatin being 14 microM and 306 mumol/min per mg protein, respectively. Ketopantoyl lactone is also a good substrate (Km = 333 microM and Vmax = 481 mumol/min per mg protein). Reverse reaction was not detected with pantoyl lactone and NADP+. The enzyme is inhibited by quercetin, several polyketones and SH-reagents. 3,4-Dihydroxy-3-cyclobutene-1,2-dione, cyclohexenediol-1,2,3,4-tetraone and parabanic acid are uncompetitive inhibitors for the enzyme, the Ki values being 1.4, 0.2 and 3140 microM, respectively, with isatin as substrate. Comparison of the enzyme with the conjugated polyketone reductase of Mucor ambiguus (S. Shimizu, H. Hattori, H. Hata and H. Yamada (1988) Eur. J. Biochem. 174, 37-44) and ketopantoyl-lactone reductase of Saccharomyces cerevisiae suggested that ketopantoyl-lactone reductase is a kind of conjugated polyketone reductase.

Molecular genetics of the platelet serotonin system in first-degree relatives of patients with autism.

PubMed

Cross, Sarah; Kim, Soo-Jeong; Weiss, Lauren A; Delahanty, Ryan J; Sutcliffe, James S; Leventhal, Bennett L; Cook, Edwin H; Veenstra-Vanderweele, Jeremy

2008-01-01

Elevated platelet serotonin (5-hydroxytryptamine, 5-HT) is found in a subset of children with autism and in some of their first-degree relatives. Indices of the platelet serotonin system, including whole blood 5-HT, 5-HT binding affinity for the serotonin transporter (K(m)), 5-HT uptake (V(max)), and lysergic acid diethylamide (LSD) receptor binding, were previously studied in 24 first-degree relatives of probands with autism, half of whom were selected for elevated whole blood 5-HT levels. All subjects were then genotyped for selected polymorphisms at the SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 loci. Previous studies allowed an a priori prediction of SLC6A4 haplotypes that separated the subjects into three groups that showed significantly different 5-HT binding affinity (K(m), p=0.005) and 5-HT uptake rate (V(max), p=0.046). Genotypes at four individual polymorphisms in SLC6A4 were not associated with platelet 5-HT indices. Haplotypes at SLC6A4 and individual genotypes of polymorphisms at SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 showed no significant association with whole blood 5-HT. Haplotype analysis of two polymorphisms in TPH1 revealed a nominally significant association with whole blood 5-HT (p=0.046). These initial studies of indices of the 5-HT system with several single-nucleotide polymorphisms at loci in this system generate hypotheses for testing in other samples.

AhIRT1 and AhNRAMP1 metal transporter expression correlates with Cd uptake in peanuts under iron deficiency

PubMed Central

Xia, Shenglan; Deng, Rubo; Liu, Caifeng; Shi, Gangrong

2017-01-01

Fe deficiency may increase Cd accumulation in peanuts. However, the mechanisms are not yet fully understood. In the present study, two contrasting peanut cultivars, Luhua 8 (low seed-Cd cultivar) and Zhenghong 3 (high seed-Cd cultivar) were used to investigate the effect of Fe deficiency on the uptake and accumulation of cadmium (Cd) by hydroponic experiments. Under Fe-sufficient conditions, compared with Luhua 8, Zhenghong 3 had higher specific root length (SRL) and proportion of fine roots with a lower Km for Cd and showed slightly higher expression of AhIRT1 and AhNRAMP1 in the roots. These traits may be responsible for high capacity for Cd accumulation in Zhenghong 3. Under Fe deficiency, the increase of Cd accumulation was much larger in Zhenghong 3 than in Luhua 8. Kinetics studies revealed that the Vmax for Cd influx was 1.56-fold higher in Fe-deficient plants than in Fe-sufficient plants for Zhenghong 3, versus 0.48-fold higher for Luhua 8. Moreover, the increased expression levels of AhIRT1 and AhNRAMP1 induced by Fe deficiency was higher in Zhenghong 3 than in Luhua 8. Yeast complementation assays suggested that the AhIRT1 and AhNRAMP1 may function as transporters involved in Cd uptake. In conclusion, the different Cd accumulation between the two cultivars under Fe deficiency may be correlated with Vmax value for Cd uptake and the expression levels of AhIRT1 and AhNRAMP1 in the roots. PMID:28981520

A GUI-based Tool for Bridging the Gap between Models and Process-Oriented Studies

NASA Astrophysics Data System (ADS)

Kornfeld, A.; Van der Tol, C.; Berry, J. A.

2014-12-01

Models used for simulation of photosynthesis and transpiration by canopies of terrestrial plants typically have subroutines such as STOMATA.F90, PHOSIB.F90 or BIOCHEM.m that solve for photosynthesis and associated processes. Key parameters such as the Vmax for Rubisco and temperature response parameters are required by these subroutines. These are often taken from the literature or determined by separate analysis of gas exchange experiments. It is useful to note however that subroutines can be extracted and run as standalone models to simulate leaf responses collected in gas exchange experiments. Furthermore, there are excellent non-linear fitting tools that can be used to optimize the parameter values in these models to fit the observations. Ideally the Vmax fit in this way should be the same as that determined by a separate analysis, but it may not because of interactions with other kinetic constants and the temperature dependence of these in the full subroutine. We submit that it is more useful to fit the complete model to the calibration experiments rather as disaggregated constants. We designed a graphical user interface (GUI) based tool that uses gas exchange photosynthesis data to directly estimate model parameters in the SCOPE (Soil Canopy Observation, Photochemistry and Energy fluxes) model and, at the same time, allow researchers to change parameters interactively to visualize how variation in model parameters affect predicted outcomes such as photosynthetic rates, electron transport, and chlorophyll fluorescence. We have also ported some of this functionality to an Excel spreadsheet, which could be used as a teaching tool to help integrate process-oriented and model-oriented studies.

L-arginine transport in retinas from streptozotocin diabetic rats: correlation with the level of IL-1 beta and NO synthase activity.

PubMed

Carmo, A; Cunha-Vaz, J G; Carvalho, A P; Lopes, M C

1999-11-01

Several evidences suggest that the pro-inflammatory cytokines IL-1 beta and the radical NO are implicated as effectors molecules in the pancreatic beta-cells dysfunction; an event preceding the pathogenesis of diabetes. IL-1 beta induces the expression of the inducible isoform of NO synthase (iNOS), which use L-arginine as substrate to overproduce NO. However, it is not known whether these events may participate in the development of diabetic retinopathy, which is the main cause of blindness. In this work, we found an increased level of IL-1 beta in retinas from streptozotocin-induced (STZ) diabetic rats. We also observed that the activity of the NO synthase (NOS) and the L-arginine uptake are enhanced in retinas from STZ-induced diabetic rats as compared to retinas from control rats. We found that the uptake of L-arginine in retinas from control and diabetic rats occurs through a transporter resembling the Y + system, i.e. it is saturable, not affected over the pH range 6.5 to 7.4, and is independent of the extracellular Na+. Nevertheless, the L-arginine transport in retinas from diabetic rats occurs through a carrier with lower affinity (K(m) = 25 microM) and higher capacity (Vmax = 295 +/- 22.4 pmol L-arginine/mg protein) than in retinas from control rats (K(m) = 5 microM and Vmax = 158 +/- 12.8 pmol L-arginine/mg protein) which is correlated with the increased NOS activity and consequent depletion of the intracellular pool of L-arginine.

A Single Amphetamine Infusion Reverses Deficits in Dopamine Nerve-Terminal Function Caused by a History of Cocaine Self-Administration.

PubMed

Ferris, Mark J; Calipari, Erin S; Rose, Jamie H; Siciliano, Cody A; Sun, Haiguo; Chen, Rong; Jones, Sara R

2015-07-01

There are ∼ 1.6 million people who meet the criteria for cocaine addiction in the United States, and there are currently no FDA-approved pharmacotherapies. Amphetamine-based dopamine-releasing drugs have shown efficacy in reducing the motivation to self-administer cocaine and reducing intake in animals and humans. It is hypothesized that amphetamine acts as a replacement therapy for cocaine through elevation of extracellular dopamine levels. Using voltammetry in brain slices, we tested the ability of a single amphetamine infusion in vivo to modulate dopamine release, uptake kinetics, and cocaine potency in cocaine-naive animals and after a history of cocaine self-administration (1.5 mg/kg/infusion, fixed-ratio 1, 40 injections/day × 5 days). Dopamine kinetics were measured 1 and 24 h after amphetamine infusion (0.56 mg/kg, i.v.). Following cocaine self-administration, dopamine release, maximal rate of uptake (Vmax), and membrane-associated dopamine transporter (DAT) levels were reduced, and the DAT was less sensitive to cocaine. A single amphetamine infusion reduced Vmax and membrane DAT levels in cocaine-naive animals, but fully restored all aspects of dopamine terminal function in cocaine self-administering animals. Here, for the first time, we demonstrate pharmacologically induced, immediate rescue of deficits in dopamine nerve-terminal function in animals with a history of high-dose cocaine self-administration. This observation supports the notion that the DAT expression and function can be modulated on a rapid timescale and also suggests that the pharmacotherapeutic actions of amphetamine for cocaine addiction go beyond that of replacement therapy.

Properties and mutation studies of a bacteriophage-derived chimeric recombinant staphylolytic protein P128

PubMed Central

Saravanan, Sanjeev Rajagopalan; Paul, Vivek Daniel; George, Shilpa; Sundarrajan, Sudarson; Kumar, Nirmal; Hebbur, Madhavi; Kumar, Naveen; Veena, Ananda; Maheshwari, Uma; Appaiah, Chemira Biddappa; Chidambaran, Muralidharan; Bhat, Anuradha Gopal; Hariharan, Sukumar; Padmanabhan, Sriram

2013-01-01

P128 is a chimeric anti-staphylococcal protein having a catalytic domain from a Staphylococcus bacteriophage K tail associated structural protein and a cell wall targeting domain from the Staphylococcus bacteriocin-lysostaphin. In this study, we disclose additional properties of P128 and compared the same with lysostaphin. While lysostaphin was found to get inactivated by heat and was inactive on its parent strain S. simulans biovar staphylolyticus, P128 was thermostable and was lytic towards S. simulans biovar staphylolyticus demonstrating a difference in their mechanism of action. Selected mutation studies of the catalytic domain of P128 showed that arginine and cysteine, at 40th and 76th positions respectively, are critical for the staphylolytic activity of P128, although these amino acids are not conserved residues. In comparison to native P128, only the R40S mutant (P301) was catalytically active on zymogram gel and had a similar secondary structure, as assessed by circular dichroism analysis and in silico modeling with similar cell binding properties. Mutation of the arginine residue at 40th position of the P128 molecule caused dramatic reduction in the Vmax (∆OD600 [mg/min]) value (nearly 270 fold) and the recombinant lysostaphin also showed lesser Vmax value (nearly 1.5 fold) in comparison to the unmodified P128 protein. The kinetic parameters such as apparent Km (Km APP) and apparent Kcat (KcatAPP) of the native P128 protein also showed significant differences in comparison to the values observed for P301 and lysostaphin. PMID:24251076

Kinetic behaviour of calf intestinal alkaline phosphatase with pNPP.

PubMed

Chaudhuri, Gouri; Chatterjee, Saswata; Venu-Babu, P; Ramasamy, K; Thilagaraj, W Richard

2013-02-01

The hydrolysis of p-nitrophenyl phosphate (pNPP) by calf intestinal alkaline phosphatase (CIAP) was investigated with respect to kinetic parameters such as V(max), K(m) and K(cat) under varying pH, buffers, substrate concentration, temperature and period of incubation. Highest activity was obtained with Tris-HCl at pH 11, while in the case of glycine-NaOH buffer the peak activity was recorded at pH 9.5. The enzyme showed the following kinetic characteristics with pNPP in 50 mM Tris-HCl at pH 11 and 100 mM glycine-NaOH at pH 9.5 at an incubation temperature of 37 degrees C: V(max), 3.12 and 1.6 micromoles min(-1) unit(-1); K(m), 7.6 x 10(-4) M and 4 x 10(-4) M; and K(cat), 82.98 s(-1) and 42.55 s(-1), respectively. CIAP displayed a high temperature optimum of 45 degrees C at pH 11. The kinetic behaviour of the enzyme under different parameters suggested that the enzyme might undergo subtle conformational changes in response to the buffers displaying unique characteristics. Bioprecipitation of Cu2+ from 50 ppm of CuCl2 solution was studied where 64.3% of precipitation was obtained. P(i) generated from CIAP-mediated hydrolysis of pNPP was found to bind with copper and precipitated as copper-phosphate. Thus, CIAP could be used as a test candidate in bioremediation of heavy metals from industrial wastes through generation of metal-phosphate complexes.