Stabilization,solubilization, partial purification and some properties of cytochrome P-450 present in CaCl2-precipitated human placental microsomes

Two forms of a carbon monoxide-binding cytochrome were found to exist in CaCl₂-precipitated microsomal subfractions of human placental homogenates at term. These exhibited absorption maxima at wavelengths of 450 nm and 421–432 nm. Conversion of P-450 to the degradation product could be prevented by resuspension of the microsomes in buffered solutions containing 20% glycerol and 10^?4M dithiothreitol. Solubilization of such resuspended microsomes with sodium cholate could be effected with minimal degradation. The solubilized pigment then could be partially purified by differential fractionation with ammonium sulfate. The partially purified cytochromes did not appear to bind desmethylimpramine but did bind aniline and nicotinamide to yield atypical type II difference spectra with maxima at 435 nm and minima at 416 nm. A type I difference spectrum could be elicited with androstenedione and a spectral dissociation constant (K_S) of 4.7×10^?8M was obtained. Androstenedione also appeared to effectively prevent the binding of carbon monoxide to the cytochrome.     

Bioluminescence imaging: a shining future for cardiac regeneration

Advances in bioanalytical techniques have become crucial for both basic research and medical practice. One example, bioluminescence imaging (BLI), is based on the application of natural reactants with light‐emitting capabilities (photoproteins and luciferases) isolated from a widespread group of organisms. The main challenges in cardiac regeneration remain unresolved, but a vast number of studies have harnessed BLI with the discovery of aequorin and green fluorescent proteins. First described in the luminous hydromedusan Aequorea victoria in the early 1960s, bioluminescent proteins have greatly contributed to the design and initiation of ongoing cell‐based clinical trials on cardiovascular diseases. In conjunction with advances in reporter gene technology, BLI provides valuable information about the location and functional status of regenerative cells implanted into numerous animal models of disease. The purpose of this review was to present the great potential of BLI, among other existing imaging modalities, to refine effectiveness and underlying mechanisms of cardiac cell therapy. We recount the first discovery of natural primary compounds with light‐emitting capabilities, and follow their applications to bioanalysis. We also illustrate insights and perspectives on BLI to illuminate current efforts in cardiac regeneration, where the future is bright.     

Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication

DNA Polymerases generate pyrophosphate every time they catalyze a step of DNA elongation. This elongation reaction is generally believed as thermodynamically favoured by the hydrolysis of pyrophosphate, catalyzed by inorganic pyrophosphatases. However, the specific action of inorganic pyrophosphatases coupled to DNA replication in vivo was never demonstrated. Here we show that the Polymerase-Histidinol-Phosphatase (PHP) domain of Escherichia coli DNA Polymerase III α subunit features pyrophosphatase activity. We also show that this activity is inhibited by fluoride, as commonly observed for inorganic pyrophosphatases, and we identified 3 amino acids of the PHP active site. Remarkably, E. coli cells expressing variants of these catalytic residues of α subunit feature aberrant phenotypes, poor viability, and are subject to high mutation frequencies. Our findings indicate that DNA Polymerases can couple DNA elongation and pyrophosphate hydrolysis, providing a mechanism for the control of DNA extension rate, and suggest a promising target for novel antibiotics.     

New nitrogen mustards structurally related to (L)-carnitine

Enantiopure nitrogen mustards which mimic (L)-carnitine framework are prepared by a multi-step synthesis from the (R)-di-tert-butyl malate and their antitumor properties evaluated.     

The Impacts of Household Consumption and Options for Change

This introductory article situates the contributions that comprise this special issue within the field of sustainable consumption and production (SCP) studies. After a brief review of the policy history surrounding SCP, we organize our discussion and the subsequent collection of articles into two groups. The first suite of articles views the environmental impacts associated with household consumption from the perspectives of different consumer groups, income levels, and geographic areas. This work confirms and refines several insights that have been developing over the past several years, namely that food and beverages, mobility, housing, and energy-using products are the most critical consumption domains from the standpoint of environmental sustainability and that higher household income leads to greater (but less than proportional) impacts. The second subset of articles analyzes the potential for mitigating these impacts through behavioral changes and innovation strategies. Although the contributions to this special issue describe several noteworthy examples of information- and team-based initiatives to catalyze behavioral changes, the state of knowledge pertaining to this aspect of the consumption problem is much more inchoate. Research on the formulation and implementation of effective "change management for sustainable consumption" should be treated as an area of priority attention for industrial ecologists.     

Optimal foot shape for a passive dynamic biped

Passive walking dynamics describe the motion of a biped that is able to "walk" down a shallow slope without any actuation or control. Instead, the walker relies on gravitational and inertial effects to propel itself forward, exhibiting a gait quite similar to that of humans. These purely passive models depend on potential energy to overcome the energy lost when the foot impacts the ground. Previous research has demonstrated that energy loss at heel-strike can vary widely for a given speed, depending on the nature of the collision. The point of foot contact with the ground (relative to the hip) can have a significant effect: semi-circular (round) feet soften the impact, resulting in much smaller losses than point-foot walkers. Collisional losses are also lower if a single impulse is broken up into a series of smaller impulses that gradually redirect the velocity of the center of mass rather than a single abrupt impulse. Using this principle, a model was created where foot-strike occurs over two impulses, "heel-strike" and "toe-strike," representative of the initial impact of the heel and the following impact as the ball of the foot strikes the ground. Having two collisions with the flat-foot model did improve efficiency over the point-foot model. Representation of the flat-foot walker as a rimless wheel helped to explain the optimal flat-foot shape, driven by symmetry of the virtual spoke angles. The optimal long period foot shape of the simple passive walking model was not very representative of the human foot shape, although a reasonably anthropometric foot shape was predicted by the short period solution.     

Optimal discus trajectories

A general 3-D dynamic model for men's and women's discus flight is presented including precession of spin angular momentum induced by aerodynamic pitching moment. Dependence of pitching moment coefficient on angle of attack is estimated from experiment. Numerical integration of 11 equations of motion for nominal release speed v₀=25 m/s and axial spin p₀=42 rad/s also requires 3 other release conditions; initial discus flight path angle β₀, pitch attitude θ₀, and roll angle φ₀. Optimal values for these release conditions are calculated iteratively to maximize range and are similar for both men and women. The optimal men's trajectory and range R=69.39 m is produced by the strategy β₀=38.4°, θ₀=30.7°, and φ₀=54.4°. Initial angular velocities except spin are chosen to minimize wobble but an optimal initial spin rate p₀=25.2 rad/s exists that also maximizes range. Optimal 3-D range exceeds that predicted by 2-D models because, although angle of attack and lift are negative initially, 3-D motion allows advantageous orientation of lift later in flight, with tilt of the axis of symmetry from vertical becoming much smaller at landing. Optimal strategies are discontinuous with wind speed, resulting in slicing and kiting strategies in large head and tail winds, respectively. Sensitivity of optimal range is largest to initial β₀ and least to φ₀. Present calculations do not account for dependence of initial release angle or spin on release velocity or among other release conditions.     

Fertilization and allelopathy modify Pinus halepensis saplings crown acclimation to shade

Pinus halepensis Mill., is a Mediterranean pioneer forest species with shade-intolerant features. The purpose of this study is to better understand how stand fertility and allelopathic properties of adult trees influence shade acclimation of saplings. Crown growth and morphological plasticity were studied under different light, fertilization, and allelopathic conditions in a nursery experiment. We tested whether shade-acclimation capacity increases with fertilization, and is affected by autotoxicity due to pine leachates. We examined stem diameter, and crown characteristics (length, width, shape, and density) in a factorial experiment with two levels for each tested factor: light (full and 20% reduced light), fertilization (low and high rate of NPK fertilizer) and allelopathy (control and allelopathic leachates uptake). In our study, shading induced a significantly higher crown length, width, and surface. Fertilization strongly increased crown length and vertical expended crown shape (the ratio crown length/crown width). Leachates uptake reduced crown length and density, highlighting an autotoxicity phenomenon. We concluded that P. halepensis saplings presented a shade-avoiding syndrome and that the crown shade-acclimation response increased with fertilization but was severely compromised by autotoxicity. We finally discuss the role of fertilization and allelopathy in early P. halepensis acclimation ability.     

Musculoskeletal modeling and dynamic simulation of the thoroughbred equine forelimb during stance phase of the gallop

Because thoroughbred racehorses have a high incidence of forelimb musculoskeletal injuries, a model was desired to screen potential risk factors for injuries. This paper describes the development of a musculoskeletal model of the thoroughbred forelimb and a dynamic simulation of the motion of the distal segments during the stance phase of high-speed (18 m/s) gallop. The musculoskeletal model is comprised of segment, joint, muscle-tendon, and ligament information. The dynamic simulation incorporates a proximal forward-driving force, a distal ground reaction force model, muscle activations, and initial positions and velocities. A simulation of the gallop after transection of an accessory ligament demonstrated increased soft tissue strains in the remaining support structures of the distal forelimb. These data were consistent with those previously reported from in vitro experimental data and supported usefulness of the model for the study of distal forelimb soft tissue mechanics during the stance phase of the gallop.