Recent advances in the large-scale production of antibody fragments using lower eukaryotic microorganisms

BACKGROUND AND OBJECTIVES: Magnetic resonance microscopy (MRM) is a technique that is worthwhile for anesthesiologists because it allows spinal cord and plexus anatomy to be visualized three dimensionally and followed over time in the same animal. For example, the long-term effect of indwelling intrathecal or plexus catheters can be studied in situ, and convective and diffusive forces within intrathecal, epidural, or nerve sheath spaces can be investigated. Further, diffusion-weighted MRM, which measures an "apparent diffusion coefficient" (ADC), can be used to track the presence of ischemia, hypoperfusion, or cytotoxic edema. This study investigates problems associated with the use of in vivo MRM for spinal cord and peripheral nerve studies in the rat. METHODS: Twenty-one anesthetized female Fisher CDF rats were used. Group 1 (n=7) was used for anatomic three-dimensional studies. Groups 2 (n=4), 3 (n=4), and 4 (n=6) were used for measurements of the ADC. Group 2 served as controls, group 3 received lumbar intrathecal catheters, and group 4 received cervical intrathecal catheters. RESULTS: Cervical spine, lumbar spine, and spinal nerves and ganglia were accurately visualized with MRM. As a rule, spinal cord gray and white matter were better demonstrated using diffusion-weighted proton stains. By contrast, T2-weighted proton staining superiorly demonstrated structures surrounding the spinal cord. In groups 3 and 4, indwelling intrathecal catheters did not affect the spinal cord ADC, indicating normal blood flow and no cytotoxic edema. Contrast studies revealed nonhomogeneous distribution of contrast predominately in the lateral and ventral intrathecal space. CONCLUSION: Three-dimensional diffusion-weighted MRM displays cervical and lumbar spine anatomy accurately in vivo. Apparent diffusion coefficients measurements are feasible in rat cervical spinal cord with intrathecal catheters. Spinal cord ADCs are unaffected by intrathecal catheters, indicating normal spinal cord perfusion.     

NUMERICAL SOLUTION OF FORCED CONVECTION HEAT TRANSFER IN He II

A theoretical investigation of one-dimensional forced convection heat transfer in He II is conducted. The problem of interest involves a flow tube containing He II, which is heated at its midpoint along its length. Two modes of heating are analyzed: step function and square pulse. The one-dimensional He II energy equation is used to find the temperature distribution along the tube for both steady-state and transient situations. For the steady-state case, a numerical integration routine is used to obtain a solution, whereas for the solution of the transient case, a finite-difference scheme is developed. The numerical temperature profiles are then shown to compare well with the results of an experiment.     

Monte Carlo uncertainty quantification of the effective delayed neutron fraction

The applicability of Monte Carlo techniques, namely the Monte Carlo sensitivity method and the random-sampling method, for uncertainty quantification of the effective delayed neutron fraction β_eff is investigated using the continuous-energy Monte Carlo transport code, MCNP, from the perspective of statistical convergence issues. This study focuses on the nuclear data as one of the major sources of β_eff uncertainty. For validation of the calculated β_eff, a critical configuration of the VENUS-F zero-power reactor was used. It is demonstrated that Chiba's modified k-ratio method is superior to Bretscher's prompt k-ratio method in terms of reducing the statistical uncertainty in calculating not only β_eff but also its sensitivities and the uncertainty due to nuclear data. From this result and a comparison of uncertainties obtained by the Monte Carlo sensitivity method and the random-sampling method, it is shown that the Monte Carlo sensitivity method using Chiba's modified k-ratio method is the most practical for uncertainty quantification of β_eff. Finally, total β_eff uncertainty due to nuclear data for the VENUS-F critical configuration is determined to be approximately 2.7% with JENDL-4.0u, which is dominated by the delayed neutron yield of ²³⁵U.     

Substrate Activity Screening (SAS) and Related Approaches in Medicinal Chemistry

Substrate activity screening (SAS) was presented a decade ago by Ellman and co‐workers as a straightforward methodology for the identification of fragment‐sized building blocks for enzyme inhibitors. Ever since, SAS and variations derived from it have been successfully applied to the discovery of inhibitors of various families of enzymatically active drug targets. This review covers key achievements and challenges of SAS and related methodologies, including the modified substrate activity screening (MSAS) approach. Special attention is given to the kinetic and thermodynamic aspects of these methodologies, as a thorough understanding thereof is crucial for successfully transforming the identified fragment‐sized hits into potent inhibitors.     

Estimates of N2O and CH4 fluxes from agricultural lands in various regions in Europe

According to the revised 1996 IPCC guidelines, several emission factors are needed to calculate national inventories of N₂O emissions from agriculture. To estimate the direct N₂O emissions from mineral soils, an emission factor of 0.0125 kg N₂O-N per kg N applied is currently being used. From recent literature data it was clearly shown that real N₂O emissions could differ substantially from this value. Based on the IPCC methodology an inventory of N₂O emission from agriculture in Europe (EU-15) has been made. In 1996, the N₂O emission was estimated at 672 Gg N₂O-N. The N₂O emission per country varied between 10 and 177 Gg N₂O-N. The N₂O emission per ha agricultural land in the various countries varied between 1.7 and 14.2 kg N₂O-N ha⁻¹. Highest N₂O emissions per ha were found in countries with a high agricultural intensity, such as the Netherlands, Belgium-Luxembourg, Denmark and Germany. Agricultural soils are a sink for atmospheric methane. An oxidation capacity of 2.5 and 1.5 kg CH₄ ha⁻¹ yr⁻¹ was put forward for grasslands and arable land, respectively. Based on land use data of 1993, the CH₄ sink of agricultural lands in EU-15 was estimated at 303.5 Gg CH₄. In general, it could be concluded that N₂O emissions from soils (327 Tg CO₂ equivalents) are far more important than its sink function for CH₄ (6.3 Tg CO₂ equivalents). This revised version was published online in August 2006 with corrections to the Cover Date.     

Crystallization of vulcanizates. I. Low‐temperature crystallization as a function of extent of cure for polyisoprene vulcanized with tetramethylthiuram disulfide/sulfur and 2‐bisbenzothiazole‐2,2′‐disulfide/sulfur

The crystallization of polyisoprene, vulcanized to various degrees of cure with tetramethylthiuram disulfide/sulfur and 2‐bisbenzothiazole‐2,2′‐disulfide (MBTS)/sulfur formulations, was studied in a density column at ?25°C. The densities of vulcanizates before crystallization decrease progressively with cure time, which is ascribed to an increase in free volume occasioned by the formation of accelerator‐terminated pendent groups on the polymer chain. The induction period before the onset of crystallization increases and both the rate of and the degree of crystallization decrease with extent of cure. This is attributed primarily to the presence of residual pendent groups on the polymer chain and secondly to crosslink formation. The changes are more marked with MBTS formulations where pendent groups are more bulky. MBTS compounds fail to crystallize once vulcanized to the point where a gel has formed. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 2565–2572, 2001     

Load‐carrying ability of polystyrene products with molded‐in holes

The combined influence of stress concentration, weld line, and local distribution of molecular orientation on injection molded polystyrene (PS) plates with a moldedin hole is described. Birefringence measurements were used to investigate the molecular orientation, and three‐point flexure was used to investigate the mechanical properties. Both blunt notch tests and transverse bending tests were carried out. Blunt notch tests showed a strong variation of the material resistance to fracture around the hole. The molecular orientation increased the material resistance not only at the hole equator, but also at the flow split area. In a large domain, wela line fracture was the dominant fracture mechanism. Transverse bending tests also showed a strong influence of the molecular orientation on the load‐carrying ability. The weld line was definitely the weakest region, but the reduction caused by the weld line itself appeared to be small.     

Markerless tracking of complex human motions from multiple views

We present a method for markerless tracking of complex human motions from multiple camera views. In the absence of markers, the task of recovering the pose of a person during such motions is challenging and requires strong image features and robust tracking. We propose a solution which integrates multiple image cues such as edges, color information and volumetric reconstruction. We show that a combination of multiple image cues helps the tracker to overcome ambiguous situations such as limbs touching or strong occlusions of body parts. Following a model-based approach, we match an articulated body model built from superellipsoids against these image cues. Stochastic Meta Descent (SMD) optimization is used to find the pose which best matches the images. Stochastic sampling makes SMD robust against local minima and lowers the computational costs as a small set of predicted image features is sufficient for optimization. The power of SMD is demonstrated by comparing it to the commonly used Levenberg–Marquardt method. Results are shown for several challenging sequences showing complex motions and full articulation, with tracking of 24 degrees of freedom in ≈1 frame per second.