Retroviral display of antibody fragments; interdomain spacing strongly influences vector infectivity

Five different single-chain antibody fragments (scFv) against human cell-surface antigens were displayed on murine ecotropic retroviral vectors by fusing them to the Moloney SU envelope glycoprotein. The spacing between the scFv and the SU glycoprotein was varied by fusing the scFv to residue +7 or to residue +1 of Moloney SU and by inserting linker sequences of different lengths between the domains. All of the chimeric envelopes were efficiently incorporated into vector particles and could bind to human cells through their displayed antibody fragments, but did not infect them. The spacing between the scFvs and the SU glycoproteins had no significant effect on the efficiency of envelope expression or viral incorporation and did not affect the binding properties of the chimeric envelopes, nor did it influence the efficiency of targeted gene delivery to human cells by scFv-displaying vectors. However, on murine fibroblasts the infectivity of vectors incorporating the chimeric envelopes was strongly influenced by the length of the interdomain spacer. The titers were very low when the single-chain antibodies were fused through a tripeptide linker to SU residue +7 and were greatly enhanced (up to 10(5)-fold) when they were fused to SU residue +1 through a heptapeptide linker. These results point to the importance of steric interactions between the domains of chimeric envelope glycoproteins and may have implications for retroviral vector design for human gene therapy.     

Cell-type-specific expression of rat steroid 5 alpha-reductase isozymes

The enzyme steroid 5 alpha-reductase (EC 1.3.99.5) is a component of an intercellular signaling pathway that determines cell fate in the primordium of the mammalian reproductive tract. During male phenotypic sexual differentiation, the dihydrotestosterone product of this enzyme binds to the androgen receptor and initiates development of the external genitalia and prostate. Genes encoding two isozymes of steroid 5 alpha-reductase with different biochemical properties and tissue distributions have recently been isolated. In the current study, we utilize in situ hybridization analysis to determine cell-type-specific expression patterns of the 5 alpha-reductase isozyme mRNAs in two androgen target tissues (regenerating ventral prostate and epididymis) and a peripheral tissue (liver). In regenerating ventral prostate, the type 1 mRNA is expressed in basal epithelial cells whereas expression of the type 2 mRNA is largely confined to stromal cells. These results were confirmed by immunohistochemical analysis and are consistent with distinct roles played by the isozymes in the prostate. In the epididymis, both 5 alpha-reductase isozyme mRNAs are expressed in epithelial cells. Only the type 1 mRNA is present in the liver. This mRNA is distributed in a striking spatial gradient extending from hepatocytes surrounding the portal triad (high expression) to those surrounding the central vein (low to absent expression). These findings demonstrate cell-type-specific expression of the steroid 5 alpha-reductase isozymes and underscore their distinct and overlapping functions in androgen physiology.     

Deposition mechanisms in langmuir-blodgett films

As part of a study of the possible application of polymerisable Langmuir-Blodgett (LB) films as ultra-fine-line e-beam resists, an investigation of the variation of film structure of 22-tricosenoic acid with differing deposition conditions has been made. Unexpected effects with significant implications for deposition speed and resist sensitivity have been observed, and the new techniques for film characterisation developed during the investigation have resulted in a revised model of deposition explaining the observed independence of the disorder causing optical scattering and the macroscopic features observed by polarised microscopy.     

Energy pricing policies in developing countries

Government energy pricing policies have multiple and often conflicting objectives: economic efficiency, government revenues (for both parastatal supply companies and the treasury), maintenance or improvement of income distribution, promotion of particular sectors (such as industrial exporters and local resource development) demand management and security of supply. It is important to examine the impacts on and the trade-offs between these objectives resulting from alternative policies in order to assist in policy selection. This article discusses the more important objectives and their conflicts and outlines an approach for the quantitative examination of alternative policies.     

Community-based behavioral training approaches for people with mental retardation and mental illness.

Reviews challenges associated with behavioral training approaches for individuals with mental retardation and mental illness in the community. Family and non-family facilitated training are considered. Professional practice issues are reviewed, and justification for multifactor behavioral assessment is offered. Future research directions are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Terrestrial digital video broadcasting for mobile reception using OFDM

Performance of a system design for digital video broadcasting is examined with emphasis on mobile reception. Orthogonal frequency division multiplexing (OFDM) is used to achieve good bandwidth efficiency and to mitigate the intersymbol interference resulting from the channel delay spread. The resulting equivalent channel including OFDM can be modeled as a flat Rayleigh fading channel plus an interchannel interference (ICI) term due to the channel Doppler spread. This ICI term is analyzed and shown to result in an error floor. Performance improvements due to antenna diversity and trellis-coded modulation (TCM) are given. Finally, multiresolution modulation is discussed as a means of achieving graceful degradation and giving degrees of freedom for further performance improvement.This research was supported by the Multimedia Systems R & D Laboratory, Hitachi, Ltd.     

Laboratory evaluation of the XR-Bond Dentin/Enamel Bonding System

The shear bond strengths of the XR-Bonding System used in conjunction with Herculite composite, to the dentine of forty extracted human permanent first and second molars were determined after the test specimens were stored in physiological saline at 37 degrees C for 48 hours, one week, two weeks and four weeks, respectively. A shear load was applied to the base of the bonded composite cylinders with a knife-edged rod at a crosshead speed of 0.5 mm/minute. The shear bond strengths were expressed in megapascals (MPa). The quantitative microleakage of Class V preparations in dentine (cementum) in forty-eight extracted human maxillary permanent canines restored with the same dentinal bonding system and after storage in physiological saline at 37 degrees C for the same time intervals as for the shear bond strength tests, was determined. On the final day of each time interval the teeth were thermocycled X 500 in a 2 per cent methylene blue solution between 8 degrees C and 50 degrees C with a dwell time of 15 seconds. Microleakage was determined by a spectrophotometric dye-recovery method and expressed in microgram dye/restoration. There was a significant trend for the shear bond strengths to increase with duration of storage (p = 0.01) but the quantitative microleakage was not significantly different (p = 0.75).     

Block Copolymer Surface Reconstuction: A Reversible Route to Nanoporous Films

Thin films of block copolymers have been used as templates and scaffolds for the fabrication of arrays of nanostructured materials. In general, a chemical modification of the film or the removal of one of the components by photodegradative methods is required to produce a nanoporous film that serves as a template or scaffold. Here, however, the preferential interaction of one of the components with a solvent is shown to produce a reconstruction of the block copolymer film that, upon drying, leads to the generation of a nanoporous template. The area density of the pores is identical to that of the original copolymer thin film. Since no chemical reactions occurr, the process is fully reversible. Upon heating the copolymer film above its glass‐transition temperature, mobility is imparted to the copolymer and the original copolymer film with oriented domains is recovered. The film reconstruction significantly simplifies the generation of nanoporous templates.