Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex.

HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophysical studies of the Impβ/Rev complex.

A single-use, size-specific, nylon arthroplasty guide: a preliminary study for hip resurfacing.

Introduction: In arthroplasty surgery, positioning of the components must be accurate and reproducible to avoid complications. Conventional guides are often used to align a component, but they require surgical skill and experience, and are prone to error. To this end, a single-use, size-specific, nylon guide (single-use nylon guide) has been developed for the purpose of increasing the accuracy without adding extra cost to the operation.

Sequential molecularly imprinted solid-phase extraction methods for the analysis of resveratrol and other polyphenols.

Molecularly imprinted polymers (MIPs) templated with either the phytoalexin, (E)-resveratrol, or its structural analog, 3,5-dihydroxy-N-(4-hydroxyphenyl)benzamide, have been used in tandem for the sequential extraction of (E)-resveratrol from aqueous peanut meal extracts in high purity and in near quantitative yields. Re-processing of the (E)-resveratrol-depleted peanut meal extract with the 3,5-dihydroxy-N-(4-hydroxyphenyl)benzamide imprinted MIP yielded additional polyphenolic components, identified as A-type procyanidins. Tandem liquid chromatography-electrospray ionization mass spectrometry confirmed the identity and purity of the isolated products.

Impact on monoclonal antibody production in murine hybridoma cell cultures of adenosine receptor antagonists and phosphodiesterase inhibitors.

The effects of different adenosine receptor antagonists and cyclic nucleotide phosphodiesterase (PDE) inhibitors on monoclonal antibody (mAb) titer and cell viability of murine hybridoma cells in culture were measured as part of our investigations to discover additives that enhance mAb production. Specific adenosine receptor antagonists and PDE inhibitors were found to enhance or decrease the titer of immunoglobulin G1 (IgG1) mAbs relative to negative controls, depending on the specific compound and cell line employed. The observed enhancements or decreases in IgG1 mAb titer appeared to be mainly due to an increase or decrease in specific productivity rates (ngmAb/cell), respectively.

The Application of Template Selectophores for the Preparation of Molecularly Imprinted Polymers.

Molecularly imprinted polymers are versatile materials with wide application scope for the detection, capture and separation of specific compounds present in complex feed stocks. A major challenge associated with their preparation has been the need to sacrifice one mole equivalent of the template molecule to generate the complementary polymer cavities that selectively bind the target molecule. Moreover, template molecules can often be difficult to synthesise, expensive or lack stability.

A morphometric study of normal and varus knees.

Purpose: The aim of the study was to investigate varus and normal knee morphologies to identify differences that may affect knee replacement alignment or design for varus knees.

Methods: Computed tomography scans of varus and normal knees were analyzed, and geometric shapes, points and axes were fit to the femur and tibia independently. These points were then projected in the three anatomical planes to measure the variations between the two groups.

A local reference frame for describing the proximal human femur: application in clinical settings.

Objective: The conventional reference frame for the femur has limited relevance for the planning of hip surgery as the femoral neck axis, a crucial reference for surgeons, has to be independently derived. The purpose of this study is to develop and validate a reliable frame of reference for the proximal femur that can be applied in clinical settings.

Materials And Methods: Ten three-dimensional models of femurs were obtained.

Identification of protein binding partners of ALK-5 kinase inhibitors.

We have investigated the binding characteristics of a potent member of the bis-ortho-substituted five-membered nitrogen heterocycle class of ALK-5 kinase inhibitors with lysates of cultured HEK-293 cells to identify protein binding partners of potential biological significance. An affinity chromatographic resin containing an immobilized ALK-5 kinase inhibitor, 2-phenyl-4-[3-(pyridin-2-yl)-1H-pyrazol-4-yl]pyridine, was used to capture specific proteins from the cell lysate. The soluble inhibitor was then used to specifically elute the proteins which selectively bound to the pharmacophore ligand structure.

Mathematical representation of the normal proximal human femur: application in planning of cam hip surgery.

Precise modelling of the proximal femur can be used for detecting and planning corrective surgery for subjects with deformed femurs using robotic technology or navigation systems. In this study, the proximal femoral geometry has been modelled mathematically. It is hypothesised that it is possible to fit a quadratic surface or combinations of them onto different bone surfaces with a relatively good fit.

A method of assessing the severity of cam type femoro-acetabular impingement in three dimensions.

Femoroacetabular impingement is caused by abnormal morphology of either the femur or acetabulum or both. Diagnostic criteria currently include an alpha angle of over 50° on a lateral radiograph. In this study, CT scans of symptomatic hips (n = 37) were compared with normal hips (n = 34) obtained from CT colonoscopy procedures.

Cam type femoro-acetabular impingement: quantifying the diagnosis using three dimensional head-neck ratios.

Objective: Cam hips are commonly quantified using the two-dimensional α angle. The accuracy of this measurement may be affected by patient position and the technician's experience. In this paper, we describe a method of measurement that provides a quantitative definition of cam hips based upon three-dimensional computed tomography (CT) images.

2-Phenyl and 2-heterocyclic-4-(3-(pyridin-2-yl)-1H-pyrazol-4-yl)pyridines as inhibitors of TGF-β1 and activin A signalling.

Novel inhibitors of TGF-β1 and activin A signalling based on a 2-aryl-4-(3-(pyridin-2-yl)-1H-pyrazol-4-yl)pyridine pharmacophore have been synthesised. Compounds containing phenyl or aromatic nitrogen heterocycle substituents inhibited both types of signalling with HEK-293T cells in culture, with a selectivity preference for TGF-β1. Synthetic compounds containing pyridin-3-yl, pyrazol-4-yl, pyrazol-1-yl or 1H-imidazoyl-1-yl substituents exhibited structural and functional attributes suitable for further investigation related to the development of more potent TGF-β inhibitors.

Preparation of molecularly imprinted polymers for the selective recognition of the bioactive polyphenol, (E)-resveratrol.

(E)-Resveratrol imprinted polymers have been rationally designed with the aid of molecular modelling and NMR spectroscopic titration techniques to determine the optimal ratio of the template to functional monomer for polymer formation. Based on this approach, (E)-resveratrol imprinted polymers were prepared via non-covalent self-assembly with the functional monomer 4-vinylpyridine (4VP) in a 1:3 molar ratio. Polymerisation in the presence of a cross-linker resulted in rigid block copolymers that had selective capacities towards (E)-resveratrol (e.

Enrichment of (E)-resveratrol from peanut byproduct with molecularly imprinted polymers.

Molecularly imprinted solid phase extraction (MISPE) has been employed to isolate and concentrate bioactive polyphenols from peanut press waste. To this end, a molecularly imprinted polymer (MIP) templated with the phytoalexin (E)-resveratrol has been prepared via self-assembly with the functional monomer 4-vinylpyridine (4VP) in a 1:3 molar ratio. Subsequent molecular interrogation of the MIP binding sites demonstrated preferential structural selectivity for (E)-resveratrol with respect to other structurally related naturally occurring compounds.

Peptide mapping with mobile phases of intermediate pH value using capillary reversed-phase high-performance liquid chromatography/electrospray ionisation tandem mass spectrometry.

This investigation describes the separation of tryptic peptides by capillary reversed-phase high-performance liquid chromatography (RP-HPLC) with eluents in the intermediate pH range, followed by in-line electrospray ionisation tandem mass spectrometry (ESI-MS/MS) analysis. For these purposes, gradient elution procedures with an aqueous eluent containing 20 mM ammonium formate, and an increasing content of acetonitrile or methanol, were employed. Compared to the analysis of the same tryptic peptides under low-pH conditions with an ion-pairing reagent, the increase in the pH with the 20 mM ammonium formate mobile phase led to significant changes in both peptide retention to the reversed-phase column and the collision-induced dissociation at the MS/MS stage as a consequence of the changes in the physico-chemical properties of these peptides, such as their overall charge, polarity and relative hydrophobicity.

Antibody library-based tumor endothelial cells surface proteomic functional screen reveals migration-stimulating factor as an anti-angiogenic target.

Angiogenesis is critical for cancer development and metastasis. Here we have employed a functional antibody library-based proteomic screen to identify proteins that participate in and might be used as therapeutic targets for tumor-related angiogenesis. Mice were immunized with human esophageal cancer endothelial cells (HECEC).

Synthesis, purification and bioactivity of recombinant human activin A expressed in the yeast Pichia pastoris.

The transforming growth factor-beta (TGF-beta) superfamily member, activin A, plays a central role in the regulation of multiple physiological processes including cell differentiation, mitogenesis, embryogenesis, apoptosis and inflammation. In normal cells, activin A signalling is regulated to maintain cellular and tissue health and suppress tumour growth. Disruption of activin A signalling has been implicated in tumour formation and progression.

Effects of age and alcohol exposure during early life on pyramidal cell numbers in the CA1-CA3 region of the rat hippocampus.

We have previously shown that exposing rats to a relatively high dose of ethanol during early postnatal life can result in an alteration in spatial learning ability. The hippocampal formation is known to be involved in the control of this ability. The purpose of the present study was to determine whether exposure of rats to ethanol during early postnatal life had either immediate or delayed effects on the numbers of pyramidal cells in the CA1-CA3 subregion of the hippocampus.

Cessation of methadone maintenance treatment using buprenorphine: transfer from methadone to buprenorphine and subsequent buprenorphine reductions.

Background: Buprenorphine is used in the treatment of opioid dependence. Due to its pharmacology, the transfer from methadone to buprenorphine may precipitate withdrawal symptoms.

Methods: Methadone maintained patients with clinical indicators of stability who were seeking withdrawal from methadone were recruited from three Australian states.

Effects of alcohol exposure during early life on neuron numbers in the rat hippocampus. I. Hilus neurons and granule cells.

We previously showed that 16-day-old rats exposed to a relatively high dose of ethanol at 10-15 postnatal days of age have fewer neurons in the hilus region of the hippocampus compared with controls. Dentate gyrus granule cell numbers, however, showed no statistically significant changes attributable to the ethanol treatment. It is possible that some of the changes in brain morphology, brought about as a result of the exposure to ethanol during early life, may not be manifested until later in life.