Octaheme tetrathionate reductase is a respiratory enzyme with novel heme ligation.

We have isolated a soluble cytochrome from Shewanella oneidensis that contains eight covalently attached heme groups and determined its crystal structure. One of these hemes exhibits novel ligation of the iron atom by the epsilon-amino group of a lysine residue, despite its attachment via a typical CXXCH motif. This heme is most likely the active site for tetrathionate reduction, a reaction catalyzed efficiently by this enzyme.

Predictive adaptive responses to maternal high-fat diet prevent endothelial dysfunction but not hypertension in adult rat offspring.

Background: Population-based studies suggest that fetal adaptive responses to maternal dietary imbalance confer survival benefit when the postnatal diet remains suboptimal but increase susceptibility to cardiovascular disease when postnatal nutrition is improved. We have investigated "predictive adaptive" responses in a rodent model in which adult offspring of fat-fed dams develop characteristics of the metabolic syndrome.

Methods And Results: Sprague-Dawley rats were fed a fat-rich diet or normal chow throughout pregnancy and weaning.

Proteomic, functional, and domain-based analysis of in vivo 14-3-3 binding proteins involved in cytoskeletal regulation and cellular organization.

Background: 14-3-3 proteins are abundant and conserved polypeptides that mediate the cellular effects of basophilic protein kinases through their ability to bind specific peptide motifs phosphorylated on serine or threonine.

Results: We have used mass spectrometry to analyze proteins that associate with 14-3-3 isoforms in HEK293 cells. This identified 170 unique 14-3-3-associated proteins, which show only modest overlap with previous 14-3-3 binding partners isolated by affinity chromatography.

Severe toxicity associated with a markedly elevated mycophenolic acid free fraction in a renal transplant recipient.

A 58-year-old man with end-stage renal failure secondary to polycystic kidney disease developed a profoundly elevated mycophenolic acid (MPA) free fraction and associated severe toxicity after cadaveric renal transplantation. Initial immunosuppressive therapy was 4 mg/kg body weight bid cyclosporin (Neoral; Novartis Pharmaceutical Co Ltd, Sydney, Australia) given orally with 1 g bid mycophenolate mofetil (MMF) (CellCept; Roche Products Pty Ltd, Sydney, Australia). In the first 5 days posttransplantation, the serum creatinine concentration fell, and the patient developed profound hypoalbuminemia (serum albumin <20 g/L) and hyperbilirubinemia (serum bilirubin >150 micromol/L) that resulted from progressing biliary obstruction.

Therapeutic drug monitoring of immunosuppressant drugs by high-performance liquid chromatography-mass spectrometry.

The currently expanding range of immunosuppressant agents has placed new challenges on therapeutic drug-monitoring (TDM) services. Many of these drugs require the measurement of concentrations with subsequent dosage adjustment to maximize efficacy while minimizing toxicity. HPLC-mass spectrometry (HPLC-MS) is a relatively new technique for drug quantification and thus TDM of immunosuppressant drugs.

Free mycophenolic acid should be monitored in renal transplant recipients with hypoalbuminemia.

The current approach for therapeutic drug monitoring in renal transplant recipients receiving mycophenolate mofetil (MMF) is measurement of total mycophenolic acid (MPA) concentration. Because MPA is highly bound, during hypoalbuminemia the total concentration no longer reflects the free (pharmacologically active) concentration. The authors investigated what degree of hypoalbuminemia causes a significant change in protein binding and thus percentage free MPA.

The effect of sulfur fertilizer on glucoraphanin levels in broccoli (B. oleracea L. var. italica) at different growth stages.

Three sulfur (S) treatements were imposed by applying gypsum to three broccoli cultivars (Claudia, Marathon, and TB-234) known to differ in glucoraphanin content of mature seeds. The S treatments were control (very low added S), low S (23 kg S ha(-)(1)), and high S (92 kg S ha(-)(1)). The gypsum applications during the early vegetative phase of the three broccoli cultivars increased S uptake and the glucoraphanin content in each plant organ.

Expression, purification and preliminary crystallographic studies of a single-point mutant of Mos1 mariner transposase.

A soluble single-point mutant of full-length Mos1 mariner transposase (MW = 40.7 kDa) has been overexpressed in Escherichia coli, purified to 95% homogeneity and crystallized. This provides the first example of the crystallization of a eukaryotic transposase.

Comparison of the ADVIA Centaur and Abbott AxSYM immunoassay systems for a routine diagnostic virology laboratory.

Background: Routine diagnostic laboratories are confronted with an ever-increasing workload with limited resources. Automation has provided some solutions to these challenges particularly high through put analysers such as the Abbott AxSYM.

Objective: The aim of the present study was to compare the performance of two automated immunoassay systems in a diagnostic virology laboratory.

Doryporella smirnovi sp. nov. (Bryozoa: Cheilostomata) and its impact on phylogeny and classification.

A new species of Doryporella, D. smirnovi sp. nov.

A pharmacodynamic investigation of tacrolimus in pediatric liver transplantation.

Although monitoring of tacrolimus blood concentrations is standard clinical practice following liver transplantation, a greater understanding of the relationship between trough concentrations and clinical outcome is required. The aim of this study was to perform a pharmacodynamic investigation of tacrolimus in pediatric liver transplant recipients. A retrospective analysis was performed on 35 pediatric liver recipients who received oral tacrolimus as the primary immunosuppressant.

Enzymatic and structural characterization of non-peptide ligand-cyclophilin complexes.

Piperidine ligands are described that provide the first examples of non-peptidic ligand structures for the cyclophilin family of proteins. Crystal structures of two ligand complexes are compared with the unliganded protein and show ligand-induced changes in side-chain conformation and water binding. A peptidylprolyl cis-trans-isomerase assay showed the dissociation constants of the two ligands to be 320 and 25 mM.

Kinetic inhibitor of hydrate crystallization.

We present the results of a combined theoretical/experimental study into a new class of kinetic inhibitor of gas hydrate formation. The inhibitors are based on quaternary ammonium zwitterions, and were identified from a computational screen. Molecular dynamics simulations were used to characterize the effect of the inhibitor on the interface between a type II hydrate and natural gas.

The effect of oral iron admiinistration on mycophenolate mofetil absorption in renal transplant recipients: a randomized, controlled trial.

Background: Oral iron supplements are frequently prescribed to renal transplant recipients in the early posttransplant period. A recent trial in seven healthy volunteers demonstrated a significant 91% reduction in mycophenolate mofetil (MMF) absorption when coadministered with oral iron. However, the effect of iron on MMF absorption in renal transplant patients has not been studied.

Effect of atorvastatin on cyclosporine pharmacokinetics in liver transplant recipients.

Background: The development of hyperlipidemia after liver transplant is frequently treated with hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) such as atorvastatin. As atorvastatin and the primary immunosuppressant drug, cyclosporine, are metabolized by the same pathway, there is the potential for an interaction.

Objective: To determine the effect of atorvastatin on cyclosporine pharmacokinetics in liver transplant recipients.

A screen for drugs that protect against the cytotoxicity of polyglutamine-expanded androgen receptor.

Spinobulbar muscular atrophy is a neurodegenerative disorder caused by expansion of a CAG triplet repeat sequence encoding a polyglutamine tract in the androgen receptor. It has been shown that the mutant protein is toxic in cell culture and triggers an apoptotic cascade resulting in activation of caspase-3. We developed an assay of caspase-3 activation in cells expressing the mutant androgen receptor.

Proteomics in Vaccinology and Immunobiology: An Informatics Perspective of the Immunone.

The postgenomic era, as manifest, inter alia, by proteomics, offers unparalleled opportunities for the efficient discovery of safe, efficacious, and novel subunit vaccines targeting a tranche of modern major diseases. A negative corollary of this opportunity is the risk of becoming overwhelmed by this embarrassment of riches. Informatics techniques, working to address issues of both data management and through prediction to shortcut the experimental process, can be of enormous benefit in leveraging the proteomic revolution.

Effectiveness of electric stimulation of the vastus medialis muscle in the rehabilitation of patients after total knee arthroplasty.

Objective: To investigate the possible effect of electric muscle stimulation (EMS) of the vastus medialis on the walking speed, Hospital for Special Surgery (HSS) knee score, and Physiological Cost Index (PCI) of patients during rehabilitation after total knee arthroplasty (TKA).

Design: Prospective, randomized controlled trial.

Setting: Various departments at a district general hospital in the United Kingdom.

Quantification of free mycophenolic acid and its glucuronide metabolite in human plasma by liquid-chromatography using mass spectrometric and ultraviolet absorbance detection.

The immunosuppressant drug mycophenolic acid (MPA) and its major metabolite, mycophenolic acid glucuronide (MPAG), are highly bound to albumin. An HPLC-tandem-MS (HPLC/MS/MS) and an HPLC-UV assay were developed to measure free (unbound) concentrations of MPA and MPAG, respectively. Ultrafiltrate was prepared from plasma (500 microl) by ultrafiltration at 3000 x g for 20 min (20 degrees C).

Stage of chronic kidney disease predicts seroconversion after hepatitis B immunization: earlier is better.

Background: Hepatitis B virus (HBV) infection remains a concern in dialysis populations, and vaccination programs have been less successful than those in the general population. Reasons for poor response include malnutrition, uremia, and the generalized immunosuppressive state of patients with chronic kidney disease (CKD). This prospective cohort study evaluated factors impacting on the effectiveness of a vaccination program before dialysis therapy initiation, including level of kidney function.

Handedness and schizotypy in a Japanese sample: an association masked by cultural effects on hand usage.

Previous research has shown a robust association between schizotypy and mixed/ambiguous-handedness, but little is known about the universality of this relationship outside Western cultures. The present paper examines this issue in a sample of 413 Japanese students administered (in Japan) the Annett handedness questionnaire and a schizotypy scale (STA). Conventional analyses of current hand preference, using several indices derived from the Annett scale, mostly failed to replicate previous findings.

Intravenous ascorbic acid: protocol for its application and use.

High dose intravenous(i.v.) ascorbic acid (AA) has been used as therapy for infectious disease from bacterial and viral origin and adjuvant therapy for cancer.

A standard operating procedure for assessing liquid handler performance in high-throughput screening.

The thrust of early drug discovery in recent years has been toward the configuration of homogeneous miniaturized assays. This has allowed organizations to contain costs in the face of exponential increases in the number of screening assays that need to be run to remain competitive. Miniaturization brings with it an increasing dependence on instrumentation, which over the past several years has seen the development of nanodispensing capability and sophisticated detection strategies.

Valproic acid increases SMN levels in spinal muscular atrophy patient cells.

Spinal muscular atrophy (SMA) is an inherited motor neuron disease caused by mutation of the telomeric copy of the survival motor neuron gene (SMN1). Although a centromeric copy of the survival motor neuron gene (SMN2) is retained in all patients with SMA, it differs from SMN1 at a critical nucleotide such that the majority of SMN2 transcripts lack exon 7 and encode an unstable, truncated protein. Here, we show that valproic acid increases levels of exon 7-containing SMN transcript and SMN protein in type I SMA patient-derived fibroblast cell lines.

Production of nuclear transfer llama (lama glama) embryos from in vitro matured llama oocytes.

To date, there have been no reports of somatic cell nuclear transfer in llamas. The application of this methodology to the camelid industry could be helpful in the propagation of genetically valuable animals. The objective of this study was to produce nuclear transfer llama embryos comparing the development of these llama embryos cultured in either CR1aa medium (treatment A) or G1.