Moonlighting Proteins: Diverse Functions Found in Fungi.

Moonlighting proteins combine multiple functions in one polypeptide chain. An increasing number of moonlighting proteins are being found in diverse fungal taxa that vary in morphology, life cycle, and ecological niche. In this mini-review we discuss examples of moonlighting proteins in fungi that illustrate their roles in transcription and DNA metabolism, translation and RNA metabolism, protein folding, and regulation of protein function, and their interaction with other cell types and host proteins.

The value of evaluating the abdominal aorta in patients <50 years of age presenting for abdominal ultrasound.

INTRODUCTION: Practitioners of US routinely include a survey of the abdominal aorta during abdominal US in accordance with international guidelines. Such practice is of uncertain value in younger patients. METHODOLOGY: This study was a retrospective review of 2000 abdominal US examinations which included visualisation of the aorta in patients <50 years of age. Patient demographics and referral details were recorded, and US images and reports were reviewed for the presence of aortic and periaortic pathology. RESULTS: The most common indications for US were abdominal pain (1337, 44%), deranged liver function tests (453, 15%), nausea and/or vomiting (229, 8%), elevated inflammatory markers (146, 5%), pancreatitis (134, 4%) and pyrexia (127, 4%). Fewer than half (977, 49%) of the reports contained a comment regarding the aorta. Aortic pathology was reported in 2 (0.1%) cases. Both were reported as aortic ectasia and both represented a false-positive diagnosis. One male patient had a known abdominal aortic aneurysm with endovascular aortic repair. No new aortic aneurysms were found. All cases of atherosclerotic disease were ignored, and none were reported. Periaortic pathology was encountered on 1 patient, but this was known. No case of new periaortic pathology was detected. CONCLUSION: Routine and indiscriminate imaging of the abdominal aorta during abdominal US in patients <50 years of age is not evidence based. No new case of abdominal aortic aneurysm or new para-aortic pathology was detected, all cases of atherosclerosis were ignored, and two false-positive diagnoses of aortic ectasia were made.

The expanding world of metabolic enzymes moonlighting as RNA binding proteins.

RNA binding proteins play key roles in many aspects of RNA metabolism and function, including splicing, transport, translation, localization, stability and degradation. Within the past few years, proteomics studies have identified dozens of enzymes in intermediary metabolism that bind to RNA. The wide occurrence and conservation of RNA binding ability across distant branches of the evolutionary tree suggest that these moonlighting enzymes are involved in connections between intermediary metabolism and gene expression that comprise far more extensive regulatory networks than previously thought. There are many outstanding questions about the molecular structures and mechanisms involved, the effects of these interactions on enzyme and RNA functions, and the factors that regulate the interactions. The effects on RNA function are likely to be wider than regulation of translation, and some enzyme-RNA interactions have been found to regulate the enzyme's catalytic activity. Several enzyme-RNA interactions have been shown to be affected by cellular factors that change under different intracellular and environmental conditions, including concentrations of substrates and cofactors. Understanding the molecular mechanisms involved in the interactions between the enzymes and RNA, the factors involved in regulation, and the effects of the enzyme-RNA interactions on both the enzyme and RNA functions will lead to a better understanding of the role of the many newly identified enzyme-RNA interactions in connecting intermediary metabolism and gene expression.

How to set-up and perform contrast-enhanced ultrasound.

Contrast-enhanced ultrasound (CEUS) is an important part of current ultrasound imaging practice. Sonographers, radiologists and other sonologists should consider CEUS as a standard tool in the diagnostic toolbox of ultrasound and utilise it liberally to solve a wide range of imaging problems whilst reducing the need to resort to CT or MRI. Setting up a CEUS service is within easy reach of all motivated practitioners. The initial process requires assessment of the demand for CEUS, ensuring staff readiness, preparing administrative processes and obtaining CEUS supplies. The CEUS examination includes gaining informed consent, ensuring authorisation to administer contrast agent (preferably by means of a standing order), conventional pre-scan of the area of interest, insertion of a peripheral IV cannula, preparation of the contrast agent, initiation of the contrast imaging mode, administration of the contrast agent, performance of the examination and aftercare. A number of other important considerations are discussed including cannulation and IV certification, scopes of practice for sonographers performing CEUS, contrast dosing, scheduling, training, interpretation, reporting and quality control.

The clinical implications of adopting new criteria for the grading of internal carotid artery stenosis.

INTRODUCTION: A patient's eligibility for carotid endarterectomy (CEA) is determined primarily by the degree of carotid stenosis detected on duplex ultrasound. The Australasian Society for Ultrasound in Medicine (ASUM) criteria are widely used to grade carotid stenoses in many practices throughout Australasia. METHODS: We sought to investigate the potential impact on the grading of carotid artery stenosis if practitioners switched from the ASUM criteria to the United Kingdom's joint recommendation (UKJR) criteria by reviewing 100 patients with a haemodynamically significant carotid artery stenosis. RESULTS: We found agreement between the criteria in 100% of cases for stenoses <50%, in 80% of cases for stenoses 50-69%, in 89% of cases for stenoses ≥70% and in 100% of cases for stenoses ≥80%. While there was variation in grading of stenoses in 16% of cases, reclassification resulted in no change in the number of patients eligible for CEA. The UKJR guideline enabled more precise categorisation of haemodynamically significant stenosis into deciles. DISCUSSION: Because the UKJR guideline is more comprehensive, we believe that adopting this guideline would enable the ultrasound practitioner to grade carotid stenoses more precisely, better understand the nuances of carotid duplex imaging and more successfully navigate and interpret complex carotid examinations, without impacting the number of patients eligible for CEA.

Inhibition of Glycolate Oxidase With Dicer-substrate siRNA Reduces Calcium Oxalate Deposition in a Mouse Model of Primary Hyperoxaluria Type 1.

Primary hyperoxaluria type 1 (PH1) is an autosomal recessive, metabolic disorder caused by mutations of alanine-glyoxylate aminotransferase (AGT), a key hepatic enzyme in the detoxification of glyoxylate arising from multiple normal metabolic pathways to glycine. Accumulation of glyoxylate, a precursor of oxalate, leads to the overproduction of oxalate in the liver, which accumulates to high levels in kidneys and urine. Crystalization of calcium oxalate (CaOx) in the kidney ultimately results in renal failure. Currently, the only treatment effective in reduction of oxalate production in patients who do not respond to high-dose vitamin B6 therapy is a combined liver/kidney transplant. We explored an alternative approach to prevent glyoxylate production using Dicer-substrate small interfering RNAs (DsiRNAs) targeting hydroxyacid oxidase 1 (HAO1) mRNA which encodes glycolate oxidase (GO), to reduce the hepatic conversion of glycolate to glyoxylate. This approach efficiently reduces GO mRNA and protein in the livers of mice and nonhuman primates. Reduction of hepatic GO leads to normalization of urine oxalate levels and reduces CaOx deposition in a preclinical mouse model of PH1. Our results support the use of DsiRNA to reduce liver GO levels as a potential therapeutic approach to treat PH1.

Discovery and Characterization of a Potent Interleukin-6 Binding Peptide with Neutralizing Activity In Vivo.

Interleukin-6 (IL-6) is an important member of the cytokine superfamily, exerting pleiotropic actions on many physiological processes. Over-production of IL-6 is a hallmark of immune-mediated inflammatory diseases such as Castleman's Disease (CD) and rheumatoid arthritis (RA). Antagonism of the interleukin IL-6/IL-6 receptor (IL-6R)/gp130 signaling complex continues to show promise as a therapeutic target. Monoclonal antibodies (mAbs) directed against components of this complex have been approved as therapeutics for both CD and RA. To potentially provide an additional modality to antagonize IL-6 induced pathophysiology, a peptide-based antagonist approach was undertaken. Using a combination of molecular design, phage-display, and medicinal chemistry, disulfide-rich peptides (DRPs) directed against IL-6 were developed with low nanomolar potency in inhibiting IL-6-induced pSTAT3 in U937 monocytic cells. Targeted PEGylation of IL-6 binding peptides resulted in molecules that retained their potency against IL-6 and had a prolongation of their pharmacokinetic (PK) profiles in rodents and monkeys. One such peptide, PN-2921, contained a 40 kDa polyethylene glycol (PEG) moiety and inhibited IL-6-induced pSTAT3 in U937 cells with sub-nM potency and possessed 23, 36, and 59 h PK half-life values in mice, rats, and cynomolgus monkeys, respectively. Parenteral administration of PN-2921 to mice and cynomolgus monkeys potently inhibited IL-6-induced biomarker responses, with significant reductions in the acute inflammatory phase proteins, serum amyloid A (SAA) and C-reactive protein (CRP). This potent, PEGylated IL-6 binding peptide offers a new approach to antagonize IL-6-induced signaling and associated pathophysiology.