ESCRT-0 assembles as a heterotetrameric complex on membranes and binds multiple ubiquitinylated cargoes simultaneously.

The ESCRT machinery consists of multiple protein complexes that collectively participate in the biogenesis of multivesicular endosomes (MVEs). The ESCRT-0 complex is composed of two subunits, Hrs and STAM, both of which can engage ubiquitinylated substrates destined for lysosomal degradation. Here, we conduct a comprehensive analysis of ESCRT-0:ubiquitin interactions using isothermal titration calorimetry and define the affinity of each ubiquitin-binding domain (UBD) within the intact ESCRT-0 complex.

Novel protein genes in animal mtDNA: a new sex determination system in freshwater mussels (Bivalvia: Unionoida)?

Mitochondrial (mt) function depends critically on optimal interactions between components encoded by mt and nuclear DNAs. mitochondrial DNA (mtDNA) inheritance (SMI) is thought to have evolved in animal species to maintain mito-nuclear complementarity by preventing the spread of selfish mt elements thus typically rendering mtDNA heteroplasmy evolutionarily ephemeral. Here, we show that mtDNA intraorganismal heteroplasmy can have deterministic underpinnings and persist for hundreds of millions of years.

The complexities of p97 function in health and disease.

p97 is a homohexameric, toroidal machine that harnesses the energy of ATP binding and hydrolysis to effect structural reorganization of a diverse and primarily uncharacterized set of substrate proteins. This action has been linked to endoplasmic reticulum associated degradation (ERAD), homotypic membrane fusion, transcription factor control, cell cycle progression, DNA repair, and post-mitotic spindle disassembly. Exactly how these diverse processes use p97 is not fully understood, but it is clear that binding sites, primarily on the N- and C-domains of p97, facilitate this diversity by coordinating a growing collection of cofactors.

Advances in CMOS Solid-state Photomultipliers for Scintillation Detector Applications.

Solid-state photomultipliers (SSPMs) are a compact, lightweight, potentially low-cost alternative to a photomultiplier tube for a variety of scintillation detector applications, including digital-dosimeter and medical-imaging applications. Manufacturing SSPMs with a commercial CMOS process provides the ability for rapid prototyping, and facilitates production to reduce the cost. RMD designs CMOS SSPM devices that are fabricated by commercial foundries.

SV2 mediates entry of tetanus neurotoxin into central neurons.

Tetanus neurotoxin causes the disease tetanus, which is characterized by rigid paralysis. The toxin acts by inhibiting the release of neurotransmitters from inhibitory neurons in the spinal cord that innervate motor neurons and is unique among the clostridial neurotoxins due to its ability to shuttle from the periphery to the central nervous system. Tetanus neurotoxin is thought to interact with a high affinity receptor complex that is composed of lipid and protein components; however, the identity of the protein receptor remains elusive.

Assessing appropriate parenteral nutrition ordering practices in tertiary care medical centers.

Background: Parenteral nutrition (PN) is an essential feeding route for specific patient populations. Despite its utility, PN is invasive, costly, and associated with clinical complications. In most U.

Control of exocytosis by synaptotagmins and otoferlin in auditory hair cells.

In pre-hearing mice, vesicle exocytosis at cochlear inner hair cell (IHC) ribbon synapses is triggered by spontaneous Ca(2+) spikes. At the onset of hearing, IHC exocytosis is then exclusively driven by graded potentials, and is characterized by higher Ca(2+) efficiency and improved synchronization of vesicular release. The molecular players involved in this transition are still unknown.

Otoferlin is a calcium sensor that directly regulates SNARE-mediated membrane fusion.

Otoferlin is a large multi-C2 domain protein proposed to act as a calcium sensor that regulates synaptic vesicle exocytosis in cochlear hair cells. Although mutations in otoferlin have been associated with deafness, its contribution to neurotransmitter release is unresolved. Using recombinant proteins, we demonstrate that five of the six C2 domains of otoferlin sense calcium with apparent dissociation constants that ranged from 13-25 µM; in the presence of membranes, these apparent affinities increase by up to sevenfold.

Rat and Drosophila synaptotagmin 4 have opposite effects during SNARE-catalyzed membrane fusion.

Synaptotagmins (Syt) are a large family of proteins that regulate membrane traffic in neurons and other cell types. One isoform that has received considerable attention is SYT4, with apparently contradictory reports concerning the function of this isoform in fruit flies and mice. SYT4 was reported to function as a negative regulator of neurotrophin secretion in mouse neurons and as a positive regulator of secretion of a yet to be identified growth factor from muscle cells in flies.

VEGF stimulates RCAN1.4 expression in endothelial cells via a pathway requiring Ca2+/calcineurin and protein kinase C-delta.

Background: Vascular endothelial growth factor (VEGF) has previously been shown to upregulate the expression of the endogenous calcineurin inhibitor, regulator of calcineurin 1, variant 4 (RCAN1.4). The aim of this study was to determine the role and regulation of VEGF-mediated RCAN1.

Regulation of exocytosis and fusion pores by synaptotagmin-effector interactions.

Synaptotagmin (syt) serves as a Ca(2+) sensor in the release of neurotransmitters and hormones. This function depends on the ability of syt to interact with other molecules. Syt binds to phosphatidylserine (PS)-containing lipid bilayers as well as to soluble N-ethylmaleimide sensitive factor receptors (SNAREs) and promotes SNARE assembly.

Use of multimedia as an educational tool to improve human papillomavirus vaccine acceptability--a pilot study.

Objective: To determine, through a pilot study, whether use of a video-based educational tool can influence overall human papillomavirus (HPV) vaccine acceptability, views on vaccine mandates, school vaccination, and acceptable age for vaccination.

Methods: Written surveys addressing HPV knowledge and vaccine acceptability were administered to study participants from January to March 2007. An initial 32-question survey was completed, followed by an 8-minute educational video, and a post-video assessment.

MicroRNA gene evolution in Arabidopsis lyrata and Arabidopsis thaliana.

MicroRNAs (miRNAs) are short regulatory RNAs processed from partially self-complementary foldbacks within longer MIRNA primary transcripts. Several MIRNA families are conserved deeply through land plants, but many are present only in closely related species or are species specific. The finding of numerous evolutionarily young MIRNA, many with low expression and few if any targets, supports a rapid birth-death model for MIRNA evolution.

Lanthanum carbonate is an effective hypophosphatemic agent for hemodialysis patients intolerant of other phosphate binders.

Background: Lanthanum carbonate (LC) is a noncalcium-containing phosphate binder of proven utility in treating hyperphosphatemia in dialysis patients, and displays a good tolerability profile.

Objective: This study addressed whether this tolerability profile could result in improved phosphate control in patients intolerant of other phosphate binders, and who remain hyperphosphatemic despite intensive dietary advice and adequate dialysis provision.

Design, Setting, And Patients: This 18-month study, from a large urban hemodialysis unit, studied 55 patients identified by the multidisciplinary team as fulfilling the following criteria: persistent hyperphosphatemia; intolerance of, or admitted nonadherence to, prescribed phosphate binders (either calcium-based or sevelamer); and provision of adequate dialysis (urea reduction ratio, >65%).

Applying global frameworks to assessment in medical education: an example of a nationally produced curriculum for cancer education.

The past decade has seen an increased effort to standardized medical curricula internationally. Despite these efforts, a lack of standardization remains evident, most likely owing to the lack of specificity with which such frameworks are often (out of necessity) constructed. As such, inconsistencies may arise owing to differences in adopted definitions and approaches to assessment.

Crystal structure of the botulinum neurotoxin type G binding domain: insight into cell surface binding.

Botulinum neurotoxins (BoNTs) typically bind the neuronal cell surface via dual interactions with both protein receptors and gangliosides. We present here the 1.9-A X-ray structure of the BoNT serotype G (BoNT/G) receptor binding domain (residues 868-1297) and a detailed view of protein receptor and ganglioside binding regions.

Detection of siRNA induced mRNA silencing by RT-qPCR: considerations for experimental design.

Background: RNA interference (RNAi) has been one of the most rapidly expanding areas of biological research in the past decade, revolutionizing the ability to analyze gene function. Thorough validation of siRNA duplexes is required prior to use in experimental systems, ideally by western blotting to show a reduction in protein levels. However, in many cases good antibodies are not available, and researchers must rely on RT-qPCR to detect knockdown of the mRNA species.

Arabidopsis RNA-dependent RNA polymerases and dicer-like proteins in antiviral defense and small interfering RNA biogenesis during Turnip Mosaic Virus infection.

Plants respond to virus infections by activation of RNA-based silencing, which limits infection at both the single-cell and system levels. Viruses encode RNA silencing suppressor proteins that interfere with this response. Wild-type Arabidopsis thaliana is immune to silencing suppressor (HC-Pro)-deficient Turnip mosaic virus, but immunity was lost in the absence of DICER-LIKE proteins DCL4 and DCL2.

Ecotoxicological risk assessment of undisturbed metal contaminated soil at two remote lighthouse sites.

Ecotoxicological risk assessments of contaminated soil are commonly completed using guideline values based on total concentrations. However, only certain fractions of contaminants are bioavailable and pose a hazard to the environment. This paper investigates the relationship between measured metal concentrations in soil and soil leachate, and the effects in organisms exposed to intact, undisturbed soil cores (wheat, Tricum aestivum) and soil leachate (lettuce, Lactuca sativa, and water flea, Daphnia magna).

Mitochondrial phylogenomics of the Bivalvia (Mollusca): searching for the origin and mitogenomic correlates of doubly uniparental inheritance of mtDNA.

Background: Doubly uniparental inheritance (DUI) is an atypical system of animal mtDNA inheritance found only in some bivalves. Under DUI, maternally (F genome) and paternally (M genome) transmitted mtDNAs yield two distinct gender-associated mtDNA lineages. The oldest distinct M and F genomes are found in freshwater mussels (order Unionoida).

Loss of synaptotagmin IV results in a reduction in synaptic vesicles and a distortion of the Golgi structure in cultured hippocampal neurons.

Fusion of synaptic vesicles with the plasma membrane is mediated by the SNARE (soluble NSF attachment receptor) proteins and is regulated by synaptotagmin (syt). There are at least 17 syt isoforms that have the potential to act as modulators of membrane fusion events. Synaptotagmin IV (syt IV) is particularly interesting; it is an immediate early gene that is regulated by seizures and certain classes of drugs, and, in humans, syt IV maps to a region of chromosome 18 associated with schizophrenia and bipolar disease.

Enzymatic processing of platinated RNAs.

The broadly prescribed antitumor drug cisplatin coordinates to DNA, altering the activity of cellular proteins whose functions rely upon sensing DNA structure. Cisplatin is also known to coordinate to RNA, but the effects of RNA-Pt adducts on the large number of proteins that process the transcriptome are currently unknown. In an effort to address how platination of an RNA alters the function of RNA processing enzymes, we have determined the influence of [Pt(NH(3))(2)](2+)-RNA adducts on the activities of 3'-->5' and 5'-->3' phosphodiesterases, a purine-specific endoribonuclease, and a reverse transcriptase.

Molecular diagnosis of a previously unreported predator-prey association in coffee: Karnyothrips flavipes Jones (Thysanoptera: Phlaeothripidae) predation on the coffee berry borer.

The coffee berry borer, Hypothenemus hampei, is the most important pest of coffee throughout the world, causing losses estimated at US $500 million/year. The thrips Karnyothrips flavipes was observed for the first time feeding on immature stages of H. hampei in April 2008 from samples collected in the Kisii area of Western Kenya.

Pulling force generated by interacting SNAREs facilitates membrane hemifusion.

In biological systems, membrane fusion is mediated by specialized proteins. Although soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptors (SNAREs) provide the minimal molecular machinery required to drive membrane fusion, the precise mechanism for SNARE-mediated fusion remains to be established. Here, we used atomic force microscope (AFM) spectroscopy to determine whether the pulling force generated by interacting SNAREs is directly coupled to membrane fusion.

Cytotype regulation by telomeric P elements in Drosophila melanogaster: variation in regulatory strength and maternal effects.

Strains carrying the X-linked telomeric P elements TP5 or TP6 varied in their ability to repress hybrid dysgenesis. The rank ordering of these strains was consistent across different genetic assays and was not related to the type of telomeric P element (TP5 or TP6) present. Strong repression of dysgenesis was associated with weak expression of mRNA from the telomeric P element and also with a reduced amount of mRNA from a transposase-producing P element contained within a transgene inserted on an autosome.