JADAS: a customizable automated data acquisition system and its application to ice-embedded single particles.

The JEOL Automated Data Acquisition System (JADAS) is a software system built for the latest generation of the JEOL Transmission Electron Microscopes. It is designed to partially or fully automate image acquisition for ice-embedded single particles under low dose conditions. Its built-in flexibility permits users to customize the order of various imaging operations.

Structure of bacterial glutathione-S-transferase maleyl pyruvate isomerase and implications for mechanism of isomerisation.

Maleyl pyruvate isomerase (MPI) is a bacterial glutathione S-transferase (GST) from the pathway for degradation of naphthalene via gentisate that enables the bacterium Ralstonia to use polyaromatic hydrocarbons as a sole carbon source. Genome sequencing projects have revealed the presence of large numbers of GSTs in bacterial genomes, often located within gene clusters encoding the degradation of different aromatic compounds. This structure is therefore an example of this under-represented class of enzymes.

Early postoperative care for free flap head & neck reconstructive surgery--a national survey of practice.

There is considerable variation in the post-operative management of head and neck free flaps in the UK. We undertook a national postal survey of maxillofacial surgical units in the UK who perform free flap reconstruction following ablative head and neck surgery. Questions were asked about the routine postoperative care of a hypothetical, straightforward patient undergoing free flap reconstruction to determine whether there were any trends in managing these patients.

CD63 is not required for production of infectious human immunodeficiency virus type 1 in human macrophages.

During the assembly of human immunodeficiency virus type 1 (HIV-1) particles, the tetraspanin CD63 can be incorporated into the viral membrane. Indeed, cell surface tetraspanin microdomains that include CD63 have been proposed as sites for virus release. In addition, antibodies against CD63 can inhibit HIV infection of macrophages.

Modular software platform for low-dose electron microscopy and tomography.

Transmission electron microscopy imaging protocols required by structural scientists vary widely and can be laborious without tailor-made applications. We present here the jeol automated microscopy expert system (james) api integrator, a programming library for computer control of transmission electron microscopy operations and equipment. james has been implemented on JEOL microscopes with Gatan CCDs but is designed to be modular so it can be adapted to run on different microscopes and detectors.

ALIX catches HIV.

ESCRT and ESCRT-associated proteins are required during the assembly and release of many RNA viruses, including HIV. Two new papers provide structures for the ESCRT-associated protein ALIX/AIP1 and demonstrate how this protein interacts with HIV Gag. One of these studies provides the clearest evidence to date that ESCRT-III mediates key events in virus release and indicates that there are cellular proteins involved in this process still to be discovered.

The ESCRT machinery is not required for human cytomegalovirus envelopment.

The human cytomegalovirus (HCMV) has been proposed to complete its final envelopment on cytoplasmic membranes prior to its release to the extracellular medium. The nature of these membranes and the mechanisms involved in virus envelopment and release are poorly understood. Here we show by immunogold-labelling and electron microscopy that CD63, a marker of multivesicular bodies (MVBs), is incorporated into the viral envelope, supporting the notion that HCMV uses endocytic membranes for its envelopment.

In macrophages, HIV-1 assembles into an intracellular plasma membrane domain containing the tetraspanins CD81, CD9, and CD53.

In macrophages, HIV-1 has been shown to bud into intracellular structures that contain the late endosome marker CD63. We show that these organelles are not endosomes, but an internally sequestered plasma membrane domain. Using immunofluorescence microscopy and immunoelectron microscopy, we find that HIV-1 buds into a compartment that contains the tetraspanins CD81, CD9, and CD53.

Vision for a global registry of anticipated public health studies.

In public health, the generation, management, and transfer of knowledge all need major improvement. Problems in generating knowledge include an imbalance in research funding, publication bias, unnecessary studies, adherence to fashion, and undue interest in novel and immediate issues. Impaired generation of knowledge, combined with a dated and inadequate process for managing knowledge and an inefficient system for transferring knowledge, mean a distorted body of evidence available for decisionmaking in public health.

An automated microseed matrix-screening method for protein crystallization.

A microseed-matrix procedure has been established with the aim of influencing the nucleation event in standard crystallization screens. The method is based on the original description of matrix seeding described by Ireton & Stoddard (2004, Acta Cryst. D60, 601-605).

Electron microscopy analysis of viral morphogenesis.

This chapter reviews the use of standard transmission electron microscopy (TEM) of plastic-embedded material, as well as protocols for the immunolabeling of cryosections, in the analysis of viral interactions with cells. It focuses particularly on the assembly of two types of enveloped viruses: (1) the beta herpesvirus—human cytomegalovirus (HCMV), and (2) the primate lentiviruses—the simian and human immunodeficiency viruses (SIV and HIV). The chapter discusses the ways EM is used to identify morphological features of the various stages in the assembly of virus particles, to distinguish immature and mature particles, or to analyze steps involved in the acquisition of lipid membranes by enveloped viruses.

Bluetongue virus entry into cells.

Bluetongue virus (BTV) is a member of the Orbivirus genus within the Reoviridae family. Like those of other members of the family, BTV particles are nonenveloped and contain two distinct capsids, namely, an outer capsid and an inner capsid or core. The two outer capsid proteins, VP2 and VP5, are involved in BTV entry into cells and in the delivery of the transcriptionally active core to the target cell cytoplasm.

A conserved dileucine motif mediates clathrin and AP-2-dependent endocytosis of the HIV-1 envelope protein.

During the assembly of enveloped viruses viral and cellular components essential for infectious particles must colocalize at specific membrane locations. For the human and simian immunodeficiency viruses (HIV and SIV), sorting of the viral envelope proteins (Env) to assembly sites is directed by trafficking signals located in the cytoplasmic domain of the transmembrane protein gp41 (TM). A membrane proximal conserved GYxxØ motif mediates endocytosis through interaction with the clathrin adaptor AP-2.

Cryoelectron microscopy of protein IX-modified adenoviruses suggests a new position for the C terminus of protein IX.

Recombinant human adenovirus is a useful gene delivery vector for clinical gene therapy. Minor capsid protein IX of adenovirus has been of recent interest since multiple studies have shown that modifications can be made to its C terminus to alter viral tropism or add molecular tags and/or reporter proteins. We examined the structure of an engineered adenovirus displaying the enhanced green fluorescent protein (EGFP) fused to the C terminus of protein IX.

Addressing sexual violence in humanitarian emergencies.

Sexual violence is a by-product of conflict commonly seen, but poorly addressed, in humanitarian emergencies. Reports reveal that extraordinary numbers of women and girls suffer physical, psychological, and social consequences of sexual violence during conflict, when fleeing conflict, and during displacement. All sectors of the humanitarian community have a role to play in the prevention of and response to sexual violence.

The effects of a control-enhancing intervention for nursing home residents: cognition and locus of control as moderators.

Objective: This study aimed to clarify which older adults benefit most from a control-enhancing intervention; in particular, whether cognitive functioning or locus of control might moderate the benefit derived.

Methods: Nursing home residents were randomly assigned to two conditions: (1) a control-enhanced condition that provided the option of caring for a plant (n=10), and (2) a comparison condition that monitored any change under the standard of care (n=10). Comparison group participants were subsequently offered the intervention, which led to a total of 17 intervention participants.

Virus entry: open sesame.

Detailed information about the replication cycle of viruses and their interactions with host organisms is required to develop strategies to stop them. Cell biology studies, live-cell imaging, and systems biology have started to illuminate the multiple and subtly different pathways that animal viruses use to enter host cells. These insights are revolutionizing our understanding of endocytosis and the movement of vesicles within cells.

HIV interaction with endosomes in macrophages and dendritic cells.

The human immunodeficiency virus type 1 (HIV-1) is an enveloped retrovirus that undergoes assembly at specific sites in infected cells. In macrophages, at least, this assembly occurs primarily on a subset of endocytic organelles that contain some of the markers found in late endosomes or multivesicular bodies (MVBs), in particular CD63. The budding of virus particles into endosomes has many features in common with the formation of exosomes and some limited biochemical comparison of HIV-1 particles produced from macrophages with exosomes suggests that the two have similar cellular origins.