Tagged actin mRNA dysregulation in IGF2BP1[Formula: see text] mice.

Gene expression is tightly regulated by RNA-binding proteins (RBPs) to facilitate cell survival, differentiation, and migration. Previous reports have shown the importance of the Insulin-like Growth Factor II mRNA-Binding Protein (IGF2BP1/IMP1/ZBP1) in regulating RNA fate, including localization, transport, and translation. Here, we generated and characterized a knockout mouse to study RBP regulation.

CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X.

Fragile X syndrome (FXS) is a leading cause of inherited intellectual disability and autism. Whereas dysregulated RNA translation in Fmr1 knockout (KO) mice, a model of FXS, is well studied, little is known about aberrant transcription. Using single-molecule mRNA detection, we show that mRNA encoding the AMPAR subunit GluA2 (but not GluA1) is elevated in dendrites and at transcription sites of hippocampal neurons of Fmr1 KO mice, indicating elevated GluA2 transcription.

Sam68 Enables Metabotropic Glutamate Receptor-Dependent LTD in Distal Dendritic Regions of CA1 Hippocampal Neurons.

The transport and translation of dendritic mRNAs by RNA-binding proteins (RBPs) allows for spatially restricted gene expression in neuronal processes. Although local translation in neuronal dendrites is now well documented, there is little evidence for corresponding effects on local synaptic function. Here, we report that the RBP Sam68 promotes the localization and translation of Arc mRNA preferentially in distal dendrites of rodent hippocampal CA1 pyramidal neurons.

Glutamate-induced RNA localization and translation in neurons.

Localization of mRNA is required for protein synthesis to occur within discrete intracellular compartments. Neurons represent an ideal system for studying the precision of mRNA trafficking because of their polarized structure and the need for synapse-specific targeting. To investigate this targeting, we derived a quantitative and analytical approach.

Quantifying Protein-mRNA Interactions in Single Live Cells.

Specific binding proteins are crucial for the correct spatiotemporal expression of mRNA. To understand this process, a method is required to characterize RNA-protein interactions in single living cells with subcellular resolution. We combined endogenous single RNA and protein detection with two-photon fluorescence fluctuation analysis to measure the average number of proteins bound to mRNA at specific locations within live cells.

Single-molecule insights into mRNA dynamics in neurons.

Targeting of mRNAs to neuronal dendrites and axons plays an integral role in intracellular signaling, development, and synaptic plasticity. Single-molecule imaging of mRNAs in neurons and brain tissue has led to enhanced understanding of mRNA dynamics. Here we discuss aspects of mRNA regulation as revealed by single-molecule detection, which has led to quantitative analyses of mRNA diversity, localization, transport, and translation.

In the right place at the right time: visualizing and understanding mRNA localization.

The spatial regulation of protein translation is an efficient way to create functional and structural asymmetries in cells. Recent research has furthered our understanding of how individual cells spatially organize protein synthesis, by applying innovative technology to characterize the relationship between mRNAs and their regulatory proteins, single-mRNA trafficking dynamics, physiological effects of abrogating mRNA localization in vivo and for endogenous mRNA labelling. The implementation of new imaging technologies has yielded valuable information on mRNA localization, for example, by observing single molecules in tissues.

Taking knowledge for health the extra mile: participatory evaluation of a mobile phone intervention for community health workers in Malawi.

In Malawi, where the majority of the population resides in rural areas, community health workers (CHWs) are the first, and often only, providers of health services. An assessment of health information needs, however, found that these frontline workers often lacked essential health information. A pilot project, implemented in 2 rural districts of Malawi between 2010 and 2011, introduced a mobile phone system to strengthen knowledge exchange within networks of CHWs and district staff.

Single β-actin mRNA detection in neurons reveals a mechanism for regulating its translatability.

The physical manifestation of learning and memory formation in the brain can be expressed by strengthening or weakening of synaptic connections through morphological changes. Local actin remodeling underlies some forms of plasticity and may be facilitated by local β-actin synthesis, but dynamic information is lacking. In this work, we use single-molecule in situ hybridization to demonstrate that dendritic β-actin messenger RNA (mRNA) and ribosomes are in a masked, neuron-specific form.

Transcription factor/microRNA axis blocks melanoma invasion program by miR-211 targeting NUAK1.

Melanoma is one of the deadliest human cancers, responsible for approximately 80% of skin cancer mortalities. The aggressiveness of melanoma is due to its capacity to proliferate and rapidly invade surrounding tissues, leading to metastases. A recent model suggests melanoma progresses by reversibly switching between proliferation and invasion transcriptional signatures.

mRNA on the move: the road to its biological destiny.

Cells have evolved to regulate the asymmetric distribution of specific mRNA targets to institute spatial and temporal control over gene expression. Over the last few decades, evidence has mounted as to the importance of localization elements in the mRNA sequence and their respective RNA-binding proteins. Live imaging methodologies have shown mechanistic details of this phenomenon.

Three-dimensional chemical imaging of embedded nanoparticles using atom probe tomography.

Analysis of nanoparticles is often challenging especially when they are embedded in a matrix. Hence, we have used laser-assisted atom probe tomography (APT) to analyze the Au nanoclusters synthesized in situ using ion-beam implantation in a single crystal MgO matrix. APT analysis along with scanning transmission electron microscopy and energy dispersive spectroscopy (STEM-EDX) indicated that the nanoparticles have an average size ~8-12 nm.

Spatial arrangement of an RNA zipcode identifies mRNAs under post-transcriptional control.

How RNA-binding proteins recognize specific sets of target mRNAs remains poorly understood because current approaches depend primarily on sequence information. In this study, we demonstrate that specific recognition of messenger RNAs (mRNAs) by RNA-binding proteins requires the correct spatial positioning of these sequences. We characterized both the cis-acting sequence elements and the spatial restraints that define the mode of RNA binding of the zipcode-binding protein 1 (ZBP1/IMP1/IGF2BP1) to the β-actin zipcode.

Single mRNA tracking in live cells.

Asymmetric distribution of mRNA is a prevalent phenomenon observed in diverse cell types. The posttranscriptional movement and localization of mRNA provides an important mechanism to target certain proteins to specific cytoplasmic regions of their function. Recent technical advances have enabled real-time visualization of single mRNA molecules in living cells.

Macrophages create an acidic extracellular hydrolytic compartment to digest aggregated lipoproteins.

A critical event in atherogenesis is the interaction of macrophages with subendothelial lipoproteins. Although most studies model this interaction by incubating macrophages with monomeric lipoproteins, macrophages in vivo encounter lipoproteins that are aggregated. The physical features of the lipoproteins require distinctive mechanisms for their uptake.

Comparative activities of antibiotics against intracellular non-typeable Haemophilus influenzae.

Introduction: Non-typeable Haemophilus influenzae (NTHi) is a major bacterial pathogen of community-acquired respiratory tract infection and is usually found extracellularly, although studies have revealed that NTHi may possess the ability to invade human epithelial cells where it is then protected against attack by the local immune system and partly against the effect of antibiotics. The aim of the present study was to assess the ability of ampicillin, azithromycin, telithromycin, ciprofloxacin and moxifloxacin, five antibiotics in common clinical use, to kill NTHi within bronchial epithelial cells.

Methods: Confluent human bronchial epithelial cells were infected with NTHi 77, a particularly invasive clinical strain.

Activation of microglia acidifies lysosomes and leads to degradation of Alzheimer amyloid fibrils.

Microglia are the main immune cells of the brain, and under some circumstances they can play an important role in removal of fibrillar Alzheimer amyloid beta peptide (fAbeta). Primary mouse microglia can internalize fAbeta, but they do not degrade it efficiently. We compared the level of lysosomal proteases in microglia and J774 macrophages, which can degrade fAbeta efficiently, and we found that microglia actually contain higher levels of many lysosomal proteases than macrophages.

Leading a change process to improve health service delivery.

In the fields of health and development, donors channel multiple resources into the design of new practices and technologies, as well as small-scale programmes to test them. But successful practices are rarely scaled up to the level where they beneficially impact large, impoverished populations. An effective process for change is to use the experiences of new practices gained at the programme level for full-scale implementation.

Validation of AKACID plus as a room disinfectant in the hospital setting.

AKACID Plus, a novel polymeric guanidine with broad antimicrobial activity against multiantibiotic-resistant bacterial strains, was used in the present study as a room disinfectant. Disinfection of closed rooms experimentally contaminated with antibiotic-susceptible and multiresistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, and Escherichia coli was performed using AKACID Plus at concentrations of 0.1, 0.

Antimicrobial and toxicological profile of the new biocide Akacid plus.

Objectives: Akacid plus is a new member of the polymeric guanidine family of disinfectants. It was especially developed to enhance the antimicrobial activity of this class with significantly less toxicity. The in vitro activity of Akacid plus compared with chlorhexidine digluconate and mupirocin was tested against a total of 369 recent clinical isolates.

In vitro antimicrobial activity of the novel polymeric guanidine Akacid plus.

The bactericidal and fungicidal activity of Akacid plus, a novel polymeric compound of the cationic family of disinfectants, was compared with chlorhexidine digluconate using quality control strains of Staphylococcus aureus, Enterococcus hirae, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. In vitro activity was determined using the quantitative suspension tests described by the European Committee for Standardization. These use concentrations of 0.

The India Local Initiatives Program: a model for expanding reproductive and child health services.

The India Local Initiatives Program adapted a model used in Indonesia and Bangladesh to implement the government's reproductive and child health strategy. From 1999 to 2003, three Indian nongovernmental organizations (NGOs) provided services for 784,000 people in four northern states. The program established health committees in 620 villages, recruited and trained 1,850 community health volunteers, and added 232 sites to extend government services.

Development of macrolide-resistance and comparative activity of telithromycin in streptococci in Austria, 1996-2002.

The increase detected in macrolide resistance in streptococci in various parts of the world has brought into question the usefulness of macrolides as first line therapy for respiratory tract infections. In a nationwide study, a total of 3012 Streptococcus pneumoniae and 499 Streptococcus pyogenes isolates were collected from 1996 to 2002 and tested for their susceptibility to penicillin, azithromycin, clarithromycin and telithromycin (2002 only). Penicillin-intermediate and -resistant isolates of S.

Antimicrobial susceptibility and macrolide resistance genes in Streptococcus pyogenes collected in Austria and Hungary.

A total of 341 clinical isolates of Streptococcus pyogenes from Vienna, Austria and three Hungarian cities were tested for susceptibility to four macrolides and 12 other antibiotics. All isolates were fully susceptible to penicillin and the other beta-lactams tested. A high level of tetracycline resistance was found in Austria (26.