Breaching the diffusion barrier that compartmentalizes the transmembrane glycoprotein CE9 to the posterior-tail plasma membrane domain of the rat spermatozoon

CE9 is a posterior-tail domain-specific integral plasma membrane glycoprotein of the rat testicular spermatozoon. During epididymal maturation, CE9 undergoes endoproteolytic processing and then redistributes into the anterior-tail plasma membrane domain of the spermatozoon (Petruszak, J. A. M., C. L. Nehme, and J. R. Bartles. 1991. J. Cell. Biol. 114:917-927). We have determined the sequence of CE9 and found it to be a Type Ia transmembrane protein identical to the MRC OX-47 T-cell activation antigen, a member of the immunoglobulin superfamily predicted to have two immunoglobulin-related loops and three asparagine-linked glycans disposed extracellularly. Although encoded by a single gene and mRNA in the rat, the majority of spermatozoal CE9 is of smaller apparent molecular mass than its hepatocytic counterpart due to the under-utilization of sites for asparagine-linked glycosylation. By fluorescence recovery after photobleaching, CE9 was determined to be mobile within the posterior- tail plasma membrane domain of the living rat testicular spermatozoon, thus implying the existence of a regional barrier to lateral diffusion that is presumed to operate at the level of the annulus. Through the development of an in vitro system, the modification of this diffusion barrier to allow for the subsequent redistribution of CE9 into the anterior-tail domain was found to be a time-, temperature-, and energy- dependent process.     

Breaching the great wall: peptidoglycan and microbial interactions

Once thought to be a process that occurred only in a few human pathogens, release of biologically active peptidoglycan fragments during growth by Gram-negative bacteria controls many types of bacterial interaction, including symbioses and interactions between microorganisms. This Perspective explores the role of peptidoglycan fragments in mediating a range of microbial-host interactions, and discusses the many systems in which peptidoglycan fragments released during bacterial growth might be active.     

Breaching the Barrier: Quantifying Antibiotic Permeability across Gram-negative Bacterial Membranes

The double-membrane cell envelope of Gram-negative bacteria is a sophisticated barrier that facilitates the uptake of nutrients and protects the organism from toxic compounds. An antibiotic molecule must find its way through the negatively charged lipopolysaccharide layer on the outer surface, pass through either a porin or the hydrophobic layer of the outer membrane, then traverse the hydrophilic peptidoglycan layer only to find another hydrophobic lipid bilayer before it finally enters the cytoplasm, where it typically finds its target. This complex uptake pathway with very different physico-chemical properties is one reason that Gram-negative are intrinsically protected against multiple classes of antibiotic-like molecules, and is likely the main reason that in vitro target-based screening programs have failed to deliver novel antibiotics for these organisms. Due to the lack of general methods available for quantifying the flux of drugs into the cell, little is known about permeation rates, transport pathways and accumulation at the target sites for particular molecules. Here we summarize the current tools available for measuring antibiotic uptake across the different compartments of Gram-negative bacteria.     

Breaching Self-Tolerance to Alu Duplex RNA Underlies MDA5-Mediated Inflammation

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Nightmares Among Cambodian Refugees: The Breaching of Concentric Ontological Security

This article explores the nightmares of Cambodian refugees in a cultural context, and the role of nightmares in the trauma ontology of this population, including their role in generating post-traumatic stress disorder (PTSD). Among Cambodian refugees attending a psychiatric clinic, we found that having a nightmare was strongly associated with having PTSD (χ² = 61.7, P < 0.001, odds ratio = 126); that nightmares caused much distress upon awakening, including panic attacks, fear of bodily dysfunction, flashbacks and difficulty returning to sleep; that nightmare content was frequently related to traumatic events; that nightmares resulted in a decrease in the sense of “concentric ontology security” (i.e., in an increased sense of physical and spiritual vulnerability in a culture that conceives of the self in terms of concentric, protective layers), including fears of being attacked by ghosts; and that nightmares frequently led to the performance of specific practices and rituals aiming to extrude and repel attacking forces and to create “protective layers.” Cases are presented to illustrate these findings. The Discussion considers some treatment implications of the study.

血脑屏障与脑药物转运

血脑屏障的存在使大分子药物难以进入脑中发挥疗效。成为中枢神经系统疾病治疗的瓶颈。本就血脑屏障的结构特点、大分子药物转运入脑的途径及药物与载体间的连接策略等问题进行了综述。     

Breaking the language barrier

English has undoubtedly become the universal language of science. But how did this come about and what are the potential benefits?     

Effects of the Sand Bar Breaching on Typha domingensis (PERS.) in a Tropical Coastal Lagoon

Coastal lagoons are usually subjected to several kinds of human impacts, especially eutrophication. The breaching of the sand bar, which separates the lagoon from the ocean, by human action, is a common process used to decrease the negative effects of eutrophication. The aims of this research were to evaluate the effects of the artificial sand bar breaching on the populations of the aquatic macrophyte Typha domingensis and the subsequent effects on nutrients concentration in a tropical coastal lagoon. Samplings were carried out monthly from February/01 to January/02 in a monospecific stand of T. domingensis at Imboassica lagoon (Rio de Janeiro/Brazil). Two sampling sites, in the middle and in the border of the stand, were marked in three different transects. Water depth was measured and the aerial biomass sampled with a 0.25 m² quadrat. The macrophyte samples were separated into live and dead material and the shoot length, shoot density and number of leaves were analyzed. All plant material was oven dried till constant weight and net primary production, dead stand crop production and the nutrients release through decomposition were estimated at each site. The decrease in water level due to sand bar opening affected negatively T. domingensis populations, but the most intense effects were observed in the middle of the stand. The shoot mortality was highly enhanced after the sand bar breaching and the nutrients were released through decomposition to the water column. It reflected on an input of 11.5 kg C m⁻², 0.22 kg N m⁻² and 0.13 kg P m⁻² into the lagoon, which represent from 22.5 to 44.8 ton P and 35.9 to 71.8 ton N to the lagoon. The decrease of nutrients concentration after the sand bar breaching was not successfully accomplished. The decay of T. domingensis stands due to the sand bar breaching neutralized the exportation of nutrients to the ocean, and contributed to the phosphorous increase in the water column. Thus, to a better management of aquatic ecosystems subjected to human eutrophication, the role of aquatic macrophytes decomposition on internal fertilization of aquatic ecosystems should be accounted.     

Breaching biological barriers: protein translocation domains as tools for molecular imaging and therapy

The lipid bilayer of a cell presents a significant barrier for the delivery of many molecular imaging reagents into cells at target sites in the body. Protein translocation domains (PTDs) are peptides that breach this barrier. Conjugation of PTDs to imaging agents can be utilized to facilitate the delivery of these agents through the cell wall, and in some cases, into the cell nucleus, and have potential for in vitro and in vivo applications. PTD imaging conjugates have included small molecules, peptides, proteins, DNA, metal chelates, and magnetic nanoparticles. The full potential of the use of PTDs in novel in vivo molecular probes is currently under investigation. Cells have been labeled in culture using magnetic nanoparticles derivatized with a PTD and monitored in vivo to assess trafficking patterns relative to cells expressing a target antigen. In vivo imaging of PTD-mediated gene transfer to cells of the skin has been demonstrated in living animals. Here we review several natural and synthetic PTDs that have evolved in the quest for easier translocation across biological barriers and the application of these peptide domains to in vivo delivery of imaging agents.     

Proteases and the gut barrier

Serine proteases, cysteine proteases, aspartic proteases and matrix metalloproteinases play an essential role in extracellular matrix remodeling and turnover through their proteolytic action on collagens, proteoglycans, fibronectin, elastin and laminin. Proteases can also act on chemokines, receptors and anti-microbial peptides, often potentiating their activity. The intestinal mucosa is the largest interface between the external environment and the tissues of the human body and is constantly exposed to proteolytic enzymes from many sources, including bacteria in the intestinal lumen, fibroblasts and immune cells in the lamina propria and enterocytes. Controlled proteolytic activity is crucial for the maintenance of gut immune homeostasis, for normal tissue turnover and for the integrity of the gut barrier. However, in intestinal immune-mediated disorders, pro-inflammatory cytokines induce the up-regulation of proteases, which become the end-stage effectors of mucosal damage by destroying the epithelium and basement membrane integrity and degrading the extracellular matrix of the lamina propria to produce ulcers. Protease-mediated barrier disruption in turn results in increased amounts of antigen crossing into the lamina propria, driving further immune responses and sustaining the inflammatory process.     

Breaking the MapReduce stage barrier

The MapReduce model uses a barrier between the Map and Reduce stages. This provides simplicity in both programming and implementation. However, in many situations, this barrier hurts performance because it is overly restrictive. Hence, we develop a method to break the barrier in MapReduce in a way that improves efficiency. Careful design of our barrier-less MapReduce framework results in equivalent generality and retains ease of programming. We motivate our case with, and experimentally study our barrier-less techniques in, a wide variety of MapReduce applications divided into seven classes. Our experiments show that our approach can achieve better job completion times than a traditional MapReduce framework. This is due primarily to the interleaving of I/O and computation, and forgoing disk-intensive work. We achieve a reduction in job completion times that is 25% on average and 87% in the best case.     

Development of the blood-brain barrier

The endothelial cells forming the blood-brain barrier (BBB) are highly specialized to allow precise control over the substances that leave or enter the brain. An elaborate network of complex tight junctions (TJ) between the endothelial cells forms the structural basis of the BBB and restricts the paracellular diffusion of hydrophilic molecules. Additonally, the lack of fenestrae and the extremely low pinocytotic activity of endothelial cells of the BBB inhibit the transcellular passage of molecules across the barrier. On the other hand, in order to meet the high metabolic needs of the tissue of the central nervous system (CNS), specific transport systems selectively expressed in the membranes of brain endothelial cells in capillaries mediate the directed transport of nutrients into the CNS or of toxic metabolites out of the CNS. Whereas the characteristics of the mature BBB endothelium are well described, the cellular and molecular mechanisms that control the development, differentiation and maintenance of the highly specialized endothelial cells of the BBB remain unknown to date, despite the recent explosion in our knowledge of the growth factors and their receptors specifically acting on vascular endothelium during development. This review summarizes our current knowledge of the cellular and molecular mechanisms involved in the development and maintenance of the BBB.